Key result
The addition of ciliary neurotrophic factor (CNTF) to the culture medium significantly increased the neuronal connectivity rate of rat superior cervical ganglion neurons from 12% to 66%.
Absolute Event Rate: 66% vs 12%
p-value: p=<0.001
Optimized dissociation and culture conditions using sequential collagenase/trypsin and CNTF significantly improve the yield and cholinergic connectivity of rat superior cervical ganglion neurons for dual patch clamp recordings.
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.
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Amendola et al. (2015) studied this question. Ciliary neurotrophic factor (CNTF) vs. Nerve growth factor (NGF) alone was evaluated on Neuronal connectivity rate (p=<0.001). The addition of ciliary neurotrophic factor (CNTF) to the culture medium significantly increased the neuronal connectivity rate of rat superior cervical ganglion neurons from 12% to 66%.
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