The influence of muscle-conditioned medium on cholinergic maturation in spinal cord cell cultures

Brookes, Neville; Burt, David R.; Goldberg, Alan M.; Bierkamper, George G.

doi:10.1016/0006-8993(80)90994-4

Cited by 32 publications

(16 citation statements)

References 24 publications

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“…We also determined ChAT activity because it is a reliable marker for motor neurons, and has thus been used to follow motor neuron behavior both during spinal cell fractionation ( Sehnaar and Schaffner, 1981), and during culture of mixed spinal cells (see e.g., Giller et al, 1977;Brookes et aL, 1980;Kaufman et aL, 1985) or in motor neuron-enriched spinal cultures (Flanigan et aL, 1985).…”

Section: Discussionmentioning

confidence: 99%

“…For example it has been shown that cells dissociated from early embryonie chick, rat, or mouse spinal cords can be maintained in culture, and that media taken from cultured muscle cells or soluble extracts of muscle tissue increased the Ievels of cell survival, neurite outgrowth, and/or transmitter synthesis (see, e.g., Giller et aL, 1977;Bennett et aL, 1980;Brookes et aL, 1980;Godfrey et aL, 1980;Henderson et al, 1981;Longo et aL, 1982;Tanaka and Obata, 1982;Tanaka and Obata, 1983;Smith and Appel, 1983;Kaufman et al, 1985;Smith et aL, 1985). The fact, however, that motor neurons were not separated from other spinal cells makes the interpretation of these results ambiguous because it is not possible to decide whether the observed effects result from a direct action on the neurons in question or an indirect effect via other neuronal or nonneuronal cells (see Fukada, 1980).…”

Section: Introductionmentioning

confidence: 99%

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Muscle-derived factors that support survival and promote fiber outgrowth from embryonic chick spinal motor neurons in culture

Dohrmann

Edgar

Sendtner

et al. 1986

Developmental Biology

156

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The purpose of the experiments reported is to provide an unambiguous demonstration that embryonie skeletal muscle contains factors that act directly on embryonie spinal motor neurons both to support their survival and to stimulate the outgrowth of neurites. Cells of lumbar and brachial ventral spinal cords from 6-day-old chick embryos were separated by centrifugation in a two-step metrizamide gradient, and a motor neuron enriched fraction was obtained. Motor neurons were identified by retrogradely labeling with rhodamine isothiocyanate, and were enriched fourfold in the motor neuron fraction relative to unfractionated cells. In culture, the isolated motor neurons died within 3-4 days unless they were supplemented with embryonie chick skeletal muscle extract. Two functionally distinct entities separable by ammonium sulfate precipitation were responsible for the effects of muscle extracts on motor neurons. The 0-25% ammonium sulfate precipitate contained molecules that alone bad no effect on neuronal survival but when bound to polyornithine-coated culture substrata, stimulated neurite outgrowth and potentiated the survival activity present in muscle. Mostofthis activity was due to a laminin-like molecule being immunoprecipitated with antisera against laminin, and immunoblotting demonstrated the presence of both the A and B chains of laminin. A long-term survival activity resided in the 25-70% ammonium sulfate fraction, and its apparent total and specific activities were strongly dependent on the culture substrate. In contrast to the motor neurons, the cells from the other metrizamide fraction (including neuronal cells) could be kept in culture for a prolonged time without addition of exogenous factor(s).

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Muscle-derived factors that support survival and promote fiber outgrowth from embryonic chick spinal motor neurons in culture

Dohrmann

Edgar

Sendtner

et al. 1986

Developmental Biology

156

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“…12. Cultures were obtained from dissociated, arachnoid-free ventral spinal cord tissue prepared from rat embryos in the 14th to 15th day of The use of DNA-mediated gene transfer techniques (1) to identify the origin of transforming sequences in malignant cells has provided evidence that changes in genes in the normal cell can give rise to activated forms with oncogenic potency [(2-5); see (6) for review]. This emerging class of cellular transforming sequences has been detected by a biological assay in which mouse NIH 3T3 cells incubated with DNA from tumor cell lines of different tissue origins acquire an altered morphology and the ability to grow in soft agar.…”

Section: Morphologymentioning

confidence: 99%

“…Coculture of skeletal myotubes with spinal cord cells enhances neuronal survival, promotes outgrowth of neurites, and increases the activity of choline acetyltransferase (4). Media conditioned with myotubes can reproduce some of the effects of coculturing (5)(6)(7)(8), which indicates the existence of diffusible trophic factors in vitro but which may be of limited value for the complete characterization of these factors since only small quantities can be obtained from monolayers of cultured cells (9). Direct extraction of such factors from skeletal muscle tissue would substantially increase the amount of material available for purification and provide a direct method for assaying the role of muscle-derived factors in the survival of motoneurons during development and the sprouting of motoneurons in the denervated adult.…”

mentioning

confidence: 99%

Extracts of Skeletal Muscle Increase Neurite Outgrowth and Cholinergic Activity of Fetal Rat Spinal Motor Neurons

Smith

Appel

1983

Science

132

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A soluble extract of rat skeletal muscle increased neurite outgrowth and cholinergic activity of dissociated ventral spinal neurons in culture. The effects were concentration-dependent, saturable, and labile in the presence of heat or trypsin. The morphological enhancement was produced only by skeletal muscle extract and decreased with developmental age, whereas the cholinergic enhancement was produced by extracts of cerebral cortex and cardiac and skeletal muscle and did not change with age. These changes were specific for ventral cord neurons, but no species specificity was observed with respect to the muscle source or the neuronal target.

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“…More recent experiments. in vitro have pointed to roles for muscle-derived factors in enhancing neurite extension (9), transmitter synthesis (10,11), and cell survival (12) of spinal neurons.…”

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confidence: 99%

Neurite outgrowth from embryonic chicken spinal neurons is promoted by media conditioned by muscle cells.

Henderson¹,

Huchet²,

Changeux³

1981

Proc. Natl. Acad. Sci. U.S.A.

111

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The effect of media conditioned by muscle cells on the development in vitro ofchicken spinal neurons was studied. Neural tube cells of 4.5-day chicken embryos were dissociated after trypsinization and cultured in serum-free minimum essential medium conditioned for 4 days over cultures of fused chicken myotubes. After 20 hr in conditioned medium (protein concentration, 10-50 ftg/ml), about 50% of surviving cells had extended neurites, whereas in cultures in nonconditioned medium this value was about 10%. The active factor(s) in conditioned medium is macromolecular and its activity was completely destroyed by incubation with trypsin. Concentrated samples ofconditioned medium were analyzed by gel filtration on columns of Sepharose CL-6B. The activity was recovered in peaks with apparent molecular weights of 40,000 and 500,000 and at the exclusion volume of the column. Media conditioned by chicken liver and skin cells also contained neurite-promoting activity but at lower levels. No activity was detected in Nerve Growth Factor, insulin, fetal calf serum, or horse serum or in media conditioned by chicken lung, chicken heart, or C6 glioma cells.During the phase of embryogenesis leading up to the formation of a given synapse, a complex set of interactions between the nerve cell and its potential target is brought into play (1, 2). These interactions may be qualified as "anterograde," such as for example the effect of the nerve on the distribution and metabolism of receptors in the target cell membrane, or, conversely, as "retrograde" (3). It has been postulated for many systems (3-5) that the target cell liberates soluble retrograde factors upon which the neuron depends for its survival and morphological and biochemical differentiation.The best candidate for such a role is the nerve growth factor (NGF) (6), but there is indirect evidence that, in the embryonic spinal cord, motoneurons destined to innervate skeletal muscle may be affected by comparable retrograde factors liberated by their target tissues. Results obtained in vivo, such as the enhancement of naturally occurring cell death by limb extirpation in the chicken embryo (7) and the rescue of some motoneurons by the graft of a supernumerary limb (8), suggested that these factors might be necessary for cell survival. More recent experiments. in vitro have pointed to roles for muscle-derived factors in enhancing neurite extension (9), transmitter synthesis (10, 11), and cell survival (12) of spinal neurons.In this paper, we report the effects of media conditioned by skeletal muscle cells upon the development of dissociated cell cultures ofembryonic chicken spinal cord. We have established a rapid quantitative assay for factors affecting one crucial aspect of the development of spinal neurons: neurite extension. Investigations using this assay have provided a preliminary characterization of the substance(s) responsible for the growth-promoting effect of muscle-conditioned medium. MATERIALS AND METHODSSpinal Cord Cultures. Neural tubes were dissected from 4....

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The influence of muscle-conditioned medium on cholinergic maturation in spinal cord cell cultures

Cited by 32 publications

References 24 publications

Muscle-derived factors that support survival and promote fiber outgrowth from embryonic chick spinal motor neurons in culture

Muscle-derived factors that support survival and promote fiber outgrowth from embryonic chick spinal motor neurons in culture

Extracts of Skeletal Muscle Increase Neurite Outgrowth and Cholinergic Activity of Fetal Rat Spinal Motor Neurons

Neurite outgrowth from embryonic chicken spinal neurons is promoted by media conditioned by muscle cells.

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