A STUDY ON GENITAL SKIN ANDROGEN RECEPTOR BY DISPERSED CELL METHOD Yasukazu Sen and Masaaki Morioka Department of Urology, Kochi Medical School, Nankoku, Japan (Director: Prof. Yukitoshi Fujita) Yoshiteru Kohama Department of Urology, Okayama University, Okayama, Japan (Director: Prof. Hiroyuki Ohmori) We have measured androgen receptor in whole cell, cytosol and nuclear fractions of cultured skin fibroblasts originated from the genital area. Samples were obtained from the following patients: 10 boys with phimosis, 5 boys with cryptorchism, 4 boys and a man with hypospadia, 2 infertile men and 3 normal men. Androgen receptor was measured using methyltrienolone (R1881) as the ligand according to the assay technique of Eil et al. using dispersed cell method. The results were as follows: 1. There were no significant statistical differences in the dissociation constant among the diseases. 2. Hypospadia cases showed lower maximum binding capacities (Bmax) of whole cell than phimotic boys but the difference was not statistically significant. 3. The Bmax of normal men were statistically higher than those of phimotic boys and about 80% of these receptors were seen in the nuclear fraction. We also experienced low Bmax levels in an adult hypospadia case and two infertile men. These resutls suggest that this method is a simple and rapid method to study both nuclear and cytosol androgen receptor at the same time. It is also suggested that there are some cases with reduced androgen receptor levels in hypospadia and infertile cases.

Fig. 1 Dispersed cell assay method Fig. 2 Saturation curve and Scatchard Plot analysis in nuclear and cytosol fraction of dispersed cell nuclear Bmax(N)=19. 99x102sites/cell Kd (N) =0. 18nM cytosol Bmax(C)=9. 88x102sites/cell Kd (C) =0. 18nM

Fig. 3 Time course of 3H-R1881 specific binding in whole cell assay Table 1 Kd and Bmax of each assay method (Mean+S. D., Kd: nm, Bmax: fmol/mg protein) Fig. 4 Steroid specificity of 3H-R1881 binding in whole cell assay Table 2 Changes of R1881 binding over serial subcultures (Kd: nm, Bmax: fmol/mg protein)

Table 3 Androgen receptor in cultured genital skin fibroblast (dispersed cell assay)

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