NGF induces neonatal rat sensory neurons to extend dendrites in culture after removal of satellite cells.
Vertebrate sensory neurons have a pseudo-unipolar morphology; their somata are covered by satellite cells and lack dendrites or synaptic contacts. However, when neonatal rat sensory neurons from the nodose ganglia develop in culture in absence of satellite cells and with NGF, they form synapses among themselves. In this study, we investigated whether neonatal rat nodose neurons express dendrites under the same culture conditions. We show by Lucifer yellow injection that nodose neurons remain typically unipolar when cocultured with their ganglionic satellite cells. However, when these neurons are cultured without satellite cells, virtually all neurons acquire a multipolar morphology. Moreover, when NGF is added to satellite cell-free cultures, several neurons extend dendrites; these processes stain positively for microtubule-associated protein-2. NGF induces a 17-fold increase in dendritic outgrowth after 3 weeks but has little effect on axon number. In addition, we find that the ability of nodose neurons to extend dendrites is developmentally regulated. Furthermore, in a combined morphological and electrophysiological study, using whole-cell voltage-clamp technique with Lucifer yellow in the recording solution, we demonstrate a positive correlation between the extent of dendritic outgrowth and the density of ACh currents, suggesting that these dendrites have ACh receptors. Our results indicate that neonatal rat nodose neurons are capable of extending dendrites and that extrinsic factors can induce or suppress their extension. In addition, the results suggest that these dendrites may act as principal post-synaptic structures for synapse formation that occurs in these cultures.
De Koninck
P
P
Department of Biology, McGill University, Montréal, Québec, Canada.
Carbonetto
S
S
Cooper
E
E
D000818:Animals; D000831:Animals, Newborn; D001369:Axons / Q000502:physiology / Q000648:ultrastructure; D002478:Cells, Cultured; D003712:Dendrites / Q000502:physiology* / Q000648:ultrastructure; D007150:Immunohistochemistry; D007546:Isoquinolines; D008869:Microtubule-Associated Proteins / Q000032:analysis; D009414:Nerve Growth Factors / Q000494:pharmacology*; D009475:Neurons, Afferent / Q000502:physiology / Q000648:ultrastructure*; D009620:Nodose Ganglion / Q000166:cytology*; D051381:Rats; D011950:Receptors, Cholinergic / Q000502:physiology
D007546:Isoquinolines; D008869:Microtubule-Associated Proteins; D009414:Nerve Growth Factors; D011950:Receptors, Cholinergic; C017475:lucifer yellow