Choroid tissue supports the survival of ciliary ganglion neurons in vitro.
It is well established that during in vivo development the neurons of the avian ciliary ganglion are dependent for their survival on structures in the eye. Separate neuron populations innervate intraocular smooth and striated muscle targets. All ciliary neurons survive when cocultured with striated muscle. We demonstrate that when ciliary ganglion neurons are plated on explants of the choroid coat (a smooth muscle-containing target tissue) using a defined medium (N2), the neurons survive and grow vigorously into the tissue, forming contacts between axons and target cells identified as smooth muscle. Conditioned medium from choroid explants also rescues all the neurons, as does coculturing ciliary ganglion neurons with dissociated choroid cells. However, the presence of horse serum and chick embryo extract in the medium inhibits the choroid's ability to support ciliary neurons. The effects of these additives on the phenotypic expression of the smooth muscle may explain the inability of previous investigators to demonstrate target-derived support from smooth muscle preparations. Because the choroid contains cell types other than smooth muscle (e.g., fibroblasts and endothelial cells), we could not identify smooth muscle as the only cell type responsible for the release of the soluble trophic factor present in the target tissue. However, indirect evidence using avian primary fibroblast cultures, a fibroblast cell line, and an anatomically simple smooth muscle preparation, the avian amnion, suggests that smooth muscle cells are sufficient to account for the observed trophic activity, and that similar target-derived molecules support the survival of both types of ciliary ganglion cells.
Wentzek
L A
LA
Physiology and Neurobiology Department, University of Connecticut, Storrs 06269-3042.
Bowers
C W
CW
Khairallah
L
L
Pilar
G
G
D016475:3T3 Cells; D000818:Animals; D002450:Cell Communication; D002470:Cell Survival; D002478:Cells, Cultured; D002642:Chick Embryo; D002829:Choroid / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D017077:Culture Media, Conditioned; D005347:Fibroblasts / Q000166:cytology; D005726:Ganglia, Parasympathetic / Q000166:cytology*; D007700:Kinetics; D051379:Mice; D008854:Microscopy, Electron; D009130:Muscle, Smooth / Q000166:cytology / Q000502:physiology*; D009474:Neurons / Q000166:cytology* / Q000648:ultrastructure; D009924:Organ Culture Techniques; D012867:Skin / Q000166:cytology; D013997:Time Factors
D017077:Culture Media, Conditioned