Effects of ethylnitrosourea on expression of proto-oncogene pp60c-src and high-molecular-weight neurofilament protein in rodent embryo central nervous system cells in vitro

Toxicology and Applied Pharmacology
E M Faustman, C Sweeney

Abstract

Effects of exposure to ethylnitrosourea (ENU) on expression of proteins that play a role in neuronal differentiation were examined in central nervous system (CNS) micromass embryo cell cultures. ENU is a known developmental toxicant which affects neuronal development. The proteins selected were the protein product of the src proto-oncogene (pp60c-src) and high-molecular-weight neurofilament protein (NF). pp60c-src has marked increases in amount and kinase activity in neurons at the time of differentiation and NF is found in differentiated neurons. CNS micromass cultures are primary cells from Day 12 rat embryo midbrains which are plated at high density and differentiate into neurons during 5 days in culture. Proteins were quantitated by polyacrylamide gel electrophoresis separation of equal amounts of total cell protein followed by transfer to membranes, immunoblotting, and densitometric scanning of blots. Dose-dependent decreases in cell growth and differentiation were confirmed using endpoints of cell number, protein content of cultures, neutral red uptake, hematoxylin staining of differentiated cells, and levels of binding of the neurotransmitter [3H]gamma-aminobutyric acid. Concentrations which inhibited response by 50% com...Continue Reading

Citations

May 1, 2001·Current Protocols in Toxicology·R A Ponce

Related Concepts

Cell Differentiation Process
Cell Division Phases
Cell Survival
Densitometry
Dose-Response Relationship, Drug
SDS-PAGE
Ethylnitrosourea
Forelimb
Gamma-Aminobutyric Acid, Zinc Salt (2:1)
Midbrain Structure

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