PRIMARY CULTURES OF DISSOCIATED SYMPATHETIC NEURONS: I. Establishment of Long-Term Growth in Culture and Studies of Differentiated Properties

doi:10.1083/jcb.59.2.329

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ARTICLE

PRIMARY CULTURES OF DISSOCIATED SYMPATHETIC NEURONS

: I. Establishment of Long-Term Growth in Culture and Studies of Differentiated Properties

Richard E. Mains ¹ and Paul H. Patterson ¹

¹ From the Department of Neurobiology, Harvard Medical School, Boston, Massachusetts 02115.

Dr. Mains' present address is the Department of Biology, University of Oregon, Eugene, Oregon 97403.

Rat sympathetic ganglia were disrupted by mechanical agitationto yield dissociated primary neurons, and the conditions forlong-term growth in culture of the isolated neurons were examined.The neurons were grown with or without non-neural cells, simplyby the addition or deletion of bicarbonate during growth inculture. Fluorescence histochemistry indicated that the isolatedneurons contained catecholamines; incubations with radioactiveprecursors were used to verify the synthesis and accumulationof both dopamine and norepinephrine. The neurons also producedoctopamine using tyramine as precursor, but not with tyrosineas the precursor. In the presence of eserine, older culturessynthesized and stored small amounts of acetylcholine. The culturesdid not synthesize and accumulate detectable levels of radioactive $gamma$ -aminobutyric acid, 5-hydroxytryptamine, or histamine.

Submitted on February 21, 1973
Revised on June 7, 1973

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