April - June 2005, Volume 1, Issue 2
General Endocrinology
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Acta Endocrinologica (Buc)
Radian S, Bensaada M, Lautier C, Moles JP, Grigorescu F, Gussi I, Badiu C, Nastasia S, Hudita D, Leonte L, Marinescu B, Coculescu M
Molecular genetics strategies to identify vasotocin coding sequences in humans: family-specific approach using genomic DNA and fetal tissues mRNAs
Acta Endo (Buc) 2005, 1 (2): 131-144doi: 10.4183/aeb.2005.131
Vertebrate nonapeptide neurohormones constitute an evolutionarily conserved family, involved in vital functions, such as hydro-osmotic balance regulation, reproduction and social behaviour. Two human members of this family are known, vasopressin (AVP) and oxytocin (OXT), with their highly homologous genes closely located on Chr 20p13. Presence of vasotocin (AVT) in man has been suggested, but remains controversial, and genetic evidence is lacking. AVT activity could be explained by the presence of a third distinct gene for AVT or an RNA-processing mechanism involving products of AVP and/or OXT genes. To test the first hypothesis, we developed bioinformatics and experimental approaches using genomic DNA and fetal tissues mRNAs. Family-specific primers for AVT and neurophysin were designed based on CODEHOP strategy and used in our experiments. Results of bioinformatics and genomic DNA experiments (family-specific and Alu step-out PCR) suggest there is no evidence for an AVT gene in the genome. RNA-based techniques 3?-RACE and Family-Specific Domain Restriction Fragment RTPCR provided evidence for new transcript species that could code for AVT. Further experiments will be needed to characterize them. We discuss potential mechanisms of AVT mRNA generation based on AVP and OXT mRNAs, by alternative splicing, heterologous transsplicing or RNA-editing. While all methods we developed proved feasible, current results suggest there is no AVT gene in the genome, but specific mRNAs could be present in fetal tissues. Their full characterization may potentially allow identification of vasotocin mRNA and shed light on a subject of fundamental scientific interest.
Keywords: AVT gene, family-specific, neurohypophyseal, fetal
Correspondence: Florin Grigorescu, Molecular Endocrinology, IURC, 641 ave du Doyen Gaston Giraud, 34093 Montpelllier CEDEX5, FRANCE, florin.grigorescu@montp.inserm.fr
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