@article{c51fc1f7505a4be28c748a2ca7b849b3,
title = "Target cell stimulation of dissociated serotonergic neurons in culture",
abstract = "Dissociated mesencephalic raphe cells from fetal rats (14-18 days) were grown in culture in 96 well Linbro plates. The maturation of serotonergic cells was qualitatively studied using immunocytochemistry with a serotonin antibody and quantitatively by measuring the retention of radioactivity following incubation in the presence of a low concentration of [3H]5-hydroxytryptamine (6 × 10-8 M). The 5-hydroxytryptamine immunoreactive neurons showed specific staining in the perikaryon, nucleus, dendrities, axons and growth cones. These neurons formed varicose fibers and growth cones after 18 h in culture and survived for up to 21 days in culture. Each serotonergic neuron concentrated approximately 1 fmol of serotonin after 20 min of incubation. Maturation of mesencephalic serotonergic neurons was increased in co-cultures of both normal (hippocampus, cerebral cortex, olfactory bulb and striatum) and abnormal (spinal cord) target neurons. The best stimulation was produced by dissociated hippocampal neurons (14-18 days of gestation) on mesencephalic raphe cells (14 days of gestation) after 4 days in culture. This stimulation was seen in culture conditions which favored neuronal but not glial survival. Our results obtained using cultures of dissociated serotonergic cells are consistent with an expansive network pattern developed by this chemical transmitter system in the adult brain.",
author = "Azmitia, {E. C.} and Whitaker-Azmitia, {P. M.}",
note = "Funding Information: cationicm oleculesp rovidea good substratefo r neur- onal attachmenta nd growth.47C ultures of 8 day chick embryo telencephalona re 98% neuronal (as determinedb y cells that bind tetanust oxin) if plated on polylysine.@T hesea uthorss uggestt hat the poly- D-lysine substratei nhibits gliai proliferation. Col- lagens ubstratec ulturesa reo vergrownw ith glial cells by 12-13 days. Rat hippocampal cells plated on poly-r)-lysinet reatedc overslipss how a markeds up- pression of non-neuronal ceIls.{\textquoteleft}OI n our studies on poly-D-lysinec oatedc ulture chambers,h ippocampal cells strongly stimulateds erotonergicu ptakem atur- ation (Table 2). However, the interpretation that poly-cationic moleculess uppressg lial cells is con- foundedb y the recento bservationt hat astocyticc ells grow and proliferatei n a definedm edia on a poly- D-lysine substrate.55 The secondl ine of evidencef or the sourceo f the neuronotrophice ffectb eing neuronali s derivedf rom the useo f anti-mitoticd rugsto eliminatea ll dividing cells. This procedure is extensivelyu sed for the control of glial proliferation.{\textquoteleft}3~2O*ur3 5st udiess how a delayo f sevend aysb eforei ntroducinga ra-C results in the presenceo f a large number of post-mitotic astrocytes(i dentifiedb y GFA-positive staining, Fig. 8). Thus, in our experimentsa, ra-C is added at the time of initial plating and maintainedf or 5 days in culture. The results show poor survival of raphe culturesa lone,b ut co-culturew ith hippocampus(a nd to a lessere xtents pinal cord) resultsi n a dramatic stimulationo f the S-HT high-a~nity uptakes ystem. ~ek~owle~ge~ent.~-Trhees earchwas supported by a In the final serieso f studies,c o-culturesa re grown National Science Foundation grant from Deveiopment in a defined serum-freem edia that is selectivef or Neurobiologyp anel No. BNS-83-04704T. echnicalassis- neuronai growth.{\textquoteleft}r Hippocampal target cells are by Delia SalazarT. he authorsw ish to thankD r Alain Privat tancep rovidedb y Martha Davila and secretariaal ssistance effectivein stimulatingt hem aturationo f serotonergic for his supporti n establishingti ssuec ulturesi n our labora- cells in this serum-freem edia. Co-cultures of dopa-tory and to Philip Alvarez for his helpfuf editorial sug- minergic neurons and striatal cells grown in serum-gestions. Copyright: Copyright 2015 Elsevier B.V., All rights reserved.",
year = "1987",
month = jan,
doi = "10.1016/0306-4522(87)90005-4",
language = "English (US)",
volume = "20",
pages = "47--63",
journal = "Neuroscience",
issn = "0306-4522",
publisher = "Elsevier Ltd",
number = "1",
}