Sperm and oocyte isolation methods for biochemical and proteomic analysis.
Caenorhabditis elegans gonad is a simple model to investigate molecular mechanisms that regulate fundamental cell and developmental processes. The strength of the model is that C. elegans is amenable to genetic manipulation. The complete genome sequence, advances in mass spectrometry, and RNA-mediated interference (RNAi) are now providing a technical infrastructure that complements biochemical and proteomic approaches. This chapter describes simple methods for sperm and oocyte isolation and gonad liberation that can be performed routinely in the lab, without expensive equipment. These methods are ideal for biochemical and proteomic applications, including those aiming to identify proteins based on affinity or biological activity. Germline mRNA expression profiles, RNAi feeding clones, and Gateway-engineered ORFeome vectors are available to help validate experimental results.
Miller
Michael A
MA
Department of Cell Biology, University of Alabama at Birmingham, USA.
D000818:Animals; D002469:Cell Separation / Q000379:methods; D005260:Female; D008297:Male; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D013094:Spermatozoa / Q000166:cytology* / Q000378:metabolism*
D020543:Proteome