Localization And Different Expression Of LHR And MRNA In The Uterus Of Estrous Cycle Phases In Jining Gray Goat

The mammalian reproduction action mainly reside in the hypothalamus- pituitary-gonad axis. After receiving the signals of gonadotropin releasing hormone(GnRH) from hypothalamus ,the gonadotropic cells in pars distalis hypophyseos secrete reproductive key hormone—FSH and LH , which specially bind with the corresponding G protein-coupled receptors(GPCR) of target cell in the ovary. The membrane receptor—FSHR and LHR are mediated by GPCR which can mediate and stimulate cAMP system. Consequently, the gonadotropine signals are transferred into cell.The recent research found that LHR,not only exists in the gonads,as the granulosa cell of the ovary and the Sertoli's cell of the testis,but also exists in the uterus,fallopian tube and the other gential tract of the mammal.Knockout of the LHR in the mouse resulted in infertile males and females with markedly atrophied external and internal genitalia.These research investigate LHR in the genitial tract plays important roles in the mammalian reproduction regulation.Jining gary goat is a species of lamb leathers in China, characterized by precosity and high productivity. The initial mating of female goats happened approximately 6 months old.The average yean percentage of primiparity is 163.1, general speaking,one year two fetus or two years three fetus,average yean rate is 293.7%.To invest the high productivity mechanism of Jining gray goat , We research the localization and the gene expression of the LHR and its mRNA in the uterus,which provide the theory and breeding practice foundation in elevating goat production and utilizating economic value. Experiment 1 Different expression of LHR mRNA in the uterus of four estrous cycle phases in Jining gray goatTotal RNA extracted from the uterus of Jining gray goat was used for RT-PCR amplification of goat luteinizing hormone receptor (LHR) gene. A DNA fragment of 289 bp in size was obtained and cloned into pGEM-T vector. Positive clones were identified and sequenced. Using GAPDH as an inner control, an optimal semi-quantitative RT-PCR method was established to analyze LHR mRNA expression levels in the goat uterus of four estrous cycle phases. Sequenced analysis indicated that this fragment was a partial sequence of LHR cDNA and identical to the reported LHR mRNA (AF379199). The LHR mRNA expression was different in the uterus of Jining gray goat during the four estrous cycle phrases, being lowest in the estrus (0.454±0.06), highest in the diestrus (0.705±0.09), intermediate in the proestrus (0.553±0.07).The expression in the estrus was significantly lower than in other oestrous cycle phases (P<0.05).Experiment 2 Immunohistochemistry research of LH receptor in uterus of Jining gray goatThe uterus of Jining gray goat ,collected in estrus and dioestrous,were used to identify the localization of LHR and their changes in different oestrus stage by means of immunohistochemistry and image analysis.The result suggested LHR mainly exsited in the stroma cells of the endometrium and the epithelial cells of uterine glands,Uterine smooth muscle cells had low levels of expression.we first found that it also exited in the cells of carunculae. The amounts of LHR in the stroma cells of the endometrium had significant difference between estrus and dioestrous of Jining gray goat(P<0.05),but there was no difference in the epithelial cells of uterine glands(P>0.05).