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H T Cline

Publications and source records attributed to H T Cline.

43 records · Page 3Linked to original sources

N-methyl-D-aspartate receptor antagonist desegregates eye-specific stripes.

The optic tecta of surgically produced three-eyed tadpoles were chronically exposed to the N-methyl-D-aspartate (NMDA) receptor antagonist aminophosphonovaleric acid (APV), or to NMDA itself, to assess the influence of NMDA receptor/channels on the eye-specific segregation of retinal ganglion cell (RGC) terminals that occurs whenever two retinas innervate one tectal lobe. Exposure of the tectum to the active isomer of APV produces desegregation of the RGC terminals without blocking electrical activity in the afferents or altering their terminal arbor morphology. Exposure to the inactive isomer of APV causes no perturbation of the normal stripe pattern. APV-induced desegregation is completely reversible within 2 weeks of removal of the APV. In addition, exposure of the optic tectum to NMDA results in stripes with sharper borders and fewer forks and fusions than untreated animals. These results suggest that the NMDA receptor/channel plays a role in eye-specific segregation in the three-eyed tadpole.

2-Amino-5-phosphonovalerate↗

Development of neurotransmitter metabolism in embryos of the leech Haementeria ghilianii.

We have investigated the development of neurotransmitter metabolism in embryos of the glossiphoniid leech Haementeria ghilianii. The neurotransmitter content of dissected embryonic tissues was measured by means of radioenzymatic assays, while the presence of neurotransmitters in individual identified neurons was detected by means of immunocytochemical and monoamine histofluorescence techniques. The capacity for synthesis of neurotransmitters was measured by incubating dissected embryonic tissues in radiolabeled neurotransmitter precursors. A specific neurotransmitter uptake system present in some neurons was detected by means of an autoradiographic technique. At an early stage of development of the nervous system, when most neurons are just beginning process outgrowth, the nerve cord acquires the capacity to synthesize ACh, 5-HT, and GABA from their immediate precursors, and contains ACh. Moreover, 5-HT-immunoreactive neurons and neurons that are capable of GABA uptake can be identified. Dopamine-containing neurons are first detected by their histofluorescence at a slightly later stage, after process outgrowth is under way. As development continues, the content of and capacity for synthesis of these neurotransmitters increase, as does the number of neurons capable of GABA uptake. During the earlier stages of development, ACh content exceeds 5-HT content, which in turn exceeds dopamine content. By the end of embryogenesis, however, 5-HT and dopamine contents have greatly increased relative to ACh content, with 5-HT content exceeding ACh content by a factor of 2. Of the neurotransmitters thus far studied, 5-HT is present in the highest amount in the juvenile and adult nerve cord. Our results indicate that in the development of the leech nervous system neurotransmitter metabolism is one of the first neuronal characters to differentiate and that the subsequent levels of the different neurotransmitters are differentially regulated.

Acetylcholine↗

Evidence for GABA as a neurotransmitter in the leech.

In the leech, Hirudo medicinalis, the inhibitory motor neurons to the longitudinal muscles in the body wall, cells 1 and 2, are linked via central inhibitory synapses to the excitatory motor neurons innervating the same muscles. Examination of these synapses showed that the inhibitors are GABAergic according to several electrophysiological and pharmacological criteria. Presynaptic release of neurotransmitter during passage of depolarizing current into the inhibitors, as well as direct application of GABA to the excitor cell bodies, hyperpolarizes the postsynaptic excitor. Moreover, both synaptic and extrasynaptic GABA receptors of the excitors are specifically blocked by the GABA antagonist bicuculline methiodide. The inhibitors, dissected from the ganglion and grown in culture, synthesize GABA when exposed to the GABA precursor glutamate, whereas the excitors do not synthesize detectable levels of GABA under these same conditions. The innervation and neurotransmitter sensitivity of the longitudinal muscles in the body wall of the glossiphoniid leeches Haementeria ghilianii and H. officinalis were examined in order to determine if the inhibitory neurotransmitter at the neuromuscular junction is GABA. Individual muscle fibers are innervated by both inhibitory and excitatory motor neurons in a manner such that the inhibitory and excitatory nerve terminals and neurotransmitter receptors are spatially and electrically separate. Intracellular recordings taken from the muscle fibers reveal a resting potential of about -70 mV. The amplitude of the spontaneous inhibitory junctional potentials (IJPs) falls to zero at a polarization of about -65 mV and reverses in sign at the normal resting potential.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Identified GABAergic inhibitory motor neurons in the leech central nervous system take up GABA.

Physiologically identified inhibitory motor neurons in the leech central nervous system were double-labeled by intracellular injection of the dye Lucifer Yellow and gamma-amino-[3H]butyric acid ([3H]GABA) uptake to demonstrate that these inhibitory neurons are among the approx. 35 neurons per abdominal ganglion previously shown to take up GABA. GABA uptake into the inhibitory motor neurons is stimulated by electrical activity in the neurons.

Action Potentials↗

3H-GABA uptake selectively labels identifiable neurons in the leech central nervous system.

Segmental ganglia of the leech ventral nerve cord synthesize the neurotransmitter gamma-aminobutyric acid (GABA) when incubated in the presence of the precursor glutamate, suggesting that there may be GABA-ergic neurons in the leech nerve cord. GABA-accumulating neurons of the two taxonomically distant leech species, Haementeria ghilianii and Hirudo medicinalis, have been labeled by taking advantage of their high-affinity uptake system for the neurotransmitter. Autoradiography of sectioned segmental ganglia previously exposed to 3H-GABA reveals a reproducible pattern of about thirty 3H-GABA-labeled neuronal cell bodies per ganglion. The majority of 3H-GABA-labeled neuronal cell bodies are bilaterally paired, although some apparently unpaired cell bodies also accumulate label. Neuronal processes were reproducibly labeled by GABA uptake and could be traced in the neuropil through commissures and fiber tracts into the segmental nerve roots and interganglionic connectives, respectively.

Animals↗

Stimulus-secretion coupling in isolated adrenal chromaffin cells: calcium channel activation and possible role of cytoskeletal elements.

The catecholamine secretory function of a preparation of isolated bovine adrenal chromaffin cells has been further characterized under conditions designed to elucidate the mechanism of calcium channel activation and the possible role of cytoskeletal elements in stimulus-secretion coupling. Three related sets of data were obtained: (1) Differences in kinetics, Ca dependence, strength, and additivity of the secretory response to acetylcholine (ACh) versus excess K; (2) the effects on secretion of the Ca channel-blocking agents, Ni, Mg, and verapamil; and (3) the Ca dependence of vinblastine action on ACh- and K-evoked secretion. The results suggest that a major portion of the Ca influx required for catecholamine release enters the cell via voltage-dependent Ca channels with some additional Ca influx via the ACh receptor channel. Comparison of the present secretion data with corresponding known electrophysiological properties of isolated chromaffin cells provides added evidence for a role of chromaffin cell action potentials in regulation of Ca influx and the secretory response. Elevated Ca concentrations enhanced K-evoked secretion to levels comparable to that of Ach but did not induce a vinblastine block of K-evoked release. This provides further evidence against a role of microtubules in the common exocytosis event per se. However, a role of cytoskeletal elements in directing the movement of secretory granules, or an action of vinblastine at cholinergic receptors, remain distinct possibilities.

Acetylcholine↗