Expression of glycosphingolipids in serum-free primary cultures of mouse kidney cells: male-female differences and androgen sensitivity.
The expression of neutral glycosphingolipids was examined in primary kidney cell cultures derived from adult male and female beige mutant mice (C57BL/6J;bgj/bgj) with enrichment for proximal tubule cells during preparation of explants and using defined serum-free medium for the culture conditions. Cells proliferated for 7 days in vitro to provide confluent or nearly confluent monolayers of epithelial-type growth indicative of proximal tubule cells. The male vs female differences in neutral glycosphingolipids seen in the kidney in vivo were retained in these 7 day cultures. Cultures derived from males contained galacto- and digalactosylceramides whereas those from females did not express these types of glycolipids. Also, male cells had higher ratios of sphingosine: phytosphingosine containing species in Nfa (non-hydroxy fatty acid) globotriaosylceramide and in glucosylceramide than females. The shift in sphingosine: phytosphingosine to male ratios in Nfa globotriaosylceramide and in glucosylceramide could be stimulated in female kidney cells by treatment with 10(-5) M testosterone or 5 alpha-dihydrotestosterone. The male-specific expression of neutral glycosphingolipids, then, appears to be stable character of male-type differentiation in mouse kidney that is passed on during proliferation in culture. Female kidney cells retain an ability to respond to androgens with specific changes in neutral glycosphingolipid expression during 7 days of growth in vitro in serum-free conditions, but do not respond with the induction of the male-specific glycolipids galacto- and digalactosylceramides as seen in vivo.
Gross
S K
SK
E.K. Shriver Center for Mental Retardation, Waltham, MA 02254.
Lyerla
T A
TA
Evans
J E
JE
McCluer
R H
RH
D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D016895:Culture Media, Serum-Free; D013196:Dihydrotestosterone / Q000494:pharmacology; D004847:Epithelial Cells; D005227:Fatty Acids / Q000032:analysis; D005260:Female; D005699:Galactosylceramides / Q000032:analysis; D005786:Gene Expression Regulation / Q000187:drug effects; D006028:Glycosphingolipids / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D012727:Sex Characteristics; D013110:Sphingosine / Q000031:analogs & derivatives / Q000032:analysis; D013739:Testosterone / Q000494:pharmacology; D014281:Trihexosylceramides / Q000032:analysis
D000728:Androgens; D016895:Culture Media, Serum-Free; D005227:Fatty Acids; D005699:Galactosylceramides; D006028:Glycosphingolipids; D014281:Trihexosylceramides; D013196:Dihydrotestosterone; D013739:Testosterone; C018549:globotriaosylceramide; C012491:phytosphingosine; D013110:Sphingosine