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D L Barker

Publications and source records attributed to D L Barker.

At least 37 records · Page 2Linked to original sources

Nerve growth-promoting factor produced in culture media conditioned by specific CNS tissues of the snail Helisoma.

Medium conditioned by tissue from the CNS of the snail, Helisoma, is capable of promoting neurite outgrowth in isolated neurons from adult central ganglia. The conditioning factor(s) (CF), contained in conditioned medium (CM), is produced only by central ganglionic rings and buccal ganglia and not by other tissues, including hemolymph. CF requires a minimum of 24 h to be produced or released into the medium. At 12 h growth-promoting activity was not detectable. CF binds tightly to the polylysine substratum and its activity is not mimicked by addition of various sera, NGF or fibronectin. CF activity is abolished by chymotrypsin, trypsin or heating to 100 degrees C, but is stable to DNase and RNase treatment. The percentage of cells exhibiting neurite outgrowth is approximately linear with the amount of neural tissue used to condition the medium up to 2 ganglionic rings/ml. Addition of more ganglia fails to stimulate a greater response. This apparent plateau of CM activity appears to be a function of production and/or release of CF, rather than a saturation effect on plated cells, since dose-response curves for dilutions of CM are approximately linear regardless of the number of ganglia used for conditioning. In addition, anisomycin inhibits 35% of CF appearance under conditions of over 90% protein synthesis inhibition in the ganglia used to produce the CM. Under these conditions anisomycin has no apparent effect on the maintenance of electrical excitability. The inhibitor data suggest that 65% of CF is derived from a pre-existing storage pool and that the remainder is synthesized during the 72 h conditioning period.

Amino Acids↗

Dopamine in a molluscan nervous system: synthesis and fluorescence histochemistry.

The nervous system of the pond snail, Helisoma trivolvis, was investigated for its ability to synthesize and accumulate 3H-catecholamines from 3H-tyrosine. 3H-Dopamine, but not 3H-norepinephrine, was synthesized by several ganglia. The highest accumulations were found in the cerebral, pedal, and buccal ganglia. The Falck-Hillarp and glyoxylic acid fluorescence histochemical techniques were applied to the buccal ganglia to visualize dopamine-containing cells. Fluorescing cells were found on both dorsal and ventral sides of the ganglion. Peripheral nerves of the buccal ganglia also displayed catecholamine fluorescence and accumulated 3H-dopamine. However, no 3H-dopamine synthesis occurred in the cerebral-buccal connectives, which connect the buccal ganglia with the rest of the central nervous system. Therefore, we conclude that there is a dopaminergic system intrinsic to the buccal ganglia and their peripheral targets.

Animals↗

Activation by dopamine of patterned motor output from the buccal ganglia of Helisoma trivolvis.

The buccal ganglia of the snail, Helisoma trivolvis, contain an intrinsic system of dopamine-containing neurons (Trimble, Barker, and Bullard, 1983). Dopamine, when bath applied to the isolated buccal ganglia, activates patterned motor output in a dose-dependent fashion. Haloperidol blocks the activating effect of dopamine, but the similar activation evoked by serotonin is not blocked by haloperidol. We suggest that there are two separate mechanisms for activating patterned motor output from the buccal ganglia. One is serotonergic, emanating from identified cerebral ganglion cells (Granzow and Kater, 1977), while the other is dopaminergic, involving neurons intrinsic to the buccal ganglia.

Animals↗

A neurochemical description of the dopaminergic innervation of the stomatogastric ganglion of the spiny lobster.

The spiny lobster stomatogastric ganglion has been shown to be innervated by catecholaminergic processes which derive from cells of large central ganglia (Kushner and Maynard, 1977). Biochemical evidence had indicated that the stomatogastric system synthesizes dopamine and not norepinephrine from tritiated tyrosine (Barker, Kushner, and Hooper, 1979). Studies reported here document that the stomatogastric ganglion itself contains dopamine, as measured with a sensitive endogenous assay. Moreover, the ganglion can synthesize dopamine from tritiated tyrosine or DOPA. Additionally, when incubated in tritiated dopamine, the ganglion takes up dopamine and protects it from degradation; this process is inhibited by cocaine. When incubated with 3H-tyrosine, small but measurable amounts of tritiated dopamine were detected in the medium surrounding the ganglion.

Animals↗

Acetylcholine synthesis and accumulation in the CNS of Drosophila larvae: analysis of shibirets, a mutant with a temperature-sensitive block in synaptic transmission.

A radiochemical method is applied to the study of neurotransmitter metabolism in Drosophila. The larval CNS is a favorable system for analyzing acetylcholine (ACh) metabolism, since the pool of [3H]ACh rapidly reaches a steady state with a high ratio of intracellular [3H]ACh to [3H]choline. A temperature-sensitive paralytic mutant, shibirets, shows reduced [3H]ACh accumulation at the restrictive temperature. This reduction is not the result of decreased synthesis of [3H]ACh, but rather an abnormally rapid rate of release, which is not prevented by blocking tetrodotoxin-sensitive nerve activity.

Acetylcholine↗

Separate factors produced by the CNS of the snail Helisoma stimulate neurite outgrowth and choline metabolism in cultured neurons.

Neurons from the snail Helisoma require a brain-derived factor(s) for neurite outgrowth in both organ and isolated cell culture. This factor is released from the CNS of Helisoma when brains are incubated in defined medium, producing a conditioned medium (CM). In addition to its growth-promoting activity, CM also enhances total uptake of 3H-choline and the incorporation of 3H-choline into specific metabolites: acetylcholine, phosphorylcholine and lipid. This choline metabolism-enhancing factor(s) is distinct and separable from neurite growth-promoting factor: 1. Over 95% of neurite growth-promoting activity can be removed from CM by adsorption to a polylysine surface while there is no loss of choline metabolism-enhancing activity. 2. When central ganglia were treated with anisomycin, a potent inhibitor of molluscan protein synthesis, the choline metabolism-enhancing activity was completely absent from the resulting CM, while the growth promoting activity was reduced by only 35%. These results suggest that the Helisoma CNS produces a variety of trophic factors that are involved in regulating the interaction between neuronal growth and metabolism.

Cells, Cultured↗

Formation of novel central and peripheral connections between molluscan central neurons in organ cultured ganglia.

An in vitro organ culture system for buccal ganglia of the adult snail, Helisoma, is described. The system supports: (1) maintenance of characteristic electrophysiological parameters of identified neurons over seven days of culture; (2) choline metabolism including uptake and synthesis over the same duration; (3) sprouting and growth of neurons in response to axotomy; (4) the formation of novel central electrotonic connections between identified neurons as a result of sprouting and growth. These observations on neuronal growth and the formation of connections are similar to those made with in vivo culture. The use of in vitro culture allows precise manipulations not previously possible. When buccal ganglia are cultured in vitro with the cut distal ends of peripheral nerve trunks held closely apposed, axons of neurons 5R and 5L in the nerve trunks are capable of forming electrotonic connections similar to central connections. The capability of these neurons to form electrotonic connections via their peripheral axons implies that special structures (i.e., central neurites) are not required for the formation of connections; and neither are special environments (i.e., the central neuropile) required for these connections.

Animals↗

Synaptic mechanisms that generate network oscillations in the absence of discrete postsynaptic potentials.

Synaptic mechanisms were examined in the pyloric network of the lobster stomatogastric which generate network oscillations in the absence of discrete postsynaptic potentials (PSPs). In normal saline, the unstimulated pyloric network underwent weak bursting in only a few cells. Stimulation of the input nerve, or bath application of the input neurotransmitter dopamine, produced similar vigorous bursting in many pyloric neurons. In saline-containing tetrodotoxin (TTX) plus dopamine, action potentials and corresponding discrete PSPs were blocked, but the underlying slow wave oscillations in network neurons continued. No oscillations occurred in TTX-saline without dopamine. The generation of these nonspiking network oscillations can be explained by the interaction between two synaptic mechanisms which do not produce discrete PSPs: neurotransmitter activation of bursting pacemaker oscillations in a single network neuron, and graded inhibition between network neurons.

Action Potentials↗

[(3)H]Choline uptake and metabolism in nonsynaptic regions of a crustacean sensory nerve.

The posterior stomach nerve (PSN) is a crustacean sensory nerve containing about 60 cholinergic neurons, which are devoid of synaptic interactions. Kinetic analysis shows that the PSN takes up [(3)H]choline by both low-affinity (K(m) = 163 micron) and high-affinity (Na(¿dependent) (K(m) - 1 micron) processes. The capacity of the high-affinity system is only about 1% that of the low-affinity system. The high-affinity system is not tightly coupled to acetylcholine (ACh) synthesis, and it appears that both ACh and phosphorylcholine are formed from an intracellular pool of choline, which is fed by both uptake systems. There are differences in the rates of [(3)H]choline uptake and (3)H metabolite accumulation between regions of the PSN that contain neuronal cell bodies and those that do not. These differences may arise from differences in the relative proportion of neuronal to nonneuronal tissue in each nerve region.

Acetylcholine↗

Clinical pharmacy services in a hospital-based home care program.

The Hospital-Based Home Care (HBHC) Program at the Veterans Administration Medical Center in San Francisco, California, is a specialized medical service designed to provide comprehensive continuity of care to the veteran patient in his or her own home through the use of a multidisciplinary team approach. Professional health care services are provided by nurses, dieticians, physical therapists, pharmacists, physicians, and social workers. Professional services provided by the clinical pharmacists include: the evaluation of prescribed medication regimen; product identification; patient counseling and education; drug therapy consultant to the HBHC team; and liaison between the HBHC team and outpatient pharmacy services.

California↗

Synthesis of dopamine and octopamine in the crustacean stomatogastric nervous system.

The spiny lobster stomatogastric nervous system synthesizes dopamine and octopamine in vitro from exogenous [3H]tyrosine. Each amine accumulates with a specific distribution among 9 separately analyzed regions within the system. Synthesis of other catecholamines was not observed. [3H]Dopamine is found in nerves, ganglia, and identified commissural ganglion cell bodies in which catecholamine histofluorescence has been demonstrated. The biosynthetic and histochemical data together indicate that dopaminergic cells send axons from the commissural ganglia to the stomatogastric ganglion neuropil along the same pathway followed by fibers that activate the pylroic motor network. The results support the hypothesis that dopamine mediates activation of the pyloric system in vivo, as observed in vitro. [3H]Octopamine accumulates primarily in the commissural and stomatogastric ganglia, where it may modulate neuronal activity, but octopaminergic cells and release sites within the stomatogastric system have not been identified.

Animals↗

Synthesis of octopamine by insect dorsal median unpaired neurons.

Dorsal unpaired median (DUM) neurons of locusts and grasshoppers inhibit the instrinsic rhythm of contraction of metathoracic extensor tibia muscle fibers. This physiological action is mimicked by very low concentrations of octopamine and by higher concentrations of dopamine and noradrenaline. We have examined the synthesis of biogenic amines from tritiated tyrosine by these neurons. DUM cell bodies and a peripheral nerve containing the DUM axon which terminates in the extensor tibia both produced octopamine, but neither noradrenaline nor dopamine synthesis was detected. The observations support the suggestion that the dorsal unpaired median neurons are octopaminergic.

Animals↗

Acetylcholine and lobster sensory neurones.

Experiments are presented in support of the hypothesis that acetylcholine functions as a sensory transmitter in the lobster nervous system.1. Several different peripheral sensory structures incorporate radioactive choline into acetylcholine. The preparation most enriched in sensory as opposed to other nervous elements (the antennular sense organs of the distal outer flagellum) does not incorporate significant amounts of glutamate, tyrosine or tryptophan into any of the other major transmitter candidates.2. There is a parallel between the distribution of the enzyme choline acetyltransferase and the proportion of sensory fibres in nervous tissue from many parts of the lobster nervous system.3. Isolated sensory axons contain at least 500 times as much choline acetyltransferase per cm of axon as do efferent excitatory and inhibitory fibres.4. Abdominal ganglia and root stumps show a decline in the rate of incorporation of choline into acetylcholine 2 to 8 weeks after severing the first and second roots bilaterally (leaving the connectives and third roots intact). Extracts of the root stumps exhibit a significantly lower level of choline acetyltransferase 2 weeks after this operation.5. Curare and atropine partially block an identified sensory synapse in the lobster abdominal ganglion.

Acetylcholine↗