Apoptosis in the antral follicles of swamp buffalo and cattle ovary: TUNEL and caspase-3 histochemistry.
The present study was carried out to investigate the pattern of apoptosis in the healthy antral and atretic follicles of Philippine swamp buffaloes (BU) in comparison with Holstein-Friesian (HF) cows. Paraffin sections of healthy follicles and various stages of atretic follicles were stained using the terminal deoxynucleotidyl transferase (TdT)-mediated biotinylated deoxyuridine triphosphates (dUTP) nick end-labelling (TUNEL) method to detect DNA fragmentation and cleaved caspase-3 antibody to detect cells committed to undergo apoptosis. Five equidistant areas of a follicle were counted for the presence of TUNEL- and caspase-3-positive cells. Healthy follicles of BU and HF contained no TUNEL-positive cells in the granulosa and theca layer but showed some caspase-3 positivity. The granulosa layer of advanced atretic follicles showed a significantly higher frequency of caspase-3 positivity than the healthy and early atretic follicles in both breeds. The frequency of caspase-3-positive cells of BU was significantly higher than HF in the granulosa layer of healthy, early atretic and advanced atretic follicles. In the theca interna layer, BU and HF showed a significantly lower and higher frequency of TUNEL-positive cells in the late atretic follicles compared with advanced atretic follicle, respectively. However, the frequency of caspase-3-positive cells of both BU and HF in the late atretic follicles was significantly higher than the advanced atretic follicles in the theca interna layer. These results indicate that caspase-3 and DNA fragmentation is involved in the buffalo ovarian apoptotic process.
Feranil
Jb
J
Laboratory of Animal Science, Graduate School for International Development and Cooperation, Hiroshima University, Higashi-Hiroshima, Japan.
Isobe
N
N
Nakao
T
T
D000818:Animals; D017209:Apoptosis; D002020:Buffaloes / Q000502:physiology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D002417:Cattle / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D053938:DNA Fragmentation; D005260:Female; D005496:Follicular Atresia; D007150:Immunohistochemistry / Q000662:veterinary; D020287:In Situ Nick-End Labeling / Q000662:veterinary; D006080:Ovarian Follicle / Q000378:metabolism / Q000473:pathology / Q000502:physiology*; D013045:Species Specificity; D013799:Theca Cells / Q000473:pathology
D053148:Caspase 3; D020169:Caspases