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Early synaptogenesis in vitro: Role of axon target distance

Early synaptogenesis in vitro: Role of axon target distance In contrast to some previous reports suggesting a delay in synapse formation in vitro, we found that under ideal conditions, most hippocampal and hypothalamic rat neurons were synaptically coupled after 3 or 4 days in vitro. Synaptophysin immunocytochemistry revealed strongly stained presynaptic boutons by 3 days in vitro. Studies with time‐lapse laser confocal imaging of FM‐143 revealed that axonal boutons were recycling their synaptic vesicles, an indication of synapse formation, as early as 3 days after plating. To test the hypothesis that neurite outgrowth was enhanced in high‐density cultures, thereby increasing the probability of synapse formation, neurons were transfected with the jellyfish green fluorescent protein (GFP) gene. After 2 days in high‐density cultures, green fluorescent neurites were about three times longer than in sister neurons plated in low‐density cultures. Even in single dishes, GFP‐transfected cells in contact with other neurons had neurites that were at least three times longer and grew faster than more isolated cells. Neurons grew longer neurites (+51%) when growing on surface membranes of heat‐killed neurons than on polylysine, underlining the importance of plasma membrane contact. Calcium imaging with fura‐2 and whole cell recording showed that both GABA and glutamate presynaptic release occurred after 3 or 4 days in vitro in high‐density cultures but was absent in low‐density cultures at this time. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Journal of Comparative Neurology Wiley

Early synaptogenesis in vitro: Role of axon target distance

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References (44)

Publisher
Wiley
Copyright
"Copyright © 1998 Wiley Subscription Services, Inc., A Wiley Company"
ISSN
0021-9967
eISSN
1096-9861
DOI
10.1002/(SICI)1096-9861(19981005)399:4<541::AID-CNE7>3.0.CO;2-1
Publisher site
See Article on Publisher Site

Abstract

In contrast to some previous reports suggesting a delay in synapse formation in vitro, we found that under ideal conditions, most hippocampal and hypothalamic rat neurons were synaptically coupled after 3 or 4 days in vitro. Synaptophysin immunocytochemistry revealed strongly stained presynaptic boutons by 3 days in vitro. Studies with time‐lapse laser confocal imaging of FM‐143 revealed that axonal boutons were recycling their synaptic vesicles, an indication of synapse formation, as early as 3 days after plating. To test the hypothesis that neurite outgrowth was enhanced in high‐density cultures, thereby increasing the probability of synapse formation, neurons were transfected with the jellyfish green fluorescent protein (GFP) gene. After 2 days in high‐density cultures, green fluorescent neurites were about three times longer than in sister neurons plated in low‐density cultures. Even in single dishes, GFP‐transfected cells in contact with other neurons had neurites that were at least three times longer and grew faster than more isolated cells. Neurons grew longer neurites (+51%) when growing on surface membranes of heat‐killed neurons than on polylysine, underlining the importance of plasma membrane contact. Calcium imaging with fura‐2 and whole cell recording showed that both GABA and glutamate presynaptic release occurred after 3 or 4 days in vitro in high‐density cultures but was absent in low‐density cultures at this time.

Journal

The Journal of Comparative Neurology – Wiley

Published: May 5, 1998

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