Abstract
The rodent superior cervical ganglion (SCG) is a useful and readily accessible source of neurons for studying the mechanisms of sympathetic nervous system (SNS) development and growth in vitro. The sympathetic nervous system (SNS) of early postnatal animals undergoes a great deal of remodeling and development; thus, neurons taken from mice at this age are primed to re-grow and establish synaptic connections after in situ removal. The stereotypic location and size of the SCG make it ideal for rapid isolation and dissociation. The protocol described here details the requirements for the dissection, culture and differentiation of SCG neurons. The protocol is suitable for culturing neurons from late embryonic gestation to approximately postnatal day 3. The culture technique discussed below utilizes glass coverslips for the microscopic examination of fixed cells.
Materials and Reagents
Equipment
Procedure
Note: All solutions and equipment coming into contact with cells must be sterile. Aseptic technique is critical throughout the procedure; thus, dissection taking place in a tissue culture hood modified for the use of a stereoscopic microscope may yield the best results.
Recipes
Acknowledgments
This protocol is adapted from Quach et al. (2013) and Eldredge et al. (2008).
References
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