A technique for culturing brain nuclei from postnatal rats

Abstract

The technique for culturing brain nuclei from postnatal rats is described in detail. Key features of this simple method of culturing brain nuclei are: (a) to make brain slices of a particular brain region and to isolate the brain nucleus under direct visualization using a dissecting microscope; (b) to use papain for dissociation; (c) to grow neuron cultures over a glial feeder layer; and (d) to use rat serum (prepared in the laboratory) in the culture medium. We have developed neuronal cultures from several types of brain nuclei (such as the locus coeruleus) and identified the type of neurons immunocytochemically and histochemically. With this culture method, we can obtain high purity cultures of specific types of brain neurons. Our brain nucleus cultures are excellent materials for cellular and molecular physiological studies. Long-term changes of a single neuron belonging to a particular neuron type can be observed. Experiments using the patch clamp technique and intracellular injection of antibodies and antisense oligonucleotides are feasible.