PMID: 3760948Oct 1, 1986Paper

Primary culture of identified neurons from the visual cortex of postnatal rats

The Journal of Neuroscience : the Official Journal of the Society for Neuroscience
J E Huettner, R W Baughman

Abstract

We have examined the properties of neurons from the visual cortex of postnatal Long Evans rats in dissociated cell culture. Visual cortex from rat pups 1-15 d old was subjected to enzymatic and mechanical dissociation to yield a suspension of single cells. Neurons plated onto collagen or a feeder layer of astrocytes rapidly extended processes and survived for 4-10 weeks. Antisera to glutamic acid decarboxylase, choline acetyltransferase, and vasoactive intestinal polypeptide stained 22 +/- 2, 2.3 +/- 0.3, and 2.4 +/- 0.2% of all neurons, respectively, suggesting that different neuronal classes survived roughly in proportion to their number in vivo. In order to study a particular identified class of cortical neurons, we prelabeled cells in vivo by retrograde transport of a fluorescent tracer. Neurons in layer V of visual cortex that project to the superior colliculus were labeled after injecting fluorescent latex microspheres into the colliculus. Retrogradely labeled neurons were readily identified immediately after dissociation and throughout the period in vitro. After 2 weeks in culture, labeled cells exhibited many ultrastructural features characteristic of pyramidal neurons in vivo. Intracellular recording techniques were us...Continue Reading

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