Comparative proteomic analysis to study molecular events during gonad development in mice.
Sex determination represents a critical bifurcation in the road of embryonic development. It is based on a finely regulated network of gene activity, as well as protein-protein interactions and activation or silencing of signaling pathways. Despite the identification of a number of critical genes, many aspects of the molecular cascade that drives the differentiation of the embryonic gonad into either a testis or an ovary remain poorly understood. To identify new proteins involved in this cascade, we employed two-dimensional gel electrophoresis and mass spectrometry to compare the protein expression profiles of fetal mouse testes and ovaries. Three proteins, hnRPA1, TRA1, and HSC71, were found to be expressed in a male-specific manner and this expression was confirmed by real-time reverse transcriptase polymerase chain reaction (RT-PCR) and in situ hybridization. Moreover, HSC71 was found to be hyperphosphorylated in male compared to female gonads, emphasizing the advantage of the proteomic approach in allowing the detection of posttranslational modifications.
Wilhelm
Dagmar
D
Division of Molecular Genetics and Development, Institute for Molecular Bioscience, University of Queensland, Brisbane, Queensland, Australia.
Huang
Eugene
E
Svingen
Terje
T
Stanfield
Scott
S
Dinnis
Diane
D
Koopman
Peter
P
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D050883:HSC70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D010053:Ovary / Q000196:embryology* / Q000254:growth & development / Q000378:metabolism*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D051716:Replication Protein A / Q000235:genetics / Q000378:metabolism; D013737:Testis / Q000196:embryology* / Q000254:growth & development / Q000378:metabolism*
D050883:HSC70 Heat-Shock Proteins; C494372:Hspa8 protein, mouse; D008562:Membrane Glycoproteins; D020543:Proteome; D051716:Replication Protein A; C497168:Rpa1 protein, mouse; C053387:endoplasmin