Regulation of alpha2-macroglobulin expression in rat Sertoli cells and hepatocytes by germ cells in vitro.
Germ cells isolated from rat testes by trypsinization have been shown to yield unwanted artifacts in biological assays, since conditioned media derived from these germ cells (germ cell-conditioned media [GCCM]) can modulate Sertoli cell secretory function because of the presence of residual trypsin. To determine whether germ cells themselves can modulate Sertoli cell function, we isolated germ cells from tubules by a mechanical procedure and assessed the effect of these cells on Sertoli cell alpha2-macroglobulin (alpha2-MG) steady-state mRNA level. It was found that germ cells indeed could stimulate Sertoli cell alpha2-MG expression. This effect is probably mediated by a soluble factor(s) released from germ cells, since GCCM fractionated by HPLC contained multiple fractions that can stimulate Sertoli cell alpha2-MG expression dose-dependently. These results illustrate that germ cells play a role in regulating testicular alpha2-MG expression. Since Sertoli cells synthesize and secrete many of the serum proteins behind the blood-testis barrier that are also produced by hepatocytes, we sought to ascertain whether germ cells can affect hepatic alpha2-MG expression. When germ cells were cocultured with hepatocytes isolated from adult rats, the hepatocyte alpha2-MG steady-state mRNA level was shown to be stimulated by germ cells dose-dependently. Using different pools of fractions derived from GCCM after their fractionation by a preparative anion-exchange HPLC column, GCCM was found to contain a factor(s) that stimulated hepatocyte alpha2-MG expression dose-dependently. More importantly, the fractions that stimulated hepatocyte alpha2-MG expression had a retention time different from that of the factor(s) that affected Sertoli cell alpha2-MG expression. These data illustrate that germ cells secrete multiple biological factors capable of regulating alpha2-MG expression in the testis and the liver. In summary, this study reveals a possible physiological link between the testis and the liver in that germ cells may release a factor(s) capable of modulating alpha2-MG expression in both organs.
The Population Council, Center for Biomedical Research, New York, New York 10021, USA.
L
L
Braghiroli
B
B
Silvestrini
C
C
Sorrentino
J
J
Grima
D
D
Mruk
C Y
CY
Cheng
D000375:Aging; D000818:Animals; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D003907:Dexamethasone / Q000494:pharmacology; D005786:Gene Expression Regulation; D015850:Interleukin-6 / Q000494:pharmacology; D008099:Liver / Q000378:metabolism*; D008115:Liver Regeneration; D008297:Male; D011506:Proteins / Q000302:isolation & purification; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012708:Sertoli Cells / Q000378:metabolism*; D013094:Spermatozoa / Q000502:physiology*; D000511:alpha-Macroglobulins / Q000235:genetics* / Q000502:physiology
D017077:Culture Media, Conditioned; D015850:Interleukin-6; D011506:Proteins; D012333:RNA, Messenger; D000511:alpha-Macroglobulins; D003907:Dexamethasone