Detection of Follicular Apoptosis in Water Buffalo (Bubalus bubalis) Ovary by Histology and Nick End Labelling Technique

Abstract

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.

Follicular survival, activation of primordial follicles and DNA fragmentation after storage of goat ovaries at 35ºC in supplemented Minimal Essential Medium

“…After the preservation of ovarian tissue, in vitro culture and TUNEL can be used as reliable methods for the evaluation of follicular viability and DNA fragmentation, respectively (ABD-ALLAH, 2010;SREEJALEKSHMI et al, 2011). In the current study, the percentage of histologically normal follicles reduced when ovaries were stored in both media and preservation periods and then cultured for 5 days.…”

Immunohistochemical Localization of Fibroblast Growth Factor‐2 in the Sheep Ovary and its Effects on Pre‐antral Follicle Apoptosis and Development In Vitro

“…Although histological analysis of atresia in follicles remains the most classic and reliable method, the 3′‐end‐labelling technique (TUNEL) happens to be a very sensitive method for the in situ visualization of apoptosis at the cellular level (Sreejalekshmi et al. ). Moreover, detection of DNA fragmentation in early stage of follicular atresia using TUNEL has been more specific than the morphological features (Yang and Rajamahendran ).…”

Basic FGF Promotes Proliferation of Ovarian Granulosa Cells in the Laying Chickens Via FGFR1 and PKC Pathway

“…Moreover, bFGF led to a substantial decrease in the incidence of apoptosis. Most ovarian follicles fail to develop fully but instead undergo degeneration by apoptosis of follicle cells (Quirk et al 2006), and the initial structural changes in atretic follicles were seen primarily in the granulosa cells (Sreejalekshmi et al 2011). Available evidence suggests that bFGF prevents rat granulosa cells from undergoing apoptosis, which involves PKA and PKC signalling pathways (Aharoni et al 1995;Lynch et al 2000).…”