Alternative splicing plays an important role in the expression of genetic information. Among the best understood alternative splicing factors are transformer and transformer-2, which regulate sexual differentiation in Drosophila. Like the Drosophila genes, the recently identified mammalian homologues are subject to alternative splicing. Using an antibody directed against the major human transformer-2 beta isoform, we show that it has a widespread expression in the rat brain. Pilocarpine-induced neuronal activity changes the alternative splicing pattern of the human transformer-2-beta gene in the brain. After neuronal stimulation, a variant bearing high similarity to a male-specific Drosophila tra-2179 isoform is switched off in the hippocampus and is detectable in the cortex. In addition, the ratio of another short RNA isoform (htra2-beta2) to htra2-beta1 is changed. Htra2-beta2 is not translated into protein, and probably helps to regulate the relative amounts of htra2-beta1 to beta3. We also observe activity-dependent changes in alternative splicing of the clathrin light chain B, c-src and NMDAR1 genes, indicating that the coordinated change of alternative splicing patterns might contribute to molecular plasticity in the brain.

Alternative splice-site selection is regulated by the relative concentration of individual members of the serine-arginine family of proteins and heterogeneous nuclear ribonucleoproteins. Most of these proteins accumulate predominantly in the nucleus, and a subset of them shuttles continuously between nucleus and cytosol. We demonstrate that in primary neuronal cultures, a rise in intracellular calcium concentration induced by thapsigargin leads to a translocation of the splicing regulatory protein tra2-beta1 and a consequent change in splice-site selection. To investigate this phenomenon under physiological conditions, we used an ischemia model. Ischemia induced in the brain causes a cytoplasmic accumulation and hyperphosphorylation of tra2-beta1. In addition, several of the proteins binding to tra2-beta1, such as src associated in mitosis 68 and serine/arginine-rich proteins, accumulate in the cytosol. Concomitant with this subcellular relocalization, we observed a change in alternative splice-site usage of the ICH-1 gene. The increased usage of its alternative exons is in agreement with previous studies demonstrating its repression by a high concentration of proteins with serine/arginine-rich domains. Our findings suggest that a change in the calcium concentration associated with ischemia is part of a signaling event, which changes pre-mRNA splicing pathways by causing relocalization of proteins that regulate splice-site selection.

Le bassin versant du Narh Ibrahim est classé parmi les sites du patrimoine mondial. Les rejets sauvages solides et liquides ont un impact important sur le développement d’une contamination bactériologique tout au long de la rivière. Dans cette étude, des paramètres physico-chimiques et bactériologiques ont été suivis sur neuf sites de prélèvement qui couvrent la rivière Nahr Ibrahim de son amont jusqu’à son estuaire. L’ensemble des paramètres est étudié pendant des périodes de l’année caractérisées par un temps sec ou un temps de crue. Au cours de ces périodes, ces paramètres ont révélé une influence importante du type d’occupation des sols et des phénomènes de lessivage sur la composition bactériologique de la rivière. L’origine et le degré de la contamination bactérienne instantanée ont été également identifiés. Une approche statistique multivariée a montré que l’effet de la localisation du site masque l’effet date sur un même site et pendant la même période. Les sites en aval de la rivière sont caractérisés par une pollution en nitrate et une contamination bactériologique alors que les sites en amont sont marqués par une contamination bactériologique seulement.The Nahr Ibrahim catchment area is classified as an international heritage site. Along the Nahr Ibrahim River, solid and liquid effluents have an important impact on bacterial contamination. Physico-chemical and bacteriological parameters were studied at nine sampling sites located along the Nahr Ibrahim River from its source to its estuary. Total bacteria, total coliforms, fecal coliforms and fecal streptococci were studied during dry and wet weather periods. During these periods, the studied parameters revealed an important influence of the type of soil and leaching on the bacteriological composition of the river. Furthermore, the origin and the degree of temporary bacteriological contamination were identified. A multivariate statistical approach demonstrated that the effect of site location masked the effect of sampling date on the degree of bacteriological contamination. Downstream river sites were characterized by nitrate and bacterial pollution whereas upstream sites showed only by bacterial contamination