[3H]acetylcholine synthesis in cultured ciliary ganglion neurons: Effects of myotube membranes

Db, Gray; Jb, Tuttle

doi:10.1016/0012-1606(87)90230-2

Cited by 12 publications

(5 citation statements)

References 29 publications

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“…Our observation that the factor stimulating somatostatin expression is soluble and secreted from choroid cells is consistent with other studies in which soluble factors have been shown to influence neurotransmitter expression (Patterson and Chun, 1977;Kessler et al, 1984;Fukada, 1985;Zurn and Do, 1988;Denis-Donini, 1989;Iacovitti et al, 1989;Martinou et al, 1989;Nawa and Patterson, 1990;Nawa and Sah, 1990) and differs from membrane-contact-mediated factors that also have been reported (Hawrot, 1980;Black, 1985, 1986;Kessler et al, 1986;Gray and Tuttle, 1987;Wong and Kessler, 1987;Adler et al, 1989). Preliminary charcterization of size suggests that the somatostatin-stimulating activity (SSA) is a macromolecule and not a small metabolite (Zurn and Do, 1988) or a neuropeptide (Denis-Donini, 1989).…”

Section: Discussionsupporting

confidence: 79%

Stimulation of somatostatin expression in developing ciliary ganglion neurons by cells of the choroid layer

Coulombe

Nishi

1991

J. Neurosci.

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An important component of neuronal development is the matching of neurotransmitter expression with the appropriate target cell. We have examined how peptide transmitter expression is controlled in a simple model system, the avian ciliary ganglion (CG). This parasympathetic ganglion contains 2 distinct types of neurons: choroid neurons, which project to vasculature in the eye's choroid layer and use somatostatin as a co-transmitter with ACh, and ciliary neurons, which innervate the ciliary body and iris and use ACh but no known peptide co-transmitter. We have found that the earliest developmental stage in which neurons with somatostatinlike immunoreactivity (SOM-IR) are consistently found in vivo is stage 30 (embryonic day 6.5), a time shortly after the extension of neurites to targets in the eye's choroid layer. In cell culture, CG neurons expressed SOM-IR in co-culture with choroid cells, but not when cultured with striated muscle myotubes or with ganglion non-neuronal cells. No significant differences in neuronal survival or in ChAT activity were observed under these different co-culture conditions, which suggests that somatostatin expression is independently regulated. The stimulation of somatostatin expression was also specific in that other neuropeptides commonly found in autonomic neurons [neuropeptide Y (NPY), substance P (SP), vasoactive intestinal polypeptide (VIP)] were not induced in the presence of choroid cells. The ability to stimulate SOM-IR was not contact dependent because a macromolecule of greater than or equal to 10 kDa in choroid-conditioned medium (ChCM) was found to stimulate somatostatin expression in a dosage-dependent fashion. The somatostatin-stimulating activity induced SOM-IR in more than 90% of CG neurons, as well as in retrogradely labeled ciliary neurons, which would not normally express SOM-IR. Thus, the expression of somatostatin in cultured CG neurons is regulated by a macromolecule produced by cells in the choroid layer, a target normally innervated in vivo by CG neurons expressing somatostatin.

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Section: Discussionsupporting

confidence: 79%

Stimulation of somatostatin expression in developing ciliary ganglion neurons by cells of the choroid layer

Coulombe

Nishi

1991

J. Neurosci.

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“…Newly inserted receptors are apparently directed to the sites of nerve-muscle contact (Role et al, 1985). Contact with the muscle influences neuronal receptor metabolism (Crean et al, 1982;Tuttle, 1983) and neuronal transmitter synthesis Gray and Tuttle, 1987). These in vitro studies support the notion that neuron-target interactions are developmentally important.…”

supporting

confidence: 68%

“…Ciliary ganglia were isolated and cultured as previously described (Tuttle et al, 1980;Gray and Tuttle, 1987). The ganglia were removed by microdissection from chick embryos incubated for 9-10 d, , and placed in sterile, modified Tyrode's saline (in mM: 130 NaCl,3 KCl,20 NaHCO,,4 CaCl,, 1 MgCl,, 12 glucose, 10 HEPES, and 0.5% antibiotic-antimycotic mix; Gibco Laboratories, Grand Island, NY).…”

Section: Methodsmentioning

confidence: 99%

Cultured smooth muscle targets lack survival activity for ciliary ganglion neurons

Creedon

1988

J. Neurosci.

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Cultured neurons require specific trophic agents in order to survive. This dependence is thought to resemble the neuron-target interdependence that develops in vivo during synaptogenesis and neuronal cell death. The notion that neurons in general derive trophic support from their synaptic targets is based primarily on studies of peripheral neurons and motor neurons. To assess the general applicability of this nerve-target relationship, we tested the ability of vascular smooth muscle (VSM) to support dissociated neurons from the chick ciliary ganglion. The ciliary ganglion contains 2 distinct neuronal populations, one of which innervates striated muscle, the other VSM. Striated muscle cocultures are known to support all of the neurons in the ganglion for extended periods. Dissociated neurons were therefore cocultured in microwells containing VSM derived from the rat or chick aorta and from the choroid coat of the chick eye. Surviving neurons were counted after 1, 2, 5, and 7 d. Striated muscle is able to support full neuronal survival in the same assay. However, in no case was VSM capable of contributing to neuronal survival in vitro. The neurons in the VSM cocultures were able to form neurites and make contacts with their putative targets, as confirmed by scanning electron and light microscopy. The presence of viable and differentiated smooth muscle cells was demonstrated in the cultures by transmission electron microscopy and analysis of smooth muscle alpha-actin. The failure of VSM and even the choroid target tissue to support the survival of their innervating neurons suggests that novel mechanisms may operate to provide trophic support for neurons innervating VSM targets.

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“…The molecular mechanism of the signal that directly increases ChAT activity is not known, although both soluble and membrane-bound cholinergic neuronal differentiation factors have been described (5,14,15,21,25,30). Moreover, the nature of the increase itself is not well defined.…”

Section: Discussionmentioning

confidence: 99%

Neural cell adhesion molecule regulates cell contact-mediated changes in choline acetyltransferase activity of embryonic chick sympathetic neurons

Acheson

Rutishauser

1988

The Journal of Cell Biology

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Abstract. El0 chick sympathetic ganglion cells display a cell contact-dependent rise in choline acetyltransferase (CHAT) specific activity over the first several days in culture. This effect can be mimicked by addition of crude membrane fractions prepared from El0 retina and adult chicken brain, but not by those from El0 brain. The effects of both cell-cell and membrane-cell contact are inhibited by the addition of anti-NeAM Fab fragments. The membranes capable of increasing ChAT and those which are ineffective all contain NCAM, however their relative levels of NCAM polysialic acid differ. Whereas membranes with high polysialic acid NCAM are ineffective, selective enzymatic removal of polysialic acid renders them capable of producing an increase in CHAT. The inhibition of NCAM-mediated adhesion produced by Fab fragments can be compensated for by addition of wheat germ agglutinin, but only with membranes whose NCAM has low levels of polysialic acid. Taken together, these data suggest that NCAM can regulate cell contact-mediated increases in ChAT activity. We propose that NCAMmediated adhesion promotes contact between cell membranes to allow the transmission of an otherwise NCAM-independent signal. In addition, NCAM's polysialic acid moiety appears to influence the ability of cells to transmit this signal, even in the presence of an alternative adhesion mechanism.

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[3H]acetylcholine synthesis in cultured ciliary ganglion neurons: Effects of myotube membranes

Cited by 12 publications

References 29 publications

Stimulation of somatostatin expression in developing ciliary ganglion neurons by cells of the choroid layer

Stimulation of somatostatin expression in developing ciliary ganglion neurons by cells of the choroid layer

Cultured smooth muscle targets lack survival activity for ciliary ganglion neurons

Neural cell adhesion molecule regulates cell contact-mediated changes in choline acetyltransferase activity of embryonic chick sympathetic neurons

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