Molecular cloning, characterisation and tissues expression analysis of the goose (Anser cygnoides) vasoactive intestinal peptide (VIP) gene

Abstract

1. Vasoactive intestinal peptide (VIP) is involved in the control of prolactin (PRL) release and plays a pivotal role as a regulator of reproductive behaviour and neuroendocrine secretion in birds. 2. In this study, a 941-bp cDNA fragment covering the complete coding region (CDS) of goose VIP gene was identified. The cDNA contains a 32-bp 5'-untranslated region (UTR), a 603-bp CDS and a 306-bp 3'-UTR containing two ATTTA sequence elements, two polyadenylation signals (AATAAA) and a 25-bp poly (A) tail. 3. Seven exons and 6 introns were identified, and both the cDNA and genomic DNA sequences showed high identity with those of other species. 4. The sequence analysis indicated that there were two alternatively spliced transcripts the long transcript (VIP-1) encoded both VIP and peptide histidine isoleucine exons and the short one (VIP-2) only encoded VIP. 5. RT-PCR analysis indicates that the expression level of the VIP-1 is much lower than that of VIP-2, and that VIP-1 is negligible or absent in muscle, abdominal fat, ovary and spleen, whereas VIP-2 is widely distributed in all the examined tissues. 6. A total of 12 single nucleotide polymorphisms (SNPs), including 2 SNPs located in the coding region and 10 variations in intron regions, were identified in goose VIP gene.

Molecular characterization and tissue expression profiles of prepro-vasoactive intestinal peptide in the Chinese alligator (Alligator sinensis) during the active and hibernating periods

“…It consists of a 221‐bp 5′‐UTR, a 606‐bp complete coding region (CDS), and a 312‐bp 3′‐UTR. Only one ATTTA element was found in the 3′‐UTR of prepro‐VIP cDNA in Chinese alligator, which is different from two in the swan goose (Liu et al., ) and three in the flounder (Nam et al., ); previous reports indicated that the ATTTA elements were typically existed in cytokine mRNAs, which mediate RNA instability (Nam et al., ). The 3′‐UTR also contains two polyadenylation signals (AATAAA) in Chinese alligator, which is consistent with those observed in birds (Hiyama, Sato, Zadworny, and Kansaku, ; Liu et al., ; McFarlin et al., ; You et al., ) and fishes (Nam et al., ), but is different with mammals, in which there were three polyadenylation signals (AAUAAA) in the 3′‐UTR of their prepro‐VIP (Giladi et al., ; Itoh et al., ; Tsukada et al., ).…”

“…PHI/PHM is structurally related to VIP and shares many of VIP's biological actions but is generally less potent than VIP (Henning, & Sawmiller, ). The molecular structure of prepro‐VIP had been extensively studied in mammals and birds, such as human Homo sapiens (Itoh, Obata, Yanaihara, & Okamoto, ; Tsukada et al., ), rat Rattus norvegicus (Giladi, Shani, & Gozes, ), chicken Gallus gallus (McFarlin, Lehn, Moran, MacDonald, & Epstein, ), turkey Meleagris gallopavo (Pitts et al., ), and goose Anser cygnoides (Liu et al., ), wherein structural conservation of the prepro‐VIP was found, whereas some variations were also reported especially in the PHI region. The reptiles are in a special phylogenetic position, which have a sister group relationship with birds and from which mammals have evolved (Janke, & Arnason, ), so it is of particular interest to investigate the molecular structure of reptilian prepro‐VIP.…”

“…Many studies have shown that VIP can be produced in neuronal and various peripheral tissues (Gomariz, Leceta, Garrido, Garrido, & Delgado, ; Lamperti, Rosen, & Villa‐Komaroff, ; Liu et al., ; Nam et al., ; Wang, & Conlon, ; Wang, Zhang, Zhou, Wang, & Wu, ; You, Silsby, Farris, Foster, and El Halawani, ). VIP mRNA tissue‐specific expression in turkey analyzed by Northern blots indicated that VIP is expressed in hypothalamus, small intestine, large intestine, uterus, and oviduct (You et al., ).…”

“…VIP mRNA tissue‐specific expression in turkey analyzed by Northern blots indicated that VIP is expressed in hypothalamus, small intestine, large intestine, uterus, and oviduct (You et al., ). Tissues expression analysis by RT‐PCR technique revealed that the VIP mRNA in goose is expressed in the hypothalamus, pituitary, muscle, subcutaneous fat, duodenum, kidney, liver, heart, pancreas, oviduct, ovary, lung, and spleen (Liu et al., ). In adult flouder Paralichthys olivaceus , the prepro‐VIP mRNA was detected by RT‐PCR amplification in the intestine, spleen, stomach, and heart, whereas not in the muscle, liver, and kidney (Nam et al., ).…”

“…Many studies have shown that VIP can be produced in neuronal and various peripheral tissues (Gomariz, Leceta, Garrido, Garrido, & Delgado, 1994;Lamperti, Rosen, & Villa-Komaroff, 1991;Liu et al, 2014;Nam et al, 2009;Wang, & Conlon, 1993;Wang, Zhang, Zhou, Wang, & Wu, 2014;You, Silsby, Farris, Foster, and El Halawani, 1995).…”