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J A Freeman

Publications and source records attributed to J A Freeman.

At least 127 records · Page 7Linked to original sources
PubMed1981

Analysis of alpha-bungarotoxin binding in the goldfish central nervous system.

We report here the equilibrium, kinetic, and pharmacological analysis of alpha-125I-bungarotoxin (alpha-125I-Bgt) binding to a Triton X-100-solubilized goldfish brain synaptosomal fraction. In addition, a refined analysis of equilibrium binding to a particulate synaptosomal fraction is presented. Equilibrium binding from both particulate and soluble fractions revealed an apparent heterogeneity of binding sites. Kinetic analysis of the soluble receptor revealed linear association kinetics and nonlinear dissociation kinetics. The dissociation curve suggested the presence of at least two rate constants. Potential sources of the binding heterogeneity found in both the equilibrium binding and dissociation kinetics experiments are (1) multiple receptor species, (2) multiple ligand species, and (3) different, possibly interconvertible, states of a single receptor type. No evidence for the first two alternatives was found. Support for the third alternative was obtained by observing the effect of cholinergic ligands on alpha-125I-Bgt dissociation. Carbamylcholine and d-tubocurarine increased the apparent proportion of rapidly dissociating sites, suggesting that the two binding affinities can be interconverted and may arise from a single receptor type. Evidence concerning the identity of the alpha-Bgt binding protein as a nicotinic acetylcholine receptor is discussed.

Animals↗
PubMed1980

Localization of alpha-bungarotoxin binding sites to the goldfish retinotectal projection.

The optic tectum of the goldfish Carassius auratus is a rich source of alpha-bungarotoxin (alpha-Btx) binding protein. In order to determine whether some fraction of these receptors is present at retinotectal synapses, we have compared the histological distribution of receptors revealed by the use of [125I]alpha-Btx radioautography to the distribution of optic nerve terminals revealed by the use of cobalt and horseradish peroxidase (HRP) techniques. The majority of alpha-Btx binding is concentrated in those tectal layers containing primary retinotectal synapses. The same layers contain high concentrations of acetylcholinesterase (AChE), revealed histochemically. Following enucleation of one eye, there is a loss of alpha-Btx binding in the contralateral tectum, observed both by radioautography and by a quantitative binding assay of alpha-Btx binding. Approximately 40% of the alpha-Btx binding sites are lost within two weeks following enucleation. By contrast, no significant change in AChE activity could be demonstrated up to 6 months following enucleation. These results are discussed in light of recent studies which show that the alpha-Btx binding protein and the nicotinic acetylcholine receptor are probably identical in goldfish tectum. We conclude that the 3 main classes of retinal ganglion cells projecting to the goldfish tectum are nicotinic cholinergic and that little or no postdenervation hypersensitivity due to receptor proliferation occurs in tectal neurons following denervation of the retinal input.

Acetylcholine↗
PubMed1980

Electrophysiologic evidence that retinotectal synaptic transmission in the goldfish is nicotinic cholinergic.

A previous study identified, by conduction velocity following optic nerve shock, 3 classes of retinal fibers which project to 3 distinct laminae of the goldfish optic tectum. In the present study, the effect of various pharmacological agents on the synaptic efficacy of each of the 3 classes of retinal fibers was assessed by the use of current source-density analysis. All 3 classes of optic fibers appear to be nicotinic cholinergic. Six different nicotinic antagonists were tested. All 6 were effective in decrementing the responses of all 3 classes to a criterion level: alpha-bungarotoxin (10-8 M), alloferin (10-5 M), curare (10-4 M), metocurine (10-4 M), hexamethonium (10-4 M) and gallamine (10-3 M). Atropine, a muscarinic antagonist, had only a slight effect even at 10-3 M. Five nicotinic agonists tested also decremented synaptic responses: nicotine (10-5 M), carbamylcholine (10-4 M), acetylcholine (10-4 M), succinyl choline (10-4 M) and decamethonium (10-3 M), presumably via cellular depolarization and receptor desensitization. Two inhibitors of acetylcholinesterase prolonged the response at 10-4 M and decremented it as well at 10-3 M. Hemicholinium 3, an inhibitor of the high affinity uptake of choline, produced a gradual activity-dependent decrement in the responses. Beta-bungarotoxin, a presynaptically-acting toxin, abolished not only the postsynaptic components but also the presynaptic components at 10-6 M. In all other cases the presynaptic deflections were generally unaffected, and with the exception of the toxins, a return to at least 90% of the control value was achieved. In contrast, GABA (10-3 M) and bicuculine (10-4 M) both produced no discernible effect on the 3 classes of responses, and glutamate (10-3 M) produced only a slight decrement, which probably represents a non-specific effect.

Animals↗
PubMed1980

Magnetic field of a nerve impulse: first measurements.

The magnetic field of the action potential from an isolated frog sciatic nerve was measured by a SQUID magnetometer with a novel room-temperature pickup coil. The 1.2 x 10(-10) tesla field was measured 1.3 millimeters from the nerve with a signal-to-noise ratio of 40 to 1.

Action Potentials↗
PubMed1979

Characterization of the nicotinic acetylcholine receptor isolated from goldfish brain.

We have studied the binding of alpha-bungarotoxin to a particulate fraction of goldfish brain enriched in synaptosomes. The binding is specific and saturable and exhibits the pharmacological properties of a nicotinic cholinergic receptor. Equilibrium binding measurements yield a single dissociation constant (KD) of 0.92 nM. Kinetic analysis revealed one association rate constant and two dissociation rate constants. Dissociation constants calculated from kinetic measurements were 1.9 nM and 12.5 pM. The toxin . receptor complex is readily solubilized in nonionic detergent. The isoelectric point of the toxin . receptor complex was found to be 5.00 +/- 0.01. Sedimentation velocity analysis in sucrose/H2O and sucrose/D2O gradients in conjunction with Sepharose 4B chromatography and diffusion experiments yielded a sedimentation constant of 11.45, a partial specific volume of 0.79 cm3/g for the toxin . receptor . detergent complex, and a molecular weight of approximately 340,000 for the toxin . receptor complex.

Animals↗
PubMed1979

Responses to stimulation of marginal fibers in the teleostean optic tectum.

The marginal fibers (mf) constitute a major fiber component of the teleostean optic tectum, and this report deals with the physiological properties of these fibers and their postsynaptic elements. The mf are unmyelinated axons which originate at the torus longitudinalis and run lateralwards, parallel to one another, along the tectum's most superficial layer (stratum marginale). Here they synapse upon the dendritic arborizations of the pyramidal (p) neurons. These arborizations originate from a single apical dendritic shaft which, near the soma, receives retinofugal axon terminals. The p-neurons also have a basal dendritic shaft and a descending axon, both of which branch out horizontally at the stratum griseum centrale (SGC). The mf were stimulated through surface microelectrodes, and field potentials were recorded on-beam throughout the tectal thickness by means of micropipettes. The mf action potential (M-spike) may show two negative subpeaks which propagate at 0.20 and 0.16 m/s. Its refractory period is followed by a period of increased amplitude and decreased latency. The M-spike is followed by a series of slow waves, namely: (a) The S-wave, which probably represents the monosynaptic depolarization of the p-neuron's apical arbor; (b) The N-wave which possibly represents an active current sink at the point where these dendritic arborizations merge into the apical dendritic shaft; (c) The D-wave, recorded at the SGC, which possibly represents the activation of the p-neuron's axon and terminals (and perhpas also basal dendritic shaft and branches); and (d) the L-wave, which might represent a later depolarization of the p-neuron's apical arborizations. Morphologically and electrophysiologically, there are several similarities between the tectal mf/p-neuron system and the cerebellar parallel fiber/Purkinje cell system.

Animals↗
PubMed1979

Pulmonary hemodynamics in systemic hypertension. Long-term effect of minoxidil.

To assess the effect of minoxidil on pulmonary hemodynamics, we performed cardiac catheterization on ten patients, prior to the administration of minoxidil, at six months after its addition to their existing drug regimens, and on seven patients following 12 or more months of minoxidil therapy. Systemic blood pressure and systemic vascular resistance were significantly reduced at six months and following 12 or more months of minoxidil therapy. Before receiving minoxidil, nine of ten patients had elevated pulmonary vascular resistance. There were no significant changes in pulmonary vascular resistance following the initiation of minoxidil. The data suggest that pulmonary hypertension is common in patients with long-standing poorly controlled systemic hypertension, and that minoxidil, in doses sufficient to significantly reduce systemic blood pressure, does not cause or aggravate pulmonary hypertension.

Adult↗
PubMed1977

Possible regulatory function of acetylcholine receptor in maintenance of retinotectal synapses.

alpha-Neurotoxins bind to cholinergic receptor, block transmission, and induce sprouting of retinal terminals in the toad tectum. New connections retain an orderliness that suggests a selective affinity between presynaptic terminals. The results suggest that postsynaptic cells exert a control, associated with receptors, on the growth of presynaptic terminals and on the maintenance of their synaptic connections.

Acetylcholine↗
PubMed1977

Bilateral ganglion cell branches in the normal rat: a demonstration with electrophysiological collision and cobalt tracing methods.

The application of microelectrode recording and anatomical tracing methods to the subcortical optic projections of the normal rat shows that the uncrossed pathway from the retina of this mammal is substantially composed of branches of crossed axons. In the recording experiments, paired stimuli to the optic tracts produces an attenuated antidromic response in the optic nerve which is best explained by the collision of impulses which travel in branches of the same parent axon. Cobalt injection of one optic tract results in retrograde filling of axons in the entire contralateral optic nerve and filling of axons in restricted regions of the ipsilateral optic nerve. This procedure also results in the filling of axons in the opposite optic tract. The locations of the filled axons in the opposite tract correspond to the positions of crossed and uncrossed projections from the temporal retinae: ventrolateral and dorsomedial, respectively. The position of the temporal retinal projections in both optic tracts was determined by applying silver degeneration methods after small lesions of the retina.

Animals↗