Molecular characterization of three gonad cell lines.
To facilitate the study of the regulation and downstream interactions of genes involved in gonad development it is important to have a suitable cell culture model. We therefore aimed to characterize molecularly three different mouse gonad cell lines. TM3 and TM4 cells were originally isolated from prepubertal mouse gonads and were tentatively identified as being of Leydig cell and Sertoli cell origin, respectively, based upon their morphology and hormonal responses. The third line is a conditionally immortalized cell line, derived from 10.5-11.5 days post-coitum (dpc) male gonads of transgenic embryos carrying a temperature-sensitive SV40 large T-antigen. We studied by reverse transcription-polymerase chain reaction (RT-PCR) the expression profiles of a number of genes known to be important for early gonad development. Moreover, we assessed these cell lines for their capacity to induce SOX9 transcription upon expression of SRY, a key molecular event occurring during sex determination. We found that all three cell lines were unable to upregulate SOX9 expression upon transfection of SRY-expression constructs, even though these cells express many of the studied embryonic gonad genes. These observations point to a requirement for SRY cofactors for direct or indirect upregulation of SOX9 expression during testis determination.
Beverdam
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A
Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia.
Wilhelm
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D
Koopman
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P
D000818:Animals; D002460:Cell Line; D057137:DAX-1 Orphan Nuclear Receptor; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D053823:Hedgehog Proteins; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D060850:LIM-Homeodomain Proteins; D007985:Leydig Cells / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009687:Nuclear Proteins; D000072079:Patched Receptors; D011956:Receptors, Cell Surface; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012708:Sertoli Cells / Q000378:metabolism; D019849:Sex Determination Processes; D012733:Sex Differentiation; D051876:Sex-Determining Region Y Protein; D013737:Testis / Q000166:cytology* / Q000196:embryology / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors; D025721:WT1 Proteins / Q000235:genetics / Q000378:metabolism
D057137:DAX-1 Orphan Nuclear Receptor; D004268:DNA-Binding Proteins; D053823:Hedgehog Proteins; D018398:Homeodomain Proteins; D060850:LIM-Homeodomain Proteins; C117058:Lhx9 protein, mouse; D008565:Membrane Proteins; C539966:Nr0b1 protein, mouse; D009687:Nuclear Proteins; D000072079:Patched Receptors; D011956:Receptors, Cell Surface; D018168:Receptors, Retinoic Acid; D012097:Repressor Proteins; D051876:Sex-Determining Region Y Protein; C497746:Sry protein, mouse; D015534:Trans-Activators; D014157:Transcription Factors; D025721:WT1 Proteins