Search PubMedSearch

Biomedical subjects

M Cynader

Publications and source records attributed to M Cynader.

At least 19 recordsLinked to original sources

Morphology and distribution of neurons and glial cells expressing beta-adrenergic receptors in developing kitten visual cortex.

The morphology and distribution of cells expressing beta-adrenergic receptors has been studied in developing kitten visual cortex using a monoclonal antibody which recognizes both beta-1 and beta-2 adrenergic receptors. We found specific populations of neurons and glial cells which express beta-adrenergic receptor immunoreactivity in the kitten visual cortex. In adult animals, the receptors are most concentrated in the superficial and deep cortical layers (layers I, II, III and VI). About 50% of the stained neural cells in adult cat visual cortex are glial cells. Most of the immunoreactive neurons in layers III and V are pyramidal cells while those in layers II and IV are more likely to be nonpyramidal cells. In neonatal kittens, staining is weaker than that in adult cats and it appears to be concentrated in neurons of the deep cortical layers and in the subcortical plate and white matter. Only a few immunoreactive glial cells were found at this age. Receptor numbers increase after birth and by 24 days of age, the laminar distribution of beta-adrenergic receptors approaches that of adult animals. Immunoreactive glial cells in the white matter show a progressive increase in number throughout postnatal development.

Aging

Preferential innervation of immunoreactive choline acetyltransferase synapses on relay cells of the cat's lateral geniculate nucleus: a double-labelling study.

Relationships between cholinergic axons and GABA cells in the lateral geniculate and the perigeniculate nuclei were quantitatively assessed by combining pre-embedding immunocytochemical visualization of choline acetyltransferase with post-embedding immunogold labelling of GABA at the electron microscopic level. In the lateral geniculate nucleus, vesicle-containing profiles immunoreactive for choline acetyltransferase made synapses exclusively with dendritic profiles both within and outside synaptic glomeruli. Only 7.0% (seven of 100 dendrites) of the target dendrites tested were positive for GABA. Of these, the majority (85.7%; 6/7) were dendritic profiles containing synaptic vesicles. This is in contrast with the overall population of geniculate synapses where 21.3% of the targets were GABA-immunopositive, a three-fold significant difference. In the perigeniculate nucleus, immunoreactive choline acetyltransferase synapses targeted mainly dendritic profiles (93.9%; 31/33) and to a lesser extent somata (6.1%). In this nucleus, all unequivocally defined targets were GABA-immunopositive. Cholinergic inputs thus show a preference to target geniculate relay neurons without a strong innervation of GABA-immunoreactive interneurons. In the perigeniculate, however, cholinergic synapses provide a powerful input to GABAergic cells. This suggests that the facilitatory effects of acetylcholine on the excitability of relay cells would be mediated through (i) a direct innervation of the geniculate relay cells, and (ii) a cholinergic inhibitory innervation of the recurrent inhibition to the lateral geniculate nucleus via the perigeniculate nucleus.

Animals

Enrichment of glutamate in zinc-containing terminals of the cat visual cortex.

The presence of glutamate and GABA was examined in zinc-containing terminals of the cat visual cortex using a post-embedding immunogold method. The surface density of immunogold-labelling was also evaluated in morphologically defined ultrastructural elements, namely terminals having round synaptic vesicles and making asymmetrical synapses (RA boutons), terminals with flat vesicles and symmetrical synapses (FS) and glial cell processes. Glutamate immunoreactivity was highest in RA terminals and in zinc-containing boutons. It was lower in FS terminals and lowest in glial cell processes. GABA immunoreactivity was highest in FS terminals and low in all other ultrastructural elements analysed, including zinc-containing terminals. Therefore, zinc-containing terminals show an enrichment of glutamate and they are likely to use this amino acid as their neurotransmitter. Moreover, the fact that many RA terminals that are negative for zinc show an enrichment of immunoreactive glutamate suggests that zinc-containing fibres represent a subpopulation of the glutamate axonal network.

Animals

Calcium calmodulin dependent kinase II in cat visual cortex and its development.

A monoclonal antibody against the alpha-subunit of calcium/calmodulin-dependent protein kinase II (CAM-K II) was used to visualize the kinase in developing kitten visual cortex. CAM-K II was first expressed in neurons of the deep cortical layers (V and VI) at postnatal day 1-4 and appeared in the remaining cortical layers within the first 2 weeks. The level of immunoreactivity declined in cells of layer V and upper layer VI at about 30-40 days of age. By postnatal day 90, the most densely labelled neurons were concentrated in cortical layers II, III, lower layer IV and in layer VI. This laminar pattern remained constant into adulthood. EM studies showed that the kinase was found in both pre- and postsynaptic locations. About twice as many immunopositive neurons were found in cortical layers II-IV and VI in young adult cats when geniculate input was removed by an unilateral thalamic lesion performed early in life. These results indicate that expression of CAM-K II is developmentally regulated in visual cortical neurons; the alteration of immunoreactivity after early LGN lesions suggests that the level of the kinase (or its alpha-subunit) is also regulated by cortical input.

Animals

Pre- and postnatal development of GABA receptors in Macaca monkey visual cortex.

GABA is a putative inhibitory neurotransmitter in adult mammalian visual cortex but also has been implicated as playing a crucial role in cortical information processing during development. In order to understand better the role of GABA during primate visual cortex development, we have examined the time course of GABAA and GABAB receptor ontogenesis in 18 Macaca nemestrina monkeys ranging from fetal day 61 (F61d) to adulthood. The GABA and benzodiazepine binding sites of the GABAA receptor were detected by 3H-muscimol (3H-MS) and 3H-flunitrazepam (3H-FZ), respectively. GABAB receptors were detected by 3H-baclofen (3H-BA). All ligands were visualized by in vitro autoradiography. Quantitative analysis of film density was done to compare laminar changes during pre- and postnatal development. Saturation binding experiments were done for MS and FZ binding sites to determine receptor number (Bmax) and affinity (Kd) at selected pre- and postnatal ages. Both MS and FZ binding sites were present at F61d-72d throughout the cortical plate and marginal zone. FZ binding sites were more dense than MS binding sites over the cortical plate at young ages and were especially dense over the marginal zone. FZ binding sites also were present in lesser amounts over the subplate and intermediate zone, but not over the subventricular zone. By F119d-126d, layer 4 could be distinguished by its higher density for both ligands. The basic adult laminar pattern was established for both MS and BZ binding sites by birth (birth = F165d-170d). After birth, MS density increases dramatically in all layers, but layer 4C remains most dense to adulthood. FZ labeling is heavy in both layers 4 and 3 at birth but after 4 weeks after birth (P4 wk) it declines somewhat in the supragranular layers so that layer 4C now predominates. Labeling in layers 5/6 virtually disappears after birth. BA binding sites were present at F126d, at which time layer 4 was slightly lighter than the remainder of striate cortex; this laminar pattern remained basically the same throughout our series to adulthood. Competitive binding of agonist and antagonists for the GABAA receptor showed that MS binding characteristics were similar at F126d and P8.5 years (yr). MS binding site Bmax was about 8% of adult values at F72d, 24% by F126d, and 56% at F152d. Bmax then rose rapidly after birth to peak at P18wk at 169% of adult values, and then declined to P1yr. A second peak of 143% was found around P3.5yr, with adult values reached by P8.5yr.(ABSTRACT TRUNCATED AT 400 WORDS)

Aging

Effect of the richness of the environment on neurons in cat visual cortex. I. Receptive field properties.

In a recent study, it was demonstrated that the number of synaptic contacts associated with flat vesicles (FS synapses) is higher in the visual cortex of cats raised in an enriched environmental condition (EC) compared to those reared in an impoverished condition (IC). Moreover, the size of the FS synaptic contacts is also affected by the richness of the animal's environment during development. Based on evidence that the vast majority of FS synapses are GABAergic (gamma-aminobutyric acid) and that many of the properties of visual cortex neurons are influenced by GABA-dependent mechanisms, it has been suggested that these morphological synaptic changes induced by the richness of the environment correlate with differences in cortical receptive field properties. In the present study, this has been explored by recording visual responses of area 17 cells in cats raised either in isolation (IC) or in a colony with ample environmental stimulation (EC). Enriched visual cortex contains a higher proportion of orientation selective cells and a lower proportion of orientation biased and unoriented cells. In addition, orientation tuning is significantly sharper in EC animals (mean bandwidth of responsive units is equal to 32 degrees) than in IC cats (mean bandwidth is equal to 38 degrees; P less than 0.001). This is mostly due to the greater incidence of orientation biased units in impoverished cortex (23% in EC and 41% in IC animals; P less than 0.01). Unit responsivity is significantly affected by the richness of the environment. We found that all units of the EC cortex were responsive to light stimuli. In contrast, 14% of the impoverished cells studied fail to increase their response to at least twice the standard deviation of the spontaneous activity and were judged as unresponsive. We suggest that the lower responsivity in IC visual units is related to the higher number of GABAergic synapses per IC neuron, while the broader selectivity in IC cortex might be due to a more diffuse distribution of the GABAergic inhibitory connections.

Animals

Effect of the richness of the environment on neurons in cat visual cortex. II. Spatial and temporal frequency characteristics.

The quantitative distribution of synaptic contacts in the cat cerebral cortex is affected by the richness of the environment: the number of round-asymmetrical (RA) synapses per neuron is higher while the number of flat-symmetrical (FS) contacts is lower in the visual cortex of cats raised in an enriched environmental condition (EC) compared to those reared in an impoverished condition (IC). The size of FS synaptic contacts is also affected by the complexity of the animal's environment. It has been suggested that these morphological synaptic changes induced by the richness of the environment correlate with differences in physiological properties of the visual cortex. This question has been explored by assessing the cortical unit contrast sensitivity as a function of spatial and temporal frequency of area 17 cells in cats raised either in isolation (IC) or in a colony with ample environmental stimulation (EC). Contrast sensitivity is affected by the richness of the environment: at the preferred spatial frequency, an average enriched unit can detect lower contrasts (mean = 0.6%) than an average impoverished cell (0.9%; P less than 0.002). In addition, the mean highest spatial frequency that can be resolved (acuity) is greater in EC than in IC cells (1.7 and 1.2 cycles per degree, respectively, P less than 0.0001), whereas the spatial frequency at which units respond best is not statistically affected by the environment. The preferred temporal frequency also show a significant difference in EC (1.0-6.5 Hz) and IC units (0.9-4.0 Hz; P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Differential effects of quinolinic acid lesions on muscarinic acetylcholine receptors in cat visual cortex during postnatal development.

Quinolinic acid (QA) lesions of neurons in cat visual cortex were combined with conventional in vitro autoradiographic methods in order to define the cellular locus of the muscarinic acetylcholine receptor (mAChR). Animals of various postnatal ages had QA unilaterally injected into the visual cortex. Four to fourteen days later they were sacrificed and processed for electron microscopy (EM) or in vitro autoradiography. QA lesions at the various postnatal ages were found to eliminate intrinsic cortical neurons and their processes while leaving intact glia, fibers of passage and axon terminals from outside the lesion zone. Autoradiograms of visual cortex labelled with [3H]QNB (which labels M1 and M2 subtypes) showed an age-dependent loss of binding sites, with the greatest decreases occurring after 65 days postnatal. Examined separately, only the M1 mAChRs labelled with [3H]pirenzepine exhibited these age-dependent alterations. The results indicate a differential distribution of the M1 mAChRs during postnatal development. The loss of receptors late in postnatal life following QA suggests a dominantly neuronal locus; the relatively small loss early in postnatal life suggests a locus on other cellular elements.

Aging

Development of phorbol ester (protein kinase C) binding sites in cat visual cortex.

Tritiated phorbol-12,13-dibutyrate [( 3H]PDBu), a phorbol ester, was utilized to autoradiographically localize protein kinase C (PKC) in the cat visual cortex. Thin, slide-mounted sections of adult cat brain were used to characterize binding of [3H]PDBu. This was found to be saturable, reversible, and more readily displaced by phorbol ester than by synthetic diacylglycerols. Binding sites displayed a tissue concentration of 20 pmol/mg protein, and a dissociation constant of 8.0 nM. [3H]PDBu was slow to associate with its receptor, requiring 9.5 h to reach equilibrium. Autoradiograph revealed that PKC is heterogeneously distributed in the cat brain, and displays a laminar-specific pattern in the visual cortex. This laminar distribution undergoes marked changes during the first two months of postnatal life. In the visual cortex of neonatal kittens, [3H]PDBu binding is confined to layers I and V. Layer III acquires high levels of binding by postnatal day 15, layer II by 28 days, and layer VI becomes labelled by 40 days of age. Adult animals exhibit high levels of binding in all laminae except layer IV. Age-dependent changes in PKC's laminar distribution do not seem to be correlated with specific anatomical, neurochemical, or behavioural events during development. PKC appears to be associated with cell bodies or processes intrinsic to the visual cortex, and is probably not located on the terminals of cortical afferents.

Aging

Unilateral eyelid suture increases GABAA receptors in cat visual cortex.

We have examined the number, characteristics and distribution of GABAA receptor in the visual cortex of normal cats and in cats monocularly deprived by unilateral eyelid suture from early in postnatal life. Receptor densities were about 100% higher in the deprived animals than in their normal counterparts. No changes in receptor affinity were noted. The GABAA receptor increase appeared to affect all laminae in the visual cortex. The results suggest that an increase in GABAA receptors may underlie or result from the physiological consequences of early monocular deprivation.

Age Factors

Sodium channel toxins veratrine and veratridine modify opioid and muscarinic but not beta-adrenergic binding sites in brain slices.

We have examined the influence of the sodium channel toxins veratrine and veratridine on mu-opioid ([3H]-DAGO), muscarinic ([3H] NMS) and beta-adrenergic ([3H] CGP) receptors in rat brain slices. These drugs reduce opioid and muscarinic binding while leaving beta-receptors unaffected. Veratrine is inhibitory at 0 degree or at 30 degrees C whereas veratridine is without effect at 0 degree C. These data suggest that some factor contained in the mixture of drugs (veratrine) can block opioid and muscarinic receptors independently of depolarization. Veratridine does not affect muscarinic receptors at ice temperature. Similar observations were made in thin sections of cat brain at 0 degree C. The concentrations of the toxins which cause 50% inhibition of binding are well within the range (5 x 10(-5) M-10(-4) M) routinely used for depolarization experiments. We suggest that caution be used in the interpretation of results obtained from veratrum alkaloid-induced depolarizations. It would not be surprising if the binding of other ligands to their receptors was also affected.

Animals

Surgical undercutting prevents receptor redistribution in developing kitten visual cortex.

Recent studies have shown that several receptor populations in cat visual cortex undergo alterations in their laminar distributions during postnatal development (Shaw et al., 1984a,b; 1986b). These redistributions occur during the first few months of postnatal life, coincident with the physiologically defined critical period for cortical plasticity. In the present communication, we demonstrate that receptor redistributions can be prevented from occurring, or progressing once started, by surgically isolating the visual cortex at appropriate postnatal ages. These data suggest that the maturation of the chemical circuitry of the visual cortex is dependent on factors of extrinsic origin.

Animals

Benzodiazepine ([3H]flunitrazepam) binding in cat visual cortex: ontogenesis of normal characteristics and the effects of dark rearing.

[3H]Flunitrazepam (FNZ) binding sites were characterized in homogenates of cat visual cortex during normal postnatal development and following dark rearing from birth. In parallel experiments, the distribution and density of [3H]FNZ binding sites were examined by in vitro autoradiographic or 'scrape' methods. In homogenates, Bmax measurements showed low early values, rising to a peak in receptor density at about 60 days postnatal, followed by a decline in adulthood. At all ages, gamma-aminobutyric acid (GABA) altered the Kd, but not the Bmax of [3H]FNZ binding sites. Kd values showed a general increase with age, parallelled by an increased sensitivity to GABA. Receptor autoradiography revealed that the highest density of [3H]FNZ binding sites was in layer IV of cats of all ages. Deafferentation of extrinsic inputs to the visual cortex by surgical undercutting did not alter this pattern of laminar distribution, indicating that the receptors were associated with intrinsic cortical elements rather than subcortical inputs. Dark rearing had no effect on [3H]FNZ laminar distribution in the visual cortex. The Bmax was higher at 30 days postnatal, but did not differ significantly thereafter. Modulation by GABA was concomitantly higher at 30 days, but lower than normal in dark-reared animals at ages greater than 30 days postnatal. The results are discussed in relation to the normal and abnormal development of GABA receptors in the cat visual cortex.

Animals

Anomalous 2-deoxyglucose uptake and acetylcholinesterase activity in cat LGN after optic chiasm section.

Sectioning the optic chiasm in young kittens denervates layer A of the lateral geniculate nucleus (LGN) on both sides of the brain. We removed one eye of these split chiasm kittens in adulthood, rendering the ipsilateral LGN devoid of primary retinal input. Nonetheless, one day after the eye removal, consumption of 14C-2-deoxyglucose (2-DG) was much higher in layer A than in layer A1 of the LGN. The anomalous 2-DG uptake was not due to enhanced cortico-geniculate or other visual input to layer A, but instead appears related to a marked increase in acetylcholinesterase activity in this layer following the early chiasm section.

Acetylcholinesterase

Laminar distribution of receptors in monkey (Macaca fascicularis) geniculostriate system.

We have examined the laminar distributions of eight types of receptor in the primary visual cortex (area 17) and the lateral geniculate nucleus (LGN) of the macaque monkey. The receptor populations and subpopulations examined included those selective for gamma-aminobutyric acid (GABA) (using [3H]-muscimol as ligand), L-glutamate-related receptors (using [3H]-L-glutamate and [3H]-AMPA), muscarinic acetylcholine (using [3H]-quinuclidinyl benzilate--QNB and [3H]-N-methyl scopolamine--NMS), cholecystokinin (CCK) (using [3H] pentagastrin), benzodiazepine (using [3H]-flunitrazepam), and adenosine (using [3H]-cyclohexyladenosine--CHA). Each of the receptors examined exhibited characteristic and differing laminar patterns of binding in the striate cortex. Perhaps reflecting the high density of cell bodies and synapses in layer 4C, most receptors, except those labelled by [3H]-L-glutamate or [3H]-AMPA, showed dense concentrations in this layer. Layers 4B and 5, which contain relatively few cell bodies and heavy myelin concentrations, were in general lightly labelled. Layer 6 showed relatively heavy labelling when [3H]-AMPA (quisqualate) or [3H]-pentagastrin (CCK) were used as ligands. The superficial layers of the cortex were zones of relative concentration of GABA, benzodiazepine, acetylcholine, glutamate-related, and adenosine receptors. In general, the binding patterns resembled those previously described for cat visual cortex, but there were also some clear differences. The distributions of all of these receptors likely reflect the differential input substances to different laminae of the visual cortex. Of the receptors examined, only those for GABA, benzodiazepine, and acetylcholine were found in substantial concentration in the LGN. Of these, GABA and benzodiazepine receptors showed especially dense binding in the magnocellular layers of the LGN compared to the parvicellular layers.

Animals

The laminar distributions and postnatal development of neurotransmitter and neuromodulator receptors in cat visual cortex.

We review efforts to further understand the development and nature of sensory processing mechanisms in the cat visual cortex. In vitro autoradiographic and homogenate assay techniques have been employed to determine the laminar distribution and characteristics of various neurotransmitter and neuromodulator receptor populations during postnatal development. Each receptor population shows a distinct laminar-specific pattern of binding, which, in most cases, is age-dependent. Changes in receptor number and affinity are also observed during postnatal development. These findings indicate that major alterations in the basic chemical circuitry of cat visual cortex are a normal feature of postnatal maturation and may play a role in plasticity mechanisms.

Aging

Characterization, distribution, and ontogenesis of adenosine binding sites in cat visual cortex.

In vitro autoradiographic techniques were used to characterize binding sites for 3H-cyclohexyladenosine (CHA) and 3H-5'-N-ethylcarboxamidoadenosine (NECA) in cat and kitten visual cortex. 3H-CHA binding sites in adult cat have a Bmax of 1,363 fmol/mg protein and a Kd of 6.8 nM. Displacement experiments indicate that 3H-CHA binds to an adenosine receptor similar to the A1-adenosine receptor described by other investigators. 3H-NECA binding sites in adult cat have a Bmax of 518 fmol/mg protein and a Kd of 15.4 nM. Displacement experiments do not allow us to identify this binding site unambiguously. Bmax values increase during postnatal development for both binding sites, peaking in adulthood for 3H-CHA and at 30 d for 3H-NECA. Kd values show neither consistent nor significant differences during postnatal development for either binding site. 3H-CHA and 3H-NECA binding sites are concentrated in layers 1-3 and upper layer 5 in the visual cortex of adult cats. These laminar patterns, however, change during postnatal development, showing the densest binding in the deep cortical layers (5 and 6) in kittens younger than 30 d of age and a fairly homogeneous binding in older kittens before achieving the adult distribution.

Animals