贮精能力差异母鸡的贮精腺形态、激素水平及激素受体基因表达量的研究

The Sperm Storage Tubules Morphology,Hormones Concentration,and Hormone Receptor Genes Expression of Laying Hens with Divergent Sperm Storage Ability

旨在探究不同贮精能力母鸡的贮精腺形态、主要性激素水平以及激素受体基因表达量的差异,以期进一步揭示母鸡贮精能力差异产生的原因。本研究以27周龄158只的白来航母鸡和28只公鸡为试验材料,混精连续输精2 d,第3天开始按照个体收集种蛋孵化,根据输精后21 d内每天的种蛋受精情况统计个体受精率作为母鸡贮精能力;挑选高、低贮精能力极端个体各4只,分别为高、低贮精能力组;采血测定血清孕酮、雌激素、睾酮和催乳素激素浓度;解剖获取富含贮精腺的子宫阴道连接部组织,并沿纵向分为两份,一份制作石蜡切片并HE染色,用于贮精腺形态观察,另一份采用荧光定量PCR检测相应激素受体基因的表达量。结果表明,高、低贮精能力组子宫阴道连接部的黏膜面积、贮精腺数量、贮精腺密度均差异不显著(P>0.05),但高贮精能力组母鸡的贮精腺平均横截面积显著高于低贮精能力组(P<0.05);高贮精能力组母鸡的血清孕酮激素浓度显著高于低贮精能力组(P<0.05),雌激素、睾酮和催乳素激素浓度在组间均差异不显著(P>0.05);与低贮精能力组相比,高贮精能力组睾酮受体基因和催乳素受体基因表达上调,雌激素α、β受体基因和孕酮受体基因下调,但未达显著水平(P>0.05)。结果提示,母鸡的贮精能力可能与贮精腺横截面积有关,此外,孕酮激素可能对诱导贮精腺中精子的激活起重要作用,从而影响母鸡持续受精能力。

The aim of this study was to explore the differences in sperm storage tubules morphology,hormone concentration,and hormone receptor genes expression in hens with divergent sperm storage ability,and to further reveal the mechanism underlying sperm storage.A total of 158 White Leghorn laying hens and 28 rosters of 27 weeks of age were used in this study.After the artificial insemination with mixed semen for two consecutive days,the eggs were collected for each laying hen starting from the 3rd day for 21 days and incubated.According to the fertilization status of the eggs within 21 days after insemination,the individual fertilization rate was counted as the hen’s sperm storage ability.The top 4 hens with the highest sperm storage ability and the 4 with the lowest sperm storage ability were selected and named high and low sperm storage ability groups,respectively.The concentrations of progesterone,estrogen,testosterone,and prolactin hormone in serum were detected.The uterine-vaginal junction area of the 8 laying hens was dissected and divided into two parts in the longitudinal direction.One was made into paraffin sections and HE stained for the morphology observation,and the other one was used to detect the expression of the corresponding hormone receptor genes using qRT-PCR assay.The results showed that there was no significant difference in mucosal area of uterine-vaginal junction,sperm storage tubules numbers and density of hens between high and low sperm storage ability groups(P>0.05).The average cross section area of sperm storage tubules of hens in high sperm storage ability group was higher than that of the low sperm storage ability group(P<0.05).The concentration of progesterone in serum of hens in high sperm storage ability group was higher than that of hens in low sperm storage ability group(P<0.05),but there was no significant difference in concentration of estrogen,testosterone or prolactin in serum between groups(P>0.05).Compared with the low sperm storage ability group,hens in the high sperm storage ability group had up-regulated expression of testosterone and prolactin receptor genes,and down-regulated expression of estrogenα/βreceptor and progesterone receptor genes,but the differences were not significant(P>0.05).In conclusion,the cross section area of sperm storage tubules may be related to the sperm storage ability of laying hens;in addition,progesterone hormone may play an important role in sperm activation in the sperm storage tubules,which affects the hen’s ability to continuous fertilization.