Immunolocalization of Retinol-binding Protein and Profiles of Its mRNA as Related to Testicular Development in the Beef Bull

Abstract

A study was conducted to identify the cell types that express retinol-binding protein (RBP) in the bovine testis and to compare relative steady-state levels of RBP mRNA expression at different times of testicular development. At the ages of 10 ( n = 3), 20 ( n = 8), and 34 ( n = 7) wk, Angus bulls were bled three times at 1.5-hr intervals, then surgically castrated. Blood samples were analyzed for follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T), by radioimmunoassay and the degree of seminiferous tubule development was evaluated histologically in sections of fixed tissue samples stained with hematoxylin and eosin. Immunolocalization of RBP was based on the biotin-strepavidin-horseradish peroxidase method. Testis weight and concentrations of LH and T increased with age (P < 0.05), but those of FSH did not change (P > 0.05) between 10 and 34 wk. Seminiferous tubules at 10 wk contained immature Sertoli cells and gonocytes whereas, at 20 wk, spermatogonia and few spermatocytes were detected. At 34 wk, Sertoli cells appeared differentiated and spermatids were observed. RBP was immunolocalized in Sertoli, Leydig, and peritubular cells at the ages of 10, 20, and 34 wk. Furthermore, no differences in staining between Sertoli cells from tubules with or without germ cells were detected. Northern hybridization of testicular RNA with an RBP cDNA probe revealed the presence of a 1.4-Kb mRNA, which was similar to previous RBP transcripts found in other bovine tissues. Quantitative slot blot analysis revealed that steady-state RBP mRNA levels were 50% higher at 10 wk (P < 0.05) than at 20 and 34 wk of age.