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Nitric oxide synthase in rat visual cortex: an immunohistochemical study.

The aim of the present study was to identify the distribution of two isoforms of the nitric oxide synthase (NOS), the neuronal (nNOS) and the endothelial (eNOS) form, in rat visual cortex. Immunohistochemical localisation of each NOS isoform was studied with three tissue-processing protocols. In the first one, immunohistochemical reactions were made on 30-microm-thick sections with membrane detergents, Triton or Saponin, used to increase the permeability of the tissue for the antibodies. In the second protocol, we excluded these detergents from all solutions to avoid a destruction of the cellular membrane. In the third protocol, we used thin paraffin sections (5 microm thick) to assure delivery of the antibodies to intracellular structures. Our data demonstrate, that both neuronal and endothelial isoforms of the NOS are present in the visual cortex. Among the neurones labelled by the antibodies against eNOS or nNOS, some excitatory cells were definitely present. nNOS immunopositive were neurones and a dense network of fibres, presumably axons. Some of the neurones were heavily labelled in a Golgi-like manner, while others showed only weak labelling. eNOS immunopositivity was found in the blood vessels and in neurones. eNOS positive neurones were much more numerous than nNOS-containing cells, and represent about 60% of the cortical cells. However, with antibodies against eNOS, we never observed neurone-specific cell features. The NOS-containing cells found in our present study represent a possible morphological substrate for production of nitric oxide (NO).

Animals↗

The organization of corticocortical projections from area 17 to area 18 of the cat's visual cortex.

Retrogradely transported tracers were injected into area 18 of the visual cortex of the adult cat to study the organization of corticocortical projections from area 17 to area 18. All injections, whether very small or relatively large, and irrespective of their exact location in area 18, produced a discontinuous, clustered distribution of labelled cells, mainly in layers II, III and upper IV, in a topographically related region of area 17. The mean centre-centre distance between neighbouring patches was about 750 microns. We conclude that the overall population of cells projecting to area 18 is genuinely distributed in a patchy fashion and that they provide an efficient spatial sample of information from area 17. Comparison of the dimensions of each injection site and of the retrogradely labelled territory suggested that each region in area 18 receives a convergent input from a zone in area 17 whose visual field representation is about 0.8 M-1 deg larger in all directions (where M is the magnification factor in millimetres per degree at the termination site in area 18). Pairs of injection were made in area 18 by placing small volumes of two fluorescent tracers, fast blue and diamidino yellow, side-by-side in either a rostrocaudal or a mediolateral plane, with different distances between them. When the boundaries of the dense central cores of two injection sites were separated, at their closest points, by about 1.6 mm, the two corresponding distributions of labelled cells in area 17 were just non-overlapping, suggesting that each group of cells in area 17 sends a divergent projection in innervate a zone about 0.8 mm larger in all directions in area 18. More closely spaced injections led to overlap of the distributions of labelling by the two dyes, with shared clusters containing a mixture of labelled cells. The proportion of double-labelled cells in these shared clusters never exceeded 4.4% (but was 70% after sequential injection of the two dyes at a single point). We conclude that, although each cluster of cells sends a divergent projection to area 18, the majority of individual axons terminate more discretely, perhaps providing specific inter-connections between functionally corresponding 'columns' in the two areas.

Animals↗

Ocular dominance in layer IV of the cat's visual cortex and the effects of monocular deprivation.

1. The relation between the physiological pattern of ocular dominance and the anatomical distribution of geniculocortical afferents serving each eye was studied in layer IV of the primary visual cortex of normal and monocularly deprived cats. 2. One eye was injected with radioactive label. After allowing sufficient time for transeuronal transport, micro-electrode recordings were made, and the geniculocoritcal afferents serving the injected eye were located autoradiographically. 3. In layer IV of normal cats, cell were clustered according to eye preference, and fewer cells were binocularly driven than in other layers. Points of transition between groups of cells dominated by one eye and those dominated by the other were marked with electrolytic lesions. A good correspondence was found between the location of cells dominated by the injected eye and the patches of radioactively labelled geniculocortical afferents. 4. Following prolonged early monocular deprivation, the patches of geniculocortical afferents in layer IV serving the deprived eye were smaller, and those serving the non-deprived eye larger, than normal. Again there was a coincidence between the patches of radioactively labelled afferents and the location of cells dominated by the injected eye. 5. The deprived eye was found to dominate a substantial fraction (22%) of cortical cells in the fourth layer. In other cortical layers, only 7% of the cells were dominated by the deprived eye. 6. These findings suggest that the thalamocortical projection is physically rearranged as a consequence of monocular deprivation, as has been demonstrated for layer IVc of the monkey's visual cortex (Hubel, Wiesel & Le Vay, 1977).

Animals↗

Spatial and temporal analysis by neurons in the representation of the central visual field in the cat's lateral suprasylvian visual cortex.

We studied quantitatively the receptive-field properties of 74 units recorded from the representation of the central visual fields in the cat's lateral suprasylvian (LS) visual cortex. In agreement with previous workers, we found that LS receptive fields tended to be large and to lack discernible spatial structure. They resembled the complex receptive fields of areas 17 and 18 in their general organization. We examined the responses of these neurons to moving optimally oriented sinusoidal gratings that varied in spatial and temporal frequency of drift. Most LS neurons were selective for the spatial frequency of sinusoidal gratings; 7% responded to all spatial frequencies below a cutoff value. In agreement with previous reports, the optimal spatial frequencies for LS neurons covered a wider range than is seen in either area 17 or 18 alone (0.05-1 cycle/deg), but are certainly included in the range covered by both these afferent areas. Individual neurons in LS responded to a range of spatial frequencies broader than is typical for neurons in areas 17 and 18. The effect of varying the drift rate of otherwise optimal gratings was similar in LS to that reported for areas 17 and 18. Most neurons were optimally responsive to drift rates between 0.5 and 4 Hz, and resolved frequencies as high as 10-30 Hz. A few neurons had optima higher than 6 Hz and resolved frequencies in excess of 30 Hz. We conclude that the receptive fields of LS neurons reflect rather closely the properties of their afferents from areas 17 and 18.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Kindling of the visual cortex in cats: comparison with amygdaloid kindling.

Kindling of the primary visual cortex (VC) was compared with that of the amygdala in cats. VC kindling was basically similar to kindling of the amygdala in that daily electrical stimulation can lead to the development of a generalized convulsion in most subjects, a growth of afterdischarges in their configuration and duration, and a reduction of the afterdischarge threshold. The kindling response of the VC differed from that of the amygdala in a number of respects, i.e., a high afterdischarge threshold, a different pattern of behavioral seizure development, an abrupt growth of electroclinical seizures coincident with the onset of a generalized convulsion, an intersubject variability in seizure susceptibility, and a marked seizure instability. In VC kindling the afterdischarge propagation into the amygdala was not observed until the generalized convulsion developed, and the early involvement of afterdischarge was seen in the pulvinar, lateral geniculate body, and superior colliculus. These data suggest that a neural mechanism different from amygdaloid kindling may participate in VC kindling, and that the subcortical structures of the visual system are involved in the preferential pathway for a seizure generalization from the VC.

Amygdala↗

Geniculo-cortical afferents form synaptic contacts with vasoactive intestinal polypeptide (VIP) immunoreactive neurons of the rat visual cortex.

The lateral geniculate nucleus of the rat was injected with the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) to see if geniculo-cortical axons terminate on vasoactive intestinal polypeptide immunoreactive (VIP-IR) neurons of the primary visual cortex. PHA-L-labelled boutons attached to VIP-IR perikarya and dendrites were identified as presynaptic parts of asymmetrical synapses. This geniculo-cortical projection to VIP-IR cells in the visual cortex and comparable findings in the somatosensory cortex suggest that sensory input from specific thalamic nuclei may influence local circuit inhibition and the metabolic state within the cortical domain via VIP-IR neurons.

Animals↗

[Structural differences of spatial-frequency filters in the cat visual cortex].

Sensitivity profiles of receptive fields (RF) of the cat's visual cortex along the width and length of the field were determined. They served as criteria for determining structural differences of the earlier discovered types of spatial-frequency filters. It has been shown that RF with spatial-frequency selectivity in a narrow range of orientations close to the preferable one, have the "one-dimensional" sensitivity profiles in the same range. For RF showing spatial-frequency selectivity along the RF length and width two-dimensional sensitivity profiles along these RF dimensions were revealed. A group of fields was found whose pattern of sensitivity profiles depended on the geometrical properties of images which stipulated the functional changes o RF filtering properties. Possible role of different types of spatial-frequency filters in image processing is discussed.

Animals↗

Induction of LTP and LTD in visual cortex neurones by intracellular tetanization.

Neurones from supragranular layers of rat visual cortex slices were activated by intracellular tetanization (IT) without concomitant presynaptic stimulation. The effect of IT was examined on EPSPs evoked at low stimulation intensity from two subsets of afferents by electrodes positioned in layers II and IV, respectively. In 17 of 23 inputs to 15 cells IT led to changes in EPSP amplitudes which persisted throughout the recording period (from at least 40 min to 3 h). For 10 potentiated inputs (nine cells) and eight depressed inputs (seven cells), EPSP amplitudes measured 30 min after tetanization were 167 +/- 14% and 55 +/- 14% of the pretetanic controls, respectively. In seven cells both inputs changed, in five cases modifications were of the opposite and in two cases of the same polarity.

Animals↗

Experience-dependent development of feedforward and feedback circuits between lower and higher areas of mouse visual cortex.

Using whole cell recordings, we studied excitatory and inhibitory postsynaptic currents (EPSCs, IPSCs) in feedforward (FF) and feedback (FB) circuits between areas V1 and LM (lateromedial) in developing mouse visual cortex. We found that in mice reared with normal visual experience, FF and FB synapses onto layer 2/3 pyramidal neurons develop equal but submaximal strengths whose EPSCs are increased by monocular lid suture. In contrast, the development and experience-dependence of FF- and FB-IPSCs is pathway-specific. The difference develops during the critical period by strengthening FF-IPSCs, while keeping FB-IPSC amplitudes constant. Monocular lid suture increases FB-IPSCs but does not affect FF-IPSCs.

Animals↗

A morphological correlate of synaptic scaling in visual cortex.

We studied the response of dendritic spines in the thalamic-recipient zone of rat visual cortex to simple manipulations of the visual environment. We measured the morphologies of a total of 3824 spines located on the basal dendrites of 60 layer 3 pyramidal cells. As expected from previous studies, we found a significantly lower spine density in dark-reared animals at postnatal day 30 (P30) compared with light-reared controls. Additional analysis revealed that the spines in dark-reared animals were significantly shorter and more bulbous than in light-reared animals. When these two results were combined, we found that the total synaptic area per unit length of dendrite was conserved, compatible with the phenomenon of "synaptic scaling." We also found that the increase in average spine head diameter is reversed by 10 d of light exposure (starting at P20), but surprisingly, the decrease in spine density is not. Thus, not all effects of dark rearing can be reversed by subsequent visual experience, even when the experience occurs during the third postnatal week.

Animals↗

Noradrenergic effects on rat visual cortex: single-cell microiontophoretic studies of alpha-2 adrenergic receptors.

The catecholamine noradrenaline has been proposed to modulate the excitability of cortical neurons, and such a regulation may be mediated by specific adrenergic receptors. We characterized, using electrophysiological recordings, the types of responses of single cells in the rat visual cortex (areas 17 and 18) to the iontophoretic application of adrenergic agents. For the majority of spontaneous and visually-driven cells sampled, noradrenaline decreased the firing frequency, and in some cases of visually-driven cells could increase the signal/noise ratio. These effects were also documented after the application of the alpha-2 adrenergic agonists clonidine and oxymetazoline, and could be reduced or blocked by a previous ejection of the specific alpha-2 antagonist idazoxan. The present study supports a role for alpha-2 adrenoceptors in the modulation of sensory inputs to the visual cortex.

Adrenergic alpha-Agonists↗

Golgi-like staining of visual cortex cells obtained by extracellular biocytin application in vitro.

We report here the application of biocytin (a biotin-lysine complex) as an extracellular tracer in vitro. Biocytin was applied extracellularly, revealing Golgi-like staining of cells in the adult in vitro rat visual cortex. Micropipettes were filled with a solution of 2.3-2.6% biocytin dissolved in 0.05 M Tris buffer, pH 7.4. Biocytin was applied by one of 3 methods: diffusion, pressure injection or drop application. Cell bodies and dendrites around the application site and their efferent axonal processes were stained; dendritic spines were often visible. The injection sites varied in size from a single cell to a diameter of 400 microns. When applied in layer I-III, few filled cells were also seen in layers IV and V, outside the application site. The drop application (5-10 microliters) of biocytin resulted in filling of cells throughout the cortex. The combination of biocytin and the slice preparation was found to be very useful in revealing cell morphology and tracing interlaminar connections in the visual cortex. The advantages of this technique are its ease of application, the precise and restricted injection sites, and Golgi-like morphological detail.

Animals↗

Neurons with perineuronal sulfated proteoglycans in the human visual cortex, with special reference to their reactions to lectins.

The human visual cortex, especially its ganglionic lamina, was found to contain many neurons with perineuronal sulfated proteoglycans which were stained with cationic iron colloid and aldehyde fuchsin. It also contained many neurons with surface glycoproteins labeled with lectin Vicia villosa agglutinin (VVA) or Glycine max agglutinin (SBA). Double staining frequently showed that the neurons stained with cationic iron colloid were not labeled with lectin VVA or SBA. Hyaluronidase and chondroitinase ABC/heparitinase/keratanase digestions eliminated the perineuronal cationic iron colloid reaction, but never interfered with the cell surface lectin labeling. These findings indicate that the cell surface glycoproteins reactive to lectin VVA or SBA are neither structural elements nor adhesive molecules of the proteoglycans. Double staining further demonstrated that in some neurons with perineuronal sulfated proteoglycans, the cytoplasm was labeled with lectin Arachis hypogaea agglutinin (PNA). It was further noticed that the lectin VVA-labeled neurons were not always identical with the neurons labeled with lectin SBA or with lectin PNA.

Colloids↗

Reverse sequencing syllables of spoken words activates primary visual cortex.

Using fMRI, we investigated the neural correlates for sequencing the individual syllables of spoken words in reverse order. The comparison of this task to a control task requiring subjects to repeat identical syllables given acoustically revealed the activation of the primary visual cortex. Because one syllable is generally expressed by one kana character (Japanese phonogram), most subjects used a strategy in which the kana character string corresponding to the word was imagined visually and then read mentally in reverse order to perform the task effectively. Such strategy was not used during a control condition. These results suggest that the primary visual cortex plays a role in the generation of an imagined string.

Acoustic Stimulation↗

MR imaging signal response to sustained stimulation in human visual cortex.

The response of signal intensity to transient (on-off) motor and sensory stimulation has been well studied; however, the dependence of signal response on the duration of stimulus requires further characterization. The objective of this study was to determine the time course of signal response in the human visual cortex to prolonged, sustained stimulation and to examine possible contributory physiologic mechanisms. Nine healthy volunteers underwent magnetic resonance (MR) imaging during sustained visual stimulation with light-proof binocular goggles. With photic stimulation, activation was observed in all subjects as an increase in signal intensity of the visual cortex. With sustained stimulation, a gradual decrease in signal intensity was subsequently observed, with progression toward an apparent steady state. Correlation with positron emission tomographic, MR spectroscopic, and visual evoked-potential data suggests that the initial uncoupling of cerebral blood flow and oxidative metabolism with a neuronal activation burst may represent a transient phenomenon. This quick-response phase may proceed to an equilibrium coupling of flow and oxidative metabolism, with a gradual normalization of venous deoxyhemoglobin levels and signal intensity.

Adult↗

Synaptic connections of an interneuron with axonal arcades in the cat visual cortex.

A single, isolated interneuron with axonal arcades in the cat visual cortex was analysed in detail by both light and electron microscopy. The neuron was impregnated by the Golgi-Kopsch method, gold-toned, and processed for electron microscopy using the ethanolic phosphotungstic acid (PTA) staining method of Bloom & Aghajanian (1968). These methods, in combination, resulted in the successful identification of a large number of synaptic boutons arising from the axon of the cell under study. We examined serially at the electron microscope level 210 boutons of the axonal arborization of the cell. Of these, 152 formed identifiable symmetrical synaptic contacts with a variety of postsynaptic elements. The vast majority of the postsynaptic targets were dendritic profiles, which represented 95.7% of all the synaptic contacts identified. Only one example was observed of two labelled boutons making contacts with the same postsynaptic element; the rest were apparently on different elements. This distribution of synapses, characterized by the lack of convergence, is very similar to that reported by other authors for a certain kind of double bouquet cell which, in turn, shares some morphological features with the neurons with axonal arcades. It is suggested that fine details of the geometry of the axonal arborization of a given cell are an important reflection of the distribution of its synapses.

Animals↗

Scanning electron microscopy of the development of layer I of the human visual cortex (area 17).

The development of layer I of the human visual cortex was studied. Tissues were obtained from 14 aborted and stillborn human fetuses, ranging in age from 13 to 32 fetal weeks. The middle third of the rostrocaudal area 17 above the left calcarine sulcus was selected for observation. The sections were cut perpendicular to the pial surface of the cortex. The specimens were processed for scanning electron microscopy. At 13-16 weeks the Cajal-Retzius neurons were the predominant neuronal elements of layer I, with a small amount of fibers present. In addition, it was found that at 13 weeks the folding plasma membranes of the glial cells formed primary myelin sheaths which wrapped around the nerve cell bodies and processes. Axosomatic and axodendritic contacts were present in the cortices of the fetuses. The nerve fibers of layer I increased progressively with age. However, at 17 weeks, some degenerating apical dendrites were occasionally found. These degenerating dendrites appeared fragmented and beaded. At 21-23 weeks, layer I was well developed with all its essential components present. However, the upper half of the cortical plate was still undifferentiated at this age. At 26-32 weeks, layer I had a prominent plexiform lamina. This lamina was poor in neurons and rich in fibers. In summary, during layer I ontogenesis, there were the following major changes: (1) a change from higher to lower neuronal density: (2) changes in morphology of neurons; (3) initial appearance of synapses early in development; (4) early onset of glial wrapping; (5) presence of degenerating fibers during ontogenesis, and (6) increase in fiber proportion as fetus aged.

Humans↗

[Types of neurons in the visual cortex of the adult and young rat].

The neurons of the visual cortex of adult und juvenile rats (Wistar) were studied with Golgi Rapid and Golgi Kopsch methods at the age of one to twenty days, 4 months and 18 months. The nerve cells were classified according to the form and size of their soma, the branching and surface of their dendrites and their axonal ramification. The neuron types show great similarities with those described in other species. We distinguish pyramidal neurons in L II-L VI, pyramid-like neurons--e.g. horizontally orientated neurons--neurons with a thick basal dendrite and multiangular neurons in L I as well as the spiny multipolar neurons in L IV all showing spiny dendrites and long axons. Neurons with spiny or sparsely spined dendrites and short axons are spine-free multipolar neurons with horizontal axons, sparsely spined polarized neurons with vertical axons, spine-free bipolar neurons with vertical axons and chandelier neurons. In general, all described cell types are nearly undifferentiated in the first postnatal week. The axons are in a more advanced stage than soma and dendrites at this time. During the process of differentiation the soma looses its irregular shape and the somaspines disappear. The immature dendrites show varicosities, which disappear during dendritic lengthening. The dendritic surface becomes either smooth or spiny. The axonal growth is connected with a loss of varicosities and finally with the formation of terminal boutons. At the time of eye-opening all described cell types have reached their differentiated state, but spine density and distal dendritic length have not yet achieved adult values.

Age Factors↗