Department of Animal and Rangeland Sciences, Oregon State University, OR, USA.
Department of Fisheries, Wildlife, And Conservation Sciences, Oregon State University, OR, USA.
Theriogenology. 2023 Nov;211:198-202. doi: 10.1016/j.theriogenology.2023.08.022. Epub 2023 Aug 28.
The purpose of this research was to quantify sperm acrosome associated 3 protein expression in the ovaries of young (3.0 ± 0.9 months, n = 11) and adult (10.4 ± 2.8 months, n = 11) queens. Immunohistochemistry was performed on formalin-fixed, paraffin-embedded feline ovarian sections. Ovaries were obtained following routine ovariohysterectomy of queens. Cellular expression of sperm acrosome associated 3 protein was measured in primordial, primary, secondary, and tertiary follicles using an image-analysis software's red, green, and blue stack and manual thresholding functions. The oocyte nucleus, ooplasm, granulosa cells, and theca cells were outlined using the freehand selection tool and mean grey value was recorded. Results from each cellular location were compared between age groups using a Student's t-test and between follicle stages using an analysis of variance. Compared to adult queens, younger queens had significantly greater sperm acrosome associated 3 protein expression in granulosa cells of primary, secondary, and tertiary follicles. Also, theca cells of secondary and tertiary follicles had significantly greater sperm acrosome associated 3 protein expression in younger queens compared to adult queens. The oocyte nucleus of primordial, primary, and secondary follicles had significantly greater sperm acrosome associated 3 protein expression in younger queens compared to adult queens. However, sperm acrosome associated 3 protein expression within the ooplasm did not differ significantly between age groups of any follicle type. More research is needed to determine what role sperm acrosome associated 3 protein may play in female fertility in animals as well as what mechanisms regulate ovarian sperm acrosome associated 3 protein expression over time.
本研究的目的是定量分析年轻(3.0±0.9 个月,n=11)和成年(10.4±2.8 个月,n=11)母猫卵巢中精子顶体相关蛋白 3 的表达。对福尔马林固定、石蜡包埋的猫卵巢切片进行免疫组织化学染色。这些卵巢是在常规卵巢切除术时从母猫身上获得的。使用图像分析软件的红色、绿色和蓝色堆栈和手动阈值功能,在原始卵泡、初级卵泡、次级卵泡和三级卵泡中测量精子顶体相关蛋白 3 的细胞表达。使用徒手选择工具勾勒出卵母细胞核、卵质、颗粒细胞和膜细胞,并记录平均灰度值。使用学生 t 检验比较两组年龄组之间的每个细胞位置的结果,使用方差分析比较卵泡阶段之间的结果。与成年母猫相比,年轻母猫的初级、次级和三级卵泡的颗粒细胞中,精子顶体相关蛋白 3 的表达显著更高。此外,与成年母猫相比,二级和三级卵泡的膜细胞中,年轻母猫的精子顶体相关蛋白 3 的表达也显著更高。与成年母猫相比,原始卵泡、初级卵泡和次级卵泡的卵母细胞核中的精子顶体相关蛋白 3 表达显著更高。然而,任何类型卵泡的卵质中,精子顶体相关蛋白 3 的表达在两组年龄之间均无显著差异。需要进一步的研究来确定精子顶体相关蛋白 3 在动物的雌性生育能力中可能扮演的角色,以及随着时间的推移,哪些机制调节卵巢中精子顶体相关蛋白 3 的表达。