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Neo-sex chromosomes often originate from sex chromosome-autosome fusions and constitute an important basis for the study of gene degeneration and expression in a sex chromosomal context. Neo-sex chromosomes are known from many animal and plant lineages, but have not been reported in birds, a group in which genome organization seems particularly stable. Following indications of sex linkage and unexpected sex-biased gene expression in warblers (Sylvioidea; Passeriformes), we have conducted an extensive marker analysis targeting 31 orthologues of loci on zebra finch chromosome 4a in five species, representative of independent branches of Passerida. We identified a region of sex linkage covering approximately the first half (10 Mb) of chromosome 4a, and associated to both Z and W chromosomes, in three Sylvioidea passerine species. Linkage analysis in an extended pedigree of one species additionally confirmed the association between this part of chromosome 4a and the Z chromosome. Markers located between 10 and 21 Mb of chromosome 4a showed no signs of sex linkage, suggesting that only half of the chromosome was involved in this transition. No sex linkage was observed in non-Sylvioidea passerines, indicating that the neo-sex chromosome arose at the base of the Sylvioidea branch of the avian phylogeny, at 47.4-37.6 millions years ago (MYA), substantially later than the ancestral sex chromosomes (150 MYA). We hypothesize that the gene content of chromosome 4a might be relevant in its transition to a sex chromosome, based on the presence of genes (for example, the androgen receptor) that could offer a selective advantage when associated to Z-linked sex determination loci. [ABSTRACT FROM AUTHOR]

The endangered minnow Iberochondrostoma almacai is an endemic Iberian cyprinid with a restricted and fragmented distribution. Here, we describe the genetic structure of the species and infer its demographic history from six nuclear-encoded microsatellite loci and mitochondrial cytochrome b sequences. Genetic diversity was low (microsatellite He<0.45; mtDNA π<0.0015), and both markers resolved two groups: one from the northern Mira drainage and one from the Arade and Bensafrim drainages. The relatively low differentiation between these groups (0.09< FST<0.29; 0.31< ΦST <0.57) suggests past headwater captures and/or that populations were large until recently. The genetic diversity and differentiation estimates were compared with those for other three endangered cyprinids inhabiting similar intermittent rivers. Microsatellite data indicate a population decrease in the last 100–2400 years, probably as a result of anthropogenic disturbance. Human activities together with an intermittent flow of these rivers apparently led to local extinctions with consequent fragmentation and contraction in range. We recognize two management units corresponding to the two genetic groups identified. To maintain/increase genetic diversity, we recommend habitat restoration actions and measures to increase gene flow within and/or between the two units, under controlled reproductive programmes. Ecological experiments should be performed to ensure the success of supplementation among the two units. Moreover, the reintroductions in unoccupied drainages are suggested if further data confirm the presence of I. almacai in the recent past. [ABSTRACT FROM AUTHOR]

Abstract: In fish of the Squalius alburnoides complex, hybridisation and polyploidy have affected sex ratios, resulting in strong correlations between sex and genotype. The preponderance of females among triploids and the occurrence of an all male lineage among diploids seem to imply that sex ratio deviations should have a strong genetic basis. Until now, no information has been gathered regarding the molecular basis of sex determination in this intricate hybrid system. Thus, putative regulatory elements of the cascade that potentially are involved in sex determination in S. alburnoides have to be investigated. Being reported to have an important role in teleost sex determination, and more particularly in male gonad development, the anti-Müllerian hormone, amh was a good initial candidate. Here we report the isolation, cloning and characterization of the amh ortholog in S. alburnoides and the ancestral species S. pyrenaicus. In adult S. alburnoides and S. pyrenaicus of both sexes, amh shows a gonad specific expression pattern, restricted to the Sertoli cell lineage in testis and to granulosa cells in ovaries. During development, it plays an early role in male gonad differentiation in S. alburnoides. Overall the observed patterns are similar to what has been reported in other teleost species. This suggests a conserved role of amh and implies that its expression dynamics cannot be directly responsible for the sex ratio deviations reported in S. alburnoides. It is possible that a conjunction of other factors could be contributing for sex ratio imbalance. The present results constitute the starting point in the characterization of the S. alburnoides sex determination cascade, a process that we expect to shed some light on the molecular basis of sex distribution, within the context of hybrid system evolution. [Copyright &y& Elsevier]

Several studies have demonstrated the hybrid origin of Squalius alburnoides, a complex of diploid, triploid and tetraploid fish, with Squalius pyrenaicus as the maternal ancestor and an Anaecypris hispanica-like species as the paternal ancestor. The aim of this study was to assess patterns of genetic diversity and similarity between sympatric forms of the different ploidy levels of S. alburnoides and related parental species in the two largest basins that are part of its southern Iberian distribution, the Tejo and Guadiana, using microsatellites. High levels of genetic diversity were found in populations of S. alburnoides from both basins, reflecting the hybrid origin, the high diversity of forms in these river basins and the different modes of reproduction of the complex. Microsatellites indicated low values of genetic differentiation between the non-hybrid all-male form of S. alburnoides and A. hispanica, supporting the view that these forms share a common ancestor. In some instances, diploid and triploid hybrids from the same river basin were more closely related to each other than to their counterparts in the other basins, suggesting an ancient origin for the complex in these Iberian southern basins. [ABSTRACT FROM AUTHOR]

Abstract: The Squalius alburnoides complex, of hybrid origin, comprises diploid, triploid and tetraploid forms and has a widespread distribution in the Iberian waters. The southern populations of this complex, sympatric with S. pyrenaicus, show high genetic variability, diversity of forms and reproductive modes which create pathways that may allow for the establishment of a new species in the future. Here we report a contrasting view over the S. alburnoides complex: in the Mondego River basin (northern Portugal), nuclear “non-hybrid” and tetraploid forms are absent and a clearly impoverished genetic diversity is observed, contributing to a general scarcity of possibilities of generating novel genetic material. Moreover, the bisexual species involved in the maintenance of the complex in this basin (S. carolitertii) exhibits a considerably lower genetic variability, when compared with S. pyrenaicus. The observed differences suggest that, despite being originated by similar hybridization events and maintained by analogous reproductive mechanisms, different populations of the complex were exposed to distinct evolutionary constrains, which in some cases resulted in diversification and speciation while in others led to a compromising situation in terms of evolutionary potential. Additionally, and for the first time all forms were used in the calculation of genetic distances and diversity indices, widening the possibilities of analysis of the complex by allowing the inclusion of a large part of the available data, irrespective of ploidy level. [Copyright &y& Elsevier]

Calmodulin (CaM), a calcium-binding protein, is found in high concentrations in mammalian brain where it plays a pivotal role in a large number of cellular functions. Protein kinase C (PKC), a multifunctional cytosolic enzyme, in the presence of both Ca2+ and phospholipids, transduce extracellular signals into intracellular events. Both CaM and PKC are partially involved in maintaining Ca2+ homeostasis in the cell. Any fluctuations in the intracellular Ca2+ can modulate cellular functions and may contribute to neuronal dysfunction. Hence, the present investigation was initiated to study the effects of some selected penicillium (naturally occurring tremorgenic) mycotoxins like secalonic acid, citreoviridin, and verruculogen on CaM activity, active conformation of CaM and PKC activity. Stimulation of CaM-deficient bovine brain 3-5 phosphodiesterase (PDE) indicated CaM activity. The modification of CaM active conformation was studied by the binding of fluorescent probe N -phenyl-l-napthylamine (NPN) to CaM. Alterations in the fluorescence of dansyl-CaM was used to study the effect of these compounds on complex formation between CaM and PDE. Rat brain cytosolic PKC was studied using 32P-ATP as a measure of altered protein phosphorylation. The concentrations of mycotoxins used were in the range of 10 to 50 muM. All three mycotoxins inhibited CaM-stimulated PDE activity in a concentration-dependent manner. Citreoviridin and secalonic acid inhibited NPN fluorescence and Ca2+-dependent complex formation of dansyl-CaM and PDE. The IC values for NPN fluorescence of citreoviridin and 50 secalonic acid were 13 muM and 19 muM respectively. However, verruculogen showed little effect on NPN fluorescence and the Ca2+dependent complex formation of dansyl-CaM and PDE. These mycotoxins also inhibited PKC activity in a concentration-dependent manner with IC50 values of 19.8, 25.7, and 38.4 muM for secalonic acid, citreoviridin, and verruculogen, respectively. The results of our study suggest that these mycotoxins at very low concentrations are interacting with CaM and PKC. Such an effect could lead to impairment of neurotransmission and result in neurotoxicity. [ABSTRACT FROM AUTHOR]

Teachers in five primary schools were surveyed for their knowledge and attitudes about epilepsy. An analysis of 113 teacher responses revealed knowledge deficits, especially regarding the prevalence of epilepsy, the future of children with the disorder and E.E.G. as an aid to diagnosis. Misconceptions regarding first aid were also common. More than two-thirds of the teachers believed that children with epilepsy have academic problems more often, although about half of the respondents were in favour of normal schools for such children. Most respondents recommended a teacher's career for these children in future but considered that of a bus driver, as unsuitable. Few teachers had attended any educational programme on epilepsy. Only about one-fifth of the teachers were confident in dealing with an epiletic child. Communication about epilepsy among the teachers, parents and doctors was virtually non-existent. [ABSTRACT FROM AUTHOR]