Expression of kisspeptin and its receptor in different functional classes of ovarian follicle in the buffalo (Bubalus bubalis).

Recently, we reported the differential expression of kisspeptinergic system in the bubaline hypothalamus and corpus luteum. Here, we document the expression of kisspeptin (Kp) and its receptor (Kiss1r) in the ovarian follicles of the buffalo with respect to the functional status. Follicles of ≥10 to ≤13 mm diameter (n = 45) were retrospectively categorized into active (n = 18), intermediate (n = 16) and atretic (n = 11) follicles based on the concentrations of intrafollicular progesterone (P4) and estradiol (E2). The P4:E2 ratio was significantly lower in the active follicle (0.43 ± 0.08) than that of the intermediate (3.46 ± 0.53) and atretic (28.4 ± 10.6) follicles (P < 0.05). Relative fold change in the transcripts of kisspeptin (Kiss1), Kiss1r, gonadotrophin receptors, steroid acute regulatory protein (StAR), cytochrome P450 family 11 subfamily A member 1 (CYP11A1), cytochrome P450 Family 19 subfamily A member 1 (CYP19A1), insulin like growth factor -1 (IGF-1), apoptotic factors (caspase 3 and B-cell lymphoma 2, BCL2) was calculated using qPCR in the follicular wall of the three categories of follicle (n = 8/group). In another experiment, histological sections of the ovary (n = 41) were used to group the follicles as described above and immunostaining of Kp, Kiss1r and aromatase was done. A significant upregulation of StAR, CYP11A1 and CYP19A1 in the active follicles supported the endocrine basis of follicular classification. The transcripts of Kiss1 and Kiss1r were upregulated by 19.45 fold and 4.25 fold, respectively in the active follicle as compared to other groups. Immunolocalization studies revealed that Kp and Kiss1r were localized to the basal and antral granulosa cells (GC) of the active and intermediate follicles; however, the staining intensity was stronger in the former group. Strong expression of CYP19A1 in the GC layer of active follicle supported the histological basis of defining the functional status of the follicle. It is concluded that the follicular compartment of the bubaline ovary expressed the constituents of kisspeptinergic system. The expression of Kp and Kiss1r was influenced by the functional status of the follicle with intense localization in the GC layer of the active follicles.