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Neuromodulatory afferents to thalamic nuclei are key for information transmission and thus play critical roles in sensory, motor, and limbic processes. Over the course of the last decades, diverse attempts have been made to map and describe subcortical neuromodulatory afferents to the primate thalamus, including axons using acetylcholine, serotonin, dopamine, noradrenaline, adrenaline, and histamine. Our group has been actively involved in this endeavor. The published descriptions on neuromodulatory afferents to the primate thalamus have been made in different laboratories and are not fully comparable due to methodological divergences (for example, fixation procedures, planes of cutting, techniques used to detect the afferents, different criteria for identification of thalamic nuclei…). Such variation affects the results obtained. Therefore, systematic methodological and analytical approaches are much needed. The present article proposes reproducible methodological and terminological frameworks for primate thalamic mapping. We suggest the use of standard stereotaxic planes to produce and present maps of the primate thalamus, as well as the use of the Anglo-American school terminology (vs. the German school terminology) for identification of thalamic nuclei. Finally, a public repository of the data collected under agreed-on frameworks would be a useful tool for looking up and comparing data on the structure and connections of primate thalamic nuclei. Important and agreed-on efforts are required to create, manage, and fund a unified and homogeneous resource of data on the primate thalamus. Likewise, a firm commitment of the institutions to preserve experimental brain material is much needed because neuroscience work with non-human primates is becoming increasingly rare, making earlier material still more valuable, Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. CC and IP-S were the recipients of grants from Chair in Neuroscience UAM-Fundación Tatiana Pérez de Guzmán el Bueno, Spain. MAG-C was the recipient of a Beatriz Galindo senior research position in the School of Medicine, Universidad Autónoma de Madrid (BEAGAL18/00098) and of a Grant for I+D Projects to the Beatriz Galindo Program Researchers at Universidad Autónoma de Madrid (SI2/PBG/2020–00014) from the Madrid Government (Comunidad de Madrid-Spain) under the Multiannual Agreement with Universidad Autónoma de Madrid in the line of action encouraging young research doctors, in the context of the V PRICIT (Regional Programme of Research and Technological Innovation)

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 13-12-2022, La presente tesis ha sido realizada gracias a la concesión de la ayuda FI19/00287 “Contratos Predoctorales de Formación en Investigación en Salud” (Contratos PFIS) (2020 – 2023) financiada a partir de la Acción Estratégica en Salud 2017 – 2020 del Instituto de Salud Carlos III (ISCIII) y cofinanciada por el Fondo Social Europeo (FSE)

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 04-11-2022

Sixty years ago, Friedrich Sanides traced the origin of the tangential expansion of the primate neocortex to two ancestral anlagen in the allocortex of reptiles and mammals, and proposed the Hypothesis on the Dual Origin of the Neocortex. According to Sanides, paraolfactory and parahippocampal gradients of laminar elaboration expanded in evolution by addition of successive concentric rings of gradually different cortical types inside the allocortical ring. Rodents had fewer rings and primates had more rings in the inner part of the cortex. In the present article, we perform cortical type analysis of the neocortex of adult rats, Rhesus macaques, and humans to propose hypotheses on homology of cortical areas applying the principles of the Hypothesis on the Dual Origin of the Neocortex. We show that areas in the outer rings of the neocortex have comparable laminar elaboration in rats and primates, while most 6-layer eulaminate areas in the innermost rings of primate neocortex lack homologous counterparts in rats. We also represent the topological distribution of cortical types in simplified flat maps of the cerebral cortex of monotremes, rats, and primates. Finally, we propose an elaboration of the Hypothesis on the Dual Origin of the Neocortex in the context of modern studies of pallial patterning that integrates the specification of pallial sectors in development of vertebrate embryos. The updated version of the hypothesis of Sanides provides explanation for the emergence of cortical hierarchies in mammals and will guide future research in the phylogenetic origin of neocortical areas, This work was supported by grants from the National Institute of Mental Health to BZ (Grant Nos. R01 MH101209 and R01 MH118500). MAG-C was the recipient of a Beatriz Galindo senior research position in the Faculty of Medicine at Universidad Autónoma de Madrid (BEAGAL18/00098) and of a Grant for I+D Projects by the Madrid Government (Comunidad de Madrid-Spain) under the Multiannual Agreement with Universidad Autónoma de Madrid in the line of action encouraging youth research doctors, in the context of the V PRICIT (Regional Program of Research and Technological Innovation), reference: SI2/PBG/2020-00014

This study analyzes the dimensional changes of the glands from prostate cancer by applying stereology to estimate the variations in volume, length, surface, and cellular densities of tumor acini. Normal and tumor acini were visualized using immunohistochemistry for cytokeratin18. On immunostained sections, parameters related to the dimensions and cell population of prostate acini were measured. The immunohistochemical expression of proliferative cell nuclear antigen was also measured to correlate the quantitative changes estimated with the proliferative activity of the epithelium. The average cell volume in normal and tumor epithelium was estimated using the method of the nucleator. The relative size of the acini was similar in the carcinoma compared with the normal prostate. Within the acini, the fraction of acinar volume occupied by the epithelium was significantly higher in cancer than in the nontumor prostate. Conversely, the glandular lumen of the cancer acini is lower than in the normal acini. The significant increase of acinar length density in the carcinoma indicates that the glandular tree's growth in the carcinoma is higher and with more branches than in the case of nonneoplastic glands. The basal surface density is higher in the carcinoma than in the controls. The number of epithelial cells per unit length of acini was significantly decreased in the neoplastic glands. This "dilution" of the cell population along the cancer acinus can be explained by the significant increase in the tumor cell's mean cell volume.

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 15-06-2022

W-Tau, a new tau human-specific splicing isoform generated by intron retention, has been recently described. This isoform contains an 18-residue unique sequence corresponding to the translation of the retained region of intron 12. In this work, we have described that such 18-amino-acid peptide from the retained intron 12 can inhibit tau and β amyloid peptides aggregation under in vitro conditions. This inhibitory function is also present in smaller fragments of the 18-residue peptide, This research was funded by the Spanish Ministry of Science and Innovation (BES-2015-074405 and PGC2018-096177-BI00), the Center for Networked Biomedical Research on Neurodegenerative Diseases, and the Comunidad de Madrid (S2017/BMD). Institutional grants from the Fundación Ramón Areces and Banco de Santander are also acknowledged.

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 25-03-2022, A través de los diversos componentes del Sistema Sensorial de Vibrisas los roedores procesan características estimulares como la posición, velocidad y aceleración, movimiento angular, amplitud, frecuencia, repetición y duración. La información táctil es codificada en patrones de disparo específicos, que pueden variar dependiendo del estado de alerta del animal, y se transmite hacia estructuras centrales como el Complejo Trigeminal y el Tálamo. La actividad neuronal de los núcleos talámicos Posterior (POm) y Ventral Posterior Medial (VPM) se ha puesto a prueba a lo largo de esta tesis doctoral bajo diversas condiciones de estimulación mecánica, eléctrica y farmacológica. Dicha actividad se encuentra principalmente regulada por proyecciones descendentes que parten de la Neocorteza, cuya influencia tiende a distinguir entre núcleos de relevo primario y asociativos, en concierto con sinapsis GABAérgicas de axones provenientes del núcleo Reticular (nRt) y sistemas neurotransmisores como el colinérgico. Estos núcleos talámicos guardan una estrecha relación con la Corteza de Barriles (S1BC) en tanto en cuanto conforman circuitos relativamente cerrados, bien para transmitir información táctil de alta resolución en el caso de VPM, bien para ejercer de integrador y/o mediador entre regiones corticales sensoriales y motoras, como es el caso de POm. Las distintas poblaciones neuronales encontradas en S1BC procesan e integran la información táctil en microcircuitos tanto horizontales como verticales; en esta tesis doctoral se ha observado que las agrupaciones de neuronas supragranulares, granulares e infragranulares muestran un comportamiento diferencial en pruebas de competición entre estímulos somestésicos y auditivos bajo condiciones de inhibición de la Corteza Prefrontal medial (CPFm), sugiriendo una modulación córtico – cortical. El reclutamiento de diversas regiones prefrontales genera a corto plazo efectos de potenciación y depresión de la actividad talámica a través de S1BC. En esta línea, la estimulación a altas frecuencias de la Corteza Infralímbica (IL) facilita las respuestas evocadas táctiles tanto de POm como de VPM, mientras que la estimulación de la Corteza Prelímbica (PL) induce un decremento de dichas respuestas únicamente en el núcleo Posterior del Tálamo bajo las mismas condiciones experimentales. Dado que la inhibición ejercida por nRt podría no resultar suficiente para explicar este fenómeno en su totalidad, abordamos otros núcleos inhibitorios extra-talámicos que guardasen una relación estrecha entre la CPFm, S1BC y el Tálamo Somatosensorial. Una gran parte de las neuronas GABAérgicas de la región ventral de la Zona Incerta (ZIv) respondieron ante la estimulación de CPFm y el Colículo Superior como control; entorno al 40 % de las respuestas evocadas incertales se vieron facilitadas a corto plazo ante la estimulación de PL, modificando el estrecho rango temporal en el que esta estructura puede ser activada eficientemente. En este sentido, ZI juega un papel esencial en la modulación prefronto-talámica a través de sus proyecciones inhibitorias hacia POm. La inhibición incertal desenmascaró un efecto de potenciación a corto plazo en POm tras la activación sostenida de CPFm tanto en potenciales de campo, como en unidades

Tau protein is a microtubule-associated protein encoded by the MAPT gene that carries out a myriad of physiological functions and has been linked to certain pathologies collectively termed tauopathies, including Alzheimer’s disease, frontotemporal dementia, Huntington’s disease, progressive supranuclear palsy, etc. Alternative splicing is a physiological process by which cells generate several transcripts from one single gene and may in turn give rise to different proteins from the same gene. MAPT transcripts have been proven to be subjected to alternative splicing, generating six main isoforms in the central nervous system. Research throughout the years has demonstrated that the splicing landscape of the MAPT gene is far more complex than that, including at least exon skipping events, the use of 3′ and 5′ alternative splice sites and, as has been recently discovered, also intron retention. In addition, MAPT alternative splicing has been showed to be regulated spatially and developmentally, further evidencing the complexity of the gene’s splicing regulation. It is unclear what would drive the need for the existence of so many isoforms encoded by the same gene, but a wide range of functions have been ascribed to these Tau isoforms, both in physiology and pathology. In this review we offer a comprehensive up-to-date exploration of the mechanisms leading to the outstanding diversity of isoforms expressed from the MAPT gene and the functions in which such isoforms are involved, including their potential role in the onset and development of tauopathies such as Alzheimer’s disease, This research received no external funding

Resumen: Introducción: La enfermedad de Hirschsprung está causada por un defecto de la migración celular desde la cresta neural hasta el tracto gastrointestinal, resultando en la ausencia de neuronas en el plexo mientérico. Mutaciones en varios genes han sido asociadas a la enfermedad de Hirschsprung, la mayoría afectando a la vía del protooncogén RET. El objetivo de este estudio es la descripción de mutaciones tanto descritas como nuevas asociadas a la enfermedad de Hirschsprung, así como sus implicaciones pronósticas. Material y métodos: Análisis retrospectivo de pacientes con enfermedad de Hirschsprung y resultados genéticos positivos desde 1970 hasta 2013. Resultados: En la serie global, 21 pacientes tenían resultados genéticos positivos, 17 de ellos afectando la vía del protooncogén RET. Dos de las mutaciones son nuevas y no han sido previamente descritas en la literatura médica. Conclusiones: El protooncogén RET es el principal gen asociado a la enfermedad de Hirschsprung. Todavía hay múltiples mutaciones desconocidas relacionadas con la patogenia de la enfermedad. El estudio genético del gen RET debe formar parte del estudio diagnóstico de todos los pacientes con enfermedad de Hirschsprung, así como de sus familiares de primer grado en caso de que las mutaciones estén asociadas a los síndromes MEN2A y MEN2B. Abstract: Introduction: Hirschsprung disease is caused by an impairment in cell migration from the neural crest to the gastrointestinal tract, resulting in an absence of neurons in the myenteric plexus. Many mutations in several genes have been associated to Hirschsprung disease; most of them affecting the RET proto-oncogen pathway. The purpose of this study is the description of novel and known mutations in genes associated to Hirschsprung disease and their prognostic implications. Material and methods: Retrospective analysis of patients with Hirschsprung disease and positive genetic studies evaluated from 1970 to 2013. Results: We found 21 positive genetic studies in the global series, 17 of them involving the RET proto-oncogene. Two of the mutations are novel and they have not been reported in the medical literature. Conclusions: The RET protooncogene is the main gene associated with Hirschsprung disease. There are still multiple unknown mutations related to the pathogenesis of the disease. The study of this gene must be part of the work-up of all patients with Hirschsprung disease, as well as their first degree relatives if the mutation is associated with MEN2A and MEN2B syndromes.

© The Author(s) 2020, Pancreatic ductal adenocarcinoma (PDAC), the fourth leading cause of cancer death, has a 5-year survival rate of approximately 7–9%. The ineffectiveness of anti-PDAC therapies is believed to be due to the existence of a subpopulation of tumor cells known as cancer stem cells (CSCs), which are functionally plastic, and have exclusive tumorigenic, chemoresistant and metastatic capacities. Herein, we describe a 2D in vitro system for long-term enrichment of pancreatic CSCs that is amenable to biological and CSC-specific studies. By changing the carbon source from glucose to galactose in vitro, we force PDAC cells to utilize OXPHOS, resulting in enrichment of CSCs defined by increased CSC biomarker and pluripotency gene expression, greater tumorigenic potential, induced but reversible quiescence, increased OXPHOS activity, enhanced invasiveness, and upregulated immune evasion properties. This CSC enrichment method can facilitate the discovery of new CSC-specific hallmarks for future development into targets for PDAC-based therapies., We acknowledge and thank Dr. Nuria Malats and Jaime Villarreal from the Spanish National Cancer Research Center (CNIO) for RNA sequencing and analysis, funded by Fondo de Investigaciones Sanitarias (FIS) grant PI18/01347. We thank Patricia Sánchez-Tomero and Marina Ochando-Garmendia for technical assistance and support and Dr. Raúl Sánchez Lanzas for assistance with autophagy experiments. We want to particularly acknowledge the patients and the BioBank Hospital Ramón y Cajal-IRYCIS (PT13/0010/0002) integrated in the Spanish National Biobanks Network for its collaboration and, in particular, Adrián Povo Retana for macrophage isolation. We would also like to thank the Transmission Electron Microscopy Unit Laboratory, part of the UAM Interdepartmental Investigation Service (SIdI); Coral Pedrero for exceptional help with in vivo experiments; and the laboratories of Dr. Amparo Cano and Dr. José González Castaño for reagents and helpful discussions. S.V. was a recipient of an Ayuda de Movilidad del Personal Investigador del IRYCIS, a mobility grant from the Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, Spain, and a pre-doctoral fellowship from the Comunidad de Madrid, Ayudas Para La Contratación De Investigadores Predoctorales Y Posdoctorales (PEJD-2017-PRE/BMD-5062), Madrid, Spain. This study was supported by a Rámon y Cajal Merit Award (RYC-2012-12104) from the Ministerio de Economía y Competitividad, Spain (to B.S.); funding from la Beca Carmen Delgado/Miguel Pérez-Mateo from AESPANC-ACANPAN Spain (to B.S.); a Conquer Cancer Now Grant from the Concern Foundation (Los Angeles, CA, USA) (to B.S.); a Coordinated grant (GC16173694BARB) from the Fundación Asociación Española Contra el Cáncer (AECC) (to B.S.); FIS grants PI18/00757 (to B.S.), PI16/00789 (to M.A.F.-M.), PI18/00267 (to L.G.-B.; co-financed through Fondo Europeo de Desarrollo Regional (FEDER) “Una manera de hacer Europa”); a Miguel Servet award (CP16/00121) (to P.S.); a Max Eder Fellowship of the German Cancer Aid (111746) (to P.C.H.); and the German Research Foundation (DFG, CRC 1279 “Exploiting the human peptidome for Novel Antimicrobial and Anticancer Agents”; to P.C.H.).

Klinefelter syndrome (KS) is the most common sex chromosome aneuploidy. A distinctive characteristic of KS is oligozoospermia. Despite multiple studies that have described the natural history of the degenerative process of germ cells in patients with KS, the molecular mechanisms that initiate this process are not well characterized. MicroRNA (miRNA)-mediated post-transcriptional control mechanisms have been increasingly recognized as important regulators of spermatogenesis; however, only a few studies have evaluated the role of miRNAs in the gonadal failure of these patients. Here, we describe a differential expression profile for the miRNAs in testicular tissue samples taken from KS patients. We analysed testicular tissue samples from 4 KS patients and 5 control patients (obstructive azoospermia) through next-generation sequencing, which can provide information about the mechanisms involved in the degeneration of germ cells. A distinctive differential expression profile was identified for 166 miRNAs in the KS patients: 66 were upregulated, and 100 were downregulated. An interactome analysis was performed for 7 of the upregulated and the 20 downregulated miRNAs. The results showed that the target genes are involved in the development, proliferation, and differentiation processes of spermatogenesis, which may explain their role in the development of infertility. This is the first report of a miRNA expression profile generated from testicular tissue samples of KS patients.

To evaluate the relationship of the immune-checkpoint PD-1/PD-L1 with the clinical evolution of OSCC; to assess survival in OSCC based on the characteristics of TME and histologic risk score; to evaluate the clinical and histopathological relationship of OSCC with immunological TME. Material and Methods: A retrospective study was carried out on 65 samples from patients with OSCC on the floor of the mouth or tongue. Clinicopathological variables and the expression of the biomarkers PD-1, PD-L1, FoxP3, CD4, CD8, CSF1R, and p16 were recorded. The relationship of the clinical and histological variables with the expression of the biomarkers and survival was studied. Results: The univariate and multivariate analysis indicated that positive PD-1 expression was an independent protective factor for survival (overall, disease-free, disease-specific survival) and that high PD-L1 also improved survival. Poorly differentiated histological grades and metastasis were associated with a worse prognosis. PD-1 is a protective survival factor that is maintained independently of PD-L1 expression. High values of PD-L1 expression also improve survival. Higher expression of PD-1 is observed in smaller tumors, and higher expression of PD-L1 is more likely in women. No relationship between the tumor microenvironment and histologic risk score was found to influence the survival patterns studied in the OSCC. There is no evidence of a relationship between the histopathological features and the studied markers, although the positive PD-1 and PD-L1 cases have a lower risk of a high WPOI score, and positive PD-1 expression was associated with a lower DOI, This research was funded by the Fundación para la investigación Biomédica Hospital Universitario La Paz with the project number: EC_5474. The research has been partially funded through a predoctoral research grant awarded by the Official College of Dentists of Madrid (Madrid, Spain) with not project number associated

Shikonin is an ointment produced from Lithospermun erythrorhizon which has been used in traditional medicine both in Europe and Asia for wound healing and is associated with anti-inflammatory properties. The goal of this work is to assess the analgesic properties of Shikonin in the CFA-induced inflammation model of pain. Rats were subjected to inflammation of the hind paw by CFA injection with a preventive injection of Shikonin and compared to either a control group or to a CFA-inflamed group with the vehicle drug solution. Inflammation of the hind paw by CFA was assessed by measurement of the dorsal to plantar diameter. Mechanical thresholds were established by means of the Von Frey filaments which are calibrated filaments that exert a defined force. Finally, the spinal cord of the studied animals was extracted to analyse the microglia population through immunohistochemistry using the specific marker Iba-1. Our results show that Shikonin reduces the paw oedema caused by CFA inflammation. Subsequently, there is a concomitant restoration of the mechanical thresholds reduced by CFA hind paw injection. Additionally, spinal microglia is activated after CFA-induced inflammation. Our results show that microglia is inhibited by Shikonin and has concomitant restoration of the mechanical thresholds. Our findings demonstrate for the first time that Shikonin inhibits microglia morphological changes and thereby ameliorates pain-like behaviour elicited by mechanical stimulation, This work was supported by the Universidad Europea de Madrid (Grant number: 2020/UEM38). We would like to specially thank to Dr Arenillas, Hospital Universitario de Getafe (Spain) for his support and advice in animal welfare

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia . Fecha de Lectura: 02-02-2022, El síndrome de piernas inquietas (SPI) es un trastorno neurológico frecuente, con una prevalencia en nuestro medio estimada en 2-5%. En las últimas décadas se ha investigado más acerca del papel del hierro esta enfermedad, descubriéndose la existencia de un déficit de hierro en determinadas regiones cerebrales de estos pacientes. De éstas, la más consistentemente estudiada es la sustancia negra (SN) mesencefálica, estructura anatómicamente relacionada con control motor. La ecografía transcraneal (TCS) es una técnica de neuroimagen que permite cuantificar de una forma fiable, económica y no invasiva los depósitos de hierro en la SN, obteniendo el índice de ecogenicidad de SN (SNEI). Inicialmente, se presentarán dos artículos de revisión, el primero acerca del SPI y la evidencia actual sobre este trastorno, y el segundo acerca de las terapias de reposición de hierro recomendadas para tratar el SPI. A continuación se presentan tres artículos originales: un primer artículo acerca de la posible correlación entre parámetros de ferrocinética en suero y valores de hierro cerebral mediante SNEI en SPI. El siguiente acerca de la utilidad del SNEI para predecir respuesta clínica al tratamiento con hierro intravenoso en SPI, y finalmente un tercero acerca del riesgo de producir sobrecarga férrica tras el uso de hierro intravenoso en pacientes SPI

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 08-07-2022

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 26-05-2022, La aparición de los signos cardinales de la Enfermedad de Parkinson (EP) correlaciona con la pérdida de las neuronas dopaminérgicas de la sustancia negra compacta (SNc) y la consecuente depleción dopaminérgica en el estriado. El conocimiento sobre la integridad celular del estriado y de los distintos grupos dopaminérgicos mesencefálicos, así como sus conexiones, es imprescindible para entender la fisiopatología de la EP. Esta Tesis Doctoral aporta una caracterización de las poblaciones neuronales del estriado y de la SNc del cerebro de monos control y en el modelo de EP por intoxicación con MPTP (1-metil-4-fenil-1,2,3,6 tetrahidropiridina); incluye la descripción y densidad de las diferentes poblaciones neuronales del estriado, así como la integridad de estas poblaciones tras la depleción dopaminérgica estriatal. También se incluye una caracterización de diferentes subpoblaciones dopaminérgicas del mesencéfalo y sus proyecciones diferenciales. Las neuronas de proyección en el mono comprenden un 86% de la población estriatal mientras que las interneuronas comprenden el 14% restante. Las interneuronas siguen un gradiente de densidad, siendo las más abundantes las que expresan calretinina (Cr+), seguido de las parvalbúminas (Pv+), las nitrérgicas (NADPH+), las colinérgicas (ChAT+) y las que expresan tirosina hidroxilasa (TH+). Además, el tratamiento con MPTP no afectó a la densidad de ningún tipo neuronal en el caudado, putamen o estriado ventral, a excepción de las interneuronas TH+, que aumentaron hasta el doble. En el mesencéfalo de mono existen diversas subpoblaciones de neuronas dopaminérgicas. La expresión de calbindina (Cb+) y el canal GIRK2 permite distinguir entre las neuronas de la SNc dorsal y el área tegmental ventral (VTA), y las de la SNc ventral. Las neuronas Cb+ apenas se pierden tras el tratamiento con MPTP, mientras que las neuronas GIRK2 de la SNc ventral son muy vulnerables al mismo. La mayor parte las neuronas GIRK2+ co-expresan la enzima aldehído deshidrogenasa (ALDH1A1). La enzima ALDH1A1 se expresa característicamente en las columnas de neuronas que dirigen sus dendritas ventralmente hacia la SN reticulata, formando unas estructuras que se conocen como “dendron-bouquets”, implicadas en la conexión con las fibras estriadonigrales. La proyección nigroestriada se define molecularmente por la expresión de ALDH1A1 en las neuronas de origen, localizadas en la zona más ventral de la SNc. En cambio, la proyección nigropalidal tiene su origen en neuronas diferentes a las de la proyección nigroestriada, ya que la mayoría no expresan ALDH1A1. Al contrario que la nigroestriada, la inervación dopaminérgica del globo pálido en monos parkinsonianos está muy preservada. Los resultados de este trabajo apoyan la hipótesis de una segregación de proyecciones en el que las neuronas Cb+ dan origen a la vía nigropalidal, mientras que las neuronas GIRK2+/ALDH1A1+ dan lugar a la vía nigroestriada, siendo este fenotipo el que se asocia con mayor vulnerabilidad neuronal en este modelo de EP

Aging is accompanied by a decline in cognition that can be due to a lower IGF-I level. We studied response facilitation induced in primary somatosensory (S1) cortical neurons by repetitive stimulation of whiskers in young and old mice. Layer 2/3 and 5/6 neurons were extracellularly recorded in young (≤ 6 months of age) and old (≥ 20 month of age) anesthetized mice. IGF-I injection in S1 cortex (10 nM; 0.2 µL) increased whisker responses in young and old animals. A stimulation train at 8 Hz induced a long-lasting response facilitation in only layer 2/3 neurons of young animals. However, all cortical neurons from young and old animals showed long-lasting response facilitation when IGF-I was applied in the S1 cortex. The reduction in response facilitation in old animals can be due to a reduction in the IGF-I receptors as was indicated by the immunohistochemistry study. Furthermore, a reduction in the performance of a whisker discrimination task was observed in old animals. In conclusion, our findings indicate that there is a reduction in the synaptic plasticity of S1 neurons during aging that can be recovered by IGF-I. Therefore, it opens the possibility of use IGF-I as a therapeutic tool to ameliorate the effects of heathy aging., This work was supported by Grants from Ciberned and the Spanish Ministerio de Ciencia e Innovación (PID2019-107809RB-I00; PID2019-104376RB-I00; AEI/FEDER; MICINN).

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 14-07-2021, Esta tesis tiene embargado el acceso al texto completo hasta el 14-01-2023, El presente trabajo ha sido posible gracias a la financiación del I SCIII [CP13-0 0234, CP19 -0 010]; MINECO [SAF2013 - 4 5570 -R, SAF2016 -7 8666R] , FEDER; Proyecto N DG09/014 de la A gencia Laín Entralgo; Proyecto V EXEM; I RYCIS; Fondo Social Europeo a través del Programa Operativo de Empleo Juvenil y la I niciativa de Empleo Juvenil (YEI) d e la Comunidad A utónoma de Madrid [ P EJ16/MED/AI -1 153 , P EJD -2 018 -P RE/SAL -8 532 ].

Craniofacial neuropathic pain affects millions of people worldwide and is often difficult to treat. Two key mechanisms underlying this condition are a loss of the negative control exerted by inhibitory interneurons and an early microglial reaction. Basic features of these mechanisms, however, are still poorly understood. Using the chronic constriction injury of the infraorbital nerve (CCI-IoN) model of neuropathic pain in mice, we have examined the changes in the expression of GAD, the synthetic enzyme of GABA, and GlyT2, the membrane transporter of glycine, as well as the microgliosis that occur at early (5 days) and late (21 days) stages post-CCI in the medullary and upper spinal dorsal horn. Our results show that CCI-IoN induces a down-regulation of GAD at both postinjury survival times, uniformly across the superficial laminae. The expression of GlyT2 showed a more discrete and heterogeneous reduction due to the basal presence in lamina III of ‘patches’ of higher expression, interspersed within a less immunoreactive ‘matrix’, which showed a more substantial reduction in the expression of GlyT2. These patches coincided with foci lacking any perceptible microglial reaction, which stood out against a more diffuse area of strong microgliosis. These findings may provide clues to better understand the neural mechanisms underlying allodynia in neuropathic pain syndromes., Ayudas a Proyectos de Investigación UFV2021-16” from Universidad Francisco de Vitoria, and Project “797011—Introducción Técnicas Estereológicas”, from the Fundación General de la Universidad Autónoma de Madrid

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 21-01-2021, The hippocampus plays a crucial role in spatial orientation, learning and memory, and many pathological conditions (e.g., epilepsy and Alzheimer’s disease (AD)) are closely associated with synaptic alterations in this region. Numerous studies have described the ultrastructural characteristics of hippocampal synapses in experimental animals. However, there is very little information about the synaptic organization of the human hippocampus and the brain in general, which is a major problem when it comes to ap-propriately interpreting alterations in disease. A major goal in neuroscience is to di-rectly study the human brain. In the present study, we started to address this issue by analyzing the synaptic organization of the hippocampus, focusing on the CA1 field. The CA1 field receives and integrates a massive amount of information in a laminar-specific manner, and sends projections mainly to the subiculum and to subcortical nuclei and polymodal association cortices. This region is severely affected by amyloid and tau pa-thologies in AD, which are related to neuronal degeneration and synaptic alterations that seem to constitute the major neurobiological basis of cognitive dysfunction in AD. Studying the human brain via electron microscopic techniques presents certain prob-lems and the scarcity of human brain tissue suitable for ultrastructural studies is one of the most important issues to overcome. Recently, we have shown that Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) technology can be applied to study in detail the 3D synaptic organization of the human brain obtained from autopsies, yield-ing excellent results. A variety of synaptic parameters can be studied, including synap-tic density, type and spatial distribution, postsynaptic targets and the shape and size of the synaptic junctions. Using this technology, 33,705 synapses were fully reconstructed in the neuropil in all CA1 layers in five control cases with no apparent neurological al-teration and in CA1 stratum pyramidale and radiatum in five AD cases with different stages of the disease. In controls, we found that most synapses are excitatory, targeting dendritic spines and displaying a macular shape, regardless of the layer examined. However, remarkable differences were observed between layers. In AD, cases with ear-ly stages of the disease had morphologically normal-looking synapses and we observed no significant differences with regard to many synaptic parameters, including synaptic density. However, differences in the distribution of postsynaptic targets and synaptic shapes, and a lower proportion of larger excitatory synapses were found in AD cases. Individuals in late stages of the disease suffered the most severe synaptic alterations, including a decrease in synaptic density and morphological alterations of the remaining synapses. Since AD cases showed cortical atrophy, our data indicate a reduction in the total number (but not the density) of synapses at early stages of the disease, with this reduction being much more accentuated in subjects with late stages of AD. The ob-served synaptic alterations may represent a structural basis for the progressive learn-ing and memory dysfunctions seen in AD cases. Taken together, these data constitute the first extensive description of the synaptic organization of the neuropil of the human CA1 field and its possible alterations in AD, which is a necessary step for better under-standing its functional organization in both health and disease, This study was funded by grants from the Spanish “Ministerio de Ciencia e Innovacio n” (grant PGC2018-094307-B-I00 I00 and the Cajal Blue Brain Project [the Spanish partner of the Blue Brain Project initiative from EPFL, Switzerland]), the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement No. 785907 (Human Brain Project, SGA2), Centro de Investigacio n Biome dica en Red sobre Enfermedades Neurodegenerativas (CIBERNED, Spain, CB06/05/0066), the Alzheimer’s Association (ZEN-15-321663) and the UNED (Plan de Promocio n de la Investigacio n, 2014-040-UNED-POST). Marta Montero Crespo was awarded a research fellowship from the Spanish “Ministerio de Educacio n, Cultura y Deporte” (contract FPU14/02245)

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 15-10-2021, Esta tesis tiene embargado el acceso al texto completo hasta el 15-04-2023

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de lectura: 11-05-2021, Se desconoce en gran medida cómo el sistema somestésico extrae información del flujo de señales sensoriales. La información procedente de las vibrisas es procesada principalmente por dos vías ascendentes paralelas hacia la corteza cerebral. Sin embargo, se desconoce la implicación funcional de las diferentes estructuras que componen dichas rutas. Mediante la combinación de técnicas electrofisiológicas y farmacológicas in vivo en ratas, encontramos diferencias significativas entre estas vías. Aunque está bien asumido que el POm y el VPM responden a la estimulación de las vibrisas contralaterales, encontramos que el primero es capaz de responder también a las ipsilaterales. Mediante la integración de señales simultaneas procedentes de vibrisas en ambos lados de la cara, está implicado en la representación de eventos táctiles bilaterales. Esto demuestra la implicación de los núcleos talámicos sensoriales de tipo 'higher-order' en la percepción bilateral. Encontramos que los núcleos POm están mutuamente conectados a través de la corteza formando un bucle o 'loop' funcional. Revelamos la naturaleza y el contenido de los mensajes transmitidos a través de este circuito mostrando que dichos mensajes son 'patrones estructurados de actividad sostenida'. Estos mensajes son transmitidos preservando su estructura integrada. La implicación de diferentes áreas fue investigada descubriendo que S1 juega un papel protagonista en dicho 'loop' POm-POm. También encontramos diferente implicación laminar en esta área en el procesamiento de actividad sostenida y en su transmisión entre hemisferios. Proponemos un modelo teórico en el que dichos 'patrones estructurados de actividad sostenida' generados por el POm pueden jugar un papel relevante en las funciones perceptivas, motoras y cognitivas. Además, demostramos que el POm está involucrado en la representación de patrones sensoriales complejos. Este núcleo es muy sensible a la activación simultanea de las vibrisas y a las complejas interacciones espaciotemporales que se producen entre ellas. La estructura espaciotemporal de dichos patrones y la complejidad de sus partes son reflejados en precisos cambios de actividad en el POm. Nuestros resultados sugieren que este núcleo podría ser un codificador general de patrones. La naturaleza (estructurada versus discreta), el tipo (sostenido versus transitorio) y el contenido (integrado versus segregado) de la actividad neural procesada y transmitida por estos núcleos determina su implicación funcional y puede permitir clasificarlos. Proponemos la hipótesis de los Componentes Complementarios para explicar estas diferencias. Además, revelamos la capacidad del POm para ajustar el procesamiento en las cortezas S1 y S2 mediante la inducción de una precisa inhibición en determinadas capas corticales. Esta modulación está mediada por neuronas GABAérgicas de la capa 1. La hipótesis de Computación Cortical por Resultados Discretos propuesta aquí puede explicar la implicación funcional de dicho ajuste

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 21-01-2021 The hippocampus plays a crucial role in spatial orientation, learning and memory, and many pathological conditions (e.g., epilepsy and Alzheimer’s disease (AD)) are closely associated with synaptic alterations in this region. Numerous studies have described the ultrastructural characteristics of hippocampal synapses in experimental animals. However, there is very little information about the synaptic organization of the human hippocampus and the brain in general, which is a major problem when it comes to ap-propriately interpreting alterations in disease. A major goal in neuroscience is to di-rectly study the human brain. In the present study, we started to address this issue by analyzing the synaptic organization of the hippocampus, focusing on the CA1 field. The CA1 field receives and integrates a massive amount of information in a laminar-specific manner, and sends projections mainly to the subiculum and to subcortical nuclei and polymodal association cortices. This region is severely affected by amyloid and tau pa-thologies in AD, which are related to neuronal degeneration and synaptic alterations that seem to constitute the major neurobiological basis of cognitive dysfunction in AD. Studying the human brain via electron microscopic techniques presents certain prob-lems and the scarcity of human brain tissue suitable for ultrastructural studies is one of the most important issues to overcome. Recently, we have shown that Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) technology can be applied to study in detail the 3D synaptic organization of the human brain obtained from autopsies, yield-ing excellent results. A variety of synaptic parameters can be studied, including synap-tic density, type and spatial distribution, postsynaptic targets and the shape and size of the synaptic junctions. Using this technology, 33,705 synapses were fully reconstructed in the neuropil in all CA1 layers in five control cases with no apparent neurological al-teration and in CA1 stratum pyramidale and radiatum in five AD cases with different stages of the disease. In controls, we found that most synapses are excitatory, targeting dendritic spines and displaying a macular shape, regardless of the layer examined. However, remarkable differences were observed between layers. In AD, cases with ear-ly stages of the disease had morphologically normal-looking synapses and we observed no significant differences with regard to many synaptic parameters, including synaptic density. However, differences in the distribution of postsynaptic targets and synaptic shapes, and a lower proportion of larger excitatory synapses were found in AD cases. Individuals in late stages of the disease suffered the most severe synaptic alterations, including a decrease in synaptic density and morphological alterations of the remaining synapses. Since AD cases showed cortical atrophy, our data indicate a reduction in the total number (but not the density) of synapses at early stages of the disease, with this reduction being much more accentuated in subjects with late stages of AD. The ob-served synaptic alterations may represent a structural basis for the progressive learn-ing and memory dysfunctions seen in AD cases. Taken together, these data constitute the first extensive description of the synaptic organization of the neuropil of the human CA1 field and its possible alterations in AD, which is a necessary step for better under-standing its functional organization in both health and disease This study was funded by grants from the Spanish “Ministerio de Ciencia e Innovacio n” (grant PGC2018-094307-B-I00 I00 and the Cajal Blue Brain Project [the Spanish partner of the Blue Brain Project initiative from EPFL, Switzerland]), the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement No. 785907 (Human Brain Project, SGA2), Centro de Investigacio n Biome dica en Red sobre Enfermedades Neurodegenerativas (CIBERNED, Spain, CB06/05/0066), the Alzheimer’s Association (ZEN-15-321663) and the UNED (Plan de Promocio n de la Investigacio n, 2014-040-UNED-POST). Marta Montero Crespo was awarded a research fellowship from the Spanish “Ministerio de Educacio n, Cultura y Deporte” (contract FPU14/02245)

[EN] A robotic exoskeleton is an electromechanical device that can be worn by a person to increase its physical capacity, to assist locomotion or for gait rehabilitation processes. In the case of rehabilitation exoskeletons, the control system is required to be smooth and capable to match accurately with the patients’ evolution in order to optimize the eciency of their recovery, this implies the design of robust and precise controllers. In this paper, kinematic analysis, dynamic analysis and control system evaluation for ALICE rehabilitation exoskeleton is presented. Among the control techniques used are: the PD controller, adaptive PD, and the sliding mode controller. In addition, a stability analysis using the Lyapunov criterion is performed. To test the performance of the controllers, gait data obtained by the ONCE School of Physiotherapy in Madrid, which correspond to healthy people and people with multiple sclerosis, are used. MATLAB as simulation software and programming language is used., [ES] Un exoesqueleto robótico es un dispositivo electromecánico utilizado para aumentar la capacidad física de una persona, como ayuda a la locomoción o para procesos de rehabilitación de la marcha. En el caso de los exoesqueletos de rehabilitación se requiere que el sistema de control sea capaz de adaptarse adecuadamente a la evolución del paciente con el fin de optimizar su recuperación, esto implica el diseño de controladores robustos y precisos. En este trabajo se presenta el análisis cinemático, análisis dinámico y evaluación del sistema de control del exoesqueleto de rehabilitación ALICE. Dentro de las técnicas de control presentadas se encuentran: el controlador PD, PD adaptativo, y el controlador en modo deslizante. Además, se realiza un análisis de estabilidad utilizando el criterio de Lyapunov. Para probar el rendimiento de los reguladores, se utiliza un conjunto de datos de la Escuela de Fisioterapia de la ONCE de Madrid, correspondiente a personas sanas y personas con esclerosis múltiple. Se utiliza MATLAB como software de simulación y lenguaje de programación., Manuel Cardona agradece a la Fundación Carolina y a la Universidad Politécnica de Madrid, España, por el apoyo para la realización de esta investigación gracias a la beca de Doctorado otorgada en el marco del convenio con la Universidad Don Bosco, El Salvador.

Nervous systems respond with structural changes to environmental changes even in adulthood. In recent years, experience-dependent structural plasticity was shown not to be restricted to the cerebral cortex, as it also occurs at subcortical and even peripheral levels. We have previously shown that two populations of trigeminal nuclei neurons, trigeminothalamic barrelette neurons of the principal nucleus (Pr5), and intersubnuclear neurons in the caudal division of the spinal trigeminal nucleus (Sp5C) that project to Pr5 underwent morphometric and topological changes in their dendritic trees after a prolonged total or partial loss of afferent input from the vibrissae. Here we examined whether and what structural alterations could be elicited in the dendritic trees of the same cell populations in young adult rats after being exposed for 2 months to an enriched environment (EE), and how these changes evolved when animals were returned to standard housing for an additional 2 months. Neurons were retrogradely labeled with BDA delivered to, respectively, the ventral posteromedial thalamic nucleus or Pr5. Fully labeled cells were digitally reconstructed with Neurolucida and analyzed with NeuroExplorer. EE gave rise to increases in dendritic length, number of trees and branching nodes, spatial expansion of the trees, and dendritic spines, which were less pronounced in Sp5C than in Pr5 and differed between sides. In Pr5, these parameters returned, but only partially, to control values after EE withdrawal. These results underscore a ubiquity of experience-dependent changes that should not be overlooked when interpreting neuroplasticity and developing plasticity-based therapeutic strategies., This study was supported by Grants BFU2012-39960 from Spain’s Ministerio de Economía y Competitividad/Fondo Europeo para el Desarrollo Regional (MINECO/FEDER), and “Ayudas a Proyectos de Investigación UFV2021-16” from Universidad Francisco de Vitoria

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de Lectura: 04-10-2021, Esta tesis tiene embargado el acceso al texto completo hasta el 04-04-2023, La enfermedad de Alzheimer (EA) es una enfermedad neurodegenerativa que constituye la causa más común de demencia entre las personas de edad avanzada cuyas principales alteraciones neuropatológicas son la acumulación de β-amiloide (Aβ) y la hiperfosforilación de la proteína tau. Numerosos estudios han propuesto que el péptido Aβ podría ser responsable de alteraciones en la función mitocondrial en la EA. En esta tesis se ha estudiado el efecto de Aβ sobre diversos aspectos de la dinámica mitocondrial a lo largo de la evolución de esta patología. Para ello se utilizaron modelos de EA in vivo (ratones APP/PS1) e in vitro. Nuestros resultados indicaron que la sobrecarga de Aβ provocaba alteraciones en la dinámica mitocondrial, incluyendo una disminución de la biogénesis y un desbalance de las proteínas de fusión y fisión mitocondrial que culmina en un incremento de la mitofagia. Estas alteraciones son precedidas por una interacción directa del péptido Aβ en el metabolismo y en la función mitocondrial. Nuestros resultados muestran que, además de mitofagia, la sobrecarga de Aβ produce un aumento de la autofagia, tal como mostró el incremento en los niveles de sus principales marcadores en los diferentes modelos experimentales. El mecanismo de autofagia en la EA juega un importante papel tanto en la eliminación del péptido amiloide como en la eliminación de orgánulos dañados, incluyendo las mitocondrias, cuya acumulación puede resultar tóxica para las células. Una reducción excesiva en la cantidad de mitocondrias por el efecto tóxico del péptido Aβ puede tener consecuencias fatales para las neuronas, resultando en una reducción en la producción de energía confiada plenamente a las mitocondrias. Finalmente, se analizó el efecto de un inhibidor de fosfodiesterasa (PDE) 7, el compuesto S14, sobre las alteraciones previamente atribuidas a la sobrecarga de Aβ, relacionadas con la dinámica mitocondrial y con los procesos de autofagia. Nuestros datos indicaron que el compuesto S14 revertía los efectos del péptido Aβ sobre la dinámica mitocondrial y autofagia tanto en los modelos animales como celulares. Este trabajo sugiere que el tratamiento con S14 podría ser beneficioso como herramienta terapéutica para la EA, El trabajo aquí presentado ha sido realizado con cargo a los siguientes proyectos: Ministerio de Sanidad/Instituto de Salud Carlos III • PI18/00118 (2019-2021): “Validación de la proteína salival Lactoferrina como biomarcador de la enfermedad de Alzheimer: alternativa al diagnóstico y mecanismo patogénico” • PI15/00780 (2016-2018): “Inflamación periférica como factor de riesgo en la patología de la enfermedad de Alzheimer: nuevas herramientas diagnósticas y dianas terapéuticas”. • CIBERNED (PI2016/01) (2017-2018): “Alteraciones del metabolismo glucolipídico y desarrollo de la demencia de Alzheimer”. • PI12/00486 (2013-2015): “Asociación de alteraciones moleculares y funcionales de los plexos coroideos con la severidad, patología y progresión de la enfermedad de Alzheimer”. Comunidad de Madrid • S2017/BMD-3700 (NEUROMETAB-CM) (2018-2021): “Bases Metabólicas de la Neurodegeneración”

The human cerebral cortex is parcellated in hundreds of areas using neuroanatomy and imaging methods. Alternatively, cortical areas can be classified into few cortical types according to their degree of laminar differentiation. Cortical type analysis is based on the gradual and systematic variation of laminar features observed across the entire cerebral cortex in Nissl stained sections and has profound implications for understanding fundamental aspects of evolution, development, connections, function, and pathology of the cerebral cortex. In this protocol paper, we explain the general principles of cortical type analysis and provide tables with the fundamental features of laminar structure that are studied for this analysis. We apply cortical type analysis to the micrographs of the Atlas of the human cerebral cortex of von Economo and Koskinas and provide tables and maps with the areas of this Atlas and their corresponding cortical type. Finally, we correlate the cortical type maps with the T1w/T2w ratio from widely used reference magnetic resonance imaging scans. The analysis, tables and maps of the human cerebral cortex shown in this protocol paper can be used to predict patterns of connections between areas according to the principles of the Structural Model and determine their level in cortical hierarchies. Cortical types can also predict the spreading of abnormal proteins in neurodegenerative diseases to the level of cortical layers. In summary, cortical type analysis provides a theoretical and practical framework for directed studies of connectivity, synaptic plasticity, and selective vulnerability to neurologic and psychiatric diseases in the human neocortex., This work was supported by the National Institute of Mental Health (grant nos. R01 MH101209 and R01 MH118500). MG-C was the recipient of a Beatriz Galindo senior research position in the Faculty of Medicine at Universidad Autónoma de Madrid (BEAGAL18/00098) and of a Grant for ICD Projects for the Beatriz Galindo Program Researchers at Universidad Autónoma de Madrid (SI2/PBG/2020-00014)

Insulin-like growth factor-I (IGF-I) signaling plays a key role in learning and memory. IGF-I increases the spiking and induces synaptic plasticity in the mice barrel cortex (Noriega-Prieto et al., 2021), favoring the induction of the long-term potentiation (LTP) by Spike Timing-Dependent Proto-cols (STDP) (Noriega-Prieto et al., 2021). Here, we studied whether these IGF-I effects depend on endocannabinoids (eCBs) and nitric oxide (NO). We recorded both excitatory postsynaptic currents (EPSCs) and inhibitory postsynaptic currents (IPSCs) evoked by stimulation of the basal dendrites of layer II/III pyramidal neurons of the Barrel Cortex and analyzed the effect of IGF-I in the presence of a CB1 R antagonist, AM251, and inhibitor of the NO synthesis, L-NAME, to prevent the eCBs and the NO-mediated signaling. Interestingly, L-NAME abolished any modulatory effect of the IGF-I-induced excitatory and inhibitory transmission changes, suggesting the essential role of NO. Surprisingly, the inhibition of CB1Rs did not only block the potentiation of EPSCs but reversed to a depression, highlighting the remarkable functions of the eCB system. In conclusion, eCBs and NO play a vital role in deciding the sign of the effects induced by IGF-I in the neocortex, suggesting a neuromodulatory interplay among IGF-I, NO, and eCBs, This research was funded by MINECO and MICINN grants number BFU2016-80802-P AEI/FEDER, UE (MINECO) and PID2020-119358GB-I00/AEI/10.13039/501100011033 (MICINN).The APC was funded by PID2020-119358GB-I00/AEI/10.13039/501100011033 (MICINN)

Classification of tissues is an important problem in biomedicine. An efficient tissue classification protocol allows, for instance, the guided-recognition of structures through treated images or discriminating between healthy and unhealthy regions (e.g., early detection of cancer). In this framework, we study the potential of some polarimetric metrics, the so-called depolarization spaces, for the classification of biological tissues. The analysis is performed using 120 biological ex vivo samples of three different tissues types. Based on these data collection, we provide for the first time a comparison between these depolarization spaces, as well as with most commonly used depolarization metrics, in terms of biological samples discrimination. The results illustrate the way to determine the set of depolarization metrics which optimizes tissue classification efficiencies. In that sense, the results show the interest of the method which is general, and which can be applied to study multiple types of biological samples, including of course human tissues. The latter can be useful for instance, to improve and to boost applications related to optical biopsy., Agència de Gestió d'Ajuts Universitaris i de Recerca, Grant/Award Number: 2017-SGR-001500; Ministerio de Economía y Competitividad, Grant/Award Numbers: Fondos FEDER, RTI2018-097107-B-C31

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de lectura: 28-02-2020, Esta tesis tiene embargado el acceso al texto completo hasta el 28-08-2021

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de lectura: 28-02-2020, Esta tesis tiene embargado el acceso al texto completo hasta el 28-08-2021, La pérdida de dopamina en la enfermedad de Parkinson (EP) afecta intensamente al sistema nigroestriatal. Esclarecer cómo la pérdida de dopamina afecta a las estructuras cerebrales más allá del estriado es, sin embargo, necesario para comprender mejor una serie de mecanismos patogénicos y manifestaciones clínicas de la enfermedad aun mal definidos. El tálamo en primates está densa y heterogéneamente inervado por axones dopaminérgicos, la mayoría de los cuales expresan el transportador de dopamina (DAT), al igual que los del sistema nigroestriatal. Nuestra hipótesis es que en el tálamo del cerebro parkinsoniano hay déficit de dopamina, que es variable en las diversas fases de la enfermedad, y que es heterogéneo en los diversos núcleos del tálamo. Utilizamos tejido cerebral de macacos adultos sometidos o no a un protocolo de intoxicación lenta con la toxina 1-metil-4-fenil-1,2,3,6-tetrahidropiridina (MPTP). Los macacos tratados con MPTP se distribuyeron en cuatro grupos según su estado motor: asintomático, recuperado, parkinsoniano leve y parkinsoniano grave. La inervación dopaminérgica fue estudiada mediante inmunohistoquímica frente a DAT. Realizamos mapas topográficos de la distribución de axones immunoreactivos para DAT (DAT-ir) en el tálamo y estimamos la longitud total y la densidad de longitud de los axones DATir utilizando un fraccionador 3D en los núcleos dorsomediano (MD), centromedianoparafascicular (CnMd-Pf), ventrales y reticular. En comparación con los macacos control, los macacos tratados con MPTP mostraron una menor densidad de longitud axónica DAT-ir en los núcleos MD y CnMd-Pf. En MD la denervación de dopamina ya estaba presente en macacos del grupo asintomático. En el núcleo reticular, los macacos tratados con MPTP tuvieron un aumento de la densidad de longitud axónica DAT-ir en relación con los controles. La densidad de longitud de axones DAT-ir en los núcleos ventrales no mostró diferencias entre grupos. Los resultados de nuestro trabajo muestran una reacción heterogénea al MPTP de los axones dopaminérgicos que inervan el tálamo. Los cambios en la inervación dopaminérgica talámica pueden contribuir a una disfunción en la transmisión talamocortical, talamoestriada, e intratálamica, y ayudan a explicar algunas manifestaciones motoras y no motoras de la EP, como defectos de función ejecutiva y de atención, así como trastornos del sueño, Dopamine loss in Parkinson’s disease (PD) affects intensely the nigrostriatal system. Understanding how dopamine loss affects brain structures beyond the striatum is, nonetheless, an unmet need to better understand a series of ill-defined pathogenic mechanisms and clinical manifestations in PD. The primate thalamus is densely and heterogeneously innervated with dopaminergic axons, most of which express the dopamine transporter (DAT), as do the axons in the nigro-striatal system. We hypothesized that dopamine depletion may be present in the thalamus of the parkinsonian brain, that it may change over the course of parkinsonism, and that there may be regional differences among the thalamic nuclei. The toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was administered to adult macaque monkeys using a slow intoxication protocol. The treated macaques were classified into four groups according to their motor status: asymptomatic, recovered, mild parkinsonian and severe parkinsonian. Dopamine innervation was studied with immunohistochemistry for DAT. Topography maps of DAT-ir axon distribution were generated and the total length and length density of DAT-ir axons in the mediodorsal (MD), centromedian-parafascicular (CnMd-Pf), ventral and reticular nuclei of the thalamus were stereologically estimated using a 3D fractionator. Compared to controls, parkinsonian macaques exhibited less DAT-ir axonal length in the the MD and CnMd-Pf nuclei. In MD the dopamine denervation was already present in macaques that were asymptomatic at the time of sacrifice. Parkinsonian macaques had an increase of DAT-ir axon length density in the reticular nucleus compared to controls. DAT-ir axon length density in the ventral nuclei did not show differences between groups. The above results show a diverse reaction to MPTP of the dopaminergic axons innervating the thalamus. This may result in dysfunction of thalamocortical, thalamostriatal and intrathalamic transmission, and may be explain some poorlyunderstood non-motor manifestations of PD, such as attention and executive deficits, and sleep impairment, Este trabajo ha sido financiado por la Cátedra UAM - Fundación Tatiana Pérez de Guzmán el Bueno de Docencia e Investigación en Neurociencia

Studies on the thalamus have mostly focused on sensory relay nuclei, but the organization of pathways associated with emotions is not well understood. We addressed this issue by testing the hypothesis that the primate amygdala acts, in part, like a sensory structure for the affective import of stimuli and conveys this information to the mediodorsal thalamic nucleus, magnocellular part (MDmc). We found that primate sensory cortices innervate amygdalar sites that project to the MDmc, which projects to the orbitofrontal cortex. As in sensory thalamic systems, large amygdalar terminals innervated excitatory relay and inhibitory neurons in the MDmc that facilitate faithful transmission to the cortex. The amygdala, however, uniquely innervated a few MDmc neurons by surrounding and isolating large segments of their proximal dendrites, as revealed by three-dimensional high-resolution reconstruction. Physiologic studies have shown that large axon terminals are found in pathways issued from motor systems that innervate other brain centers to help distinguish self-initiated from other movements. By analogy, the amygdalar pathway to the MDmc may convey signals forwarded to the orbitofrontal cortex to monitor and update the status of the environment in processes deranged in schizophrenia, resulting in attribution of thoughts and actions to external sources., Pathways from the amygdala target specific neurons in the thalamic magnocellular mediodorsal nucleus to enable speedy and faithful transmission of emotional information to prefrontal cortex.

Microglia (MG) are the first cells to react to the abnormal incoming signals that follow an injury of sensory nerves and play a critical role in the development and maintenance of neuropathic pain, a common sequel of nerve injuries. Here we present population data on cell number, soma size, and length of processes of MG in the caudal division of the spinal trigeminal nucleus (Sp5C) in control mice and at the peak of microgliosis (7 days) following unilateral transection of the infraorbital nerve (IoN). The study is performed combining several bias- and assumption-free imaging and stereological approaches with different immunolabeling procedures, with the objective of tackling some hard problems that often hinder proper execution of MG morphometric studies. Our approach may easily be applied to low-density MG populations, but also works, with limited biases, in territories where MG cell bodies and processes form dense meshworks. In controls, and contralaterally to the deafferented side, MG cell body size and shape and branching pattern matched well the descriptions of “resting” or “surveillant” MG described elsewhere, with only moderate intersubject variability. On the superficial laminae of the deafferented side, however, MG displayed on average larger somata and remarkable diversity in shape. The number of cells and the length of MG processes per mm3 increased 5 and 2.5 times, respectively, indicating a net 50% decrease in the mean length of processes per cell. By using specific immunolabeling and cell sorting of vascular macrophages, we found only a negligible fraction of these cells in Sp5C, with no differences between controls and deafferented animals, suggesting that blood-borne monocytes play at most a very limited role in the microgliosis occurring following sensory nerve deafferentation. In sum, here we present reliable morphometric data on MG in control and deafferented trigeminal nuclei using efficient methods that we propose may equally be applied to any morphometric population analysis of these cells under different physiological or pathological conditions, This work was supported by the Spain’s Ministerio de Economía, Industria y Competitividad/Fondo Europeo de Desarrollo Regional Europeo de Desarrollo Regional (MINECO/FEDER), Grant BFU2015􀀀66941R. NG-M received a Boehringer- Ingelheim Fonds Travel Grant for a short-term stay at Prof. Helmut Kettenmann’s laboratory in the Max-Delbrück Center (Berlin).

The incidence of metabolic syndrome (MetS) is increasing worldwide which makes necessary the finding of new strategies to treat and/or prevent it. The aim of this study was to analyze the possible beneficial effects of a carob fruit extract (CSAT+&reg, ) on the cardiometabolic alterations associated with MetS in mice. 16-week-old C57BL/6J male mice were fed for 26 weeks either with a standard diet (chow) or with a diet rich in fats and sugars (HFHS), supplemented or not with 4.8% of CSAT+&reg, CSAT+&reg, supplementation reduced blood glucose, Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) and circulating levels of total cholesterol, low-density lipoprotein (LDL) cholesterol (LDL-c), insulin, and interleukin-6 (IL-6). In adipose tissue and skeletal muscle, CSAT+&reg, prevented MetS-induced insulin resistance, reduced macrophage infiltration and the expression of pro-inflammatory markers, and up-regulated the mRNA levels of antioxidant markers. Supplementation with CSAT+&reg, prevented MetS-induced hypertension and decreased the vascular response of aortic rings to angiotensin II (AngII). Moreover, treatment with CSAT+&reg, attenuated endothelial dysfunction and increased vascular sensitivity to insulin. In the heart, CSAT+&reg, supplementation reduced cardiomyocyte apoptosis and prevented ischemia-reperfusion-induced decrease in cardiac contractility. The beneficial effects at the cardiovascular level were associated with a lower expression of pro-inflammatory and pro-oxidant markers in aortic and cardiac tissues.

In the paper “The Association Between Pain Relief Using Video Games and an Increase in Vagal Tone in Children With Cancer: Analytic Observational Study With a Quasi-Experimental Pre/Posttest Methodology” (J Med Internet Res 2020;22(3):e16013) the authors noticed that one of the authors was not listed on the original published manuscript. The missing author was Mario Alonso Puig; they are first author on the corrected manuscript. Mario Alonso Puig’s affiliation will be listed as “Juegaterapia Foundation, Madrid, Spain” and will appear as affiliation 1 in the corrected manuscript. Author affiliations 1-4 in the original published manuscript will be renumbered to affiliations 2-5 in the corrected manuscript. The correction will appear in the online version of the paper on the JMIR website on July 7, 2020, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories.

Thalamocortical posterior nucleus (Po) axons innervating the vibrissal somatosensory (S1) and motor (MC) cortices are key links in the brain neuronal network that allows rodents to explore the environment whisking with their motile snout vibrissae., Thalamocortical posterior nucleus (Po) axons innervating the vibrissal somatosensory (S1) and motor (MC) cortices are key links in the brain neuronal network that allows rodents to explore the environment whisking with their motile snout vibrissae. Here, using fine-scale high-end 3D electron microscopy, we demonstrate in adult male C57BL/6 wild-type mice marked differences between MC versus S1 Po synapses in (1) bouton and active zone size, (2) neurotransmitter vesicle pool size, (3) distribution of mitochondria around synapses, and (4) proportion of synapses established on dendritic spines and dendritic shafts. These differences are as large, or even more pronounced, than those between Po and ventro-posterior thalamic nucleus synapses in S1. Moreover, using single-axon transfection labeling, we demonstrate that the above differences actually occur on the MC versus the S1 branches of individual Po cell axons that innervate both areas. Along with recently-discovered divergences in efficacy and plasticity, the synaptic structure differences reported here thus reveal a new subcellular level of complexity. This is a finding that upends current models of thalamocortical circuitry, and that might as well illuminate the functional logic of other branched projection axon systems. SIGNIFICANCE STATEMENT Many long-distance brain connections depend on neurons whose branched axons target separate regions. Using 3D electron microscopy and single-cell transfection, we investigated the mouse Posterior thalamic nucleus (Po) cell axons that simultaneously innervate motor and sensory areas of the cerebral cortex involved in whisker movement control. We demonstrate significant differences in the size of the boutons made in each area by individual Po axons, as well as in functionally-relevant parameters in the composition of their synapses. In addition, we found similarly large differences between the synapses of Po versus ventral posteromedial thalamic nucleus axons in the whisker sensory cortex. Area-specific synapse structure in individual axons implies a new, unsuspected level of complexity in long-distance brain connections.

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de lectura: 30-04-2020, En la enfermedad de Alzheimer (EA) la pérdida sináptica en el lóbulo temporal medial (LTM) ha sido postulada como el sustrato principal del déficit cognitivo inicial observado en los pacientes con dicha enfermedad. El objetivo principal de la presente tesis doctoral es describir la ultraestructura detallada de dos regiones corticales en donde comienzan a aparecer los cambios patológicos en la EA: la corteza transentorrinal (CTE) y la corteza entorrinal (CE). La importancia de este estudio radica en que conocer las alteraciones sinápticas en las primeras regiones corticales afectadas por la EA es crucial para entender mejor el progreso y los mecanismos responsables de la misma. Además de proporcionar datos sobre la EA, desentrañar la organización sináptica en condiciones normales es fundamental para conocer mejor la microorganización del cerebro humano. Para alcanzar el objetivo principal, se ha realizado un estudio cuantitativo del número y distribución de las sinapsis del neuropilo de la capa II de la CTE y de las capas II y III de la CE humana, así como de las características morfológicas de las mismas, y su distribución en los diferentes elementos postsinápticos. Se han utilizado técnicas estereológicas y de microscopía electrónica tridimensional (3D) en muestras humanas de sujetos sin alteraciones neurológicas y psiquiátricas aparentes (controles) y de sujetos diagnosticados de EA. Los resultados obtenidos muestran que, en condiciones normales, la mayoría de las sinapsis (92- 96%) son de tipo excitador (sinapsis asimétricas (SA)), siendo además estas sinapsis más grandes que las de tipo inhibidor (sinapsis simétricas (SS)). La mayoría de las sinapsis presentan una morfología macular (65-80%), y son de menor tamaño. Las SA tienen preferencia por establecerse en las cabezas de las espinas dendríticas, mientras que las SS tienen preferencia por los tallos dendríticos. Además, las SS presentan preferencia por una morfología en herradura. En las muestras procedentes de los pacientes con EA se encontró una reducción del grosor cortical y de la fracción cortical ocupada por neuronas, una disminución de la densidad sináptica, así como cambios morfológicos y de distribución en los elementos postsinápticos. Así, la presente tesis doctoral representa el primer análisis exhaustivo realizado mediante microscopio electrónica 3D de la organización sináptica del cerebro humano y sus alteraciones en la EA a nivel ultraestructural. Los resultados obtenidos pretenden contribuir a una mejor comprensión de la relación entre las alteraciones de los circuitos sinápticos y el deterioro cognitivo en la EA

PURPOSE: To study the outcome of botulinum toxin (BTX) treatment (group 1) in partially accommodative esotropia with high accommodative convergence/accommodation (AC/A) ratio, in comparison with bilateral medial rectus muscles recessions and posterior fixation (group 2). METHODS: In a retrospective comparative study, children aged 3-8 years old treated between 2011 and 2016, with partially accommodative esotropia with high AC/A ratio, deviation at distance of 10 prism diopters or more, and at least 1 year of follow-up, were included. Visual acuity, alternate prism and cover test, stereoacuity, biomicroscopy, and cycloplegic retinoscopy were carried out at initial, baseline visit, 6 months and 1 year after BTX injection or surgery. Main outcome variables were deviation at distance and near, improvement in stereoacuity, and percentage of success. We used multiple regression or proportional odds analysis to control for potential confounding variables. RESULTS: Of 95 patients, 84 were eligible, 48 children in group 1 and 36 in group 2. Deviation and stereoacuity were similar in the two groups at 6 months, but significantly better in the BTX group at 1 year (median distance deviation 0 prism diopters vs 5 prism diopters, p, JT received grant MINECO UAMA13-4E-2192

Neuroinflammation is one of the main physiopathological mechanisms of amyotrophic lateral sclerosis (ALS), produced by the chronic activation of microglia in the CNS. This process is triggered by the persistent activation of the ATP-gated P2X7 receptor (P2RX7, hereafter referred to as P2X7R). The present study aimed to evaluate the effects of the chronic treatment with the P2X7R antagonist JNJ-47965567 in the development and progression of ALS in the SOD1G93A murine model. SOD1G93A mice were intraperitoneally (i.p.) injected with either 30 mg/kg of JNJ-47965567 or vehicle 4 times per week, from pre-onset age (here, postnatal day 60; P60) until study endpoint. Body weight, motor coordination, phenotypic score, disease onset and survival were measured throughout the study, and compared between vehicle- and drug-injected groups. Treatment with the P2X7R antagonist JNJ-47965567 delayed disease onset, reduced body weight loss and improved motor coordination and phenotypic score in female SOD1G93A mice, although it did not increase lifespan. Interestingly, neither beneficial nor detrimental effects were observed in males in any of the analyzed parameters. Treatment did not affect motor neuron survival or ChAT, Iba-1 and P2X7R protein expression in endpoint individuals of mixed sexes. Overall, chronic administration of JNJ-47965567 for 4 times per week to SOD1G93A mice from pre-onset stage altered disease progression in female individuals while it did not have any effect in males. Our results suggest a partial, yet important, effect of P2X7R in the development and progression of ALS., Summary: This study implicates the P2X7 receptor (P2X7R) in amyotrophic lateral sclerosis progression in mice and shows that the effectiveness of treatment with P2X7R antagonists is sex dependent.

Background: Stimulation of the occipital or trigeminal nerves has been successfully used to treat chronic refractory neurovascular headaches such as migraine or cluster headache, and painful neuropathies. Convergence of trigeminal and occipital sensory afferents in the 'trigeminocervical complex' (TCC) from cutaneous, muscular, dural, and visceral sources is a key mechanism for the input-induced central sensitization that may underlie the altered nociception. Both excitatory (glutamatergic) and inhibitory (GABAergic and glycinergic) mechanisms are involved in modulating nociception in the spinal and medullary dorsal horn neurons, but the mechanisms by which nerve stimulation effects occur are unclear. This study was aimed at investigating the acute effects of electrical stimulation of the greater occipital nerve (GON) on the responses of neurons in the TCC to the mechanical stimulation of the vibrissal pad. Methods: Adult male Wistar rats were used. Neuronal recordings were obtained in laminae II-IV in the TCC in control, sham and infraorbital chronic constriction injury (CCI-IoN) animals. The GON was isolated and electrically stimulated. Responses to the stimulation of vibrissae by brief air pulses were analyzed before and after GON stimulation. In order to understand the role of the neurotransmitters involved, specific receptor blockers of NMDA (AP-5), GABAA (bicuculline, Bic) and Glycine (strychnine, Str) were applied locally. Results: GON stimulation produced a facilitation of the response to light facial mechanical stimuli in controls, and an inhibition in CCI-IoN cases. AP-5 reduced responses to GON and vibrissal stimulation and blocked the facilitation of GON on vibrissal responses found in controls. The application of Bic or Str significantly reduced the facilitatory effect of GON stimulation on the response to vibrissal stimulation in controls. However, the opposite effect was found when GABAergic or Glycinergic transmission was prevented in CCI-IoN cases. Conclusions: GON stimulation modulates the responses of TCC neurons to light mechanical input from the face in opposite directions in controls and under CCI-IoN. This modulation is mediated by GABAergic and Glycinergic mechanisms. These results will help to elucidate the neural mechanisms underlying the effectiveness of nerve stimulation in controlling painful craniofacial disorders, and may be instrumental in identifying new therapeutic targets for their prevention and treatment., This research has been supported by Grant BFU2015-66941R from Spain’s Ministerio de Economía Industria y Competitividad/Fondo Europeo de Desarrollo Regional (MINECO/FEDER) and SAF2016–76462 AEI/FEDER

Reelin is a large extracellular matrix (ECM) glycoprotein secreted by several neuronal populations in a specific manner in both the developing and the adult central nervous system. The extent of Reelin protein distribution and its functional role in the adult neocortex is well documented in different mammal models. However, its role in the adult spinal cord has not been well characterized and its distribution in the rodent spinal cord is fragmentary and has not been investigated in carnivores or primates as of yet. To gain insight into which neuronal populations and specific circuits may be influenced by Reelin in the adult spinal cord, we have conducted light and confocal microscopy study analysis of Reelin-immunoreactive cell types in the adult spinal cord. Here, we describe and compare Reelin immunoreactive cell type and distribution in the spinal cord of adult non-human primate (macaque monkeys, Macaca mulatta), carnivore (ferret, Mustela putorius) and rodent (rat, Rattus norvegicus). Our results show that in all three species studied, Reelin-immunoreactive neurons are present in the intermediate gray matter, ventricular zone and superficial dorsal horn and intermedio-lateral nucleus, while positive cells in the Clarke nucleus are only found in rats and primates. In addition, Reelin intermediolateral neurons colocalize with choline acetyltransferase (ChAT) only in macaque whilst motor neurons also colocalize Reelin and ChAT in macaque, ferret and rat spinal cord. The different expression patterns might reflect a differential role for Reelin in the pathways involved in the coordination of locomotor activity in the fore- and hind limbs., This work was financially supported by a grant from Ministerio de Ciencia y Tecnologia (MICIINN; BFU 2005-07857) to FC and by Mapfre Foundation (Fundación Mapfre;2011; Hospital 12 de Octubre) to AK and Kungliga Fysiografiska Sällskapet i Lund to TR-M covered costs associated to publication

Background: Dopamine loss beyond the mesostriatal system might be relevant in pathogenic mechanisms and some clinical manifestations in PD. The primate thalamus is densely and heterogeneously innervated with dopaminergic axons, most of which express the dopamine transporter, as does the nigrostriatal system. We hypothesized that dopamine depletion may be present in the thalamus of the parkinsonian brain and set out to ascertain possible regional differences. Methods: The toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine was administered to adult macaque monkeys using a slow intoxication protocol. The treated macaques were classified into 2 groups according to their motor status: nonsymptomatic and parkinsonian. Dopamine innervation was studied with immunohistochemistry for the dopamine transporter. Topographic maps of the dopamine transporter-immunoreactive axon distribution were generated and the total length and length density of these axons stereologically estimated using a 3-dimensional fractionator. Results: Parkinsonian macaques exhibited lower dopamine transporter-immunoreactive axon length density than controls in mediodorsal and centromedianparafascicular nuclei. Dopamine denervation in the mediodorsal nucleus was already noticeable in nonsymptomatic macaques and was even greater in parkinsonian macaques. Reticular nucleus dopamine transporter-immunoreactive axon length density presented an inverse pattern, increasing progressively to the maximum density seen in parkinsonian macaques. No changes were observed in ventral thalamic nuclei. Dopamine transporter-immunoreactive axon maps supported the quantitative findings. Conclusions: Changes in the dopamine innervation of various thalamic nuclei are heterogeneous and start in the premotor parkinsonian stage. These changes may be involved in some poorly understood nonmotor manifestations of PD., Chair in Neuroscience UAM-Fundación Tatiana Pérez de Guzmán el Bueno directed by C.C. J.A.O. and J.B. are currently funded by MINECO/AEI/FEDER-UE (SAF2015-67239-P), grant S2017/BMD-3700 (NEUROMETAB-CM) from Comunidad de Madrid cofinanced with Structural Funds from the European Union, La Caixa Foundation, Fundación BBVA, and Fundación Tatiana Pérez de Guzmán el Bueno.

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de lectura: 25-10-2019, Esta tesis tiene embargado el acceso al texto completo hasta el 25-04-2021, IGF‐I es un péptido con funciones tróficas bien estudiadas. Sin embargo, su papel como neuromodulador es objeto de debate. Algunas investigaciones han estudiado la acción del IGFI sobre la transmisión sináptica tanto excitadora como inhibidora y sobre la excitabilidad neuronal. De hecho, IGF‐I afecta a la extensión de los campos receptivos, y reduce el umbral de generación de potenciales de acción en respuesta a estímulos en la corteza somatosensorial, además de aumentar de la actividad del electrocorticograma (ECG) en esta corteza. Se ha demostrado que, en la modulación de la transmisión sináptica, los astrocitos juegan un papel muy importante. Asimismo, estos pueden sintetizar IGF‐I y presentan en su membrana plasmática receptores de IGF‐I (IGF‐IR). Aquí se muestra que el IGF‐I incrementa la excitabilidad e induce una potenciación a corto plazo (STP) de las corrientes postsinápticas excitadoras (EPSC) en neuronas piramidales de capa 2/3 de la corteza de barriles del ratón. Sin embargo, IGF‐I genera una modulación dual del EPSC ya que se observa una depresión a largo plazo (LTD) presináptica del EPSC cuando la acción del IGF‐I es en ausencia de potenciales de acción o de incrementos de calcio citosólico postsináptico. Adicionalmente, también induce una LTD de las corrientes postsinápticas inhibidoras (IPSC). Aunque la potenciación del EPSC mediada por IGF‐I requiere la activación de los receptores de adenosina A1 y A2A y los receptores metabotrópicos de glutamato (mGluR) del grupo I, solo la activación de los A2A es necesaria para la depresión del EPSC y del IPSC. En adición, tanto la modulación dual del EPSC como del IPSC son dependientes de aumentos de calcio citosólito astrocitario. Por tanto, IGF‐I podría tener una funcionalidad en procesos de memoria asociativa tanto in vitro como in vivo al inducir metaplasticidad favoreciendo la plasticidad sináptica inducida por protocolos de Spike Timing dependent plasticity (STDP). Por otro lado, los niveles de IGF‐I se han relacionado con procesos emocionales, estando disminuidos en la corteza prefrontal medial (mPFC) de ratas con ansiedad asociada a diabetes mellitus. Esta corteza es fundamental en la memoria del miedo condicionado, habiéndose demostrado que los aumentos de excitabilidad neuronal y eficacia sináptica en neuronas piramidales de capa 5 facilitan la memoria de extinción. En esta tesis denostamos que IGF‐I tiene la capacidad de disminuir las corrientes de potasio dependientes de calcio que median la posthiperpolarización de cinética lenta (sIAHP), lo que produce un aumento de la excitabilidad. Además, IGF‐I deprime a largo plazo tanto las corrientes sinápticas excitadoras como la inhibidoras, traduciéndose en una potenciación a largo plazo (LTP) de los potenciales postsinápticos. A nivel comportamental, estos efectos moduladores de IGF‐I en la actividad de la mPFC resulta en una facilitación de la extinción del miedo condicionado, Este estudio ha sido financiado con los siguientes proyectos: Ministerio de Economía y Competitividad, España (MINECO; BFU2013‐43668‐P y BFU2016‐80802‐P AEI/FEDER, UE).

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de lectura: 24-10-2019, Esta tesis tiene embargado el acceso al texto completo hasta el 24-04-2021, The metabolism of fatty acids and glucose are critical for cerebral energy homeostasis and maintenance of cellular functions. Two proteins involved in these processes are apolipoprotein E (apoE) which is the primary transporter of cholesterol in the brain, and insulin-like growth factor-I (IGF-I), involved in glucose handling by astrocytes. Both are related with the neurodegenerative Alzheimer’s disease (AD). The latter is considered a metabolic disorder because brain insulin resistance is a characteristic in AD, as in peripheral type II diabetes. Additionally, the apoE genotype account for the main genetic risk factor for developing the disease where the isoform apoE2 is protective and the apoE4 is detrimental. ApoE and IGF-I are locally produced in the brain, although the principal source of cerebral IGF-I is from the periphery, crossing the blood brain barrier by a receptor-specific transcytosis mechanism. Thus, we assessed possible interactions between IGF-I and two of the human apoE isoforms: apoE2 and apoE4, at several cellular processes. Firstly, we found that apoE modulates IGF-I uptake by brain endothelial cells, but not in choroid plexus epithelial cells, in an isoform specific manner. Accordingly, high fat diet, that is associated to increased circulating ApoE, impaired hippocampal IGF-I signaling in mice. We also observed that apoE4 mitigates intracellular IGF-I signaling, but it does not have any effect on the protective action of IGF-I on brain cells. In the same way, we revealed that IGF-I and apoE in an isoform-dependent way, modulate amyloid-β burden by modifying its production in astrocytes, cellular uptake and clearance from the plasma. Finally, we observed that IGF-I stimulates apoE release by astrocytes. We conclude that both proteins interact at different cellular levels within the brain and this fact may be an important link between cerebral IGF-I resistance and apoE genotype in the pathology of Alzheimer’s disease, El metabolismo de los ácidos grasos y de la glucosa son críticos en la homeostasis energética cerebral y buen funcionamiento celular. Dos de las proteínas implicadas en estos procesos son la apolipoproteína E (apoE), considerada el principal transportador de colesterol en el cerebro, y el factor de crecimiento de tipo insulínico I (IGF-I), implicado en el aporte de glucosa por los astrocitos. Ambos están relacionados con la enfermedad neurodegenerativa de Alzheimer ya que se piensa que se trata de un trastorno metabólico al ser característica la resistencia a insulina en el cerebro, de manera similar a la diabetes tipo II periférica. Por otra parte, el genotipo de apoE representa el principal factor de riesgo genético en el desarrollo de la enfermedad considerándose protectora a la isoforma apoE2 y perjudicial la isoforma apoE4. ApoE e IGF-I se producen localmente en el cerebro, sin embargo, el principal aporte de IGF-I procede de la periferia, tras cruzar la barrera hematoencefálica mediante mecanismos de transcitosis mediada por receptores específicos. Por ello, hemos evaluado las posibles interacciones entre IGF-I y las dos isoformas humanas de apoE: apoE2 y apoE4, en varios procesos celulares. En primer lugar, encontramos que apoE, de manera isoforma dependiente, modula la internalización de IGF-I por células endoteliales, pero no en células epiteliales del plexo coroideo. Así, la dieta con alto contenido en grasa, que incrementa los niveles periféricos de apoE, altera la señalización por IGF-I hipocampal en ratones. También observamos que apoE4 atenúa la señalización intracelular por IGF-I, pero no ejerce efecto alguno sobre la acción protectora del IGF-I en las células cerebrales. De la misma forma, demostramos que el IGF-I y la apoE de manera dependiente de isoforma, modulan la cantidad de β-amiloide al modificar su producción en astrocitos, internalización celular y el aclaramiento del plasma. Por último, observamos que el IGF-I estimula la secreción de apoE por astrocitos. Por tanto, concluimos que ambas proteínas interactúan en el cerebro a diferentes niveles, y esta conexión puede ser importante en el desarrollo de resistencia a IGF-I en el cerebro y el genotipo de apoE en la patología de la enfermedad de Alzheimer., La presente Tesis Doctoral ha sido realizada disfrutando de una ayuda para la formación de doctores (BES-2014-070574) asociada al proyecto SAF2013-40710-R, y posterior proyecto SAF2016-76462-C2-1-P, ambos financiados por el Ministerio de Economía y Competitividad

Fundamentos: El tratamiento de la población infantojuvenil con dolor ha mejorado considerablemente en los últimos 30 años. En España, el progreso también es evidente, y han surgido unidades o programas especializados como parte de la atención a esta población, aunque no existen estudios sobre las características o la actividad de estas unidades o programas especializados. Esta falta de información impide una evaluación adecuada de los procedimientos vigentes y dificulta una adecuada administración y gestión de estos recursos. El objetivo de este trabajo fue conocer las características de estas unidades y programas para el tratamiento de la población infantojuvenil con dolor en España. Sujetos y métodos: Se identificaron un total de 10 unidades especializadas o programas de tratamiento específicos del dolor infantil en España y se contactó con la persona responsable. A través de una encuesta autoadministrada online se recogió información sobre la organización del programa, su implicación en ámbitos relevantes como la investigación, la formación de los profesionales, la sensibilización y visibilidad del problema en la sociedad y entre los profesionales, las especialidades implicadas, el tipo de tratamientos que se ofrecen, la evaluación de resultados y los servicios prestados. Se han utilizado medias y porcentajes para describir los resultados. Resultados: De las diez unidades o programas contactados, ocho contestaron a la encuesta (80%). Los programas de tratamiento del dolor infantojuvenil demostraron ser fundamentalmente farmacológicos, también en los casos de dolor crónico. Informaban de que se preocupaban por impulsar la sensibilización de los especialistas y la sociedad sobre el problema (88%), de contribuir también a la formación de especialistas (88%) y, en menor medida, de investigar y generar conocimiento (75%). Conclusiones: Esta encuesta proporciona información valiosa sobre las características actuales de los programas y unidades especializadas para el tratamiento de la población infantojuvenil con dolor en España. Puede servir para proponer acciones que mejoren la gestión y administración actual, Background: The treatment of children and adolescents with pain has improved considerably in the last 30 years. In Spain, progress is also evident, and specialized units or programs have emerged as part of the care provided for this population. However, there are no studies on the characteristics or activities of these specialized units or programs. This lack of information prevents an adequate evaluation of the current procedures, and hinders an adequate administration and management of these resources. The objective of this work was to study the characteristics of these units and programs for the treatment of children and adolescents with pain in Spain. Methods: A total of 10 specialized units or specific treatment programs for pediatric pain in Spain were identified, and the person responsible was contacted. Through a self-administered online survey, information was collected on the organization of the program, the involvement in relevant fields such as research, training of professionals and the raising of awareness and visibility of the problem, the specialties involved, the type of treatments offered, the evaluation of results and the services provided. Results: Of the ten units or programs contacted, eight answered the survey (80%). Pain programs for children and adolescents proved to be primarily pharmacological, also for chronic pain. They reported to promote awareness about the problem (88%), to contribute to the training of specialists (88%) and to a lesser extent to research and knowledge generation (75%). Conclusions: This survey provides valuable information on the current characteristics of specialized programs and units for the treatment of children and adolescents with pain in Spain, and can be used to propose actions that improve its current management and administration, Este trabajo ha sido financiado, en parte, por el Ministerio de Ciencia y Competitividad (MINECO; PSI2015-70966-P; PSI2016-82004-REDT), la Generalitat de Catalunya (AGAUR), la Universitat Rovira i Virgili (Programa PFR), la Obra Social de Caixabank y el Fondo Europeo de Desarrollo Regional (FEDER). Jordi Miró cuenta con el apoyo de la Institució Catalana de Recerca i Estudis Avançats (ICREA-Acadèmia) y la Fundación Grünenthal

Aimof the study: Postoperative intra-abdominal abscess (PIAA) is a frequent and severe complication of acute appendicitis (AA) with peritonitis. The neutrophil-to-lymphocyte ratio (NLR) is an inflammatory marker that has been related to the development of peritonitis; however, its diagnostic role in predicting PIAA has not been evaluated. This is the first study that analyzes the usefulness of NLR as a predictor of PIAA in children operated for AA. Material and Methods: Retrospective observational study in children operated for AA in our institution during 2017–2018. Patients aged under 5 years or with incomplete laboratory determinations at hospital admission (blood count, C-reactive protein, and fibrinogen) were excluded. Demographic and laboratory parameters and the development of PIAA were analyzed. NLR was calculated by dividing the absolute number of neutrophils by the absolute number of lymphocytes. Bymeans of ROC curves, we determined the sensitivity and specificity of the different laboratory parameters to predict the development of PIAA. Results: A total of 388 patients aged 10.5 ± 2.9 year were included. Twenty (5.2%) developed PIAA. NLR presented an area under the curve (AUC) of 0.85, significantly higher than the determination of leukocytes (AUC 0.69, p < 0.001), neutrophils (AUC 0.74, p < 0.001), fibrinogen (AUC 0.68, p < 0.001) and C-reactive protein (AUC 0.73, p < 0.001). We estimated the optimal cut-off point of NLR > 10.5, with a sensitivity of 85% and a specificity of 75.2%. Conclusions: NLR is the laboratory parameter with the highest sensitivity and specificity for predicting the development of PIAA in children operated for AA. It can be useful as a predictor of worse postoperative course.

Gait is a basic cognitive purposeful action that has been shown to be altered in late stages of neurodegenerative dementias. Nevertheless, alterations are less clear in mild forms of dementia, and the potential use of gait analysis as a biomarker of initial cognitive decline has hitherto mostly been neglected. Herein, we report the results of a study of gait kinematic time series for two groups of patients (mild cognitive impairment and mild Alzheimer&rsquo, s disease) and a group of matched control subjects. Two metrics based on permutation patterns are considered, respectively measuring the complexity and irreversibility of the time series. Results indicate that kinematic disorganisation is present in early phases of cognitive impairment, in addition, they depict a rich scenario, in which some joint movements display an increased complexity and irreversibility, while others a marked decrease. Beyond their potential use as biomarkers, complexity and irreversibility metrics can open a new door to the understanding of the role of the nervous system in gait, as well as its adaptation and compensatory mechanisms.

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Anatomía, Histología y Neurociencia. Fecha de lectura: 12-04-2019, El proyecto ha sido financiado en su mayoría por la compañía Novartis farmacéutica con el código PI 110/13 JGM-INM. Adicionalmente se han usado para este proyecto fondos del proyecto PI15/02099 del Fondo de Investigación Sanitaria FIS integrado en el Plan Nacional de I+D+I 2013-2016 financiado por el Instituto de Salud Carlos III (ISCIII) y cofinanciado por el Fondo Europeo de Desarrollo Regional (FEDER).

The amygdaloid complex (AC) is a heterogeneous aggregate of nuclei located in the rostromedial region of the temporal lobe. In addition to being partly connected among themselves, the AC nuclei are strongly interconnected with the cerebral cortex, striatum, basal forebrain, hypothalamus and brainstem. Animal and human functional studies have established that the AC is a central hub of the neuronal networks supporting emotional responsivity, particularly its negative/aversive components. Dysfunction of AC circuits in humans has been implicated in anxiety, depression, schizophrenia and bipolar disorder. The small New-World marmoset monkey (Callithrix jacchus) has recently become a key model for neuroscience research. However, the nuclear and fiber tract organization of marmoset AC has not been examined in detail. Thus, the extent to which it can be compared to the AC of Old-World (human and macaque) primates is yet unclear. Here, using Nissl and acetylcholinesterase (AChE) histochemical stains as a reference, we analyzed the cytoarchitecture and nuclear parcellation of the marmoset AC. In addition, given the increasing relevance of tractographic localization for high-resolution in vivo imaging studies in non-human primates, we also identified the myelin fiber tracts present within and around the AC as revealed by the Gallyas method. The present study provides a detailed atlas of marmoset AC. Moreover, it reveals that, despite phylogenetic distance and brain size differences, every nucleus and myelinated axon bundle described in human and macaque studies can be confidently recognized in marmosets., This work was supported by funding from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-Brazil) Programa de Ciencia sem Fronteiras—Pesquisador Visitante Especial 024.028/2017, PVE 2014 and 400730/2014-6, PVE 2014 and by a grant from the Ministerio de Educación y Cultura, programa hispano brasileño de cooperación interuniversitaria, PHB H2012-0011 (Spain).