| The hormone secreted by the mammalian hypothalamic-pituitary-ovary axis(HPOA),which played an important role in regulating animal reproductive activities.Gonadootropic hormone(GTH)secreted by basal cells of the anterior pituitary,including follicle stimulating hormone(FSH)and luteinizing hormone(LH),they all belonged to the biological macromolecular glycoprotein,and can’t pass the cell membrane.The regulation of the two hormones on reproductive organ activity can only be achieved through the mediating of follicle stimulating hormone receptor(FSHR)and luteinizing hormone receptor(LHR).GTH and its receptors played an important role in regulating the reproductive and physiological activities of animal reproductive organs and reproductive regulation.To investigate the regulation of gonadotropin receptor(GTHR)on the reproductive activity of Ganjia Tibetan sheep during the estrus cycle.In this study,the distribution of follicle stimulating hormone receptor(FSHR)and luteinizing hormone receptor(LHR),and the relative expression of FSHR mRNA and LHR mRNA in oviduct and uterus tissues of Ganjia Tibetan sheep during the estrus cycle were studied.Thirty-two normal development Ganjia Tibetan sheep aged 2.5 to 3.5 years were selected and divided into four groups according to the four-point method:proestrus,estrus,metaestrus and diestrus.The oviduct and uterus of Ganjia Tibetan sheep were collected during the estrus cycle.(1)The relative expression of FSHR mRNA and LHR mRNA in the oviduct and uterus were studied by real-time fluorescence quantitative PCR(RT-PCR),GAPDH and β-actin were used as internal reference genes,respectively.(2)HE staining was used to observe the structure of oviduct and uterus of Ganjia Tibetan sheep during the estrus cycle.(3)The distribution of FSHR and LHR in oviduct and uterus were detected by the immunohistochemical Streptomyces anti-biotin protein-peroxidase ligation method,and the integral optical density values of FSHR and LHR immuno-positive productsin oviduct and uterus of Ganjia Tibetan sheep during the estrus cycle were analyzed by the computer Image pro-plus.The results of the study were as follows:1.The FSHR mRNA and LHR mRNA were expressed in the oviduct and uterus during the whole estrus cycle of Ganjia Tibetan sheep,and showed a dynamic trend.(1)The relative expression of FSHR mRNA in oviduct of Ganjia Tibetan in the proestrus,estrus,metaestrus and diestrus were respectively 0.98±1.39,1.55±0.87,0.25±0.17 and 1±0.65,it was highest in estrus,which was very significantly different from that in metaestrus(P<0.01),and was different from proestrus and diestrus(P<0.05).The relative expression of LHR mRNA in oviduct of Ganjia Tibetan in the proestrus,estrus,metaestrus and diestrus were respectively 1.44±0.02,1.89±0.03,0.60±0.75 and 1±0.45,it was the highest in estrus,which was very significantly different from that in metaestrus(P<0.01),and was different from proestrus and diestrus(P<0.05).(2)The relative expression of FSHR mRNA in uterus of Ganjia Tibetan in the proestrus,estrus,metaestrus and diestrus were respectively 0.11±0.03,1.12±0.21,0.40±0.20 and 1±0.2,it was the highest in estrus,which was very significantly different from that in proestrus and metaestrus(P<0.01).The relative expression of LHR mRNA in uterus of Ganjia Tibetan in the proestrus,estrus,metaestrus and diestrus were respectively 0.57±0.01,0.35±0.01,0.75±0.01 and 1±0.11,it was the highest in diestrus,which was very significantly different from that in estrus(P<0.01),and different from proestrus(P<0.05).2.(1)FSHR and LHR expressed in the infundibulum,ampulla and Isthmus of the oviduct during the estrus cycle of Ganja Tibetan sheep,and mainly distributed in the mucosal epithelium and lamina propria cells.The number of FSHR and LHR positive cells was the highest in the isthmus in the proestrus,metaestrus and diestrus,and it was highest in infundibulum in the estrus.The distribution of FSHR and LHR positive cells in the mucosal epithelium of the infundibulum was higher than that in the lamina propria of the infundibulum throughout the estrus cycle;the number of FSHR positive cells was the highest in the mucosal epithelium of the ampulla in theproestrus and estrus,and the cells are mostly oval.The number of LHR positive cells was the highest in the lamina propria of the ampulla in the proestrus and diestrus,and it was highest in the mucosal epithelium of the ampulla in the estrus and metaestrus;the number of FSHR and LHR positive cells was the highest in the lamina propria of the isthmus in the proestrus,estrus and diestrus,and in was highest in the mucosal epithelium of the isthmus in the metaestrus.Using computer image analysis software(IPP)show that: The expression of FSHR protein was highest in the mucosal epithelium and lamina propria cells of the infundibulum during the diestrus;the expression of LHR protein was highest in the mucosal epithelium and lamina propria cells of the infundibulum during the estrus.The expression of FSHR protein was highest in the mucosal epithelium cells of the ampulla during the diestrus,and it was highest in lamina propria cells of ampulla during the metaestrus;the expression of LHR protein was highest in the mucosal epithelium cells of ampulla during the estrus,and it was higtest in lamina propria cells of the ampulla during proestrus.The expression of FSHR protein was highest in the mucosal epithelium and lamina propria cells of the isthmus during the metaestrus;the expression of LHR protein was highest in the mucosal epithelium cells of isthmus during the metaestrus,it was highest in lamina propria cells of isthmus during the proestrus.(2)The FSHR and LHR positive cells expressed in the cervix,corpus uteri and cournual uteri of the uterus during the estrus cycle of Ganjia Tibetan sheep,and mainly distributed in stromal cells,glandular cells,vascular endothelial cells and myometrial smooth muscle cells.The number of FSHR positive cells was the highest in the cervix in the proestrus and estrus,and it was highest in the corpus uteri in the metaestrus and diestrus;the number of LHR positive cells was the highest in the corpus uteri in the proestrus and estrus,it was highest in the cournual uteri in the metaestrus,and it was highest in the cervix in the diestrus.The number of FSHR positive cells was the highest in the myometrial smooth muscle cells in the proestrus and estrus,and most of the cells were long strips,it was highest in the stromal cells in the metaestrus and diestrus,and mostly of the cells were round;the number of LHRpositive cells was the highest in the myometrial smooth muscle cells in the proestrus,most of the cells were long strips,and it was highest in stromal cells in the estrus,metaestrus and diestrus,most of the cells were round.Computer image analysis software(IPP)was used to obtain FSHR immunoreactive cells in the cervix during the estrus cycle of Ganjia Tibetan sheep.The expression of FSHR protein was highest respectively in stromal cells,glandular cells and myometrial smooth muscle cells during the estrus,and it was highest in vascular endothelial cells during the diestrus.The expression of LHR protein was highest respectively in stromal cells,glandular cells and myometrial smooth muscle cells during the diestrus,and it was highest in vascular endothelial cells during the metaestrus.Conclusion:(1)The expressions of FSHR mRNA and LHR mRNA in the oviduct of Ganjia Tibetan sheep were highest in estrus and lowest in metaestrus.(2)The expression of FSHR mRNA in uterus was the highest in estrus,and the lowest in proestrus,and the expression of LHR mRNA was highest in diestrus,lowest in estrus.(3)The expression of FSHR and LHR positive cells were expressed in the infundibulum,ampulla and isthmus of the oviduct during the estrus cycle of Ganja Tibetan sheep,the number of FSHR and LHR positive cells was the highest in the isthmus in the proestrus,metaestrus and diestrus,and it was highest in infundibulum in the estrus,and mainly distributed in the mucosal epithelium and lamina propria cells.(4)The expression of FSHR and LHR positive cells were expressed in the cervix,corpus uteri,and cournual uteri of the uterus during the estrus cycle of Ganjia Tibetan sheep,the number of FSHR positive cells was the highest in the cervix in the proestrus and estrus,and it was highest in the corpus uteri in the metaestrus and diestrus;the number of LHR positive cells was the highest in the corpus uteri in the proestrus and estrus,it was highest in the cournual uteri in the metaestrus,and it was highest in the cervix in the diestrus,and mainly distributed in stromal cells,glandular cells,vascular endothelial cells and myometrial smooth muscle cells.This study can reveal the distribution of FSHR and LHR in the oviduct and uterus of the Ganjia Tibetan sheep,and the expression of FSHR mRNA and LHRmRNA in the oviduct and uterus of the Ganjia Tibetan sheep during the estrus cycle.The results may provide experimental data including molecular and cellular levels for exploring the effects of GTH and its receptors on the function and reproductive physiological activities of oviduct and uterus.The study provided a scientific theoretical basis for studying reproductive endocrine regulation in female mammals and improving the production performance. |
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