Intracellular recordings from ganglia of the thoracic sympathetic chain of the guinea‐pig

Blackman, J. G.; Purves, Robert D.

doi:10.1113/jphysiol.1969.sp008858

Cited by 123 publications

(74 citation statements)

References 24 publications

(32 reference statements)

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“…The latter value is similar to that of sympathetic neurons in situ (24,25) and cultured in vitro (8). The electrical properties and chemosensitivity of NX-31 hybrid cells are described in detail elsewhere (26).…”

Section: For Methods) the Resultssupporting

confidence: 66%

Neuronal properties of hybrid neuroblastoma X sympathetic ganglion cells.

Greene¹,

Shain²,

Chalazonitis³

et al. 1975

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

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The neuroblastoma parent was N18TG2 (1, 2). This cell line is resistant to 1 X 10-4 M 6-thioguanine and lacks hypoxanthine phosphoribosyltransferase (HPRTase) activity. The cells do not multiply in media supplemented with 1 X 10-4 M hypoxanthine, 1 X 10-6 M aminopterin, and 1.2 X 10-5 M thymidine (HAT). The N18TG2 cells were grown in the presence of 1 X 10-4 M 6-thioguanine for at least four generations prior to use.Primary cells were obtained from superior cervical and paravertebral sympathetic ganglia dissected from 12-to 14-day-old mouse embryos (C57BL/6) and dissociated with trypsin as described (7). Single cells were suspended in complete F-14 supplemented with 10 units/ml of 2.5S nerve growth factor. Cells were seeded into 100-mm plastic tissue culture dishes (Falcon, cat. no. 3003) and incubated for 4 hr at 37'. Attached cells, which consisted predominately of fibroblast-like cells, were discarded or were used for karyotype analysis. Floating cells and cells not firmly attached to the substratum were harvested by gentle washing and used immediately for hybridization. Aliquots of the cell suspension enriched with respect to neurons were seeded in separate dishes; after 36 hr 95% of the cells that attached resembled neurons in morphology.Hybridization. Trypsinized neuroblastoma cells and normal cells from sympathetic ganglia were fused in suspension for 2 hr at 370 in 1 ml of F-14 supplemented with 10 units of nerve growth factor per ml in the absence of serum. The cell suspension contained 500 hemaglutinating units of Sendai virus inactivated with f,-propriolactone, 5 X 105 N18TG2 neuroblastoma cells, and 2.5 X 105 cells from sympathetic ganglia (the fraction enriched with neurons). The cells were gently resuspended every 20 min during incubation. After incubation, cells were suspended again and were plated into 100-mm plastic culture dishes in complete F-14 medium supplemented with 10 units of nerve growth factor per ml at concentrations of 0.25, 1.25, and 5 X 104 cells per dish. After two days of incubation, the medium was changed to complete medium plus HAT. After 15-30 days in the selective medium, discrete colonies of greater than 100 cells were observed. The medium was removed from the plates and the colonies were individually isolated with cloning cylinders and harvested with the aid of glass pipettes with fine tips. After the cell lines were isolated and established in culture, the growth medium was changed to DMEM supplemented with 5% fetal bovine serum and HAT. Characterization of Cell Lines. Karyotype analysis was performed by counting total numbers of chromosomes and

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Section: For Methods) the Resultssupporting

confidence: 66%

Neuronal properties of hybrid neuroblastoma X sympathetic ganglion cells.

Greene¹,

Shain²,

Chalazonitis³

et al. 1975

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

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“…Two factors may be considered as possible explanations for the above discrepancy: (1) the deviation from the expected amplitude distribution may be due to non-linear summation of the unit potentials making up the i.p.s.p. (del Castillo &E Katz, 1954a;Martin, 1955;Kuno &k Miyahara, 1969) or (2) the release probability of individual quanta of inhibitory transmitter may be too high to be approximated by the Poisson equation (del Castillo & Katz, 1954a;Kuno, 1964a;Blackman & Purves, 1969;Johnson & Wernig, 1971). These two possibilities were tested by the relationship plotted in Fig.…”

Section: Resultsmentioning

confidence: 99%

Quantal components of the inhibitory synaptic potential in spinal motoneurones of the cat

Kuno¹,

Weakly²

1972

The Journal of Physiology

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1. Monosynaptic i.p.s.p.s were produced in spinal motoneurones of the cat by stimulation of a pool of interneurones following chronic degeneration of descending tracts and primary afferent fibres in the lumbosacral cord. 2. Monosynaptic i.p.s.p.s so evoked by supramaximal stimuli often showed a fluctuation in amplitude with occasional failures of response. 3. When two successive stimuli were applied at a short interval, the mean amplitude of the second i.p.s.p.s was greater than that of the first. This facilitation was associated with a decrease in the number of failures, a decrease in the coefficient of variation of the amplitude distribution and an increase in the probability of occurrences of large i.p.s.p.s. 4. A statistical analysis of the i.p.s.p. amplitude fluctuation showed that the monosynaptic i.p.s.p. is composed of discrete unit potentials evoked with a certain probability in a manner described by a binomial law. 5. The application of strychnine decreased the mean amplitude of i.p.s.p.s with little change in the coefficient of variation of the i.p.s.p. amplitude distribution. 6. It is concluded that the release of inhibitory transmitter occurs in quantal steps and that strychnine blocks primarily the post‐synaptic receptors for the inhibitory transmitter.

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“…Other cells were also impaled during these experiments which failed to respond to direct stimulation. These may have been Schwann cells or the glial cells surrounding ganglion cell soma (Blackman & Purves, 1969).…”

Section: Resultsmentioning

confidence: 99%

Effect of iontophoretic application of cholinergic agonists and their antagonists to guinea‐pig pelvic ganglia

Holman

Muir

Szurszewski

et al. 1971

British J Pharmacology

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Summary1. The electrical activity of guinea-pig pelvic ganglion cells following iontophoretically applied cholinergic drugs, alone and during orthodromic nerve stimulation via the hypogastric nerve, has been recorded intracellularly. 2. lontophoretic application of nicotine (Nic) and acetylcholine (ACh) reduced membrane resistance and caused a depolarization which in approximately 80%/0 of cells led to the firing of action potentials. In the remainder, depolarization was unaccompanied by firing. 3. lontophoretic application of Nic and ACh reduced or abolished the amplitude of successively evoked orthodromic responses. 4. ACh-induced depolarization, unlike that caused by tetanic stimulation, was not followed by a subsequent increase in the frequency of synaptic potentials. 5. Di-hydroperythroidine (DH/E) and atropine (Atr) inhibited the response to both orthodromic stimulation and iontophoretic application of Nic and ACh. 6. There was no evidence for the existence of muscarinic receptors in guineapig pelvic ganglia. Iontophoretic application of muscarinic agonists alone and after tetanic stimulation of the hypogastric nerve produced no significant depolarization of the ganglion cell membrane.

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Intracellular recordings from ganglia of the thoracic sympathetic chain of the guinea‐pig

Cited by 123 publications

References 24 publications

Neuronal properties of hybrid neuroblastoma X sympathetic ganglion cells.

Neuronal properties of hybrid neuroblastoma X sympathetic ganglion cells.

Quantal components of the inhibitory synaptic potential in spinal motoneurones of the cat

Effect of iontophoretic application of cholinergic agonists and their antagonists to guinea‐pig pelvic ganglia

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