Molecular Cloning Of PRLR,PRLH And PRLHR Gene In Ovineand Theirs Expression In The Hypothalamus, Pituitary, Uterus And Ovary

Prolactin is a polypeptide hormone, anterior cells of pituitary are synthesize and secrete the prolactin, it’s play many physiological functions by bound to its receptor. While many studies have found that prolactin releasing hormone related with the secretion of PRL. The study used sheep as the experimental material, and the hypothalamus pituitary gonad axis was applied as the research object, used the molecular biology, immunohistochemistry and fluorescence quantitative method, analysed the specific expression of PRLR and PRLHR gene. the results were as follows:1. We successfully cloned the PRLR,PRLH and PRLHR, and compared the sequences respectively with the sequence of other mammals, found that the PRLR sequences were approached with Pantholops hodgsoni. A protein with 581 amino acid sequence encoded by the PRLR gene has been predicted, it has a molecular weight of 65.23 k Da, the isoelectric point of protein was p I5.12, and this protein was not stable; and another protein with 581 amino acid sequence encoded by the PRLHR gene has been predicted as well, it has a molecular weight of 41.05 k Da, the isoelectric point of protein was p I9.41, and this protein was not stable. The PRLH and PRLHR had blasted there sequence with related genes of other species found that the PRLH and PRLHR sequences were approached with Capra hircus and Pantholops hodgsoni respectively.2.We examined the m RNA expression level of PRLR,PRLH and PRLHR using Real time PCR in tissues at gonadal axis of sheep.The relative expression of PRLR in the ovary was significantly more than other tissues(P<0.05), the expression of PRLR in tissues from high to low was ovary, hypothalamus, pituitary and uterus successively. The expression of PRLH gene in uterus was significantly more than other tissues(P<0.05), the expression of PRLH in tissues from high to low was uterus ovary, pituitary, and hypothalamus successively. Expression of PRLHR in ovary and uterus were significantly different(P<0.05), the expression of PRLHR in tissues from high to low was pituitary, ovary, hypothalamus and uterus successively.3. The PRLR and PRLHR had located at the Pars distalis adenohypophysis; the cells were all round or oval, with more distribution;while they are had positive reaction in the hypothalamus, but the reaction less than the pituitary; PRLR has a positive reaction in the uterine cavity, uterus and uterine gland, while the positive reaction of PRLHR in the endometrium was higher than other parts of uterus;PRLR positive cells in the ovary was mainly distributed in the oocytes and granulosa cells, and it has more reaction than the PRLHR; In addition, the positive reaction area of PRLR including the vascular of these tissue and red blood cells.