Identification of alternatively spliced isoforms of neurotransmitter receptors from mouse genome

Abstract

Alternative splicing, the process by which the exons of primary transcripts (pre-mRNAs) from genes can be spliced in different arrangements to produce structurally and functionally distinct mRNA and protein variants, may be one of the most extensively used mechanisms that accounts for the greater macromolecular and cellular complexity of higher eukaryotic organisms. Alternative splicing is of central importance for neuronal genes and other genes involved in information processing functions. Genomic studies of alternative splicing found that the largest group of tissue-specific alternative splice isoforms was detected in brain, retina and nerve-derived tissue sources. Action potential induction and propagation is unique to each synapse and the connecting cells. These require careful and fine tuning of the neurons and other excitable cells achievable through alternative splicing of the genes involved in neurotransmission, synaptic connectivity and nervous system development. Therefore, we studied alternative splicing in neurotransmitter receptors. The aim of the study was to identify alternatively spliced transcript variants of neurotransmitter receptors by employing various bioinformatic tools and molecular biology technique including rapid amplification of cDNA ends (RACE) and reverse-transcriptase polymerase chain reaction (RT-PCR) followed by sequencing of RT-PCR products. The general aim of this study was to use a combination of bioinformatics tools and molecular biology techniques for identifying alternatively spliced isoforms/variants of neurotransmitter receptors (5HTR4) and their subunits (CHRNG and GluR1) in mouse. The specific objectives were: 1. To survey the genomic sequences, transcripts and protein isoforms of neurotransmitter receptors and their subunits from databases and literature mining. 2. To identify novel alternative spliced variants using combination of bioinformatics tools and molecular biology approaches.