Optimization of neuronal cultures from rat superior cervical ganglia for dual patch recording

Julien Amendola; Norah Boumedine; Marion Sangiardi; Oussama El Far

doi:10.1038/srep14455

Optimization of neuronal cultures from rat superior cervical ganglia for dual patch recording

Sci Rep. 2015 Sep 24:5:14455. doi: 10.1038/srep14455.

Authors

Julien Amendola^{1

2}, Norah Boumedine^{1

2}, Marion Sangiardi^{1

2}, Oussama El Far^{1

2}

Affiliations

¹ INSERM, UMR_S 1072, Marseille, France.
² Aix-Marseille Université, Marseille, France.

Abstract

Superior cervical ganglion neurons (SCGN) are often used to investigate neurotransmitter release mechanisms. In this study, we optimized the dissociation and culture conditions of rat SCGN cultures for dual patch clamp recordings. Two weeks in vitro are sufficient to achieve a significant CNTF-induced cholinergic switch and to develop mature and healthy neuronal profiles suited for detailed patch clamp analysis. One single pup provides sufficient material to prepare what was formerly obtained from 12 to 15 animals. The suitability of these cultures to study neurotransmitter release mechanisms was validated by presynaptically perturbing the interaction of the v-SNARE VAMP2 with the vesicular V-ATPase V0c subunit.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques
Cholinergic Neurons / drug effects
Cholinergic Neurons / physiology
Electrophysiological Phenomena*
Nerve Growth Factors / pharmacology
Neurons / cytology*
Neurons / drug effects
Neurons / physiology*
Neurotransmitter Agents / metabolism
Rats
Superior Cervical Ganglion / cytology*
Synaptic Transmission

Substances

Nerve Growth Factors
Neurotransmitter Agents