Fluorescence-activated cell sorting of embryonic mouse and rat motoneurons and their long-term survival in vitro.
Motoneurons from embryonic mice and rats were labeled with retrogradely transported succinyl wheat germ agglutinin (WGA)-fluorescein isothiocyanate (FITC). After dissociation of the spinal cord, fluorescent motoneurons were isolated by flow cytometry. Sorted motoneurons were maintained for as long as 6 weeks in vitro on monolayers of astrocytes in muscle-conditioned medium. Immunocytochemical staining of the cultures for various neuronal antigens suggested that sorted motoneurons are receptive to GABAergic and glycinergic, as well as cholinergic, innervation. Many of the sorted cells were also labeled intracellularly with antibodies to choline acetyltransferase (ChAT) and GABA.
Schaffner
A E
AE
Laboratory of Neurophysiology, NINCDS-NIH, Bethesda, Maryland 20892.
St John
P A
PA
Barker
J L
JL
D015761:4-Aminopyridine; D000631:Aminopyridines; D000818:Animals; D000906:Antibodies / Q000032:analysis; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D005434:Flow Cytometry; D005968:Glutamate Decarboxylase / Q000276:immunology; D051379:Mice / Q000196:embryology; D009046:Motor Neurons / Q000166:cytology / Q000276:immunology / Q000378:metabolism / Q000502:physiology; D051381:Rats / Q000196:embryology; D018009:Receptors, Glycine; D017981:Receptors, Neurotransmitter / Q000276:immunology; D013997:Time Factors; D005680:gamma-Aminobutyric Acid / Q000276:immunology
D000631:Aminopyridines; D000906:Antibodies; D018009:Receptors, Glycine; D017981:Receptors, Neurotransmitter; D005680:gamma-Aminobutyric Acid; D015761:4-Aminopyridine; D005968:Glutamate Decarboxylase