Yehia Rezq Mohamed El-Baghdady

A better understanding of the molecular mechanisms in granulosa cells (GC) is warranted, during different follicular and luteal developmental stages in buffalo cows. We aimed to (I) study the expression of selected genes in GC during follicular and luteal phases, (II) evaluate correlations between GC gene expression and steroid concentrations {17-beta estradiol (E2) and progesterone (P4)} in follicular fluid (FF), and (III) study effect of ovarian status on follicular population as well as follicular size frequency. Ovaries were collected in pairs from buffaloes (n = 178). Ovaries bearing corpus luteum (CL) were subdivided into hemorrhagic, developing, mature, and albicans. Follicles from luteal groups were classified only into small (< 4 mm) and large (9-20 mm), while follicles from follicular groups were classified into three subgroups: small (< 4 mm), medium (5-8 mm), and large (9-20 mm). The FF and GC were collected for steroid concentrations measurement and gene expression, respectively. In the follicular phase, luteinizing hormone/choriogonadotropin receptor (LHCGR) and cytochrome P450 aromatase (CYP19) in small follicles decreased compared to medium ones. Large follicle showed an increase in LHCGR and CYP19 compared to medium ones. Follicle-stimulating hormone receptor (FSHR) decreased in large compared to medium size follicles. Proliferating cell nuclear antigen (PCNA) increased in small and large follicles. Meanwhile, anti-Mullerian hormone (AMH) and phospholipase A2 group III (PLA2G3) decreased in small and large follicles. The different stages of luteal phase had a profound impact on GC gene expression. There were strong (positive and/or negative) correlations between gene expression and steroid hormones. The different scenarios between expressed genes in GC and steroid concentrations are required for the proper growth and development of follicles and CL.

A study was conducted on 40 buffalo-cows, assigned randomly, immediately after calving into three groups: group I (n = 10) injected with saline and taken as control; group II (n = 15) received 25 mg PGF2 alpha/animal (Lutalyse); group III (n = 15) received 25 mg PGF2 alpha + 25 i.u. oxytocin/animal (Syntocinon), single i.m. dose. Oxytocin and/or PGF2 alpha significantly (P less than 0.01) shortened the interval from calving to first service (38.33 and 31.53 days for groups II and III respectively, versus 91.60 days for controls). The treatment reduced the service period (38.29 and 35.87 days for groups II and III respectively, versus 45.40 days for controls). Concomitantly a significant (P less than 0.01) decrease in the open-days post partum was achieved (76.62 and 67.40 days for groups II and III respectively, versus 137.00 days for controls). In addition, the treated buffaloes needed significantly (P less than 0.01) fewer services per conception (1.67 and 1.20 S/C for groups II and III respectively) than the untreated ones (2.70 S/C), besides a substantial improvement (P less than 0.01) in their conception rate either at 60 or 85 days post partum. Significantly improved (P less than 0.05) results were obtained in the oxytocin and PGF2 alpha treated animals, than in those receiving PGF2 alpha alone for all the previous parameters, except for the service period. Buffaloes therefore seemed to respond better to such treatment than dairy cows.

Blood samples were taken once per week for 4-7 weeks from 59 buffalo calves in 14 age groups, 1-2 months apart. Hormones were quantified by validated radioimmunoassays. Values of androstenedione and testosterone were low at birth (141.3 +/- 33.5 pg/ml and 18.0 +/- 2.9 pg/ml, respectively; mean +/- s.d.). Serum androstenedione concentrations gradually increased from birth until 8 months of age and declined (P less than 0.05) thereafter, whereas mean testosterone values were low up to 8 months and then significantly (P less than 0.05) increased as age advanced. LH concentrations averaged 2.12 +/- 0.47 ng/ml at birth. Thereafter, a decline in LH values was followed by an increase between 6 and 15 months of age. We conclude that, in buffalo bull calves, the pubertal period occurs from about 8 to 15 months of age. For pubertal buffalo bulls 15-17 months of age, serum concentrations of androstenedione, testosterone and LH were 156.9 +/- 54.6 pg/ml, 208.4 +/- 93.8 pg/ml and 2.10 +/- 0.70 ng/ml, respectively.