Vargas Karina J, Sarmiento José M, Ehrenfeld Pamela, Añazco Carolina C, Villanueva Carolina I, Carmona Pamela L, Brenet Marianne, Navarro Javier, Müller-Esterl Werner, González Carlos B
Department of Physiology, Universidad Austral de Chile, Valdivia 509-9200, Chile.
Differentiation. 2009 Apr;77(4):377-85. doi: 10.1016/j.diff.2008.11.002. Epub 2009 Jan 20.
The V(2) vasopressin receptor gene contains an alternative splice site in exon-3, which leads to the generation of two splice variants (V(2a) and V(2b)) first identified in the kidney. The open reading frame of the alternatively spliced V(2b) transcript encodes a truncated receptor, showing the same amino acid sequence as the canonical V(2a) receptor up to the sixth transmembrane segment, but displaying a distinct sequence to the corresponding seventh transmembrane segment and C-terminal domain relative to the V(2a) receptor. Here, we demonstrate the postnatal expression of V(2a) and V(2b) variants in the rat cerebellum. Most importantly, we showed by in situ hybridization and immunocytochemistry that both V(2) splice variants were preferentially expressed in Purkinje cells, from early to late postnatal development. In addition, both variants were transiently expressed in the neuroblastic external granule cells and Bergmann fibers. These results indicate that the cellular distributions of both splice variants are developmentally regulated, and suggest that the transient expression of the V(2) receptor is involved in the mechanisms of cerebellar cytodifferentiation by AVP. Finally, transfected CHO-K1 expressing similar amounts of both V(2) splice variants, as that found in the cerebellum, showed a significant reduction in the surface expression of V(2a) receptors, suggesting that the differential expression of the V(2) splice variants regulates the vasopressin signaling in the cerebellum.
血管加压素V(2)受体基因在第3外显子中含有一个可变剪接位点,这导致产生两种剪接变体(V(2a)和V(2b)),最初是在肾脏中发现的。可变剪接的V(2b)转录本的开放阅读框编码一种截短的受体,其在第六跨膜区段之前显示出与经典V(2a)受体相同的氨基酸序列,但相对于V(2a)受体,在相应的第七跨膜区段和C末端结构域显示出不同的序列。在此,我们证明了V(2a)和V(2b)变体在大鼠小脑中的出生后表达。最重要的是,我们通过原位杂交和免疫细胞化学表明,从出生后早期到晚期发育阶段,两种V(2)剪接变体均优先在浦肯野细胞中表达。此外,两种变体均在成神经的外颗粒细胞和伯格曼纤维中短暂表达。这些结果表明,两种剪接变体的细胞分布受到发育调控,并提示V(2)受体的短暂表达参与了精氨酸加压素介导的小脑细胞分化机制。最后,转染的CHO-K1细胞表达的两种V(2)剪接变体的量与小脑中发现的量相似,其V(2a)受体的表面表达显著降低,这表明V(2)剪接变体的差异表达调节了小脑中的血管加压素信号传导。