Expression of Cholinergic Markers and Characterization of Splice Variants during Ontogenesis of Rat Dorsal Root Ganglia Neurons

International Journal of Molecular Sciences

AdaMaria Tata¹  Veronica Corsetti¹  Giancarlo Poiana¹  Elisabetta Botticelli¹  MichaelSebastian Salazar Intriago¹  Stefano Biagioni¹  Gabriella Augusti-Tocco¹  Carla Perrone-Capano² 

[1] Department of Biology and Biotechnology Charles Darwin, “Sapienza” University of Rome, 00185 Rome, Italy;Department of Pharmacy, University of Naples Federico II, 80138 Naples, Italy;

关键词: cholinergic locus;    sensory neurons;    acetylcholine;    choline acetyltransferase;    vesicular acetylcholine transporter;    splice variants;   

DOI  :  10.3390/ijms22115499

来源: DOAJ

【 摘 要 】

Dorsal root ganglia (DRG) neurons synthesize acetylcholine (ACh), in addition to their peptidergic nature. They also release ACh and are cholinoceptive, as they express cholinergic receptors. During gangliogenesis, ACh plays an important role in neuronal differentiation, modulating neuritic outgrowth and neurospecific gene expression. Starting from these data, we studied the expression of choline acetyltransferase (ChAT) and vesicular ACh transporter (VAChT) expression in rat DRG neurons. ChAT and VAChT genes are arranged in a “cholinergic locus”, and several splice variants have been described. Using selective primers, we characterized splice variants of these cholinergic markers, demonstrating that rat DRGs express R1, R2, M, and N variants for ChAT and V1, V2, R1, and R2 splice variants for VAChT. Moreover, by RT-PCR analysis, we observed a progressive decrease in ChAT and VAChT transcripts from the late embryonic developmental stage (E18) to postnatal P2 and P15 and in the adult DRG. Interestingly, Western blot analyses and activity assays demonstrated that ChAT levels significantly increased during DRG ontogenesis. The modulated expression of different ChAT and VAChT splice variants during development suggests a possible differential regulation of cholinergic marker expression in sensory neurons and confirms multiple roles for ACh in DRG neurons, both in the embryo stage and postnatally.

【 授权许可】

Unknown