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V J Balcar

Publications and source records attributed to V J Balcar.

At least 55 records · Page 3Linked to original sources
PubMed1992

Glutamate decarboxylase in developing rat neocortex: does it correlate with the differentiation of GABAergic neurons and synapses?

Postnatal development of glutamate decarboxylase was studied in the rat cerebral cortex. Two methods were used: estimation of the enzymatic activity of glutamate decarboxylase in homogenates of developing cortical tissue and visualization of structures containing glutamate decarboxylase-like immunoreactivity. Glutamate decarboxylase-like immunoreactivity appeared first in perikarya and dendrites and only later in axons and axon varicosities. The most rapid increase in the glutamate decarboxylase activity took place during the second postnatal week and this coincided with a rapid increase in the density of axon varicosities containing glutamate decarboxylase-like immunoreactivity but preceded the most rapid phase in the formation of GABAergic synapses by several days. However, there was a change in the characteristics of glutamate decarboxylase which correlated with GABA synaptogenesis: two fractions of glutamate decarboxylase with different sensitivities to the activating effects of Triton X-100 could be distinguished as from about the time when most of the GABAergic synapses are formed.

Animals↗
PubMed1992

Heterogeneity of high affinity uptake of L-glutamate and L-aspartate in the mammalian central nervous system.

Characteristics of high affinity uptake of L-glutamate are examined in order to evaluate the possible use of the uptake of [3H]L-glutamate, [3H]L-aspartate or any other suitable [3H]-labelled substrate as a marker for glutamatergic and aspartergic synapses in autoradiographic studies in the mammalian brain. Review of data on substrate specificity indicates the presence of at least two high affinity uptake systems specific for acidic amino acids in the central nervous tissue; one which takes up L-glutamate and L-aspartate and the other which is selective for L-glutamate only. Studies on ionic requirements, too, point to the existence of at least two distinct uptake systems with high affinity for L-glutamate. The Na(+)-dependent uptake system(s) handle(s) both L-glutamate and L-aspartate whereas the Na(+)-independent uptake system(s) show(s) selectivity for L-glutamate only. Available data do not favour the Na(+)-dependent binding of [3H]D-aspartate to thaw-mounted sections of frozen brain tissue as a suitable marker for glutamatergic/aspartergic synaptic nerve endings. However, there are reasons--such as the results of lesion studies and the existence of uptake sites which have a higher affinity for L-aspartate than for D-aspartate--to suggest that Na(+)-dependent binding of [3H]L-aspartate, rather than that of [3H]D-aspartate, should be further investigated as a possible marker for the glutamatergic/aspartergic synapses in the autoradiographic studies using sections of frozen brain.

Animals↗
PubMed1990

Presence of a highly efficient "binding" to bacterial contamination can distort data from binding studies.

[3H]GABA at low concentrations (5-10 nM) was bound by what appeared to be a "GABA receptor binding site" in bacterial contamination originating from a batch of distilled water. Under experimental conditions similar to those usually employed in [3H]GABA binding studies, the apparent binding displayed a very high "specific" component and a high efficiency in terms of [3H]GABA bound per mg of protein. The "binding" was blocked by muscimol but not by isoguvacine, SR95531 and nipecotic acid. These characteristics suggest that the presence of such spurious binding in the experiments using 3H-labeled ligands in brain homogenates may not always be very obvious and, moreover, it can result in subtle, but serious, distortions of data from such studies, which may not be immediately recognized.

Animals↗
PubMed1990

Neurochemical correlates of gamma-aminobutyrate (GABA) inhibition in cat visual cortex.

High affinity binding of [3H]gamma-aminobutyric acid (GABA) to neuronal membranes from different parts of cat visual cortex was tested for sensitivity to GABA(A) agonists isoguvacine and THIP, GABA(A) antagonist SR95531 and GABA(B) agonist baclofen. Some of the GABA(A)-binding sites were found to have a very low affinity for THIP, suggesting the presence and, possibly, uneven distribution of "non-synaptic" GABA(A) receptors in cat visual cortex. There were no differences in Km and Vmax values of high affinity uptake of GABA and in the potency of K(+)-stimulated release of GABA, between primary and association cortices. Consequently, the present results indicate that despite the anatomical and physiological differences between the primary and association feline visual cortices the neurochemical characteristics of GABAergic inhibition are very similar in the two regions.

Animals↗
PubMed1989

Ontogeny of high affinity uptake of GABA studied in 'minislices' (tissue prisms) of rat cerebral cortex.

There has been a considerable discrepancy between the values of Vmax of high affinity uptake of gamma-aminobutyric acid (GABA) obtained in slices and homogenates, respectively, of developing rat cortex. In the present communication, it is suggested that the relatively low values observed in slices of early postnatal cortex might be explained by damage to the immature tissue during the experiment. The possible locations of high affinity uptake of GABA in developing rat cortex are discussed.

Aging↗
PubMed1989

Chemical integrity of [3H]GABA used in binding studies.

A method which is claimed to be able to determine the proportion of "true" GABA within radiolabeled GABA used in binding studies was tested using [3H]GABA. The method was found to be unsuitable for 3H-labeled GABA and, furthermore, both theoretical considerations and the present experimental data indicated that it could also produce misleading results with [14C]GABA.

Animals↗
PubMed1989

Developmental changes in high-affinity uptake of GABA by cultured neurons.

High-affinity uptake of [3H]gamma-aminobutyric acid (GABA) was studied in cultures of neonatal rat cortical neurons grown on pre-formed monolayers of non-neuronal (glial) cells. Both the maximum rate (Vmax) and, to a smaller extent, the Km of [3H]GABA uptake increased with time. In addition, in parallel with these changes, 2,4-diaminobutyric acid and cis-3-aminocyclohexane-1-carboxylic acid (ACHC), compounds which are considered typical substrate/inhibitors of GABA uptake in neurons, became progressively stronger inhibitors of [3H]GABA uptake. Consequently, the present results may mean that the studies using uptake of [3H]GABA, [3H]ACHC, or [3H]DABA as a specific marker for GABAergic neurons differentiating during the ontogenetic development of the central nervous system may have to be interpreted with caution.

Acetanilides↗
PubMed1987

Neurochemistry of GABAergic system in cerebral cortex chronically exposed to bromide in vivo.

Neurochemical correlates of GABAergic synaptic transmission [binding, uptake, metabolism, and tissue content of gamma-aminobutyric acid (GABA)] were investigated in the cortex of rats that had been given 27 mM bromide in drinking water for periods of time ranging from 1 day to 1 month. No effect of bromide on any of the parameters was found and it is concluded that chronic administration of bromide has no profound effect on GABAergic inhibitory system in the rat cortex.

4-Aminobutyrate Transaminase↗
PubMed1986

Presence of neurons with GABA-like immunoreactivity in the superior cervical ganglion of the rat.

Using antibodies raised against gamma-aminobutyric acid (GABA)-glutaraldehyde complexes, we have found neurons with GABA-like immunoreactivity in the superior cervical ganglion of adult rats. The processes of these neurons formed pericellular networks around the principal ganglion cells. Electron microscopy revealed that the immunoreactive dendrites were innervated by non-reactive axon terminals which formed asymmetrical synapses and probably originated from the preganglionic nerve. Axons with GABA-like immunoreactivity, especially axonal varicosities filled with synaptic vesicles, were found in direct apposition to principal ganglion cells. The GABA-positive axons and axon varicosities persisted in experimentally decentralized (deafferented) ganglia, suggesting that the perikarya of the immunoreactive neurons were intrinsic to the superior cervical ganglion. Taken together with data on inhibitory effects of GABA in sympathetic ganglia, these findings suggest that the superior cervical ganglion of rats contains a subpopulation of inhibitory interneurons which is GABAergic. This would indicate that GABAergic neurons do not only occur in the central but also in the peripheral nervous system.

Animals↗
PubMed1986

GABA receptor binding in rat cerebral cortex and superior cervical ganglion in the absence of GABAergic synapses.

High affinity binding of [3H]gamma-aminobutyric acid (GABA) to membrane-containing homogenates prepared from rat cerebral cortex (neonatal or adult) or from adult rat superior cervical ganglion (SCG) was tested for sensitivity to baclofen, bicuculline and several other analogues of GABA. Since no major differences were found between the tissues free of GABAergic synapses (SCG and neonatal cortex) and GABAergic synapse-rich adult rat cortex, it is suggested that the presence of GABAergic synapses does not play a decisive role in the determination of pharmacological sensitivities of high-affinity GABA receptors.

Aging↗
PubMed1983

Is there a non-synaptic component in the K+-stimulated release of GABA in the developing rat cortex?

Postnatal development of the K+-stimulated release of [3H]GABA in rat cortex slices was compared to the density of Gray type 2 synapses (which correlate with GABAergic synapses in rat cortex). The [3H]GABA release system develops earlier than the inhibitory synapses which suggests that, at least in the young cortical tissue, a large part of the release mechanism is associated with other than synaptic structures.

Aging↗
PubMed1980

Central actions of somatostatin.

Somatostatin (SRIF) was applied microiontophoretically to neurons in the frontal and parietal neocortex, the hippocampus and the striatum of rats anaesthetized with either urethane or chloral hydrate. Qualitatively identical results were obtained under both anaesthetic conditions. In urethane-treated rats SRIF elicited a dose-dependent increase of the firing rate of 74% of the neurons studied in the frontal cortex and of 46% of the neurons studied in the parietal cortex. All cortical cells identified as pyramidal cells were excited. In the hippocampus SRIF provoked excitatory responses in two thirds of all neurons. Six out of the nine cells identified as pyramidal cells were excited by SRIF. In the striatum 80% of all neurons were excited. Following repeated exposure of central neurons to SRIF, the magnitude of the excitatory response gradually diminished, indicating desensitisation. SRIF in concentrations ranging from 10(-8) to 10(-4) M did not interfere with the binding of (3H)-muscimol to GABA receptor sites. The release of GABA from synapses preloaded with (3H-GABA) was not influenced by SRIF in the concentration range from 10(-6) to 10(-4) M. These results indicated that SRIF does not evoke the excitatory responses through attenuation of GABA-mediated inhibition. In conclusion, the findings support the hypothesis that somatostatin may function as a neurotransmitter in the central nervous system.

Action Potentials↗
PubMed1979

Effect of cyclic nucleotides on high affinity uptake of L-glutamate and taurine in glial and neuroblastoma cells.

Cultured glial cells (clones NN and I6) and neuroblastoma cells (clones M1 and M1NN) were used to study the effect of cyclic nucleotides on high affinity uptake of L-glutamate and taurine. Treatment of NN cells with a mixture of dBcAMP and BrcGMP for one or two days resulted in a more than 3-fold increase in the Vmax of L-glutamate high affinity uptake. Similar but smaller change was observed in a related glial clone (I6). Effects of cyclic nucleotide derivatives on high affinity uptake of L-glutamate by neuroblastoma cells (clones M1 and M1NN) or of taurine by both glial or neuroblastoma cells were either smaller or absent. Some differences were found in kinetic parameters of uptake of L-glutamate and taurine by original and re-isolated cellls, respectively. It is possible that these differences resulted from co-cultivation of glial and neuronal cells. However, since some values reported in this paper differ to some extent from those published previously, some other factors, such as subcloning, may have been responsible.

Animals↗
PubMed1979

High-affinity uptake of gamma-aminobutyric acid in cultured glial and neuronal cells.

Both glial and neuronal cells maintained in primary culture were found to accumulate [3H]GABA by an efficient "high-affinity" uptake system (apparent Km = 9 muM, Vmax = 0.018 and 0.584 nmol/mg/min, respectively) which required sodium ions and was inhibited by 1 mM ouabain. Strychnine and parachloromercuriphenylsulfonate (pCS) (both at 1mM) also strongly inhibited uptake of [3H]GABA, but metabolic inhibitors (2,4-dinitrophenol, potassium cyanide, and malonate) were without effect. Only three structural analogs of GABA (nipecotate, beta-alanine, and 2,4-diaminobutyrate) inhibited uptake of [3H]GABA, while several other compounds with structural similarities to GABA (e.g. glycine, L-proline, and taurine) did not interact with the system. The kinetic studies indicated presence of a second uptake (Km = 92 muM, Vmax = 0.124 nmol/mg/min) in the primary cultures containing predominantly glioblasts. On the other hand, only one of the neuronal cell lines transformed by simian virus SV40 appeared to accumulate [3H]GABA against a concentration gradient. Apparent Km of this uptake was relatively high (819 muM), and it was only weakly inhibited by 1 mM ouabain and 1 mM pCS. The structural specificity also differed from that of the uptake observed in the primary cultures. Significantly, non of the nontransformed continuous cell lines of either tumoral (glioma, C6; neuroblastoma, M1; M1NN) or normal (NN;I6) origin actively accumulated [3H]GABA. It is suggested that for the neurochemical studies related to GABA and requiring homogeneous cell populations, the primary cultures offer a better experimental model than the continuous cell lines.

4-Chloromercuribenzenesulfonate↗