Kazuyuki Imamura

Neuropsin regulates an early phase of Schaffer‐collateral long‐term potentiation in the murine hippocampus

We found that neuropsin, an extracellular matrix serine protease, has a regulatory effect on Schaffer‐collateral long‐term potentiation (LTP) in the mouse hippocampus. Bath application of 1–170u2003n m recombinant neuropsin modulated early phase LTP in the Schaffer‐collateral pathway with a ‘bell‐shape’ dose–response curve. The maximum enhancing activity (134% of control LTP) was found at ∼u200a2.5u2003nm. Bath application of a neutralizing antibody against neuropsin in the hippocampal slice resulted in a marked inhibition of the tetanus‐induced early phase of LTP. The in vivo continuous intraventricular infusion of an antisense oligonucleotide against neuropsin significantly reduced the amplitude of the tetanus‐induced early phase of LTP in vitro. Neuropsin did not directly change the N‐methyl d‐aspartate (NMDA) current. Thus, neuropsin appears to act as a regulatory molecule in the early phase of LTP via its proteolytic function on extracellular matrix rather than affecting NMDA receptor‐mediated calcium increase.

Brain activation study by use of positron emission tomography in unanesthetized monkeys

A system for the measurement of brain activity in conscious monkeys by positron emission tomography (PET) was established in the present study. The signal/noise ratio was maximal around 40 s for data acquisition in the PET scan with 15O-labeled water. When the monkey was stimulated by vibration and subtraction images of the data sets from regional cerebral blood flow (rCBF) changes in paired stimulation and control were superimposed on magnetic resonance images obtained from the same specimens, a somatotopic map corresponding to the sites stimulated was clearly demonstrated. Visual stimulation with a photic stimulator activated the corresponding regions of the primary visual cortex. Comparison of the activated sites and extents under the conscious state with those under anesthesia assured that the study is controllable; there was little unpredictable activation due to unlimited subject movement or to psychological effects.

Selective suppression of horizontal propagation in rat visual cortex by norepinephrine

The release of norepinephrine in the cerebral cortex from axon terminals of locus coeruleus neurons was suggested to be involved in the control of attention. Accumulating data indicate that the responses of cortical neurons are varied when norepinephrine is applied iontophoretically in the vicinity of the cells being recorded. However, it is not known how the pattern of excitatory propagation is modified when norepinephrine is applied over a wide area in the visual cortex. By applying optical imaging to rat visuocortical slices, we found a new mode of norepinephrine action; a prominent suppression of the horizontal propagation in layers II/III. This action of norepinephrine was confirmed by the simultaneous recording of field potentials from multiple sites by use of a multi‐electrode dish. Furthermore, our electrophysiological recordings showed that this norepinephrine action is exerted through suppression of excitatory neural transmission and enhancement of inhibitory transmission to the pyramidal neurons in these layers. Because the release of norepinephrine in the visual cortex is regulated by the level of attention, the neural basis of visual attention may relate partially to the suppression of the integration of visual information by norepinephrine resulting in a state‐dependent restructuring of the receptive field.

6R-TETRAHYDROBIOPTERIN PERFUSION ENHANCES DOPAMINE, SEROTONIN, AND GLUTAMATE OUTPUTS IN DIALYSATE FROM RAT STRIATUM AND FRONTAL CORTEX

The effect of 6R-tetrahydrobiopterin (R-THBP) on neurotransmitter release was investigated using in vivo brain microdialysis in urethane-anesthetized rats. Perfusion of 1.0 mM R-THBP enhanced the level of dopamine output in dialysates collected from the striatum and frontal cortex. R-THBP perfusion also increased serotonin (striatum) and glutamate outputs (striatum and frontal cortex). Dopaminergic terminals in the striatum were destroyed unilaterally by continuous infusion of 6-hydroxydopamine (6-OHDA) using an osmotic minipump system. The effect of R-THBP administration on glutamate level was found to be almost completely suppressed in the 6-OHDA-lesioned side of striatum, while in the intact side of striatum the glutamate level in the dialysates responded normally to R-THBP perfusion. These results suggest that R-THBP may play a role in the mechanisms of release of dopamine, serotonin, and glutamate. The functioning of the catecholaminergic system probably mediates the increase in glutamate output due to R-THBP perfusion.

Differential expression of immediate-early genes, c-fos and zif268, in the visual cortex of young rats: effects of a noradrenergic neurotoxin on their expression.

We investigated the expression pattern of two immediate-early genes, zif268 and c-fos, under various visual conditions using immunohistochemical and northern blot analysis in the visual cortex of young rats. The basal expression of c-fos was low and was further reduced by dark rearing that lasted for one week. A marked and transient increase was induced upon visual stimulation applied immediately after dark rearing. Zif268 showed a relatively high basal level. Its expression was reduced by dark rearing of the animals, but returned rapidly to the basal expression level following the introduction of light. Administration of N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine, a selective noradrenergic neurotoxin, suppressed the basal expression of c-fos messenger RNA. The response of c-fos to photo-stimulation was also significantly lower in the visual cortex of N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine-treated young rats. In contrast, no significant change in zif268 expression was detected between normal and N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine-treated animals. These findings suggest that differential expression of these immediate-early genes is involved in the activity-dependent regulation of cortical function. One possibility is that the noradrenergic system controls cortical function, including plasticity, by modifying the expression of c-fos.

Functional anatomy of chemical senses in the alert monkey revealed by positron emission tomography

Functional imaging technique using positron emission tomography (PET) has made it possible to localize functional brain regions in the human brain by detecting changes in regional cerebral blood flow (rCBF). Performing PET studies in the monkey will aid in integrating monkey electrophysiological research with human PET studies. We examined changes in rCBF during olfactory or combined olfactory and gustatory (flavour) stimulation using PET in the alert rhesus monkey. Olfactory or flavour stimulation with acetic acid or apple increased rCBF in the prepyriform area, substantia innominata and amygdala. Besides these areas, flavour stimulation increased rCBF in the anterior insula and frontal operculum, orbitofrontal cortex, inferior frontal gyrus and cerebellum. Apple odour or flavour stimuli increased rCBF in the inferior occipital gyrus in addition to the above areas. These findings suggest that the increases of rCBF in response to neural activities in the primary olfactory and gustatory cortices are detectable by the use of PET. In addition, regions activated by apple stimuli suggest that higher brain function might be detected with PET in the alert monkey.

ROLES OF N-METHYL-d-ASPARTATE RECEPTORS IN OCULAR DOMINANCE PLASTICITY IN DEVELOPING VISUAL CORTEX: RE-EVALUATION

We have re-examined whether N-methyl-D-aspartate receptors play a specific role in experience-dependent plasticity in kitten visual cortex. A specific antagonist of this glutamate receptor subtype, D,L-2-amino-5-phosphonovaleric acid, was directly and continuously infused into kitten striate cortex for one week concurrently with monocular lid suture. In the hemisphere infused with 50 mM antagonist, we found the usual shift in ocular dominance toward the open eye with only a few binocular cells remaining. The changes were accompanied by an extremely high incidence (38%) of abnormal cells lacking orientation selectivity across different ocular dominance groups. In kitten cortex infused with 10 mM antagonist concurrently with monocular deprivation for a week, recording from a drug-affected region near the infusion centre, we again found the U-shaped ocular dominance distribution with the high incidence of non-selective cells. In antagonist-infused, otherwise normal striate cortex of adult cats, we found that the proportion of binocular cells decreased by one-half in two cellular populations: one recorded during the continuous infusion of 10 mM antagonist under general anaesthesia and paralysis, and the other about two days after stopping the infusion. We also established that in vivo concentrations of chronically infused 10 mM antagonist decreased, not near-exponentially, but linearly with increasing distance from the infusion site. Thus, the effects of a directly and continuously infused, concentrated antagonist of N-methyl-D-aspartate receptors on receptive-field properties of visuocortical cells are complex. The present findings strongly suggest that the antagonist effects in the developing cortex may be due primarily to blockade of normal synaptic transmission rather than specific disruption of an experience-dependent mechanism underlying ocular dominance plasticity.

l-Threo-3,4-Dihydroxyphenylserine enhanced ocular dominance plasticity in adult cats

We studied whether ocular dominance plasticity can be restored to the aplastic visual cortex of the adult cat by peripheral administration of L-threo-3,4-dihydroxyphenylserine (L-threo-DOPS), an exogenous precursor of L-noradrenaline (NA). We found that NA output in the visuocortical dialysate was significantly increased by a single administration of L-threo-DOPS (200 mg or 1 g, i.p.). Single unit recordings revealed a significant reduction of binocular cells (binocularity = 0.30) in juvenile cats (7-8 months of age) that had been monocularly deprived for one month in combination with L-threo-DOPS (200 mg/day, per os). These results suggest that peripheral administration of L-threo-DOPS enhances ocular dominance plasticity, presumably through activation of the central noradrenergic system.

Cellular and subcellular localization of alpha-1 adrenoceptors in the rat visual cortex

Noradrenaline is thought to play modulatory roles in a number of physiological, behavioral, and cellular processes. Although many of these modulatory effects are mediated through alpha-1 adrenoceptors, basic knowledge of the cellular and subcellular distributions of these receptors is limited. We investigated the laminar distribution pattern of alpha-1 adrenoceptors in rat visual cortex, using immunohistochemistry at both light and electron microscopic levels. Affinity-purified anti-alpha-1 antibody was confirmed to react only with a single band of about 70-80 kDa in total proteins prepared from rat visual cortex. Alpha-1 adrenoceptors were widely distributed though all cortical layers, but relatively high in density in layers I, II/III, and V. Immunoreactivity was observed in both neuronal perikarya and processes including apical dendrites. In double-labeling experiments with anti-microtubule-associated protein 2, anti-neurofilament, anti-glial fibrillary acidic protein, anti-glutamic acid decarboxylase 65/67, anti-2-3-cyclic nucleotide 3-phosphodiesterase, and anti-tyrosine hydroxylase antibodies, alpha-1 adrenoceptors were found mainly in dendrites and somata of microtubule-associated protein 2-immunopositive neurons. About 20% of alpha-1 adrenoceptors were in GABAergic neurons. A small number of alpha-1 adrenoceptors were also distributed in axons of excitatory neurons, astrocytes, oligodendrocytes and noradrenergic fibers. Using an immunoelectron microscopic technique, numerous regions of alpha-1 adrenoceptor immunoreactivity were found in cell somata, on membranes of dendrites, and in postsynaptic regions. Moreover, a small number of immunoreaction products were also detected in axons and presynaptic sites. These findings provide the first quantitative evidence regarding the cellular and subcellular localization of alpha-1 adrenoceptor immunoreactivity in visual cortex. Moreover, the ultrastructural distribution of alpha-1 adrenoceptor immunoreactivity suggests that alpha-1 adrenoceptors are transported mainly into dendrites and that they exert effects at postsynaptic sites of neurons.

Gliotoxin-induced suppression of ocular dominance plasticity in kitten visual cortex

We studied the role of astrocytes in the regulation of ocular dominance plasticity. A small quantity of 10 microM fluorocitrate (0.2 nmol in 20 microliters) was pressure-injected into the visual cortex of 7-9-week-old kittens (subcortical depth: 0-5 mm, 20 microliters/10 min). Immediately after injection, 1 eye contralateral to the injected cortex was closed for 3 days. Single-unit recordings revealed that the proportion of binocular cells was significantly higher in a region close (approximately 1 mm) to the fluorocitrate injection site than that in a remote region (> 4 mm) within the same hemisphere and that in the opposite hemisphere. The results suggest that reduction of glial functions by fluorocitrate retarded the usual process of shift in ocular dominance of visual cells following monocular deprivation.