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Biomedical subjects

M Kossut

Publications and source records attributed to M Kossut.

At least 19 recordsLinked to original sources

Voltage-dependent L-type calcium channels in the development and plasticity of mouse barrel cortex.

Entry of calcium ions into the neuron is a triggering signal for initiation of several processes which may lead to modification of synaptic connectivity. The developmental changes of voltage-dependent L-type calcium channel (VDLCC) were studied using [3H]PN 200 110 nifedipine displaceable binding in the barrel cortex of mice, a model structure for studying cortical plasticity. In vitro binding autoradiography was used to examine quantitatively the pattern of [3H]PN 200 110 binding to brains of animals aged from 3 to 70 days. The binding values in the somatosensory cortex rose two-fold in the period examined, reaching a plateau in the 4th postnatal week. The laminar pattern of binding changed during development, with the locus of heaviest labeling shifting from layer IV to II/III in the third postnatal week and thin bands of labeling developing in layers IV and VI. A very faint barrel-like pattern of labeling in the barrel field was observed. Neither this pattern nor the binding values were altered by unilateral neonatal removal of all vibrissal follicles. Saturation studies of binding to crude synaptosomal fractions of cerebral cortex of mice aged 3, 15, 28 and 70 days revealed the presence of a single binding site, with Bmax increasing from 48.7 +/- 5.1 fmol/mg protein at postnatal day 3 to 191.7 +/- 9.6 fmol/mg protein at day 70. No developmental changes in KD values were found. No correlation was found between the critical period for cytoarchitectonic plasticity of the barrels and the time when high values of VDLCC binding were observed.

Aging

Effects of sensory deprivation upon a single cortical vibrissal column: a 2DG study.

Effects of sensory deprivation upon ability of a vibrissa to activate a functional column in the barrel field of somatosensory cortex were examined with 2 deoxyglucose (2DG) autoradiography in the rat. After six weeks of whisker plucking started at birth, and one week of recovery, the cortical column activated by the deprived whisker and labeled with 2DG had a reduced diameter but higher labeling density than the normal vibrissal column. Loss of a peri-barrel labeling zone, possibly due to loss of intracortical activation, is suggested.

Animals

5-HT1 receptors in the structures of visual pathway of normal and monocularly deprived kittens.

The density and pattern of distribution of 5-HT1 receptor sites was examined using quantitative in vitro autoradiography with [3H]5-HT as a ligand in the visual structures of 5 weeks old kittens. One group of animals had normal binocular vision, the other was monocularly deprived during the last three days of life. The density of 5-HT1 receptor sites and the pattern of their distribution in the visual structures showed distinct regional, areal and laminar differences. In the primary visual cortex (area 17) a high labelling density was found as compared with other cortical areas investigated. Three bands of high binding density were observed corresponding to cortical layers II-III, IV c and VI. This pattern distinguished area 17 from other cortical areas investigated. In subcortical visual structures very high labelling was present in the superficial visual layers of superior colliculus, but LGN showed rather weak labelling although the lamination of LGN was also seen in the pattern of distribution of 5-HT1 sites. Neither density nor pattern of 5-HT1 sites in the visual cortex and superior colliculus were affected by 3 days of monocular deprivation. High density of labelling and the distinct pattern of 5-HT1 receptor sites in the primary visual cortex suggest the important role of serotonergic transmission in the modulation of visual afferent input activity.

Animals

Loss of gamma-aminobutyric acid (GABA) immunoreactivity from mouse first somatosensory (SI) cortex following neonatal, but not adult, denervation.

Whisker follicles were surgically ablated (lesioned) on two entire rows (B and C) of the left snout of two groups of Swiss mice, in the first 2 days after birth, and in the second group at 8 weeks of age (adults). Two months after surgery GABA-immunoreactivity (GABA-IR) was examined in tangential sections of the first somatosensory (SI) 'barrel' cortex of these two groups (at which time it was also confirmed that the follicles had not regrown). In the adult-lesioned mice all of the barrels appeared intact and visually similar in both the experimental (right) and control (left) hemispheres. The staining pattern of immunopositive cells and puncta was qualitatively similar to that which we have described previously. However, in the neonatally-lesioned mice barrel rows B and C were not visible in the right hemisphere sections and there was a marked reduction in GABA-IR, with fewer immunopositive cells. Many of those that did show GABA-IR stained only weakly and the puncta were also fewer in number than in the left (control) hemisphere where the GABA-IR pattern and staining intensity was normal.

Aging

The effect of short periods of monocular deprivation on excitatory transmission in the striate cortex of kittens: a current source density analysis.

The goal of this study was to assess changes in synaptic activity in the visual cortex of kittens following brief periods of monocular deprivation. Field potentials evoked by electrical stimulation of both optic nerves were registered in vertical penetrations through areas 17 and 18 of 4-5 week-old kittens which had been monocularly deprived for 2-7 days. In order to assess the laminar distribution of synaptic currents these field potentials were subjected to a current source density analysis. Current source density profiles elicited from the deprived eye differed from those induced from the normal eye in several respects: 1) The amplitudes of the responses showed considerable variation at different recording sites across the tangential dimension of cortex. 2) On the average, sinks and sources were markedly reduced, and this reduction was relatively more pronounced in non-granular than in granular layers. 3) However, in 30% of the tracks the layer IV sink showed no attenuation. It was reduced in 48% of the tracks and completely suppressed in 22% of the tracks. These results indicate that a substantial fraction of the deprived thalamocortical synapses remained functional, but that many of these synaptic events remained subthreshold. The significance of these findings is discussed in the context of recovery processes following reverse suture.

Animals

Neuroscience in Poland.

Earlier this year I tried to locate and count the scientists who may possibly want to join the Polish Neuroscience Society. A first rough estimate indicated about 200-250 scientists, including graduate students, who do research on the nervous system. It seemed worthwhile to attempt to highlight their work so that Polish neuroscience might share in the impetus coming from the historical changes sweeping across Europe.

Forecasting

The binding of somatostatin to the mouse retina is altered by the pearl mutation.

Pearl mutants have a night-blind phenotype and abnormal optokinetic nystagmus. Preliminary results from another study showed that the light responses of retinal ganglion cells of pearl mutant mice were affected by bathing the isolated retina with low (less than 1 nM) concentrations of either somatostatin-14 or -28, whereas the responses in wild-type retinas were affected only by somatostatin-28. In order to test the possibility that these physiological differences resulted from alterations in receptor affinity, we studied the binding of 125I-[Tyr11]-somatostatin-14 and 125I-[Leu8,D-Trp22,Tyr25]-somatostatin-28 to frozen, unfixed sections of eyes of wild-type (C57BL/6J +/+) and congenic pearl mutant (C57BL/6J-pin pe(pin)/pe(pin)) mice. As found previously for wild-type mice, specific binding occurred in 3 maxima in pearl mutants: a broad band extending from the retinal ganglion cell to the inner nuclear layer, a narrow and inconstant band over the outer plexiform layer, and a band over the pigment epithelium and choroid. We quantified the label over the inner plexiform layer and found evidence for a single saturable site in both genotypes. However, several results indicate that somatostatin-14 binds more avidly to pearl mutant retinas than to wild-type retinas. In saturation binding studies, Kd for 125I-[Tyr11]-somatostatin-14 was 600 pM in pearl mutants vs 1.5 nM in wild-type; whereas, for 125I-[Leu8,D-Trp22,Tyr25]-somatostatin-28, Kd was nearly equal in the two genotypes (500 and 625 pM, respectively). Bmax was nearly equal for both ligands in both genotypes (63-69 fmol/mg protein).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Cholinergic markers in the plasticity of murine barrel field.

Cholinergic muscarinic receptor binding and acetylcholinesterase (AChE) histochemistry were studied in the barrel cortex of adult, vibrissae deprived and vibrissae denervated mice. In the control barrel field muscarinic receptors labeled with [3H]quinuclidinyl benzilate ([3H]QNB) showed a higher density in the granular cortex and a higher accumulation of label in the barrels. AChE staining revealed a punctuated pattern corresponding to the barrels in the upper part of layer IV and a reverse-image pattern of staining showing only the walls of barrels in the lower part of layer IV. Neonatal denervation of rows of vibrissae lowered both binding of [3H]QNB to the tissue in the shrunken rows and AChE activity in the denervated rows of barrels. Deprivation and late denervation produced no effects on either pattern, or intensity, of [3H]QNB labeling and AChE staining. These observations suggest that the changes in cholinergic markers are related to the altered morphological structure and not to the abnormal functioning of the barrel cortex which received reduced sensory input from the vibrissae.

Acetylcholinesterase

Localization and characterization of somatostatin binding sites in the mouse retina.

We studied the binding of [125I]Tyr11-somatostatin-14 and [125I]Leu8,D-Trp22,Tyr25-somatostatin-28 to frozen, unfixed sections of C57BL/6J mouse eyes with autoradiography. Specific binding of both ligands occurred in 3 maxima, a broad band extending from the retinal ganglion cell to the inner nuclear layers, a narrow and inconstant band over the outer plexiform layer, and a band over the retinal pigment epithelium and choroid. We quantified the label over the inner plexiform layer and found evidence for a single, saturable binding site after Scatchard analysis of saturation binding data. With [125I]Tyr11-somatostatin-14 the dissociation constant (Kd) was 1.48 nM and the total number of binding sites (Bmax) was 68 fmol/mg protein; in competition experiments the inhibitory binding constant (Ki) was 900 pM for somatostatin-14 and 350 pM for somatostatin-28. With [125I]Leu8,D-Trp22,Tyr25-somatostatin-28, Kd was 625 pM and Bmax was 69 fmol/mg protein; in competition experiments Ki was 4.58 nM for somatostatin-14 and 710 pM for somatostatin-28. These results demonstrate the existence of somatostatin receptors in the inner plexiform layer of the retina that appear to have greater specificity for somatostatin-28 than for somatostatin-14.

Animals

Single vibrissal cortical column in SI cortex of rat and its alterations in neonatal and adult vibrissa-deafferented animals: a quantitative 2DG study.

1. A quantitative 2-[14C]deoxy-D-glucose (2DG) autoradiographic study was undertaken to determine the functional (metabolic) organization of an individual column in the first somatosensory cortex (SI) of the awake restrained rat. Manual or mechanically controlled brush stroking of a single vibrissa (typically C3) was used in order to evoke neural activity and increase glucose utilization in a single cortical barrel in lamina IV and in adjacent supra- and infragranular layers in register with the single cortical barrel labeling in lamina IV. 2. The single activated vibrissal column was found to extend from layer I to the superficial half of layer VI of the contralateral SI cortex and was fusiform in shape, being of largest diameter (650-micron) in layer IV and smallest diameter (200 micron) in layer VI. 3. This single metabolic column was not uniform in its density of labeling. The greatest local cerebral metabolic rate of glucose (LCMRG) was found in layer IV (170 mumol.100 g-1.min-1), layers II-III, and V had, on the average, similar LCMRG values (115 mumol.100 g-1.min-1), which decreased with distance from layer IV. 4. The functional (metabolic) columnar activity center situated in layer IV corresponded in size to the appropriate anatomical barrel as determined from thionin-stained sections. Increased metabolic activity was not strictly confined to the candle-pin-shaped column but extended tangentially as radiating fingerlike projections toward the neighboring barrels. 5. In cortical layers other than IV, the metabolic column was less prominently labeled, and the metabolic activity also branched in the tangential plane. The labeling pattern profile in upper layers was characterized by numerous lateral spreading fingerlike projections of increased glucose metabolism. 6. Neonatal follicle lesions sparing only C3 vibrissa caused a great increase (125%) in cortical territory driven by the spared whisker with a concomitant loss of columnar shape and decrease in intensity of metabolic activity in response to its stimulation. 7. Similar lesions in adult rats produced a significant increase (40%) in dimensions of the spared C3 cortical column, which was, however, smaller than that produced after neonatal lesions. The metabolic activation of layer Vb in the spared C3 column was increased (17%) compared with that of the control column.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways

Modifications of the single cortical vibrissal column.

The anatomy, cytoarchitectonics and unit responses of the vibrissal barrel field make it a good subject for studying the central effects of peripheral receptor manipulations. The paper reviews the properties of the barrel system and the evidence for columnar functioning of the barrel cortex. The plastic changes of the single vibrissal column visualized with 2-deoxyglucose autoradiography are described. Cortical plasticity was evoked by vibrissal receptor lesions, deprivation or repetitive stimulation in neonatal and adult rats. The possible processes participating in changing the appearance of the cortical column are discussed. The role of reshaping of intracortical connections within the barrel field is suggested as an important mechanism of plasticity of the vibrissal cortical columns.

Animals

Single vibrissal cortical column in the mouse labeled with 2-deoxyglucose.

Columnar labeling was found in the primary somatosensory cortex of mice after stimulation of a single mystacial vibrissa following 2-deoxyglucose injection. The cortical vibrissal column had a cylindrical shape, passing through all layers of the cortex and was centered upon the appropriate vibrisal barrel. Columnar labeling extended beyond this barrel onto parts of neighbouring barrels, particularly within the same row. The densest labeling was found in layer IV in the barrel hollow. Removal of the non-stimulated vibrissae resulted in a subsequent lowering of 2DG uptake in the barrelfield surrounding the activated column, but did not affect the dimension of the activated column.

Animals

Autoradiographic localization of GABAergic and muscarinic cholinergic receptor sites in the visual system of the kitten.

The distribution of binding sites for [3H] muscimol (which binds to GABA, receptor) and [3H] QNB (which binds to muscarinic cholinergic receptor) was studied in visual areas of 5 week-old kittens using contact film autoradiography after incubation in vitro of brain sections. A distinct laminar localization of [3H] muscimol receptor sites was observed in young animals with the highest density in layers II/ III and IV of all cortical areas that were investigated. For [3H] QNB binding a laminar pattern of distribution was also found with labeling in layer V exceeding that in adjacent layers. A small intracortical regional variability was noted, for both of the investigated neuroreceptors. [3H] muscimol labeling of the lateral geniculate nucleus and superior colliculus was weaker in comparison to that found in the cortex. The superficial layers of the superior colliculus revealed a high density of [3H] muscimol and [3H] QNB binding sites. The laminar and regional variations in the distribution of [3H] muscimol and [3H] QNB binding sites observed in young kittens, may be related to the possible role of GABAergic and muscarinic receptors in regulation of plastic changes that occur after visual deprivation.

Animals

Changes in thalamic projection to the barrelfield of rat after neonatal lesions sparing one whisker.

The thalamic input to the barrelfield of rats with a solitary mystacial vibrissa (C3) intact was examined using succinic dehydrogenase techniques. Rats were lesioned on postnatal day (PD) 1 and brains examined at either 7, 12, 21 or 28 days later. The resultant segmentation in the hemisphere contralateral to the lesioned muzzle consisted of fused rows (A to E). At all ages row C differed from the other rows in that the center of the row was enlarged and separated from the row on either side of it. Thus the presence of a single vibrissa is sufficient to organize the thalamic input to its barrel.

Afferent Pathways

Decrease in the number of synapses formed by subcortical inputs to the striate cortex of binocularly deprived cats.

The density of synapses was determined from electronmicrographs taken from area 17 of cats binocularly deprived of pattern vision for 6 months and in normally reared litter mates. In each cat the optic radiation was transected on one side 4 days before sacrifice and the density of synapses of subcortical origin was estimated by comparing the density of normal synapses remaining on the lesioned side with the density of synapses on the unlesioned side. In normal animals 36% of the synapses were formed by subcortical afferents, but in the binocularly deprived animals this figure was reduced to only 17%. A decrease found in the total synaptic density in the deprived visual cortex was not statistically significant. Thus, binocular deprivation seems to selectively diminish the subcortical contribution to the synaptic density in the visual cortex.

Animals

Effects of sensory denervation and deprivation on a single cortical vibrissal column studied with 2-deoxyglucose.

The plasticity of a single column in the first somatosensory cortex (SI) was studied with [C14] 2-deoxyglucose (2DG) autoradiography. Columnar labelling of cortical representation of the C3 vibrissa was obtained by stimulating this vibrissa after an injection of 2DG. Plastic changes were caused either by surgical denervation (follicle removal) or deprivation (plucking out) of all vibrissae except C3. The development of changes in the columnar profile of the spared vibrissa cortical representation was studied. Both treatments caused an expansion of the spared whisker column into the surrounding barrel field cortex. Denervation produced an immediate enlargement accompanied later by a loss of columnar shape and a decrease of metabolic activity in the core of the column. Effects of deprivation were visible later and were less pronounced, the columnar shape was retained.

Animals