Sreejalekshmi P, Raghavendra B S, Subramani T Siva, Murthy V Chandrashekara, Jamuna K V, Prasad R V, Ravindra J P, Selvaraju S
Department of Veterinary Anatomy, Veterinary College, Bangalore, India.
Reprod Domest Anim. 2011 Feb;46(1):59-65. doi: 10.1111/j.1439-0531.2009.01569.x.
The objective of this experiment was to assess the features and extent of follicular apoptosis in the water buffalo (Bubalus bubalis) ovary using classical histology and nick end labelling technique. Ovaries (n=40) procured from the slaughterhouse were used for the study. The sections (5 μm) were used for detection of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) and classical histology (H&E). Those follicles showing ≥ 5% TUNEL positivity (TUNEL assay) and pyknotic nuclei (histology) in granulosa cells were classified as atretic. Based on histology, the atretic primary and secondary follicles (%) were 93.82 and 95.62 respectively. The histology study reveals that the rates (%) of atresia in <1, 1-3, 3-5 mm and >5 mm were 36.90, 40.50, 62.84 and 74.5 respectively. Further the atretic tertiary follicles (%) were significantly lower than the primary and secondary classes of follicles. TUNEL assay reveals that the atretic rate (%) of tertiary follicles in <1, 1-3, 3-5 and ≥ 5 mm class follicles were 50.88, 53.84, 81.81 and 36.36 respectively. The percentage of atresia in >5 mm diameter follicles were significantly lower in TUNEL than histology. Percentages of granulosa and thecal cells positive for atresia by TUNEL were 30.7 ± 0.53 and 13.82 ± 0.18 respectively per follicle. The initial structural changes in atretic follicles were seen primarily in the granulosa cells. In severely atretic follicles TUNEL positive granulosa cells along with theca cells have to be considered in assessing the rate and extent of atresia.
本实验的目的是使用经典组织学和缺口末端标记技术评估水牛（Bubalus bubalis）卵巢中卵泡凋亡的特征和程度。从屠宰场获取的卵巢（n = 40）用于本研究。5μm厚的切片用于检测末端脱氧核苷酸转移酶介导的dUTP生物素缺口末端标记（TUNEL）和经典组织学（苏木精-伊红染色，H&E）。那些在颗粒细胞中显示≥5% TUNEL阳性（TUNEL检测）和固缩核（组织学）的卵泡被归类为闭锁卵泡。基于组织学，闭锁的初级和次级卵泡（%）分别为93.82和95.62。组织学研究表明，直径<1、1 - 3、3 - 5和>5mm的卵泡闭锁率（%）分别为36.90、40.50、62.84和74.5。此外，闭锁的三级卵泡（%）显著低于初级和次级卵泡类别。TUNEL检测表明，直径<1、1 - 3、3 - 5和≥5mm类别的三级卵泡闭锁率（%）分别为50.88、53.84、81.81和36.36。直径>5mm卵泡的TUNEL检测闭锁率显著低于组织学检测结果。每个卵泡中通过TUNEL检测显示闭锁的颗粒细胞和膜细胞百分比分别为30.7±0.53和13.82±0.18。闭锁卵泡的初始结构变化主要见于颗粒细胞。在严重闭锁的卵泡中，在评估闭锁率和程度时必须考虑TUNEL阳性的颗粒细胞以及膜细胞。
