The long term objective of this project is to develop Dr. R. Renee Reams into an independent biomedical investigator, who will use techniques learned in a selected research mentor's laboratory to generate scientific data the an be used to apply for R01 research funding. The research mentoring will be provided in Dr. Raymond F. Novak's laboratory at the Institute of Chemical Toxicology at Wayne State University. Techniques learned by Dr. Reams will help her define and research problems that will elucidate the mechanism of action of """"""""lead"""""""" on mammalian systems. he specific techniques that Dr. Reams will learn in Dr. Novak's laboratory include cell culture of both primary hepatocytes as well as passage and cell lines, western blot analysis, total and poly (A) + RNA isolation, northern blot analysis, nuclear run on assays, reverse-transcriptase polymerase chain reaction, restriction analysis, cloning and DNA sequence analysis. These contemporary approaches in cell and molecular biology will be used by Dr. Reams in the laboratory at Florida A&M University to examine the """"""""Effects of Lead on Tyrosine Hydroxylase Gene Expression and signal Transduction in Adrenal and Neural Cells"""""""". Expression of TH and modulation of PKC enzymes will be monitored t the level of protein using a commercially available monoclonal antibody to probe western blots. Gene expression of TH and PKC will be detected by the presence of total and poly (A) + RNA isolation using northern blot analysis. Reverse- transcriptase-polymerase chain reaction will be used to examine and confirm the enhanced expression of tyrosine hydroxylase before and after lead treatment.
|Williams, T M; Ndifor, A M; Near, J T et al. (2000) Lead enhances NGF-induced neurite outgrowth in PC12 cells by potentiating ERK/MAPK activation. Neurotoxicology 21:1081-9|