Human growth hormone co-transfection assay to study molecular mechanisms of neurosecretion in PC12 cells.
The recent introduction of transient co-transfection assays, with human growth hormone (hGH) as a marker of transfection, has allowed us to study the function of transfected proteins in neurosecretion using intact PC12 cells and adrenal chromaffin cells [J. Biol Chem. 268 (1993) 10983]. Since this assay requires several non-trivial steps (transfection of the genes, triggering of secretion, and measurement of hGH), care should be taken before attributing any observed effect to the transfected proteins. In this article, we describe a detailed procedure for the hGH transfection assay and show that this approach can be used to study the molecular events involved in neurosecretion.
Sugita
Shuzo
S
Division of Cellular and Molecular Biology, Toronto Western Research Institute, University Health Network, Toronto, Ont., Canada. ssugita@uhnres.utoronto.ca
D000818:Animals; D005822:Genetic Vectors; D019382:Human Growth Hormone / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000502:physiology; D016716:PC12 Cells; D010957:Plasmids; D050765:Qa-SNARE Proteins; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D013111:Spider Venoms / Q000494:pharmacology; D014162:Transfection
D008565:Membrane Proteins; D050765:Qa-SNARE Proteins; D011994:Recombinant Proteins; D013111:Spider Venoms; D019382:Human Growth Hormone; C020763:alpha-latrotoxin