Synaptic contact between embryonic neurons and acetylcholine receptor-fibroblast.
1. Mouse fibroblast cell lines were established that stably express Torpedo californica acetylcholine receptors (AChR) on their cell surface in quantities sufficient for biochemical and pharmacological analyses, as well as electrophysiological analysis at the single channel level. 2. Surface-expressed AChRs were shown to be assembled into proper alpha 2 beta gamma delta pentamers. 3. The distribution of surface-AChRs was uniform and identical in every cell. 4. We were able to successfully coculture AChR-fibroblasts with 1-day old Xenopus laevis embryonal neurons and maintain expression of cell surface AChRs. 5. Using the voltage-clamp technique, miniature end-plate currents were recorded from AChR-fibroblasts which were contacted by neurons. The current amplitudes of these AChRs were approximately 10-fold smaller than those observed in Xenopus myocytes, and the rise-times were slower.
Hartman
D S
DS
Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06510.
Poo
M M
MM
Green
W N
WN
Ross
A F
AF
Claudio
T
T
D000200:Action Potentials; D000818:Animals; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D015151:Immunoblotting; D051379:Mice; D009474:Neurons / Q000502:physiology*; D010784:Photomicrography; D011950:Receptors, Cholinergic / Q000032:analysis / Q000502:physiology*; D013569:Synapses / Q000502:physiology; D014101:Torpedo; D014982:Xenopus laevis / Q000196:embryology*
D011950:Receptors, Cholinergic