Synapse Formation and Neurotrophic Effects on Muscle Cell Lines

Kidokoro, Yoshiaki; Heinemann, Sabine; Schubert, D; Brandt, B. L.; Klier, F. George

doi:10.1101/sqb.1976.040.01.036

Cited by 43 publications

(33 citation statements)

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“…Cultures of striated muscle cells were prepared by dissociation of cells from the hindlimbs of 18-to 21-day-old fetal C57BL/6N mice, 18-to 21-day-old fetal or newborn Wistar rats, or 10-day-old chick embryos (White Leghorn) as described previously (6). Each collagen-coated 35 mm plastic culture dish usually was inoculated with 0.3 to 1.0 X 106 cells.…”

Section: Methodsmentioning

confidence: 99%

“…In addition, Kidokoro and Heinemann have reported that neurons in explants of chick embryo spinal cord form synapses with clonal rat muscle cells (13). Under special conditions striated muscle cells have been shown to be innervated by functionally inappropriate motor neurons (14,15), parasympathetic neurons (16), sympathetic ganglion neurons (17), cerebral cortex neurons (18), and neurons from the retina (D. Puro, F. DeMello, and M. Nirenberg, unpublished data). These results provide additional evidence there are few or no restrictions with respect to the type of cholinergic neuron that can synapse with a striated muscle cell.…”

Section: Methodsmentioning

confidence: 99%

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On the specificity of synapse formation.

Puro¹,

Nirenberg²

1976

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

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Clonal neuroblastoma X glioma hybrid cells form synapses with striated muscle cells from different muscles and from different organisms, such as chick, mouse, and rat. Under appropriate conditions 65-80% of the muscle cells exhibited syna tic responses. The results indicate that most striated muscle cells have the same specificity for synapse formation and suggest that synaptogenesis is not dependent on interactions between complementary molecules on neuron and muscle cells that code for different synaptic connections.The problem of how neurons form synaptic connections and assemble into neural circuits with precision remains one of the most challenging issues in neurobiology. In essence most of the hypotheses which relate to this problem can be divided into two categories: either that cells which form synapses possess different kinds of cell recognition molecules which must interact with specificity prior to synapse formation; i.e., a code for matching one cell with another which leads to the formation of different kinds of synaptic connections (1); or the alternative hypothesis which is not based on a cell recognition code, that modification of synaptic connections dependent upon transmission across the synapse determines the final synaptic organization (2, 3).To understand the molecular nature of the factors that determine the specificity of synapse formation it would be advantageous to have homogenous populations of neurons that form synapses. Since normal neurons do not divide, clonal lines of neuroblastoma cells and somatic cell hybrids derived from neuroblastoma cells were generated and characterized with respect to receptors, neurotransmitters, action potential ionophores, and other properties which are required for synaptic communication (4,5 METHODS AND MATERIALS Cell Culture. Neuroblastoma X glioma NG108-15 hybrid cells were derived (B. Hamprecht, M. Nirenberg, and T. Amano, submitted for publication) by fusion of mouse neuroblastoma clone N18TG-2, resistant to 6-thioguanine (5), with rat glioma clone C6BU-1, resistant to 5-bromodeoxyuridine (7). NG108-15 cells were cultured at 360 in Dulbecco-Vogt modification of Eagle's minimal essential medium (DMEM; Gibco Catalog no. H-21), 10% fetal bovine serum, 100MAM hypoxanthine, 1 MAM aminopterin, and 16MAM thymidine in a humidified atmosphere of 10% C02-90% air. The hybrid cells were shifted to a more differentiated state by culturing them for 1-3 weeks in the presence of I mM N6,02'-dibutyryl-adenosine 3':5'-cyclic monophosphate (dibutyryl cAMP) (B. Hamprecht, M. Nirenberg, and T. Amano, submitted for publication) purified as described previously (6). The NG108-15 cells which were used in this study had been subcultured 15 to 20 times.Cultures of striated muscle cells were prepared by dissociation of cells from the hindlimbs of 18-to 21-day-old fetal C57BL/6N mice, 18-to 21-day-old fetal or newborn Wistar rats, or 10-day-old chick embryos (White Leghorn) as described previously (6). Each collagen-coated 35 mm plastic culture dish usually was inocul...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

On the specificity of synapse formation.

Puro¹,

Nirenberg²

1976

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

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“…For instance, the Na channels in the heart of several species, which are otherwise very similar to those in nerve and skeletal muscle, typically show a high resistance to TTX (i.e., Brown et al, 1981;Cohen et al, 198 1). Na channels with low affinity for TTX have also been described in fetal mammalian muscle and in denervated adult muscle (Harris and Marshall, 1973;Kidokoro et al, 1975;Pappone, 1980). In muscle, the presence of Na channels with low affinity for TTX is associated with muscle development and innervation (Harris and Thesleff, 1971;Harris and Mar-shall, 1973;Sherman et al, 1983).…”

Section: Ttx-resistant Channels In Pc12 Cellsmentioning

confidence: 99%

Nerve growth factor increases the number of functional Na channels and induces TTX-resistant Na channels in PC12 pheochromocytoma cells

Rudy¹,

Kirschenbaum²,

Rukenstein³

et al. 1987

J. Neurosci.

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The PC12 clone is a line of rat pheochromocytoma cells that undergoes neuronal differentiation in the presence of NGF protein. In the absence of NGF, PC12 cells are electrically inexcitable, while after several weeks of NGF treatment they develope Na+ action potentials. Past estimates made by measuring binding of 3H-saxitoxin (STX) indicate that NGF treatment brings about a large increase in Na channel density that is of sufficient magnitude to account for the induction of excitability. We have now used 22Na uptake to measure the Na permeability of PC12 cells before and after long-term NGF treatment. Treatment with NGF does not change the resting Na+ permeability. The alkaloid toxins veratridine and batrachotoxin (BTX) and scorpion toxin were used to activate Na channels. Such studies demonstrate that these toxins induce TTX-sensitive Na uptake in both NGF-treated and untreated cells and reveal differences in functional Na channel numbers per cell and per unit of membrane area that are similar to those found in the STX binding studies. On the other hand, affinities for drugs that activate these channels are not affected by NGF treatment. We also find that NGF-treated PC12 cells contain a population of Na channels with low affinity for TTX. These channels account for 5–20% of total BTX or veratridine-stimulated flux. Thus, NGF has 2 effects regarding the Na channels of PC12 cells: it increases the number of functional Na channels that otherwise behave similarly to those present before NGF treatment, and it induces the presence of TTX-resistant Na channels. These findings indicate that the PC12 model system may serve to study the developmental regulation of Na channel expression and properties.

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“…During days 6-26, the mean input resistances ofhypothyroid and hyperthyroid rats were 102-131 % and 79-120 % of that in the normal rats of the same age, respectively. This could have caused at most 31 % higher sensitivity to ACh for hypothyroid rats and 21 % lower sensitivity for hyperthyroid rats than normal rats, if sensitivities were normalized on the assumption that input resistance and ACh-induced depolarization were in proportion (Kidokoro et al 1976). Thus, the difference in extrajunctional ACh sensitivities among the normal, hypo-and hyperthyroid animals are presumably largely due to differences in ACh receptor densities on the muscle fibres.…”

Section: Reduction Of Extrasynaptic Ach Sensitivity During Developmentmentioning

confidence: 99%

“…However, to evaluate the differences of ACh receptor densities on these muscle fibres from the ACh sensitivities above, the input resistance ofthe muscle fibres should be taken into account (Kidokoro, Heinemann, Schubert, Brandt & Klier, 1976).…”

Section: Reduction Of Extrasynaptic Ach Sensitivity During Developmentmentioning

confidence: 99%

Altered developmental changes of neuromuscular junction in hypo‐ and hyperthyroid rats

Kawakami

Obata

1982

The Journal of Physiology

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SUMMARY1. Effects of thyroid hormone on the development of neuromuscular junctions (n.m.j.s.) were investigated electrophysiologically in the diaphragms (sternal region) of normal, hypo-and hyperthyroid rats from the age of birth (day 0) to day 35.2. Hypothyroidism in new-born rats was induced either by daily administration of propylthiouracil to mothers or by subcutaneous injection of 150 1sCi l311 on day 1. Hyperthyroidism was induced by daily injection of thyroxine.3. In normal rats up to day 10, muscle fibres were innervated polyneuronally. By day 20, multiple innervation was eliminated and muscle fibres received only a single input. In hypothyroid rats elimination of polyneuronal innervation was retarded by 5-8 days, while in hyperthyroid rats the elimination was accelerated by 2-3 days.4. The frequency of miniature end-plate potentials (m.e.p.p.s) in normal rats increased from one per 40 sec on days 0-5 to 1/sec on days 25-35. The m.e.p.p. frequency in hypothyroid rats was 25-65 % of that in normal rats of the same age.In hyperthyroid rats the m.e.p.p. frequency was normal up to day 18 but subnormal afterwards. The duration ofm.e.p.p. measured on day 22-23 was slower in hypothyroid rats and faster in hyperthyroid rats, relative to m.e.p.ps in normal rats.5. The sensitivity to acetylcholine (ACh) at extrajunctional regions in normal rats was about 100 mV/nC at birth and declined to 1 mV/nC by day 26. In hypothyroid rats, the ACh sensitivity was as high as 30 mV/nC on day 26; in hyperthyroid rats, ACh sensitivity on day 26 was undetectable.6. With pairs of nerve stimuli (applied at a 50 msec interval), the second end-plate potential was facilitated until day 10 and depressed after day 16 in normal rats. This shift from facilitation to depression during development was not altered in either hypoor hyperthyroid rats.7. It is concluded that the lack and excess of thyroid hormone retards and facilitates the development of n.m.j.s. respectively. Possible mechanisms for this altered development are discussed.

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Synapse Formation and Neurotrophic Effects on Muscle Cell Lines

Cited by 43 publications

References 0 publications

On the specificity of synapse formation.

On the specificity of synapse formation.

Nerve growth factor increases the number of functional Na channels and induces TTX-resistant Na channels in PC12 pheochromocytoma cells

Altered developmental changes of neuromuscular junction in hypo‐ and hyperthyroid rats

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