The Expression In Different Tissues And Localization In Testis And Epididymidis Of Gnαs

The guanine nuecleotide-binding proteins alpha S(Gnαs) was a widely expressed G protein alpha, was the important genes involved in energy and glucose metabolism in mice, was an important signal transduction molecule in the cell, and independently regulated a series of receptors including the adenosine cyclase, phospholipase C beta and ion channels. Gnαs had tissue specificity, was mainly expressed in the pituitary gland, thyroid gland, renal proximal tubule, gonad and hypothalamus in the way of maternal allele. The Gnαs had been detected in different tissues of heart, liver, kidney and other tissues of pigs, human, cattle, mice and rats,also had been detected in human testis cells and interstitial cells. So The Gnαs may play important role in testicular development, sperm maturation and energy and glucose metabolism.To study the bioinformatics analysis, the expression in different tissues of one-year-old sheep, and to locate Gnαs in the the testis and epididymidis of sheep.This study chose dorper sheep(♂)×Mongolian sheep (♀)one year old hybrid as the test objiect, the total RNA of heart, liver, spleen, brain, testis, epididymis and other 20 tissues were extracted. The expression characteristics of Gnαs mRNA in the tissueswere detected by RT-PCR, Gnαs in testis and epididymis were located by the immunohistochemical technique, the Gnαs protein were analysed by the Western Blotting technology. The main results were as follows:1. The Gnαs sequences were amplified, sequencing results were BLAST analysed by GenBank NCBI.The results showed the homology between the sequence obtained by the experiment and that of published by the NCBI of bov, pig, human, mouse, rat and brown wolf were 98.5%,95.7%,94.47%, 94.31%,93.05% and 96.68%, it showed that the conservation of Gnαs cDNA sequencesof sheep was high, and the homology between sheep and bov was highest. The sequencing results were uploaded to GenBank (Accession Number:KP940631). The 1810bp of Gnαs cDNA in sheep was obtained. The Gnαs sequence encoded 395 amino acids. There were no signal peptide, one potential glycosylation sites and potential phosphorylation sites. The transmembrane structure of Gnαs was not found.2. The Gnαs mRNA was widely expressed in various tissues of 12-month-old sheep, The expression quantities of mRNA were Pituitary>brain>in the hypothalamus>cerebellum>abomasum>longissimus muscle>spleen>Network>stomach liver>omasum>kidney>lung>foreleg muscle>leg muscle> ABS>rumen. The results showed that the expression quantities of Gnαs in the nervous system were highest.3. The expression quantities of Gnαs mRNA and protein in testis and Epididymidis were Epididymidis tail(EpT)>Epididymidis body(EpB)>Epididymidis head(EpH)>testis. The results of the immunohistochemical technique showed the Gnαs were all detected in testis and epididymis of 12-month-old hybrid sheep, the Gnαs were mainly expressed in testicular sperm cells, sperm stem cells, supporting cells, stromal cells and sperm, in the epididymis of the primary cells and the epididymis in the interstitial cells and in the epididymis. The expression signal in Interstitial cells of epididymis was strongest, the strong positive expression in the head of epididymiswas found, the positive signal in the epididymis and epididymis was weak.The results showed Gnαs played important role in testicular development, sperm maturation and energy and glucose metabolism.We speculated that Gnαs may influence the component of luminal fluid of epididymis and micro environment of sperm maturation, influence the sperm maturation.