Effect of germ cell depletion on levels of specific mRNA transcripts in mouse Sertoli cells and Leydig cells.
It has been shown that testicular germ cell development is critically dependent upon somatic cell activity but, conversely, the extent to which germ cells normally regulate somatic cell function is less clear. This study was designed, therefore, to examine the effect of germ cell depletion on Sertoli cell and Leydig cell transcript levels. Mice were treated with busulphan to deplete the germ cell population and levels of mRNA transcripts encoding 26 Sertoli cell-specific proteins and 6 Leydig cell proteins were measured by real-time PCR up to 50 days after treatment. Spermatogonia were lost from the testis between 5 and 10 days after treatment, while spermatocytes were depleted after 10 days and spermatids after 20 days. By 30 days after treatment, most tubules were devoid of germ cells. Circulating FSH and intratesticular testosterone were not significantly affected by treatment. Of the 26 Sertoli cell markers tested, 13 showed no change in transcript levels after busulphan treatment, 2 showed decreased levels, 9 showed increased levels and 2 showed a biphasic response. In 60% of cases, changes in transcript levels occurred after the loss of the spermatids. Levels of mRNA transcripts encoding Leydig cell-specific products related to steroidogenesis were unaffected by treatment. Results indicate (1) that germ cells play a major and widespread role in the regulation of Sertoli cell activity, (2) most changes in transcript levels are associated with the loss of spermatids and (3) Leydig cell steroidogenesis is largely unaffected by germ cell ablation.
O'Shaughnessy
P J
PJ
Division of Cell Sciences, Institute of Comparative Medicine, University of Glasgow Veterinary School, Bearsden Road, Glasgow G61 1QH, UK. p.j.oshaughnessy@vet.gla.ac.uk
Hu
L
L
Baker
P J
PJ
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D000988:Antispermatogenic Agents; D001483:Base Sequence; D002066:Busulfan; D002452:Cell Count; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004706:Endodeoxyribonucleases; D004950:Esterases / Q000235:genetics; D015870:Gene Expression; D007985:Leydig Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010982:Platelet-Derived Growth Factor / Q000235:genetics; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012708:Sertoli Cells / Q000378:metabolism*; D013087:Spermatids / Q000166:cytology; D013090:Spermatocytes / Q000166:cytology; D013093:Spermatogonia / Q000166:cytology; D013094:Spermatozoa / Q000502:physiology*; D013995:Time; D000099257:Meiotic Recombination Protein SPO11
D048868:Adaptor Proteins, Signal Transducing; D000988:Antispermatogenic Agents; D002066:Busulfan; D002868:Chromosomal Proteins, Non-Histone; D017931:DNA Primers; D004706:Endodeoxyribonucleases; D004950:Esterases; D010982:Platelet-Derived Growth Factor; D011506:Proteins; D012333:RNA, Messenger; D000099257:Meiotic Recombination Protein SPO11; C102019:Stra8 protein, mouse; C075278:platelet-derived growth factor A; C053886:spermatid transition proteins