Fluctuating Gene Expression and Localized Cellular Distribution of Vasoactive Intestinal Contractor (VIC) in Mouse Uterus

Saida

et al. 2003

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ABSTRACT. Endothelin-2 (ET2) is a member of the endothelin family of 21-amino acid peptides with vasoconstrictive activity. We report here the molecular cloning of the canine full-length cDNA of the precursor form of ET2, prepro-ET2 (PPET2), from intestinal tissue by means of reverse transcription-polymerase chain reaction (RT-PCR) in conjunction with 5'-and 3'-rapid amplification of cDNA ends (RACE). Aside from the poly (A) tail the cDNA was found to be 1195 bp and included an open reading frame of 534 bp encoding a PPET2 polypeptide of 178 residues, in which the regions corresponding to bioactive mature ET2 peptide, an intermediate form big-ET2, and endothelin-like peptide are found. The organ distributions of PPET2 mRNA and a splicing variant were analyzed by RT-PCR. PPET2 transcript was detected in duodenum, colon, stomach, lung, liver, uterus, ovary, testis and kidney, but not in spleen. A splicing variant was found in none of the organs. Thus, based on the cloned cDNA sequence, we established a quantitative assay for dog P PET2 mRNA level using a real-time PCR system. Quantitative analysis by this method in various organs of the dog demonstrated that the dominant gene expression occurs in the intestine, with higher expression in large intestine than in small intestine.

Section: Discussionsupporting

confidence: 92%

Cloning of Full-Length Preproendothelin-2 cDNA and Its Expression in Dog

Saida

et al. 2003

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“…A 353-bp fragment predicted from the cloned cDNA sequence was detected in testis, bladder, kidney, colon, duodenum, stomach and lung. The finding of predominant PPET2 mRNA expression in cat gastrointestinal tract is in concordance with the results of previous studies in mouse [15], rat [11], cow [14], horse [13] and dog [2]. In addition to the PCR fragment with the predicted size, another smaller, less abundant product was found in the stomach, but not in any of the other organs examined.…”

supporting

confidence: 91%

Expression of Preproendothelin-2 Splice Variant in Cat

Temma

et al. 2006

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ABSTRACT. Previous studies on human uterine and placental tissues have found variants, derived from alternatively spliced mRNAs, of preproendothelin-2 (PPET2) that lack a post-translational proteolytic site essential for normal processing. Here we report a splice variant of cat PPET2 mRNA expressed in the stomach. After cloning the full-length cDNA of cat PPET2, organ distribution analysis of the transcript by reverse transcriptase-polymerase chain reaction (RT-PCR) was performed. In addition to the fragment with a size predicted based on the cDNA sequence obtained by cloning, an additional PCR fragment of smaller size was detected in stomach tissue. Subsequent cloning and sequence analysis of the smaller PCR product demonstrated that it derives from a splice variant with full-length deletion of a region corresponding to exon 4 of the PPET2 gene. KEY WORDS: cat stomach, feline preproendothelin-2, splice variant.

“…PPET3 mRNA is detected throughout the organs examined, including heart, lung, liver, kidney, spleen, stomach, pancreas, duodenum, colon, uterus, ovary and testis. The finding of predominant expression of PPET3 mRNA in dog lung, kidney, duodenum and uterus is consistent with results of previous studies on mouse and rat [5,16]. However there is a differential finding that dog PPET3 mRNA was expressed in liver as well as testis, whereas mouse and rat mRNAs were not detected in these organs.…”

supporting

confidence: 91%

Complete cDNA Sequence and mRNA Expression of Dog Preproendothelin-3

Temma

et al. 2004

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ABSTRACT. The full-length cDNA of dog preproendothelin-3 (PPET3) was cloned from lung tissue using RT-PCR and rapid amplification of cDNA ends. Aside from the poly (A) tail, the full-length cDNA was 1976 bp. A polyadenylation signal sequence and one copy of a consensus sequence, ATTTA, which is related to mRNA turnover, was found in the 3' noncoding region. The cDNA had a 594-bp open reading frame encoding a 198-amino acid polypeptide. Regions corresponding to a bioactive mature ET3 peptide, an intermediate form known as big-ET3, and an ET3-like peptide were observed in dog PPET3. Expression of PPET3 mRNA was detected throughout the organs examined, which included heart, lung, liver, kidney, spleen, stomach, pancreas, duodenum, colon, uterus, ovary and testis. KEY WORDS: cDNA cloning, dog preproendothelin-3, mRNA expression.