Immunolocalization of steroidogenic enzymes (P450scc, P450c17, P450arom, and 3beta-HSD) in immature and mature testes of rainbow trout (Oncorhynchus mykiss).
We examined the localization of steroidogenic cells in rainbow trout (Oncorhynchus mykiss) testis during spermatogenesis by using polyclonal antibodies generated against rainbow trout cholesterol side-chain cleavage enzyme cytochrome P450 (P450scc), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), 17alpha-hydroxylase/C17,21 lyase (P450c17), and aromatase cytochrome P450 (P450arom) as markers of steroid production. Since we had previously produced specific antibodies against 3beta-HSD and P450arom, antibodies against oligopeptides corresponding to C-terminal sequences of P450scc and P450c17, predicted from rainbow trout P450scc and P450c17 cDNAs, were produced in this study. These two antibodies recognized 54-kDa (P450scc) and 59-kDa (P450c17) bands specifically in several steroidogenic organs, i.e., testis, ovary, and interrenal tissue (head kidney) in Western blots. Immunohistochemically, immunoreactive P450scc, P450c17, and 3beta-HSD, but not P450arom, were found only in interstitial Leydig cells of immature and mature testes. Immunoreactive P450arom was not detected in either testis. This study suggests that Sertoli cells and germ cells of rainbow trout testis do not contain P450scc, P450c17, P450arom, or 3beta-HSD.
Kobayashi
T
T
Laboratory of Reproductive Biology, National Institute for Basic Biology, Okazaki 444, Japan.
Nakamura
M
M
Kajiura-Kobayashi
H
H
Young
G
G
Nagahama
Y
Y
D015096:3-Hydroxysteroid Dehydrogenases / Q000032:analysis / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001141:Aromatase / Q000032:analysis / Q000276:immunology; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000032:analysis / Q000276:immunology; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis* / Q000276:immunology; D015151:Immunoblotting; D007150:Immunohistochemistry; D007985:Leydig Cells / Q000201:enzymology*; D008297:Male; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000502:physiology*; D012708:Sertoli Cells / Q000201:enzymology*; D013091:Spermatogenesis / Q000502:physiology; D013254:Steroid 17-alpha-Hydroxylase / Q000032:analysis / Q000276:immunology; D013737:Testis / Q000166:cytology / Q000201:enzymology / Q000254:growth & development
D000906:Antibodies; D003577:Cytochrome P-450 Enzyme System; D015096:3-Hydroxysteroid Dehydrogenases; D001141:Aromatase; D013254:Steroid 17-alpha-Hydroxylase; D002786:Cholesterol Side-Chain Cleavage Enzyme