甘加藏羊发情周期输卵管和子宫中THRs的表达及分布

Expression and distribution of THRs in oviduct and uterus of Ganjia Tibetan sheep during estrus cycle

【目的】为阐明甘加藏羊(Ovis aries)发情周期甲状腺激素受体α(THRα)和甲状腺激素受体β(THRβ)在输卵管和子宫中表达,探讨其对藏羊繁殖活动中的调控作用。【方法】选取健康未孕处于发情周期和乏情期的甘加藏羊30只,应用实时荧光定量PCR、蛋白免疫印迹和免疫组织化学染色方法研究了发情周期和乏情期输卵管和子宫组织THRα和THRβmRNA及其蛋白含量的表达和分布。【结果】THRα、THRβmRNA及其蛋白在藏羊发情周期和乏情期输卵管和子宫中均有表达,输卵管中THRα、THRβmRNA表达量在发情后期最高,显著(P<0.05)高于其他3个时期和乏情期;子宫中发情期表达量最高,与其他时期差异显著(P<0.05)。THRβmRNA的平均相对表达量显著高于THRαmRNA。输卵管组织中THRα蛋白在发情后期表达量最高,子宫中发情期最高,显著(P<0.05)高于与其他时期。输卵管和子宫中THRβ蛋白在发情后期表达量最高,与其他时期差异显著(P<0.05)。乏情期THRα、THRβmRNA及其蛋白的平均表达量显著低于发情周期各时期。免疫组织化学染色结果显示:藏羊发情周期THRα和THRβ免疫反应阳性细胞主要表达在输卵管的黏膜上皮、纤毛细胞、分泌细胞和肌层,在子宫中主要表现在基质细胞和子宫腺。【结论】THRα和THRβ二者两者浓度差异性表达,表明THRs对甘加藏羊发情时期的输卵管和子宫有一定的调控作用,可能参与甘加藏羊生殖生理活动。

【Objective】The study aimed to elucidate the expressions of Thyroid hormone receptorα(THRα)and Thyroid hormone receptorβ(THRβ)in the oviduct and uterus during the estrous cycle of Ovis aries,exploring their regulation effect on breeding activities of the Tibetan sheep.【Method】Thirty healthy and non-pregnant Ganjia Tibetan sheep in estrus and anestrus were employed.Real-time PCR,Western blot,and immunohistochemical staining were used to study the expression and protein content of THRαand THRβmRNA in the oviduct and uterine tissues during estrus and anestrus.【Result】The THRαand THRβmRNA and their proteins were expressed in the fallopian tubes and uterus of Tibetan sheep during the estrus cycle and anestrus period.The expression levels of THRαand THRβmRNA in the fallopian tubes were the highest in the late estrus period,which were significantly(P<0.05)higher than those in the other three periods of the estrus cycle and anestrus period.The average relative expression level of THRβmRNA was significantly higher than that of THRαmRNA in utero during estrus(P<0.05).The expression level of THRαprotein in oviduct tissues was the highest in the late estrus period,and the highest in the uterus tissues in estrus period,which was significantly(P<0.05)higher than was in other periods.The expression level of THRβprotein in the oviduct and uterus was the highest in late estrus,which significantly differed from the other periods(P<0.05).The average expression levels of THRαand THRβmRNA and their protein in the anestrous period were significantly lower than those in each estrous period.The results of immunohistochemical staining showed that THRαand THRβimmunoreactive cells were mainly expressed in the mucosal epithelium,ciliary cells,secretory cells,and myometria of the fallopian tube,and in stromal cells and uterine glands of the uterus.【Conclusion】The different-concentration expression of THRαand THRβsuggested that THRs could regulate the fallopian tube and uterus of Ganjia Tibetan sheep during estrus,which was probably involved in the reproductive physiological activities of Ganjia Tibetan sheep.