沂蒙黑山羊性周期中GTH受体在输卵管的分布及其mRNA表达规律的研究
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摘要
分布在鲁中南泰沂山区的沂蒙黑山羊是山东省优良的地方品种,该品种黑山羊以耐粗饲、抗病能力强、绒毛质量好、肉质鲜美而著称,属肉、皮、绒兼用型品种。本实验以沂蒙黑山羊为研究对象,就性周期外周血中卵泡刺激素(FSH)、黄体生成素(LH)的分泌变化、性周期不同阶段促性腺激素受体免疫反应阳性细胞在输卵管不同部位中的位置分布以及卵泡刺激素受体(FHSR)mRNA、黄体生成素受体(LHR)mRNA在输卵管不同部位表达量的差异进行了研究,结果如下:
1.应用放射免疫测定法(RIA)对沂蒙黑山羊在性周期各阶段外周血中FSH、LH的含量进行了测定。结果表明:外周血FSH、LH含量的变化趋势相似,其基本走势为由发情前期到发情期降到最低,之后又有所升高。FSH含量在发情前期最高(1.6067±0.09955mIU/ml),与其它三个时期之间差异显著(0.010.05)。
2.运用免疫组织化学方法研究了性周期中促性腺激素受体免疫阳性细胞在输卵管的分布情况。FSHR免疫阳性细胞、LHR免疫阳性细胞在性周期不同阶段的输卵管各部位中都有存在,主要存在于输卵管的黏膜上皮细胞、固有层细胞以及血管内皮细胞中。
3.利用RT-PCR技术,以GAPDH为内参基因,研究了沂蒙黑山羊性周期中FSHR mRNA、LHR mRNA在输卵管的表达差异。结果显示:性周期中FSHR mRNA、LHR mRNA在输卵管中都有表达。FSHR mRNA在漏斗部以间情期的表达量最高,发情期最低,发情期与其它三个时期之间差异极显著(P<0.01);壶腹部在间情期的表达量最高,与其它三个时期差异极显著(P<0.01),在发情前期表达量最低。在峡部则是发情后期表达量最高,发情前期最低,发情前期与其它三个时期差异极显著(P<0.01)。LHR mRNA表达量与FSHR mRNA呈相反规律,漏斗部LHR mRNA表达量在间情期最低,发情期最高,且发情期与其它三个时期之间差异极显著(P<0.01);壶腹部则是发情前期表达量最高,间情期最低,发情前期与其它三个时期之间差异极显著(P<0.01);峡部在发情前期表达量最高,发情后期最低,发情前期与其它三个时期之间差异均达极显著水平(P<0.01)。
YiMeng Black Goat from Tai-Yi mountain areas is one of the best local breeds of ShanDong, which is well known for gaining well on the roughest forage, strong inhibitory effect, high quality down and delectable flesh. YiMeng Black goat was investigated in our study for the change of FSH and LH secretion in peripheral blood during estrous cycle, distribution of GTH receptors positive cells in different phases of estrous cycle and the expression of FSHR and LHR in different parts of oviduct. The results showed that:
1. RIA (radioimmunoassay) was used to assay the concentration of FSH and LH in peripheral blood during every phases of estrous cycle. The results suggested that changes of FSH and LH concentration in peripheral blood were similar. The tend of content of GTH hormone decide to the minimum from proestrum to estrus and then rise again. Concentration of FSH in peripheral blood was highest in proestrum (1.6067±0.09955mIU/ml), which is significantly different from other three phases (0.010.05).
2. Immunohistochemical method was utilized to study the distribution of GTH receptors positive cells in oviduct during estrous cycle. FSHR positive cells and LHR positive cells were found in different parts of oviduct in different phases of estrous cycle. The majority of those positive cells were found in oviductal mucosa, lamina propria and vascular endothelial cells.
3. RT-PCR method with GAPDH as house-keeping gene was used for different expression of FSHR mRNA and LHR mRNA in oviduct during estrous cycle. The results indicated that FSHR mRNA and LHR mRNA were expressed in oviduct during estrous cycle. In infundibularis partes, the expression of FSHR mRNA was highest in diestrum and lowest in estrum. The expression of estrum has significantly different with other three phases (P<0.01). In ampulla, expression of FHSR mRNA was highest in diestrum, which is significantly different with other three phases (P<0.01); and lowest in proestrum. In isthmus the expression level of FHSR mRNA was highest in metaestrum and lowest in proestrum which is significantly different with other three phases (P<0.01). The expression of LHR mRNA is contrary to the expression of FSHR mRNA. In infundibularis partes, LHR mRNA reached the lowest level in diestrum and was highest in estrum which is significantly different with other three phases(P<0.01); In ampulla expression was highest in proestrum and lowest in diestrum, the expression in estrum has significantly different with other three phases(P<0.01); In isthmus the expression was highest in proestrum which has significantly different with other phases (P<0.01)and lowest in metaestrum.
1.应用放射免疫测定法(RIA)对沂蒙黑山羊在性周期各阶段外周血中FSH、LH的含量进行了测定。结果表明:外周血FSH、LH含量的变化趋势相似,其基本走势为由发情前期到发情期降到最低,之后又有所升高。FSH含量在发情前期最高(1.6067±0.09955mIU/ml),与其它三个时期之间差异显著(0.01
2.运用免疫组织化学方法研究了性周期中促性腺激素受体免疫阳性细胞在输卵管的分布情况。FSHR免疫阳性细胞、LHR免疫阳性细胞在性周期不同阶段的输卵管各部位中都有存在,主要存在于输卵管的黏膜上皮细胞、固有层细胞以及血管内皮细胞中。
3.利用RT-PCR技术,以GAPDH为内参基因,研究了沂蒙黑山羊性周期中FSHR mRNA、LHR mRNA在输卵管的表达差异。结果显示:性周期中FSHR mRNA、LHR mRNA在输卵管中都有表达。FSHR mRNA在漏斗部以间情期的表达量最高,发情期最低,发情期与其它三个时期之间差异极显著(P<0.01);壶腹部在间情期的表达量最高,与其它三个时期差异极显著(P<0.01),在发情前期表达量最低。在峡部则是发情后期表达量最高,发情前期最低,发情前期与其它三个时期差异极显著(P<0.01)。LHR mRNA表达量与FSHR mRNA呈相反规律,漏斗部LHR mRNA表达量在间情期最低,发情期最高,且发情期与其它三个时期之间差异极显著(P<0.01);壶腹部则是发情前期表达量最高,间情期最低,发情前期与其它三个时期之间差异极显著(P<0.01);峡部在发情前期表达量最高,发情后期最低,发情前期与其它三个时期之间差异均达极显著水平(P<0.01)。
YiMeng Black Goat from Tai-Yi mountain areas is one of the best local breeds of ShanDong, which is well known for gaining well on the roughest forage, strong inhibitory effect, high quality down and delectable flesh. YiMeng Black goat was investigated in our study for the change of FSH and LH secretion in peripheral blood during estrous cycle, distribution of GTH receptors positive cells in different phases of estrous cycle and the expression of FSHR and LHR in different parts of oviduct. The results showed that:
1. RIA (radioimmunoassay) was used to assay the concentration of FSH and LH in peripheral blood during every phases of estrous cycle. The results suggested that changes of FSH and LH concentration in peripheral blood were similar. The tend of content of GTH hormone decide to the minimum from proestrum to estrus and then rise again. Concentration of FSH in peripheral blood was highest in proestrum (1.6067±0.09955mIU/ml), which is significantly different from other three phases (0.01
2. Immunohistochemical method was utilized to study the distribution of GTH receptors positive cells in oviduct during estrous cycle. FSHR positive cells and LHR positive cells were found in different parts of oviduct in different phases of estrous cycle. The majority of those positive cells were found in oviductal mucosa, lamina propria and vascular endothelial cells.
3. RT-PCR method with GAPDH as house-keeping gene was used for different expression of FSHR mRNA and LHR mRNA in oviduct during estrous cycle. The results indicated that FSHR mRNA and LHR mRNA were expressed in oviduct during estrous cycle. In infundibularis partes, the expression of FSHR mRNA was highest in diestrum and lowest in estrum. The expression of estrum has significantly different with other three phases (P<0.01). In ampulla, expression of FHSR mRNA was highest in diestrum, which is significantly different with other three phases (P<0.01); and lowest in proestrum. In isthmus the expression level of FHSR mRNA was highest in metaestrum and lowest in proestrum which is significantly different with other three phases (P<0.01). The expression of LHR mRNA is contrary to the expression of FSHR mRNA. In infundibularis partes, LHR mRNA reached the lowest level in diestrum and was highest in estrum which is significantly different with other three phases(P<0.01); In ampulla expression was highest in proestrum and lowest in diestrum, the expression in estrum has significantly different with other three phases(P<0.01); In isthmus the expression was highest in proestrum which has significantly different with other phases (P<0.01)and lowest in metaestrum.
引文
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