308 TESTICULAR DEVELOPMENT IN PREPUBERTAL JERSEY BULL CALVES IMMUNIZED AGAINST INHIBIN

Abstract

Previous studies have shown that immunization against inhibin (INH) in bull calves increased subsequent sperm production (Martin et al. 1991 Biol. Reprod. 45, 73; Bame et al. 1996 Biol. Reprod. 54, 328). The objective of the current study was to evaluate the effectiveness of gonadotropin administration at initiation of inhibin immunization in bull calves on testicular morphology. The study was performed using the inhibin peptide (bovine inhibin α^1–26) conjugated to keyhole limpet hemocyanin (KLH). Primary treatments administered to Jersey bull calves (initial immunization at 27 ± 5 days of age; Day 1 of the experimental period) consisted of control (KLH, 250 µg, n = 9) or immunization (INH; 500 μg INH: 250 µg KLH, n = 9) with each emulsified in 2 mL of Freund's complete adjuvant (FCA). Booster immunizations (identical preparation in FCA) occurred every 21 days with the last administration on Day 84 of the trial. Subsets of calves were randomly assigned within primary treatments (TRT) to receive saline (1 mL, n = 3/TRT), FSH (20 mg, n = 3/TRT), or GnRH (50 μg, n = 3/TRT) every 8 h (0600, 1400, and 2200 h) from Day 1 to Day 3 of the study. Blood samples were obtained daily from Days 0–14 and weekly until testes collection (Day 91) for FSH, LH, testosterone (T), and determination of antibody titers. Body weight and scrotal circumference (SC) were measured at each immunization and immediately before testes removal. The right testis was weighed and used for absolute volume calculation of cell components per testis. Tissue sections were examined using a light microscope (400×). For each cell type, absolute volume of Sertoli, Leydig, and germ cells were counted according to the point counting method. Data were analyzed using MIXED procedure of SAS (SaS Institute, Inc., Cary, NC, USA). Antibody titers were increased in INH bulls compared to KLH bulls (P < 0.05) during the experimental period. Body weight (89.8 ± 14.2 kg), SC (14.6 ± 1.3 cm), and single testicular weight (19.2 ± 6.2 g) recorded at the end of the experimental period did not differ between treatments. Neither serum concentrations of FSH, LH, and T nor population of Leydig and Sertoli cells differed between treatment groups. However, a significant immunization X hormone treatment interaction was noted for germ cell volume per testis (P < 0.008). Administration of FSH at the time of initial immunization against inhibin significantly increased germ cell population (1.22 ± 0.1 cm³) compared to INH-saline bulls (0.64 ± 0.1 cm³) with INH-GnRH bulls intermediate (0.84 ± 0.1 cm³; P < 0.05). In contrast, germ cell volume was not increased following hormone administration in KLH bulls. These results suggest that gonadotropin administration at the time of inhibin immunization increases germ cell volume in the testis without altering Sertoli and Leydig cell volume.