Explant cultures of catecholamine-containing neurons from rat brain: Biochemical, histofluorescence, and electron microscopic studies

Abstract

ABSTRACr Norepinephrine (NE~producing cells of the nucleus locus ceruleus and dopamine (DA)producing cells of the substantia nigra were dissected microscopically from embryonic rat brain, explanted, and maintained in cu ture for up to 5 weeks. The cultured neurons of both brain regions showed normal maturation of axons and dendrites and formed ultrastructurali defined synaptic contacts. Fluorescence microscopy of cultured neurons from both brain regions showed typical in situ cytological features: long axonal processes with multiple varicosities for locus ceruleus cultures, and smooth, wispy nonvaricose processes in the substantia nigra cultures. All cultures processed for fluorescence microscopy contained specific catecholamine-fluorescent cells. BY radioenzyme assay for The long survival time in culture of locus ceruleus cells, the normal appearance of fluorescent cell bodies and processes, the apparent development of morphologically specialized interneuronal connections, and the ability to synthesize and store NE make these cultures ideally suited for neurophysiological recording as well as morphological, biochemical, and pharmacological experiments.Many advantages would accrue from the ability to culture specific neuronal nuclei from mammalian brain. The controlled conditions of the culture situation could yield data on the metabolism, pharmacological responsiveness, receptor specificity, and developmental characteristics of defined, homogeneous populations of neurons. To date, neuronal culture systems of mammalian brain have utilized entire brain (1, 2), individual cortical (3-6) or lower mesencephalic (7)(8)(9)(10)(11) The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

NGF and Anti-NGF: Evidence against Effects on Fiber Growth in Locus coeruleus from Cultures of Perinatal CNS Tissues

“…[7] reported negative effects of nerve growth factor on cultures of the sub stantia nigra. Locus coeruleus and substantia nigra cultures from rat fetuses have also been reported upon by Schlumpf et al [35], and explants of mouse locus coeruleus have been described by Victorov el al. [45].…”

Presynaptic transmitters and depolarizing influences regulate development of the substantia nigra in culture

“…Nevertheless, specific histochemical and biochemical markers have been used to examine the ontogeny of different neuronal populations (Coyle and Axelrod, 1972;Olson and Seiger, 1972;Coyle et al, 1973;Lauder and Bloom, 1974;Schlumpf et al, 1977;Specht et al, 1981a-c;Berger et al, 1982;Kotake et al, 1982;Verney et al, 1982;Dreyfus et al, 1986). However, specific signals governing ontogeny are largely unknown.…”

In vitro Studies on the Maturation of Mesencephalic Dopaminergic Neurons

“…Under these conditions, DA neurons within the explant do retain their histofluorescent prop erties as well as the ability to take up 'H-DA. Taken from the rat embryo, the mesencepha lon cut into very small fragments can be kept alive for several weeks in culture, allowing biochemical measurements: it exhibits cytological and ultrastructural features of well developed DA neurons (like the presence of some vesicles) [16]. Although the explant retains the original cell composition and an organotypic relationship among component cells, the latter are not readily accessible.…”