Immunohistochemcial Localization of Steroidogenic Enzymes in the Testis of Hokkaido Sika Deer (Cervus nippon yesoensis)

Abstract

ABSTRACT. The testes from 15 adult male Hokkaido Sika deer (Cervus nippon yesoensis) were collected during the rutting season (October and November). We investigated the localization of 4 kinds of steroidogenic enzymes (P450scc, 3βHSD, P450c17 and P450arom) immunohistochemically in these testicular samples. The specific immunoreactivities to these enzymes were detected only in the cytoplasm of Leydig cells. This differs to the enzyme distributions reported previously in Japanese black bear, Japanese raccoon dog, Hokkaido brown bear and American black bear, in which the same immunoreactivities were detected in Leydig cells, Sertoli cells and/or spermatogenic cells. The current study suggests that in the testes of the Hokkaido Sika deer, testosterone and estradiol-17 β may be synthesized in the Leydig cells only. KEY WORDS: Hokkaido Sika deer (Cervus nippon yesoensis), immunohistochemistry, steroidogenic enzyme.

Localization of Five Steroidogenic Enzyme mRNAs in Japanese Black Bear (Ursus thibetanus japonicus) Testes During the Mating Season by In Situ Hybridization

“…The major steroidogenic enzymes involved in androgen synthesis have been shown to localize exclusively in Leydig cells in many of the mammalian species investigated so far [19,20], leading to a general idea that androgens in the testis are produced only by Leydig cells. In accordance with the findings in other mammalian species [10,11,21,22], the present study detected the metabolic enzymes from cholesterol to androstenedione (P450scc, 3βHSD and P450c17) in interstitial tissue of the Japanese black bear testis. These results are also in agreement with the immunohistochemical localization of these three enzymes in the Japanese black bear testis [2,9].…”

“…Also, there are several reports at the protein level based on the immunohistochemical method in wild animals like brown bears [8], black bears [2, 9], raccoon dogs [10] and deer [11]. However, there have been few reports on the gene expression of steroidogenic enzymes in the testes of wild mammals.…”

Immunohistochemical Localization of Steroidogenic Enzymes in the Testis of the Sika Deer (Cervus nippon) During Developmental and Seasonal Changes

“…The primary antibodies used in this study are summarized in Table 2. The antibodies against P450scc, P450c17, 3βHSD and P450arom were used in previous studies on testicular steroidogenesis [12,15,18,25,26].…”

“…The primary antibodies used in this study are summarized in Table 2. The antibodies against P450scc, P450c17, 3βHSD and P450arom were used in previous studies on testicular steroidogenesis [12,15,18,25,26].For the ABC method, the deparaffinized sections were incubated with methanol containing 0.3% H2O2 for 10 min at room temperature (RT) to block endogenous peroxidase activity and then with normal goat serum (1:50, S-1000, Vector Laboratories) for 30 min at RT to prevent nonspecific staining. The samples were incubated with each primary antibody overnight at 4 C. After an overnight incubation, the secondary antibody, biotinylated anti-rabbit IgG raised in the goat (1:200, BA-1000, Vector Laboratories), was applied for 30 min at RT, and then the sections were incubated with ABC for 30 min at RT (1:2, PK-6100, Vectastain Elite ® ABC Kit, Vector Laboratories).…”

Immunolocalization of Steroidogenic Enzymes in the Fetal, Neonatal and Adult Testis of the Shiba Goat

“…The precise location of steroidogenic enzymes inside the testes has been studied in recent decades. Immunolocalization of various steroidogenic enzymes in the testes has been reported in several species, including humans [10], mice [18], bears [24], raccoon dogs [25] and deer [9], but data on the pattern of expression of steroidogenic enzymes necessary for androgen and estrogen production in developing testis, especially in large experimental animals, are limited.…”