Distribution of neuronal and non-neuronal spliced variants of type 1 IP 3-receptor in rat hypothalamus and brain stem

Abstract

In the nervous system, inositol 1,4,5-trisphosphate (IP 3) is one of the second messengers produced by PI hydrolysis and triggers IP 3-receptor (IP 3R) mediated calcium release from intracellular pools. Throughout the brain, the type 1 IP 3R is predominantly expressed and its mRNA is widely distributed. Alternative splicing of IP 3R1 (SI and SII) occurs in two distinct regions. SI splicing in the middle of the ligand binding domain may alter the IP 3 binding activity, while SII splicing probably affects the protein kinase A phosphorylation sites and kinetics. Selective use of IP 3-receptor subtypes may permit a tissue specific and developmentally specific expression of functionally distinct channels. The present work was focused on detection of the alternatively spliced mRNA of type 1 IP 3-receptor in individual brain structures and nuclei. Using RT-PCR we detected neuronal (535 bp) and non-neuronal (410 bp) forms. We identified both spliced variants in the majority of brain structures, except in the cerebellum and medulla. In the cerebellum, the neuronal form of type 1 IP 3R was found exclusively, while in the medulla, the non-neuronal form was much more abundant. Nevertheless, Western blot analysis and hybridization with specific antibody against IP 3R revealed no qualitative, but only quantitative differences. Similarly, IP 3 dependent calcium release did not show any differences between the cerebellum and pons. These results demonstrate the distribution of alternatively spliced S2 variants of type 1 IP 3R in selected brain structures and nuclei. The physiological relevance of these two forms remains to be elucidated by further studies.