USDA, Agricultural Research Service, Beltsville Agricultural Research Center-East, Beltsville, MD 20705, USA.
Poult Sci. 2011 Jul;90(7):1539-47. doi: 10.3382/ps.2010-01159.
The sperm storage tubules (SST) of the turkey hen, which are located in the uterovaginal junction (UVJ) of the oviduct, maintain viable sperm for up to 10 wk after a single insemination. The mechanisms of this in vivo sperm storage are poorly understood. Our objective was to evaluate mRNA and protein expression of avidin and 2 avidin-associated factors, avidin-related protein-2 (AVR2) and progesterone receptor, in the oviducts of 2 different lines to determine the extent to which they were sperm responsive and tissue specific. At 38 wk of age, Hybrid Grade Maker and Converter turkey hens were artificially inseminated with diluted semen (AI) or were sham-inseminated with extender alone (SI). Forty-eight hours after insemination, total RNA was extracted from the UVJ epithelium (containing SST) and vaginal epithelium (VGE) of SI and AI hens. Real time-polymerase chain reaction data showed a clear tissue region-specific effect on gene expression in the turkey hen oviduct, with much greater (P < 0.0001) expression in the UVJ compared with VGE region for avidin and AVR2 mRNA in both lines and for progesterone receptor mRNA in the Converter line. In contrast to real-time PCR data, in situ hybridization of SI and AI tissues showed that the presence of sperm increased avidin mRNA in the SST and UVJ surface epithelium in the Converter hens. Immunohistochemistry confirmed the presence of avidin protein in the epithelium of the UVJ in both lines; however, whereas avidin protein was localized in the SST of SI-Grade Maker hens, this protein was not detected in the SST of Converter hens. The upregulation of avidin and AVR2 mRNA within the sperm storage region indicates the involvement of avidin, and perhaps avidin analogs, in the sustained storage of sperm in the SST, possibly through the binding of biotin to avidin. The absence of avidin protein in the SST and VGE of Converter hens in the presence of increased mRNA may indicate a rapid turnover of protein.
火鸡的精子储存管（SST）位于输卵管的子宫阴道交界处（UVJ），在单次授精后可维持长达 10 周的存活精子。这种体内精子储存的机制尚不清楚。我们的目的是评估不同品系输卵管中avidin 及其 2 种avidin 相关因子（AVR2 和孕激素受体）的 mRNA 和蛋白表达，以确定它们在多大程度上对精子有反应和组织特异性。在 38 周龄时，Hybrid Grade Maker 和 Converter 火鸡母鸡用稀释精液（AI）人工授精或仅用稀释液进行假授精（SI）。授精后 48 小时，从 SI 和 AI 母鸡的 UVJ 上皮（含 SST）和阴道上皮（VGE）中提取总 RNA。实时聚合酶链反应数据显示，火鸡输卵管基因表达具有明显的组织区域特异性效应，在两个品系中，avidin 和 AVR2 mRNA 在 UVJ 中的表达明显高于 VGE 区域，而在 Converter 品系中孕激素受体 mRNA 的表达也是如此。与实时 PCR 数据相反，SI 和 AI 组织的原位杂交显示，精子的存在增加了 Converter 母鸡 SST 和 UVJ 表面上皮中的 avidin mRNA。免疫组织化学证实了两个品系 UVJ 上皮中存在 avidin 蛋白；然而，在 SI-Grade Maker 母鸡的 SST 中定位了 avidin 蛋白，而在 Converter 母鸡的 SST 中未检测到这种蛋白。avidin 和 AVR2 mRNA 在精子储存区域的上调表明 avidin 及其可能的 avidin 类似物参与了 SST 中精子的持续储存，可能是通过生物素与 avidin 的结合。在 Converter 母鸡中，尽管 mRNA 增加，但 SST 和 VGE 中不存在 avidin 蛋白，这可能表明蛋白的快速周转。
