PubMed Health⌕ Search

Biomedical subjects

M A Simmonds

Publications and source records attributed to M A Simmonds.

At least 73 records · Page 4Linked to original sources
PubMed1982

Classification of some GABA antagonists with regard to site of action and potency in slices of rat cuneate nucleus.

Compounds reported to be GABA antagonists have been studied quantitatively on dorsal funiculus fibres and terminals in the rat cuneate nucleus in vitro. The potencies of the antagonists against the GABA analogue muscimol were determined as pA2 values. Distinction was made between three different sites of antagonist action within the GABA receptor and ionophore complex. Competitive antagonists, presumed to act at the GABA receptor, and their pA2 values were bicuculline (5.98), bicuculline methochloride (5.88), strychnine (5.29) and tubocurarine (4.95). Antagonists which were not competitive and acted predominantly at the 'picrotoxin site' on the ionophore were picrotoxin (6.19), picrotoxinin (6.03), isopropylbicyclophosphate (5.82) and leptazol (2.89). A third type of antagonism was shown by frusemide. Attention is drawn to the picrotoxin site and its likely importance in the regulation of GABA-mediated inhibition by drugs.

Animals↗
PubMed1982

Effects of baclofen on the olfactory cortex slice preparation.

Baclofen has been shown to be ineffective against first order excitatory synaptic transmission in the olfactory cortex slice, whereas it is known that GABA-mediated inhibition depresses this transmission in a bicuculline-sensitive manner. In contrast, second and third order synaptic transmission, excitatory and inhibitory respectively, were depressed by baclofen. This suggests that the distribution of baclofen (GABAB) receptors differs from that of bicuculline-sensitive GABAA receptors in the olfactory cortex.

Animals↗
PubMed1982

GABA-mediated changes in excitability of the rat lateral olfactory tract in vitro.

1. Conditioning stimulation of the lateral olfactory tract (l.o.t.) in the rat olfactory cortex slice evoked a slow depolarization of the terminal regions of the l.o.t. The depolarization lasted about 150 ms and was abolished by the gamma-aminobutyric acid (GABA) antagonist bicuculline. 2. Excitability testing of the terminal regions of the l.o.t. showed an increase in excitability which lasted for about 150 ms following a conditioning stimulus. This increase in excitability was abolished by the calcium antagonist cadmium and by the GABA antagonists bicuculline and penicillin. 3. Superfused GABA caused a consistent decrease in the excitability of the terminal regions of the l.o.t. within the cortex but an increase in excitability of the axons within the tract itself. These effects were antagonized by bicuculline. K+ caused similar changes in excitability which were not antagonized by bicuculline. 4. The difference between the effects of superfused GABA and the effects of orthodromic conditioning can be explained if a more restricted location of action is assumed for the GABA released by conditioning stimulation. It is suggested that GABA causes a decrease in excitability at its locus of action and that the observed increases in excitability occur in adjacent areas of neuronal membrane.

Animals↗
PubMed1982

Leukemic-like membrane properties acquired by B lymphocytes when depleted of 185,000-dalton macromolecular insoluble cold globulin.

Human peripheral blood lymphocytes can be phenotypically identified by the presence of one or both of two proteins, 225,000-dalton macromolecular insoluble cold globulin (225-MICG) and 185,000-dalton MICG (185-MICG). T cells synthesize and insert into their plasma membrane 225-MICG, null cells 185-MICG, and B cells both 225 and 185-MICG. In contrast, the monoclonal B cells of chronic lymphocytic leukemia are characterized by the presence of 225-MICG and the absence of 185-MICG. We have recently found it possible to chemically deplete 185-MICG from viable normal B cells by treating them with diisopropylfluorophosphate (DFP), thus making normal B cells phenotypically resemble leukemic cells. In the present report we determined whether certain peculiar properties of these leukemic cells would be associated with the normal B cells chemically depleted of 185-MICG. In normal B cells, SIg diffuses in the lipid bilayer to form clusters and caps under appropriate conditions, while in chronic lymphocytic leukemia (CLL) cells this does not occur. Normal B cells depleted of 185-MICG fail to undergo capping of SIg or surface MICG under appropriate conditions. Both DFP-treated B cells and CLL cells tend to rupture when smeared on a glass slide. Both CLL cells and DFP-treated B cells fail to secrete 225-MICG after it has been synthesized intracellularly. The relationship of these findings to the mechanisms of secretion and capping are discussed.

B-Lymphocytes↗
PubMed1981

Distinction between the effects of barbiturates, benzodiazepines and phenytoin on responses to gamma-aminobutyric acid receptor activation and antagonism by bicuculline and picrotoxin.

1 Interactions of depressant and anticonvulsant drugs with the neuronal gamma-aminobutyric acid (GABA) receptor + effector system have been examined on afferent fibres to the rat cuneate nucleus in vitro. Three types of interaction have been measured: (a) potentiation of depolarizing responses to the GABA analogue, muscimol: (b) reduction in the potency of bicuculline as an antagonist of muscimol at the GABA receptor: (c) reduction in the potency of picrotoxin as an antagonist of muscimol acting on the effector mechanism. 2 Phenobarbitone reduced the potency of picrotoxin in doses which did not affect the potency of bicuculline and which caused only a small potentiation of muscimol. Pentobarbitone did not show such selectivity, a reduction in potency of picrotoxin always being accompanied by a reduction in potency of bicuculline and a substantial potentiation of muscimol. 3 Flurazepam and lorazepam both reduced the potency of picrotoxin without affecting that of bicuculline and with very little potentiation of muscimol. Phenytoin had no effect on the potency of picrotoxin whilst potentiating muscimol to the same extent as phenobarbitone. 4 The spectrum of drug activity in reducing the potency of picrotoxin correlates well with the reported anticonvulsant effects of these drugs against kindled amygdaloid seizures. Potentiation of muscimol and reduction of bicuculline potency appear more closely related to hypnotic properties.

Animals↗
PubMed1981

Chronic lymphocytic leukemia cells lack the 185,000-dalton macromolecular insoluble cold globulin present on normal B lymphocytes.

We have recently characterized two lymphocyte-associated membrane proteins which have been termed 225,000-dalton and 185,000-dalton macromolecular insoluble cold globulin (225-MICG and 185-MICG, respectively) to distinguish their major physicochemical properties. These proteins differ antigenically, structurally, and in their cellular distribution. T cells can be distinguished by the synthesis and presence in the plasma membrane of 225-MICG, Null cells by the appearance of 185-MICG, and B cells by the appearance of both 225- and 185-MICG. The characterization of these two proteins in the monoclonal B lymphocytes of chronic lymphocytic leukemia forms the basis of this report. Using immunofluorescent microscopy, we found only 225-MICG on the surface of chronic lymphocytic leukemia (CLL) cells in 15 patients, whereas control B cells from 20 individuals displayed both 225- and 185-MICG. When MICG proteins were isolated and compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, normal B cells showed two stained bands, corresponding to 225- and 185-MICG, whereas the CLL cells demonstrated only the 225-MICG band. Using labeled amino acid incorporation into cellular protein, normal B cells were shown to synthesize 225- and 185-MICG, whereas CLL cells synthesized only 225-MICG, as determined by immune or cold precipitation of labeled cell lysates. When labeled secretions from B cells and CLL cells were analyzed by immune precipitation, 225- and 185-MICG were secreted by B cells, whereas neither protein was secreted by CLL cells. When normal B cells and CLL cells were mixed, incubated, and lysed together, both 225- and 185-MICG were present, thus excluding proteolysis as a cause of the absence of 185-MICG in CLL. The lack of 185-MICG in CLL distinguishes leukemic cells from normal B lymphocytes. Furthermore, the absence of this normal cell surface protein in these leukemic cells suggests a role for 185-MICG in the malignant transformation of lymphocytes.

B-Lymphocytes↗
PubMed1980

A site for the potentiation of GABA-mediated responses by benzodiazepines.

The benzodiazepines have been well characterised as minor tranquillizers and attempts to explain their unique spectrum of activity have included suggestions that they may interact with a variety of neurotransmitter systems. Recently, a possible interaction with the gamma-aminobutyric acid (GABA) system has received most attention. Benzodiazepines potentiate the actions of both synaptically released and exogenously administered GABA on mammalian neuronal preparations but the site of action within the GABA response mechanism has not been determined. Binding studies suggest that benzodiazepines combine with highly specific sites in the neuronal membrane and that these sites have some indirect association with GABA receptors. To investigate this association further in a functioning GABA system, quantitative studies have been made in vitro on neuronal depolarisations mediated by GABA receptor activation. Evidence has already been presented that bicuculline is most probably a competitive antagonist at the GABA receptor while picrotoxin acts as an antagonist at a separate site. Here flurazepam is shown to attenuate preferentially the action of picrotoxin rather than bicuculline and a model is suggested for the site of action of these drugs within the GABA response mechanism.

Afferent Pathways↗
PubMed1978

Presynaptic actions of gamma-aminobutyric acid and some antagonists in a slice preparation of cuneate nucleus.

1 A slice preparation of the rat cuneate nucleus is described which is suitable for electrophysiological studies on the presynaptic action of drugs. 2. Superfusion of a slice with gamma-aminobutyric acid (GABA) depolarized the afferent nerves in a concentration-related manner. The responses were Cl-dependent. Depolarizations to high concentrations of GABA often faded. Glycine and L-glutamate had little effect. 3 (+)-Bicuculline antagonized GABA in an apparently competitive manner (pA2 = 5.35) at low response levels. Strychnine was 10 times less potent. Responses to high concentrations of GABA were sometimes potentiated by (+)-bicuculline and strychnine. 4 Bemegride and leptazol both antagonized GABA, but with low potency and in a manner which was clearly not competitive.

Aminobutyrates↗
PubMed1978

Effects of gamma-aminobutyric acid on nerve terminal excitability in a slice preparation of cuneate nucleus.

1 Superfusion of a slice preparation of the rat cuneate nucleus with gamma-aminobutyric acid (GABA) depolarized the afferent nerve fibres and increased their excitability. However, before the depolarization had reached its peak the increased excitability reversed to a decreased excitability, an effect which outlasted the depolarization. 2 Both components of the GABA excitability response were dose-related Cl--dependent and antagonized by bicuculline. 3 Possible mechanisms underlying the sequence of excitability changes are discussed.

Aminobutyrates↗
PubMed1978

Field potentials, inhibition and the effect of pentobarbitone in the rat olfactory cortex slice.

1. Field potentials were evoked in slices of rat olfactory cortex by stimulating the lateral olfactory tract. In addition to previously described components of the wave-form, a further distinct surface-negative potential of low amplitude and long duration (I-wave) has been described. 2. Pentobarbitone, at concentrations of 10(-5) M and above, markedly enhanced enhanced the amplitude and duration of the I-wave with only minimal effect on other components of the field potential. 3. The I-wave was reversibly reduced by the GABA antagonists bicuculline and picrotoxin and was also attenuated at rapid rates of stimulation. Low chloride medium usually caused a transient increase in amplitude of the I-wave followed by a gradual reduction, suggesting that a chloride-mediated depolarization was involved. 4. Evoked inhibition, which was most probably post-synaptic, occurred in parallel with the I-wave. This was monitored as a suppression of, or increase in latency of the population spike evoked by a second stimulus at appropriate intervals after the first. Pentobarbitone substantially increased the duration of the post-synpatic inhibition, without obvious changes in the presynaptic inhibitory phenomenon associated with antidromic firing in the lateral olfactory tract. 5. It is proposed that the I-wave is the field potential representation of a population depolarizing i.p.s.p. and that the main action of pentobarbitone is to enhance this inhibition.

Action Potentials↗
PubMed1976

Antagonism of gamma-aminobutyric acid and glycine by convulsants in the cuneate nucleus of cat.

1. Some convulsant substances have been applied to single neurones in the cat cuneate nucleus by microiontophoresis. Numerical values were derived for the effectiveness and selectivity of the substances as antagonists of gamma-aminobutyric acid (GABA) and glycine. 2. (+)-Bicuculline methochloride was the most effective GABA antagonist and it also excited many neurones. It antagonized GABA in 93% of experiments but also antagonized glycine in 41% of experiments. In most experiments the antagonism of GABA was greater than the antagonism of glycine resulting in an overall selective antagonism of GABA that was statistically significant. Nevertheless, in only about one quarter of the individual experiments was the GABA antagonism substantial and the selectivity clearcut. 3. (+)-Bicuculline and picrotoxin were less easily applied to neurones by microiontophoresis and were found to antagonize GABA in 30% and 35% of experiments, respectively. They also antagonized glycine in 25% and 30% of experiments, respectively. Overall, neither substance could be shown to be selective, statistically, although in the few individual experiments where the GABA antagonism was substantial the antagonism was clearly selective. 4. (+)-Tubocurarine antagonized GABA in 59% and glycine in 32% of experiments and it also excited many neurones. Penicillin antagonized GABA in 33% of experiments without antagonizing glycine. Neither antagonist caused any substantial antagonisms of GABA and neither showed significant selectivity overall. (-)-Bicuculline methochloride, leptazol and bemegride antagonized GABA or glycine in less than 10% of experiments, although (-)-bicuculline methochloride excited most neurones. 5. Strychnine antagonized glycine in every experiment while antagonizing GABA in only 5% of experiments. In each individual experiment the antagonism of glycine was substantial and clearly selective, resulting in a statistically significant selectivity overall. 6. It is concluded that the selective glycine antagonist strychnine is considerably better than the presently available GABA antagonists for distinguishing between responses to GABA and glycine, when the antagonists are applied by microiontophoresis.

Action Potentials↗
PubMed1976

The negative potential wave evoked in cuneate nucleus by stimulation of afferent pathways: its origins and susceptibility to inhibition.

1. The negative (N)-wave evoked at various depths in the cuneate nucleus by stimulation of afferents in the ipsilateral forepaw or dorsal column has been studied in the rat. 2. Micro-iontophoretic applications of gamma-aminobutyric acid (GABA) into the vicinity of the recording electrode markedly reduced the amplitude of the negative wave, but only when recordings were made near the base of the cuneate nucleus. Nearer the surface of the medulla, GABA was much less effective. 3. A similar depth distribution obtained for the depression of the negative wave by micro-iontophoretic Mg2+ and enhancement by Ca2+. 4. Depression of the negative wave by conditioning stimulation of the afferent pathway also showed a similar depth distribution. The conditioned depression of the negative wave was most marked during the first 30 ms after the conditioning stimulus and this early depression could be antagonized by iontophoretic (+)-bicuculline methochloride. A lesser degree of conditioned inhibition of the negative wave persisted up to 80-200 ms but this was resistant to (+)-bicuculline methochloride. Thus, conditioned depression of the negative wave appeared to be mediated only in part by a GABA-like transmitter. 5. It is concluded that the negative wave recorded near the base of the cuneate nucleus has some of the predicted properties of a post-synaptic potential. These properties are not seen when the negative wave is recorded more superficially, near the surface of the medulla.

Afferent Pathways↗
PubMed1976

Possible presynaptic inhibition in rat olfactory cortex.

1. Field potentials were evoked from rat olfactory cortex slices in vitro at room temperature by lateral olfactory tract stimulation. At very slow stimulation rates (less than 0-1 Hz) a delayed negative wave (late N-wave) was found to follow the first negative wave (N-wave). This late N-wave started 30 msec after the stimulus and lasted for 50-150 msec. 2. Low Ca2+/high Mg2+ medium abolished the late N-wave more rapidly than the N-wave, suggesting a possible multisynaptic origin. 3. The GABA antagonist bicuculline (10(-6) M) abolished the late N-wave without affecting the N-wave. 4. During the late N-wave, both the tract action potential and the N-wave to a second stimulus were reduced. This was attributed, at least in part, to collision with antidromic action potentials which could be detected during the late N-wave. 5. These findings are discussed in terms of a possible presynaptic depolarizing action of a GABA-like transmitter giving rise to presynaptic inhibition and the late N-wave.

Animals↗