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Ultrasound remains a mainstay in proximal suspensory ligament (PSL) evaluation. Despite recent improvements facilitating earlier diagnosis/treatment, needs exist for (1) characterization of normal ultrasonographic PSL cross-sectional area (CSA) in specific breeds/disciplines, (2) improved detection of early/subtle changes in the PSL, and (3) an understanding of suspensory ligament (SL) morphology change from work-related stress. The objectives of this study were to establish normal PSL CSA in Quarter Horses (QH) used for cutting via angle contrast ultrasonography, ultrasonographically monitor SL morphology as horses progress through training, and assess the impact of PSL disease on performance (Lifetime Earnings - $LTE). One hundred ten 2-year-old cutting horses had PSL ultrasound examination on all limbs, and examinations were repeated 2 years later on 51 remaining horses. PSL CSA at select locations was compared between time points and entire examinations subjectively graded by an equine radiologist. $LTE was obtained from the National Cutting Horse Association for each horse and analyzed by abnormality number, location, and development over time. A total of 267 PSL ultrasound examinations across both times points were measured and graded. Normal CSA was established by zone for forelimbs and hindlimbs. The prevalence of abnormal limbs increased between the 2-year-old and 4-year-old years, forelimb abnormalities were statistically more prevalent than hindlimb abnormalities at both time points, and osseous attachment abnormalities were statistically more prevalent than ligament abnormalities at both times points. No significant differences in PSL CSA change over time were seen in any group and there was no significant difference in $LTE among or between groups.

The invasive Japanese stiltgrass (Microstegium vimineum) affects a wide range of ecosystems and threatens biodiversity across the eastern USA. However, the mechanisms underlying rapid adaptation, plasticity, and epigenetics in the invasive range are largely unknown. We present a chromosome-level assembly for M. vimineum to investigate genome dynamics, evolution, adaptation, and the genomics of phenotypic plasticity. We generated a 1.12 Gb genome with scaffold N50 length of 53.44 Mb respectively, taking a de novo assembly approach that combined PacBio and Dovetail Genomics Omni-C sequencing. The assembly contains 23 pseudochromosomes, representing 99.96% of the genome. BUSCO assessment indicated that 80.3% of Poales gene groups are present in the assembly. The genome is predicted to contain 39,604 protein-coding genes, of which 26,288 are functionally annotated. Furthermore, 66.68% of the genome is repetitive, of which unclassified (35.63%) and long terminal repeat (LTR) retrotransposons (26.90%) are predominant. Similar to other grasses, Gypsy (41.07%) and Copia (32%) are the most abundant LTR-retrotransposon families. The majority of LTR-retrotransposons are derived from a significant expansion in the past 1-2 million years, suggesting the presence of relatively young LTR-retrotransposon lineages. We find corroborating evidence from Ks plots for a stiltgrass-specific duplication event, distinct from the more ancient grass-specific duplication event. The assembly and annotation of M. vimineum will serve as an essential genomic resource facilitating studies of the invasion process, the history and consequences of polyploidy in grasses, and provides a crucial tool for natural resource managers.SignificanceThe current lack of genomic resources for the invasive Japanese stiltgrass—and thousands of other invasive species globally—severely limits our understanding of the invasion process and hinders decision-making for effective management and control. In this study, we present a chromosome-level genome assembly and annotation of Japanese stiltgrass, a problematic weed in eastern North America, identifying a clear history of polyploidy and recent activity of transposable elements. The ultimate goal is to advance genomic studies to better understand the dynamics of non-native species during the various invasion phases, thereby providing insights into effective control strategies to manage current and future invasions.

Introduction: Dangerous behavior is considered an undesired trait, often attributed to poor training or bad-tempered horses. Unfortunately, horses with progressive signs of dangerous behavior are often euthanized due to concerns for rider safety and limitations in performance. However, this dangerous behavior may actually originate from chronic axial skeleton pain. This case series describes the medical histories and clinical presentations of horses presented for performance limitations and dangerous behavior judged to be related to intractable axial skeleton pain.Material and Methods: Fourteen horses that developed severe performance limitations resulting in euthanasia were included. A complete spinal examination and behavioral responses, gait and neurologic evaluations, diagnostic imaging, gross pathologic and histopathologic examinations of the axial skeleton were performed on all horses. A tentative diagnosis of the affected spinal region was formulated using medical records, owner and trainer complaints, and antemortem examination findings. The selected spinal regions were further examined with gross and histopathologic evaluations of the associated osseous, soft tissue and neural tissues.Results: Ten horses showed severe behavioral responses during the myofascial and mobilization examinations. Based on an aggregate evaluation, the cervicothoracic and lumbosacral regions were the most common regions believed to be the primary area of concern. All horses had moderate to severe ganglionitis present at multiple vertebral levels. Subdural and epidural hemorrhage or hematomas were a common finding (71%) in the cervicothoracic and lumbosacral regions.Discussion: In this case series, neuropathic (i.e., structural) pain was judged to be the underlying cause of dangerous behavior. The dorsal root ganglia (DRG) serve an important role in relaying peripheral sensory information to the central nervous system and ganglionitis has been associated with neuropathic pain syndromes. This series highlights the need for more in-depth understanding of pain behavior and its clinical presentation and progression in chronic or severely affected horses. Limitations of the study are the lack of age-matched control DRG and the incomplete collection of DRG from every vertebral level of interest.

Mycoheterotrophic plants derive most or all carbon and nutrients from fungal partners and represent poorly understood components of forest biodiversity. Many are rare or endangered yet can be ecological indicators of forest ecosystem function due to fungal host requirements. One such species is the IUCN red-listed (“vulnerable”), fully mycoheterotrophic orchid, Corallorhiza bentleyi. This recently described species is among the rarest plants in Appalachia, known from five counties in Virginia and West Virginia. The species has a restricted range, small population size, and is self-pollinating. Here, an integrative approach was taken in conservation genetic assessment of C. bentleyi using floral morphometrics, simple-sequence repeats, and fungal host DNA to characterize variation within and among sampling localities. Morphology reveals differentiation among individuals from six sampling localities. Surprisingly, most genetic variation is found within localities, contrary to the expectation for a selfing species. Fungal host DNA reveals extreme specificity upon a few genotypes of a single ectomycorrhizal host species, Tomentella fuscocinerea, across all localities. We discuss conservation implications of morphological, genetic, and symbiotic diversity in this vulnerable species, and recommend additional assessment of conservation status based on: an obligate reproductive mode of selfing, preventing benefits of outcrossing among genetically non-identical individuals; extreme host specificity, severely restricting niche space; and highly fragmented habitat under threat from anthropogenic disturbance. This study underscores the importance of integrative conservation assessment, analyzing multiple data sources, and reveals patterns not readily apparent from census-based assessments alone.

Precision dosing is progressing beyond the conceptual and proof-of-concept stages toward implementation. As the availability of dosing algorithms, tools, and platforms increases, so do the investment in technology services and actual implementation of clinical services offering these solutions to patients. Nowhere is this needed more than in pediatric populations, which are still reliant on adult drug development and bridging strategies to support dosing, often in the absence of actual dose-finding studies in the target pediatric population. Still, there is more work to be done to ensure that proper governance of these services is maintained, and that sustainability of these early implementations is guided by new science as it evolves and meaningful outcome data to confirm that such services deliver on both clinical and economic return on investment. In addition, the field should ensure that all approaches beyond a therapeutic drug monitoring-driven, pharmacokinetic-centric approach should be considered as the tools and services evolve, especially when pediatric-specific pharmacokinetic/pharmacodyamic and pharmacogenetic data are available and shown to be useful to guide dosing. This review evaluates current pediatric precision dosing efforts, highlighting their utility, longevity, and sustainability and assesses the current process for implementing such approaches examining current barriers that stand in the way of broader implementation and the stakeholders that must engage to ensure its ultimate success.

BACKGROUND The range of diagnostic modalities available to evaluate superficial digital flexor tendon (SDFT) injury includes magnetic resonance imaging (MRI), computed tomography (CT) and ultrasonography (US). Direct, comprehensive comparison of multi-modality imaging characteristics to end-point data has not previously been performed using a model of tendinopathy but is required to obtain a better understanding of each modality's diagnostic capabilities. OBJECTIVE To compare CT, MRI and US evaluation to outcome measures for histologic, biochemical and biomechanical parameters using an equine surgical model of tendinopathy. STUDY DESIGN Controlled experiment. METHODS Lesions were surgically created in both forelimb SDFTs of eight horses and imaged using MRI, CT and US at seven time points over 12 months. Imaging characteristics were then correlated to end point histologic, biochemical and biomechanical data using lasso regression. Longitudinal lesion size was compared between imaging modalities. RESULTS Lesion to tendon isoattenuation on CT evaluation correlated with the greatest levels of aggrecan deposition. A significant correlation between cellular density and percentage of tendon involvement on the T2-weighted sequence and signal intensity on the proton density fat saturated (PD FS) sequence was appreciated at the 12-month time point (P = .006, P = .02 respectively). There was no significant correlation between end-point data and US or contrast imaging characteristics. Cross sectional area lesion to tendon measurements were significantly largest on CT evaluation, followed by MRI and then US (P

Our understanding of epithelial-to-mesenchymal transition (EMT) has slowly evolved from a simple two state, binary model to a multi-step, dynamic continuum of epithelial-to-mesenchymal plasticity, with metastable intermediate transition states that may drive cancer metastasis. Head and neck cancer is no exception, and in this review, we use head and neck as a case study for how partial-EMT (p-EMT) cell states may play an important role in cancer progression. In particular, we summarize recent in vitro and in vivo studies that uncover these intermediate transition states, which exhibit both epithelial and mesenchymal properties and appear to have distinct advantages in migration, survival in the bloodstream, and seeding and propagation within secondary metastatic sites. We then summarize the common and distinct regulators of p-EMT as well as methodologies for identifying this unique cellular subpopulation, with a specific emphasis on the role of cutting-edge technologies, such as single cell approaches. Finally, we propose strategies to target p-EMT cells, highlighting potential opportunities for therapeutic intervention to specifically target the process of metastasis. Thus, although significant challenges remain, including numerous gaps in current knowledge, a deeper understanding of EMT plasticity and a genuine identification of EMT as spectrum rather than a switch will be critical for improving patient diagnosis and treatment across oncology.

Naegleria fowleri is a pathogenic, thermophilic, free-living amoeba which causes primary amebic meningoencephalitis (PAM). Penetrating the olfactory mucosa, the brain-eating amoeba travels along the olfactory nerves, burrowing through the cribriform plate to its destination: the brain’s frontal lobes. The amoeba thrives in warm, freshwater environments, with peak infection rates in the summer months and has a mortality rate of approximately 97%. A major contributor to the pathogen’s high mortality is the lack of sensitivity of N. fowleri to current drug therapies, even in the face of combination-drug therapy. To enable rational drug discovery and design efforts we have pursued protein production and crystallography-based structure determination efforts for likely drug targets from N. fowleri. The genes were selected if they had homology to drug targets listed in Drug Bank or were nominated by primary investigators engaged in N. fowleri research. In 2017, 178 N. fowleri protein targets were queued to the Seattle Structural Genomics Center of Infectious Disease (SSGCID) pipeline, and to date 89 soluble recombinant proteins and 19 unique target structures have been produced. Many of the new protein structures are potential drug targets and contain structural differences compared to their human homologs, which could allow for the development of pathogen-specific inhibitors. Five of the structures were analyzed in more detail, and four of five show promise that selective inhibitors of the active site could be found. The 19 solved crystal structures build a foundation for future work in combating this devastating disease by encouraging further investigation to stimulate drug discovery for this neglected pathogen.

A conditionally replication-defective human cytomegalovirus (HCMV) vaccine, V160, was shown to be safe and immunogenic in a two-part, double-blind, randomized, placebo-controlled phase I clinical trial (NCT01986010). However, the specificities and functional properties of V160-elicited antibodies remain undefined. Here, we characterized 272 monoclonal antibodies (mAbs) isolated from single memory B cells of six V160-vaccinated subjects. The mAbs bind to diverse HCMV antigens, including multiple components of the pentamer, gB, and tegument proteins. The most-potent neutralizing antibodies target the pentamer-UL subunits. The binding sites of the antibodies overlap with those of antibodies responding to natural HCMV infection. The majority of the neutralizing antibodies target the gHgL subunit. The non-neutralizing antibodies predominantly target the gB and pp65 proteins. Sequence analysis indicated that V160 induced a class of gHgL antibodies expressing the HV1-18/KV1-5 germline genes in multiple subjects. This study provides valuable insights into primary targets for anti-HCMV antibodies induced by V160 vaccination.

The capability to generate densely sampled single nucleotide polymorphism (SNP) data is essential in diverse subdisciplines of biology, including crop breeding, pathology, forensics, forestry, ecology, evolution, and conservation. However, the wet-lab expertise and bioinformatics training required to conduct genome-scale variant discovery remain limiting factors for investigators with limited resources.Here we present ISSRseq, a PCR-based method for reduced representation of genomic variation using simple sequence repeats as priming sites to sequence inter simple sequence repeat (ISSR) regions. Briefly, ISSR regions are amplified with single primers, pooled, used to construct sequencing libraries with a commercially-available kit, and sequenced on the Illumina platform. We also present a flexible bioinformatic pipeline that assembles ISSR loci, calls and hard filters variants, outputs data matrices in common formats, and conducts population analyses using R.Using three angiosperm species as case studies, we demonstrate that ISSRseq is highly repeatable, necessitates only simple wet-lab skills and commonplace instrumentation, is flexible in terms of the number of single primers used, and can generate genomic-scale variant discovery on par with existing RRS methods which require more complex wet lab procedures.ISSRseq represents a straightforward approach to SNP genotyping in any organism, and we predict that this method will be particularly useful for those studying population genomics and phylogeography of non-model organisms. Furthermore, the ease of ISSRseq relative to other RRS methods should prove useful to those lacking advanced expertise in wet lab methods or bioinformatics.

Naegleria fowleri is a pathogenic, thermophilic, free-living amoeba which causes primary amebic meningoencephalitis (PAM). Penetrating the olfactory mucosa, the brain-eating amoeba travels along the olfactory nerves, burrowing through the cribriform plate to its destination: the brain’s frontal lobes. The amoeba thrives in warm, freshwater environments, with peak infection rates in the summer months and has a mortality rate of approximately 97%. A major contributor to the pathogen’s high mortality is the lack of sensitivity of N. fowleri to current drug therapies, even in the face of combination-drug therapy. To enable rational drug discovery and design efforts we have pursued protein production and crystallography-based structure determination efforts for likely drug targets from N. fowleri. N. fowleri genes were selected if they had homology to drug targets listed in Drug Bank or were nominated by primary investigators engaged in N. fowleri research. In 2017, 178 N. fowleri protein targets were queued to the Seattle Structural Genomics Center of Infectious Disease (SSGCID) pipeline, and to date 89 soluble recombinant proteins and 19 unique target structures have been produced. Many of the new protein structures are potential drug targets and contain structural differences compared to their human homologs, which could allow for the development of pathogen-specific inhibitors. Five of the structures were analyzed in more detail, and four of five show promise that selective inhibitors of the active site could be found. The 19 solved crystal structures build a foundation for future work in combating this devastating disease by encouraging further investigation to stimulate drug discovery for this neglected pathogen.

Mycoheterotrophic plants derive most or all carbon and nutrients from fungal partners and represent poorly understood components of forest biodiversity. Many are rare or endangered yet can be ecological indicators of forest ecosystem function due to their often highly specific fungal host requirements. One such species is the IUCN red-listed (‘vulnerable’), fully mycoheterotrophic orchid,Corallorhiza bentleyi. This recently described species is among the rarest plants in Appalachia, known from five counties in Virginia and West Virginia, USA. The species has a restricted range, small population size, and is self-pollinating. Here we take an integrative approach to conservation genetic assessment inC. bentleyiusing floral morphometrics, simple-sequence repeats, and fungal host DNA to characterize variation within and among sampling localities. Morphology reveals some differentiation among individuals from six sampling localities. Surprisingly, most genetic variation is found within localities, contra to the expectation for a selfing species. Fungal host DNA reveals extreme specificity upon a few genotypes of a single ectomycorrhizal host species,Tomentella fuscocinerea,across all localities. We discuss the conservation implications of morphological, genetic, and symbiotic diversity in this vulnerable species, and recommend additional assessment of conservation status based on: an obligate reproductive mode of selfing, preventing benefits of outcrossing among genetically non-identical individuals; extreme host specificity, severely restricting niche space; and highly fragmented habitat under threat from anthropogenic disturbance. This study underscores the importance of integrative conservation assessment, analyzing multiple data sources, and reveals patterns not readily apparent from census-based assessments alone.

We present the first reported case of prosthetic joint infection caused by Clostridium bifermentans, which was treated with total joint washout and debridement allowing for the patient to retain his prosthesis and achieve full recovery. Clostridium bifermentans is a gram-positive, anaerobic, spore-forming bacterium. This organism was once considered to be non-pathogenic, but has recently been associated with cases of septic arthritis, empyema, osteomyelitis, soft tissue infection, brain abscess, bacteremia and endocarditis.

This paper examines the effect of retirement on individuals’ time spent on home production activities. Using rich longitudinal data from Australia and utilizing recent changes in the Australian Pension system which generate exogenous variation in retirement status, we show a significant increase in time devoted to home production, especially for individuals who voluntarily retire from the labour force. The empirical findings provide further evidence consistent with the majority of retirees successfully smoothing their wellbeing as they make the transition out of the labour force.

CASE DESCRIPTION A 12-year-old mixed-breed mare (horse 1) and 6-year-old Friesian gelding (horse 2) were examined for chronic lameness associated with the stifle joint. CLINICAL FINDINGS Lameness examination revealed effusion of the right (horse 1) or left (horse 2) femoropatellar and medial femorotibial joints and grade 3/5 (horse 1) or 4/5 (horse 2) lameness. A diagnosis of cranial cruciate ligament (CCL) injury with associated mineralization and avulsion (horse 1) or mineralization alone (horse 2) was facilitated in both horses with a caudomedial-craniolateral oblique radiographic view obtained 45° medial to the caudocranial line, which highlighted the origin of the ligament on the caudoaxial aspect of the lateral femoral condyle within the intercondylar fossa. These lesions were subsequently confirmed via CT. TREATMENT AND OUTCOME Arthroscopy of the medial and lateral femorotibial joints was performed for horse 1 and revealed the osseous fragment associated with the CCL, but the fragment could not be removed. Horse 2 was euthanized while anesthetized following CT owing to the poor prognosis. CONCLUSIONS AND CLINICAL RELEVANCE Radiography is typically the first imaging modality attempted for horses with CCL injury, particularly outside the hospital setting. A 45° caudomedial-craniolateral oblique radiographic view may aid in diagnosis of CCL injury when avulsion or mineralization is present. Although this view is not commonly included in the typical radiographic series for imaging of the stifle joint in horses, it should be considered when CCL injury is suspected.

Hybridization and introgression are common processes among numerous plant species that present both challenges and opportunities for studies of species delimitation, phylogenetics, taxonomy, and adaptation.Rhus integrifoliaandR. ovataare two ecologically important shrubs native to the southwestern USA and Mexico, and are known to hybridize frequently, but the morphological, genetic, and ecological implications of hybridization in these species are poorly studied on a broad geographic scale. Analyses were conducted using leaf morphology, genetic variation of plastid and nuclear loci, and species distribution models for both species and their putative hybrid introgressants across 19 localities in California and Arizona, USA. These analyses revealed evidence for morphological and genetic distinction among localities comprising putative parental species, but a high degree of morpho-genetic intermediacy among localities with putative hybrids. Comparison of morphological and genetic population structure among localities revealed evidence for putative local adaptation or widespread phenotypic plasticity. Multiple regression models identified a weak but statistically significant negative association between leaf area and precipitation. Finally, species distribution modeling inferred northward range shifts over time, with both species predicted to occupy more coastal regions in the future, possibly increasing the frequency of hybridization among them. These findings underscore the importance of integrative assessment of multiple data sources in the study of hybridizing species and highlight theRhus integrifolia-ovatacomplex as a powerful model for investigating the adaptive implications of hybridization.

Aims: To compare the outcome, in terms of lameness score or return to athletic function, of horses with acute vs. chronic digital lameness that underwent magnetic resonance imaging (MRI) of the distal limb and to compare the proportion of horses that received intra-articular therapy of the distal interphalangeal (DIP) joint and pattern of diagnostic analgesia in these groups. Methods: This is a retrospective study of horses (n = 95) with acute (≤12 weeks; n = 46) or chronic (>12 weeks; n = 49) digital lameness that underwent MRI of the distal limb from 2009–2016, at two equine referral centres in the USA. Criteria for inclusion in the study were that a majority of lameness localised distal to the fetlock, and that lameness assessments for ≥12 months following MRI could be obtained from the medical record or the owner could be interviewed regarding their horse's athletic function. Outcome was characterised by an improvement score where 2 = return to work at a previous or higher level or lameness improved by one grade or more, 1 = return to work at a lower level or lameness improved by less than one grade, and 0 = did not return to work or lameness grade worsened. Whether horses had received intra-articular therapy of the DIP joint and the pattern of diagnostic analgesia prior to MRI was also obtained from medical records or by interviewing the owner. Results: There was a difference (p = 0.004) in the proportion of horses assigned to improvement scores of 0, 1 and 2 between horses with acute or chronic lameness. There was no evidence of a difference in the likelihood of having received intra-articular therapy of the DIP joint prior to MRI between horses with chronic or acute lameness (p = 0.085). Similarly, there was no evidence of a difference in the pattern of diagnostic analgesia prior to MRI between the two groups (p = 0.94). Eighty-two percent of owners of horses with acute and 62% of those with horses with chronic lameness had a positive opinion of the utility of MRI as a diagnostic modality. Conclusion: In a population of horses with digital lameness undergoing MRI, a difference in the outcome, in terms of lameness score or return to athletic function was identified between horses with acute lameness compared to those with chronic lameness. Clinical relevance: Horses with digital lameness that undergo MRI when the lameness is acute may have an improved prognosis due to accurate diagnosis and earlier application of appropriate therapy.

A series of 3-aryl(pyrrolidin-1-yl)butanoic acids were synthesized using a diastereoselective route, via a rhodium catalyzed asymmetric 1,4-addition of arylboronic acids in the presence of (R)-BINAP to a crotonate ester to provide the (S) absolute configuration for the major product. A variety of aryl substituents including morpholine, pyrazole, triazole, imidazole, and cyclic ether were screened in cell adhesion assays for affinity against αvβ1, αvβ3, αvβ5, αvβ6, and αvβ8 integrins. Numerous analogs with high affinity and selectivity for the αvβ6 integrin were identified. The analog (S)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-((R)-3-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)pyrrolidin-1-yl)butanoic acid hydrochloride salt was found to have very high affinity for αvβ6 integrin in a radioligand binding assay (pKi = 11), a long dissociation half-life (7 h), very high solubility in saline at pH 7 (>71 mg/mL), and pharmacokinetic properties commensurate with inhaled dosing by nebulization. It wa...

High field magnetic resonance imaging (MRI) is increasingly used for horses with suspected stifle disease, however there is limited available information on normal imaging anatomy and potential incidental findings. The aim of this prospective, anatomic study was to develop an optimized high field MRI protocol for evaluation of the equine stifle and provide detailed descriptions of the normal MRI appearance of the stifle soft tissues, using ultrasound and gross pathological examination as comparison tests. Nine cadaver limbs were acquired from clinically normal horses. Stifles were evaluated ultrasonographically and then by an extensive 1.5 T MRI protocol. Subsequently, all stifles were evaluated for gross pathologic change. Findings were compared between gross evaluation and MRI imaging and described. No soft tissue structure abnormalities were identified on any evaluation. Specific descriptive findings of the meniscotibial, meniscofemoral, collateral, patellar and cruciate ligaments, and the menisci were reported. The high field MRI protocol described in this study provided high spatial and contrast resolution of the soft tissue structures, and this in turn allowed visualization of detailed structural characteristics, such as striations and variations in signal intensity. Findings supported the use of high field MRI as a modality for the evaluation of the soft tissues of the equine stifle. As clinical availability of this modality increases in the future, authors anticipate that new stifle diseases will be detected that have not previously been identified with other imaging modalities.

Inner ear spiral ganglion neurons were cultured from day 4 postnatal mice and loaded with a fluorescent Ca2+indicator (fluo-4, -5F, or -5N). Pulses of infrared radiation (IR; 1,863 nm, 200 µs, 200–250 Hz for 2–5 s, delivered via an optical fiber) produced a rapid, transient temperature increase of 6–12°C (above a baseline of 24–30°C). These IR pulse trains evoked transient increases in both nuclear and cytosolic Ca2+concentration ([Ca2+]) of 0.20–1.4 µM, with a simultaneous reduction of [Ca2+] in regions containing endoplasmic reticulum (ER). IR-induced increases in cytosolic [Ca2+] continued in medium containing no added Ca2+(±Ca2+buffers) and low [Na+], indicating that the [Ca2+] increase was mediated by release from intracellular stores. Consistent with this hypothesis, the IR-induced [Ca2+] response was prolonged and eventually blocked by inhibition of ER Ca2+-ATPase with cyclopiazonic acid, and was also inhibited by a high concentration of ryanodine and by inhibitors of inositol (1,4,5)-trisphosphate (IP3)-mediated Ca2+release (xestospongin C and 2-aminoethoxydiphenyl borate). The thermal sensitivity of the response suggested involvement of warmth-sensitive transient receptor potential (TRP) channels. The IR-induced [Ca2+] increase was inhibited by TRPV4 inhibitors (HC-067047 and GSK-2193874), and immunostaining of spiral ganglion cultures demonstrated the presence of TRPV4 and TRPM2 that colocalized with ER marker GRP78. These results suggest that the temperature sensitivity of IR-induced [Ca2+] elevations is conferred by TRP channels on ER membranes, which facilitate Ca2+efflux into the cytosol and thereby contribute to Ca2+-induced Ca2+-release via IP3and ryanodine receptors.NEW & NOTEWORTHY Infrared radiation-induced photothermal effects release Ca2+from the endoplasmic reticulum of primary spiral ganglion neurons. This Ca2+release is mediated by activation of transient receptor potential (TRPV4) channels and involves amplification by Ca2+-induced Ca2+-release. The neurons immunostained for warmth-sensitive channels, TRPV4 and TRPM2, which colocalize with endoplasmic reticulum. Pulsed infrared radiation provides a novel experimental tool for releasing intracellular Ca2+, studying Ca2+regulatory mechanisms, and influencing neuronal excitability.