| First Author | O'Shaughnessy PJ | Year | 1993 |
| Journal | J Mol Endocrinol | Volume | 11 |
| Issue | 1 | Pages | 77-82 |
| PubMed ID | 8240674 | Mgi Jnum | J:15986 |
| Mgi Id | MGI:64083 | Doi | 10.1677/jme.0.0110077 |
| Citation | O'Shaughnessy PJ, et al. (1993) Cytochrome P-450 17 alpha-hydroxylase protein and mRNA in the testis of the testicular feminized (Tfm) mouse. J Mol Endocrinol 11(1):77-82 |
| abstractText | The testicular feminized (Tfm) mouse lacks functional androgen receptors and develops with a female external phenotype and internal testes. The testes of these animals contain normal, or close to normal, numbers of Leydig cells but secrete very low amounts of androgen due to a lack of 17 alpha-hydroxylase activity. To determine whether this loss of activity is due to a lack of enzyme synthesis or a change in catalytic activity we have examined 17 alpha-hydroxylase cytochrome P-450 (P-450(17 alpha)) protein and mRNA levels in the testes of Tfm mice. Levels of P-450(17 alpha) protein were measured by immunoblotting, while mRNA was measured following reverse transcription (RT) and amplification by the polymerase chain reaction (PCR). Conditions for RT-PCR were determined which allowed semiquantification of P-450(17 alpha) mRNA relative to beta-actin mRNA. In extracts of Tfm testes P-450(17 alpha) protein was undetectable using antiserum against porcine P-450(17 alpha). In contrast, a protein of around 54 kDa was clearly detectable in extracts of control cryptorchid testes. Using RT-PCR, P-450(17 alpha) mRNA was detectable in both control and Tfm testes but, expressed in terms of beta-actin mRNA, levels of P-450(17 alpha) mRNA in control testes were 40-fold higher than those in Tfm testes. If the total amount of RNA extracted from each testis is taken into account then P-450(17 alpha) mRNA levels per testis were up to 400-fold higher in control testes. These results show that the reduced level of 17 alpha-hydroxylase activity in Tfm testes is related to reduced protein synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) |
