Study On The Developmental Expression Of Alternative Splicing MRNA Of Swine PROP1 And POU1F1 Pre-mRNA

PROP1, a pituitary transcriptional factor, is the ancestral protein of Pituitary-specific factor 1 (POU1F1) and iniate the expression of POU1F1genes in embryonic stage and maintained the postnatal expression. POU1F1 is an important transcription factor expressed in cephalic of pituitary gland and plays important role in regulation the expression of target genes of PRL, GH, TSH-βand itself by binding to the promoter of them, and regulates animal growth, development, reproduction and immunity. In this study, the developmental expression of alternative splicing mRNA of swine PROP1 and POU1F pre-mRNA and the target genes of POU1F1 (GH, PRL and TSH-β) in 1-150d Meishan pig was investigated by reverse transcription polymerase chain reaction (RT-PCR) technology. The POU1F1 cDNA was cloned and sequenced, the deduced protein was analysized as well. The main results was as follows: 1. Two alternative splicing mRNA of swine PROP1 (PROP1αand PROP1β) and four alternative splicing mRNA POU1F pre-mRNA ( POU1F1-α,POU1F1-β,△3 POU1F1and△4 POU1F1) were identified in pig pituitary by RT-PCR; and the developmental expression patterns of them and GH, PRL and TSH-βin 1-150d Meishan pig was revealed. 2. The alternative splicing mRNA of swine PROP1 showed different developmental expression patterns in the boars and gilts and the abundance of PROP1αwas higher than that of PROP1βat different develpmental stages. In male, the expression of PROP1αwas high on 1d, decreased until 60d and peaked on 90d and declined again to 90d. While the expression of PROP1αin gilts was low on 1d, peaked on 30d and decreased again on 60d, and then increased to 90d and maintained until 150d. The developmental expression of PROP1βbetween boars and gilts was similar, which was went up at first and decreased then, increased again on 90d and maintained until 150d.3. The developmental expression patterns of alternative splicing mRNA of POU1F1 were different between Meishan boars and gilts. The POU1F1αin boars decreased at first, and went up on 60d, then declined until 150d, while the POU1Fαin gilts increased on 30d and decreased on 60d, and then went up until 150d.4. The expression of GH .PRL and TSH-βgene was conducted with reverse transcription PCR in MeiShan swine.In Pituitary GH and PRLgene the expression changed differently in male and female showed sexual dimorphism during ; however, the peak appeared at different time of male and female in hypothalamus,. TSH-βgene the expression level decline after 90 day's age.5. The expression alternative messenger RNA splicing of PROP1and POU1F1 gene were showed that alternative messenger RNA splicing of PROP1 gene was similar to△4POU1F1, POU1F1-αmRNA expression was similar to PROP1 gene on 1 day,30 day ,on 90 day and 150 day had a greater difference. In male, two alternative messenger RNA splicing of PROP1 gene expression was similar to△3 POU1F1 during 1 -60 days, on 90 day and 150 day had a greater difference. In female ,△3POU1F1 expression was similar to PROP1 gene.6. Expression characteristics of POU1F1,GH and PRLin Pituitary were compared in this study, and we found that, POU1F1α,△3 POU1F1 mRNA was similar to GH mRNA and PRL mRNA had a greater difference . 7. The cDNA of swine POU1F1 sequences of 876bp size was gained by the methods of RT-PCR and sequencing in MeiShan swine Pituitary, the result sequence was analyzed by DNA Star and it demonstrated 99.7% homology with published sequence in Genbank.