Culture conditions affect the cholinergic development of an isolated subpopulation of chick mesencephalic neural crest cells
Barald, Kate F.
1989-10
Citation
Barald, Kate F. (1989/10)."Culture conditions affect the cholinergic development of an isolated subpopulation of chick mesencephalic neural crest cells." Developmental Biology 135(2): 349-366. <http://hdl.handle.net/2027.42/27739>
Abstract
Although neural crest cells are known to be very responsive to environmental cues during their development, recent evidence indicates that at least some subpopulations may be committed to a specific differentiation program prior to migration. Because the neural crest is composed of a heterogeneous mixture of cells that contributes to many vertebrate cell lineages, assessing the properties of specific subpopulations and the effect of the environment on their development has been difficult. To address this problem, we have isolated a pure subpopulation of chick mesencephalic neural crest cells by fluorescence no-flow cytometry after labeling them with monoclonal antibodies (Mabs) to a 75-kDa cell surface antigen that is associated with high affinity choline uptake. When cultures of chick mesencephalic neural crest cells are labeled with these Mabs and a fluorescent second step antibody, ~5% of the cells are antigen-positive (A+). After sorting, 100% of the resulting cultured mesencephalic neural crest cells are A+. The Mabs we used also label all of the neurons of the embryonic chick and quail ciliary ganglion in vivo and in vitro. We have compared the effect of various cell culture media on the isolated neural crest subpopulation and the heterogeneous chick mesencephalic neural crest from which it was derived. A+ cells were passaged and grown in a variety of media, each of which differently affected its characteristics and development. A+ cells proliferated in the presence of 15% fetal bovine serum (FBS) and high concentrations (10-15%) of chick embryo extract, but did not differentiate, although they retained basal levels of choline acetyltransferase (ChAT) activity. However, in chick serum and high (25 mM as opposed to 7 mM) K+, and heart-, iris-, or lung-conditioned medium, all of which are known to promote survival and/or cholinergic development of ciliary ganglion neurons, the cells ceased to proliferate and all of the cells in the culture became "neuron-like" within 10 days. No neuron-like cells were found in liver-, notocord-, or neural tube-conditioned media if FBS was used. When A+ cells were eliminated either by complement-mediated cytotoxicity or by laser-ablating A+ cells during no-flow cytometry, all ChAT activity was also eliminated, and no neuron-like cells or ChAT activity was found in cultures during a subsequent 3-week culture period. We conclude that culture conditions that normally promote ciliary ganglion neuron survival and cholinergic development also promote differentiation and development of cholinergic neuron-like cells from the isolated A+ neural crest subpopulation. These results are consistent with the hypothesis that the A+ subpopulation contributes to the formation of ciliary ganglion neurons in the embryo.Publisher
Elsevier
PMID
2776973
Types
Article
URI
http://www.sciencedirect.com/science/article/B6WDG-4DYN3FP-D/2/f66be509ae83671d25d92c1e098dc89dhttp://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=2776973&dopt=citation
Metadata
Show full item recordCollections
Remediation of Harmful Language
The University of Michigan Library aims to describe library materials in a way that respects the people and communities who create, use, and are represented in our collections. Report harmful or offensive language in catalog records, finding aids, or elsewhere in our collections anonymously through our metadata feedback form. More information at Remediation of Harmful Language.
Accessibility
If you are unable to use this file in its current format, please select the Contact Us link and we can modify it to make it more accessible to you.