High Cytochrome Oxidase Activity Research Articles

Distribution of Calcium-Binding Proteins and Cytochrome Oxidase Activity in the Projective Zone (Wulst) of the Pigeon Thalamofugal Visual Pathway: A Discussion in the Light of Current Concepts on Homology between the Avian Wulst and the Mammalian Striate (Visual) Cortex

This polemic article focuses on the current concepts of the architectonics of the hyperpallial projective zone (Wulst) in the avian thalamofugal visual pathway and the distribution of calcium-binding proteins parvalbumin (PV) and calbindin (CB), as well as cytochrome oxidase (CO) activity, in these brain regions of birds, including our histochemical and immunohistochemical data obtained on the pigeon. In the principal thalamorecipient sensory area of the Wulst, n. interstitialis hyperpallium apicale/hyperpallium intercalatum (IHA/HI), three discrete zones are identified: the two, containing punctate PV-ir neuropil and a densely packed population of mainly small PV-ir stellate and granular cells, and a single zone formed by punctate CB-ir neuropil and mainly CB-ir cells. The IHA/HI area is distinguished by a high CO activity. In the hyperpallium apicale (HA) and hyperpallium densocellulare (HD), PV-ir and CB-ir neurons of varied morphotypes are scattered diffusely throughout neuropil which is moderately immunoreactive to both calcium-binding proteins. These areas are characterized by the presence of large pyramidal-like and multipolar PV-ir and CB-ir neurons with long branched dendrites forming a cellular/fibrous layer in the superficial HA area. It is concluded that the thalamofugal visual pathway in the pigeon has two, PV- and CB-specific, channels. We draw a comparative analysis of the variability of these channels in different avian species, which is supposed to be due to variable ecological and adaptive factors. Within the current idea about the organization and evolutionary origin of the avian Wulst and the mammalian striate (visual) cortex, the homology between these brain structures is discussed.

Calcium-binding proteins and cytochrome oxidase activity in the turtle optic tectum with special reference to the tectofugal visual pathway

Using immunohistochemistry and a tracer technique we investigated the distribution in the optic tectum of turtles (Emys orbicularis and Testudo horsfieldi) of the calcium-binding proteins (CaBPr) parvalbumin (PV), calbindin (CB) and calretinin (CR) before and after labeling of the nucleus rotundus (Rot) with horseradish peroxidase. The optic tectum activity of the cytochrome oxidase (CO) was studied in parallel. In the principal link of the tectofugal visual pathway (central gray layer, SGC) in both chelonian species, the sparse PV-ir as well as CB- and CR-ir neurons were found significantly varying both in number and the intensity of immunoreactivity of their bodies and dendrites. In contrast, the superficial (SGFS) and deeper periventricular (SGP) tectal layers comprised numerous cells immunoreactive to all three CaBPr in different proportions. Only few retrogradely labeled tectorotundal SGC neurons expressed PV, CB or CR. The very large PV-ir neurons in SGC and SAC were not retrogradely labeled; morphologically they matched the efferent neurons with descending projections. SGC neurons of two chelonian species differed in the level of CO activity. Intense immunoreactivity to all three CaBPr and high CO activity were detected in both species in SGFS neuropil with some differences in sublaminar distribution patterns. The peculiarities of the CaBPr and CO activity distribution patterns in different segments of SGC neurons are discussed as related to the laminar organization of the turtle tectum and its retinal innervation. It is suggested that in the projection tectorotundal SGC neurons the CaBPr are concentrated mainly in their distal dendrites that contact retinal afferents in the superficial retinorecipient tectal layer.

An accurate method for the quantification of cytochrome C oxidase in tissue sections

Cytochrome oxidase (COX) is the enzyme that constitutes the last step of the mitochondrial electron transport chain for the production of ATP. Measurement of COX activity can be achieved by histochemistry, thus providing information about the metabolic status of the brain. Brain regions with high metabolism will present high COX activity in histochemistry assays and vice versa. Using histochemistry versus biochemistry to assess COX activity presents the advantage of providing a map of the differences in metabolism in discrete brain regions. Moreover, COX histochemistry allows quantifying the activity of a particular brain region, by converting units of optical density into units of activity. In the present work we have devised a methodology that allows not only quantifying differences in COX activity between groups, but also quantifying the amount of COX present in brain tissue sections, by directly relating optical density (OD) measurements to cytochrome C oxidase concentration, something that traditionally is achieved by the use of western blot. For this purpose we created a set of standards of known concentration of COX that were affixed to a nitrocellulose membrane, and this membrane was incubated together with the tissue sections in which COX activity was assessed. A standard curve was created using a gradient of different concentrations of purified bovine heart cytochrome oxidase (from 2μg to 0.1μg in intervals of 0.25μg). This standard curve allowed us to detect changes in optical density as low as 5%, and relate these OD differences with known concentrations of cytochrome C oxidase.

Activity of cytochrome oxidase in centers of tectofugal and thalamofugal tracts of the visual system of pigeon Columbia livia

By using a histochemical method of determination of activity of cytochrome oxidase (CO), the level of metabolic activity in pigeons has been shown to be higher in centers of the tectofugal visual channel (pretectal nuclei: Pr, SP, SP/IPS, thalamic nucleus Rot, telencephalic entopallidum) than in centers of the thalamofugal visual channel (GLd, visual area of the hyperpallium Wulst). These data agree with the concept of the dominating role of the tectofugal visual channel in organization of the bird everyday behavior. The high CO activity is also characteristic of the mesencephalic structures (EM, isthmus nuclei: IMc, IPc, SLu) modulating transduction of visual information in tectum, Rot and GLd. Similar differences in the metabolic activities between two visual system channels have been shown earlier in reptiles, which indicates the evolutionary conservatism of the tectofugal visual channel among the sauropside amniotes. However, in pigeons the level of the CO activity in some GLd nuclei approaches that in Rot, which allows us to suggest a rise in birds of the role of the thalamofugal channel in processing of information necessary for performance of complex visual functions.

Components of the pigeon tectothalamic visual pathway, revealed with aid of study of cytochrome oxidase activity and immunoreactivity to calcium-binding proteins

Distribution of activity of cytochrome oxidase (CO) and immunoreactivity to parvalbumin (Pv) and calbindin (Cb) was studied in the optic tectum of the pigeon (Columba livia). In the first link of the tectofugal pathway in the central gray layer (SGC = layer 13), small amounts of the CO-active and Pv-immunoreactive (Pv-ir) cellular bodies were revealed in its internal part (sublayer 13b). Some of these neurons located along the SGC lower boundary had long dendritic processes ascending into the superficial tectum layer (SGF). In the retinorecipient SGF sublayers and particularly in neuropil of the sublayers 4 and 7, the high CO activity correlating with Pv-immunoreactivity was found. It is suggested that a great contribution to metabolic activity of these sublayers is made by the largely branching dendritic processes of Pv-ir neurons of sublayer 13b. The projectional neurons SGC located in its external part (sublayer 13a) were CO-inactive and contained Cb. They sent long dendrites into sublayer 5b; in its neuropil, the high density of Cb-immunoreactivity and a moderate CO activity were detected. Thus, the tectal link of the pigeon tectofugal visual channel consists of two components--the Pv-specific, highly metabolically active and the Cb-specific, metabolically less active ones that transduce visual information from different retinorecipient SGF sublayers. The absence of the significant amount of CO-positive bodies of projectional neurons in SGC can be due to that metabolically more active are their dendritic arborizations in the SGF sublayers.

Core-and-belt organisation of the mesencephalic and forebrain auditory centres in turtles: Expression of calcium-binding proteins and metabolic activity

The distribution of immunoreactivity to the calcium-binding proteins parvalbumin, calbindin and calretinin and of cytochrome oxidase activity was studied in the mesencephalic (torus semicircularis), thalamic (nucleus reuniens) and telencephalic (ventromedial part of the anterior dorsal ventricular ridge) auditory centres of two chelonian species Emys orbicularis and Testudo horsfieldi. In the torus semicircularis, the central nucleus (core) showed intense parvalbumin immunoreactivity and high cytochrome oxidase activity, whereas the laminar nucleus (belt) showed low cytochrome oxidase activity and dense calbindin/calretinin immunoreactivity. Within the central nucleus, the central and peripheral areas could be distinguished by a higher density of parvalbumin immunoreactivity and cytochrome oxidase activity in the core than in the peripheral area. In the nucleus reuniens, the dorsal and ventromedial (core) regions showed high cytochrome oxidase activity and immunoreactivity to all three calcium-binding proteins, while its ventrolateral part (belt) was weakly immunoreactive and showed lower cytochrome oxidase activity. In the telencephalic auditory centre, on the other hand, no particular region differed in either immunoreactivity or cytochrome oxidase activity. Our findings provide additional arguments in favour of the hypothesis of a core-and-belt organisation of the auditory sensory centres in non-mammalian amniotes though this organisation is less evident in higher order centres. The data are discussed in terms of the evolution of the auditory system in amniotes.

Metabolic activity of pigeon thalamic and telencephalic auditory centers

Distribution of activity of mitochondrial oxidative enzyme cytochrome oxidase (CO) was studied in the thalamic (Ov) and telencephalic (field L) auditory centers of the pigeon Columbia livia. Different levels of CO activity are found in the core and belt of the centers: the high CO activity in the core of Ov (nCe) and telencephalic field L2 and the much lower or absent in the peripheral regions (Ovl, Ovm, SPO and L1 and L3). Comparison of our data with those of various avian and reptile species confirms the concept of the common plan of rostral auditory centers in sauropsid amniotes by the principle of the center-periphery (core-belt), which is characteristic of the corresponding mammalian centers. The separation of the central and peripheral parts of these centers is better pronounced in birds than in reptiles.

Distribution of metabolic activity (cytochrome oxidase) and immunoreactivity to calcium-binding proteins in the turtle brainstem auditory nuclei

Using histochemical and immunohistochemical techniques, distribution of activity of oxidative mitochondrial enzyme cytochrome oxidase (CO) and of immunoreactivity to calcium-binding proteins has been studied in spiral ganglion and auditory nuclei of brainstem in two turtle species. It has been shown that immunoreactivity to calbindin, parvalbumin, and calretinin in neurons and neuropil of nuclei of cochlear and superior olivary complexes, in nucleus of lateral lemniscus, and in spiral ganglion neurons coincides topographically with the high CO activity. The similarity of the studied metabolic and neurochemical characteristics of these auditory centers in reptiles, birds, and mammals indicates the existence of some common principles of their organization in amniotes in spite of phylogenetic differences and peculiarities of specialization of the auditory system in different species.

Metabolic activity of the thalamic and telencephalic auditory centers of reptiles

329 The lemniscal (classical projection) and extralemniscal parts of mammalian sensory systems, including the auditory system have different levels of metabolic activity. The lemniscal parts have a higher activity of cytochrome oxidase (CO) and an increased ability to accumulate 2-deoxyglucose (2-DG), whereas the extralemniscal parts are characterized by a lower level of metabolic activity [1]. A similar distribution of metabolic activity was observed in the auditory system of birds (in the mesencephalic, thalamic, and telencephalic auditory centers) [2‐4]. Earlier, we showed that the mesencephalic auditory center, torus semicircularis (TS), of some reptiles, such as turtles and lizards, has a higher CO activity in the central nucleus of the lemniscal pathway than in the peripheral extralemniscal part [5]. However, it is not clear whether the metabolic activity in the rostral centers of reptilian auditory system, viz. the auditory relay thalamic nucleus (n. reuniens, Re) and its projection zone in the telencephalon (the ventromedial part of anterior dorsal ventricular ridge, Advr), have selective distribution. In this work, we studied the distribution of the activity of the mitochondrial oxidative enzyme CO in the rostral auditory centers of two turtle species, Testudo horsfieldi and Emys orbicularis. The experiments were performed with four T. horsfieldi and four E. orbicularis. The CO activity was measured by the standard histochemical method [6] with the use of bovine heart cytochrome Σ (Sigma). The CO activity was estimated by the density of histochemical staining of frozen frontal brain slices with a thickness of 50 µ m. Thalamus. The distribution of the CO activity in the Re in both turtle species was similar and heterogeneous. In the rostral level of the thalamus, we found a CO-active band-shaped structure that continuously spread from the ventricle in the lateral direction below the ventral border of the thalamic center of the tectofugal visual system (Rot) and coincided with the location of the n. anterior (nA). The medial part of this structure is a rostral pole of Re (Figs. 1a and 2a). The middle Re level had two areas with a high CO activity. The larger dorsocentral area occupied almost the entire nucleus except for a small ventrolateral part and extended to the somatic nucleus n. ventralis (nV) below the Rot. This area was continuously connected with the rostral bandshaped structure (Figs. 1b and 2b). Like the rostral part of the Re, this area was filled with stained punctuate terminals with sparse CO-positive cells located in the lat

Involvement of the mammillary bodies in spatial working memory revealed by cytochrome oxidase activity

In view of the inconclusive findings relating the nuclei of the mammillary bodies (MB) with spatial memory, we evaluated the oxidative metabolic activity of the medial and lateral nuclei of the mammillary bodies (MB) after training young rats (30 days) of both sexes in the Morris water maze. Different groups were trained in spatial working (WM) or reference memory (RM) tasks, respectively. The corresponding naı̈ve groups swam for the same amount of time as the trained groups but without the escape platform. Control groups were added that had not been manipulated in any way. No sex-related differences were detected in the working memory task although males exhibited better reference memory than females. Cytochrome oxidase (CO) activity, an endogenous metabolic marker for neuronal activity, was measured in all the groups. CO activity increased significantly in both MB nuclei of male and female rats only in the spatial working memory group. In addition, high CO activity in the lateral nucleus of the MB was linearly correlated with lower escape latencies in both sexes after training in the working memory task. No CO activity changes were found in the basolateral amygdala (BL) in any of the experimental groups. This nucleus was used as a control brain region because of its participation in emotional behavior. The results suggest a specific role of the MB nuclei in spatial working memory in both sexes.

Histochemical identification of cortical areas in the auditory region of the human brain.

Despite numerous studies stretching over the last 100 years there is still no general agreement on the number of auditory areas in the human cortex or even how to define them by histological methods. Full definition of these areas will require a combination of functional and histological methods but, by using six complementary histological methods, of which most have been used in the monkey, we provide a clearer description of these areas. The primary auditory area was located on the posteromedial two-thirds of the first transverse temporal (Heschl's) gyrus and was distinguished by a dense band of cytochrome oxidase activity in layer IV and the base of layer III, as well as a relatively thick, pale layer V and VI. Layers V and VI together made up 40% of the cortical thickness. Acetylcholinesterase (AChE)-containing pyramidal cells were sparsely distributed within the primary auditory area. The anterolateral third of Heschl's gyrus did not have a clear band of high cytochrome oxidase activity but contained a moderately high density of AChE-containing pyramidal cells and thus appeared to be part of the auditory belt. Within Heschl's sulcus there was a third area, which had a band of high cytochrome oxidase activity and bands of high parvalbumin immunoreactivity and AChE activity in layer IV. This area appeared to be part of the auditory core. Thus the use of staining methods for cytochrome oxidase, AChE and parvalbumin provided additional information which allowed a clearer definition of auditory areas than Nissl or myelin staining alone. Our results suggest that there are two core areas surrounded by at least six belt areas in the human auditory region.

Comparative Histochemical Study of Cytochrome Oxidase Reactivity in the Rat Superior Colliculus in Norm and after Unilateral Destruction of the Inner Ear Cochlea

A comparative histochemical study of cytochrome oxidase (CO) activity in the superior colliculus (SC) of intact laboratory rats and animals with unilaterally lesioned inner ear cochlea was carried out. It was shown that distribution of CO activity in the SC of intact rats corresponded to its cytoarchitectonic laminar organization had a heterogeneous distribution between layers and homogeneous distribution inside the layers. Unilateral lesion of the inner ear cochlea results in the appearance of alternating columns with low and high CO activity in the medial complex layers and deep gray layer; these columns are directed mediolaterally and differentiated bilaterally. The appearance of the CO columns seems to be due to the spatially arranged organization of afferents coming onto SC from auditory structures.

Identification and localisation of auditory areas in guinea pig cortex.

The organisation of guinea pig auditory cortex was studied by combining histological methods with microelectrode mapping. This allowed the location of seven auditory areas to be determined in relation to the visual and primary somatosensory areas. The auditory areas were identified by single-unit recordings and their borders defined by evoked potential mapping. The visual areas were identified by their relatively high densities of myelinated fibres, while the primary somatosensory cortex was identified by its characteristic barrels of high cytochrome oxidase (CYO) activity in layer IV. The auditory region had moderate levels of CYO and myelin staining. When staining was optimal, there was a clear edge to the moderate CYO activity, which apparently corresponds to the dorsal border of the primary auditory area (AI) and the other core field that lies dorsocaudal to it (DC). Thus the primary somatosensory area and the visual and auditory regions were separated from each other by a region with lower levels of CYO and myelin staining. The ventral borders of AI and DC could not be determined histologically as there were no sharp transitions in the levels of CYO or myelin staining. The two core areas were partially surrounded by belt areas. The dorsorostral belt and most of the belt around DC responded more strongly to broad-band stimuli than pure tones, while the ventrorostral belt, small field and a belt zone ventral to the rostral part of DC responded better to pure tones. Units in the small field (S) typically had higher thresholds and broader tuning to pure tones than AI, while units in the ventrorostral belt typically had longer onset latencies and gave more sustained responses than units in AI.

Patterns of corticocortical, corticotectal, and commissural connections in the opossum visual cortex

Patterns of connections of the visual cortex of the South American opossum, Didelphis aurita, were revealed by using neuronal tracers to identify and characterize visual specializations of the peristriate cortex (PS). The visuotopy of corticotectal connections of the anterolateral portion of PS (PSal) is symmetrical to that of the striate cortex (ST or primary visual area [V1]). Three consecutive bands of commissural connections coincide, respectively, with the ST-PS border, the limit between the caudal and rostral PSal halves (PSc and PSr), and the border of PS with the parietal and temporal cortices. PSc and PSr contain regular commissural rings similar to those present in the peristriate cortex of eutherian mammals. ST projections define in PSc two strings of periodical foci consecutively concentric to V1 and a single focus in PSr. Although they were organized topographically, ascending, descending, and commissural connections between ST and PSal showed a high degree of convergence and divergence. These results conform to the model of a single area homologous to the second visual area (V2) bordering V1. Moreover, they suggest the possibility that PSal includes either one or two additional belt-like areas successively anterior to V2. Along with the finding of alternating bands of high and low cytochrome oxidase activity in PSal, the data further suggest that this region contains modular specializations similar to those of the peristriate cortex of primates and other eutherian mammals. The posterolateral peristriate cortex (PSpl) constitutes another visual area, since it consists of a distinct focus of reciprocal corticocortical and interhemispheric connections and a separate source of corticotectal projections. Finally, a visuomotor function for the orbital cortex is proposed based on its direct projections to optical tectal layers. The close cladistic relationship of opossums to mammalian ancestral forms suggests that the PSal parcelation into belt-like areas that contain modules reflects the primitive organization of the visual cortex. Moreover, a highly diffuse pattern of corticocortical connections may represent a requirement for a brain with few visual areas to perform global processing.

Patterns of corticocortical, corticotectal, and commissural connections in the opossum visual cortex

Patterns of connections of the visual cortex of the South American opossum, Didelphis aurita, were revealed by using neuronal tracers to identify and characterize visual specializations of the peristriate cortex (PS). The visuotopy of corticotectal connections of the anterolateral portion of PS (PSal) is symmetrical to that of the striate cortex (ST or primary visual area [V1]). Three consecutive bands of commissural connections coincide, respectively, with the ST-PS border, the limit between the caudal and rostral PSal halves (PSc and PSr), and the border of PS with the parietal and temporal cortices. PSc and PSr contain regular commissural rings similar to those present in the peristriate cortex of eutherian mammals. ST projections define in PSc two strings of periodical foci consecutively concentric to V1 and a single focus in PSr. Although they were organized topographically, ascending, descending, and commissural connections between ST and PSal showed a high degree of convergence and divergence. These results conform to the model of a single area homologous to the second visual area (V2) bordering V1. Moreover, they suggest the possibility that PSal includes either one or two additional belt-like areas successively anterior to V2. Along with the finding of alternating bands of high and low cytochrome oxidase activity in PSal, the data further suggest that this region contains modular specializations similar to those of the peristriate cortex of primates and other eutherian mammals. The posterolateral peristriate cortex (PSpl) constitutes another visual area, since it consists of a distinct focus of reciprocal corticocortical and interhemispheric connections and a separate source of corticotectal projections. Finally, a visuomotor function for the orbital cortex is proposed based on its direct projections to optical tectal layers. The close cladistic relationship of opossums to mammalian ancestral forms suggests that the PSal parcelation into belt-like areas that contain modules reflects the primitive organization of the visual cortex. Moreover, a highly diffuse pattern of corticocortical connections may represent a requirement for a brain with few visual areas to perform global processing. J. Comp. Neurol. 416:224–244, 2000. © 2000 Wiley-Liss, Inc.

Regional brain variations of cytochrome oxidase activity and motor co-ordination in staggerer mutant mice

A mutant mouse with cerebellar cortical atrophy, staggerer, was examined in tests of motor activity and co-ordination as well as in regional brain metabolism as assessed by cytochrome oxidase activity. Compared with non-ataxic controls, staggerer mutants had inferior performances in the open field, the wooden beam, the wooden edge, and the rotorod tests. An increase in cytochrome oxidase activity in the deep cerebellar nuclei and in some cerebellar efferent regions, such as the lateral vestibular nucleus, the parvicellular red nucleus, and the ventral tegmental area, was found in staggerer mutant mice. Abnormally high cytochrome oxidase activity in the interpositus and the dentate nuclei was linearly correlated with poor performance on the wooden beam and on the rotorod. High cytochrome oxidase activity in the lateral vestibular nucleus was also associated with poor performance on the wooden beam. Moreover, high cytochrome oxidase activity in the fastigial nucleus was associated with poor performance on the wooden beam but with high motor activity in the open field. These results indicate that a lack of innervation of Purkinje cells to the deep cerebellar nuclei is in part the cause of motor co-ordination deficits in staggerer mutant mice.

Nuclear respiratory factor-2 subunit protein: correlation with cytochrome oxydase and regulation by functional activity in the monkey primary visual cortex.

Previous studies have shown that a transcription factor of the Ets family, nuclear respiratory factor 2 (NRF-2), can activate in vitro the gene expression of cytochrome oxidase (CO), a mitochondrial enzyme of oxidative metabolism. The goals of our present study were to determine whether the distribution of NRF-2 alpha subunit proteins correlated with that of CO activity in the macaque monkey visual cortex and whether the level could be perturbed by visual deprivation. We generated polyclonal antibodies specifically against human NRF-2 alpha subunit. In normal monkeys, patterns of NRF-2 alpha distribution resembled closely that of CO activity: 1) NRF-2 alpha immunoreactivity was localized in both nuclei and cytoplasm of neurons, but the levels differed among various laminae; 2) layers IVA, IVC, and VI, which had high CO activity, were labeled more densely by NRF-2 alpha than layers I, IVB, and V, which contained lower levels of both NRF-2 alpha and CO activity; and 3) CO-rich puffs in layers II and III contained a higher level of NRF-2 alpha than CO-poor interpuffs. From 1 day to 7 days after monocular impulse blockade with tetrodotoxin, there was a progressive reduction of NRF-2 alpha in deprived ocular dominance columns, in parallel with decreases in CO activity. These results suggest that local levels of NRF-2 in the monkey visual cortex closely reflect neuronal physiological and metabolic levels revealed by CO activity and that the expression of NRF-2 alpha, like that of CO, is regulated tightly by neural functional activity.

Expression and regulation of NMDA receptor subunit R1 and neuronal nitric oxide synthase in cortical neuronal cultures: correlation with cytochrome oxidase.

Our previous studies showed a differential distribution of the glutamatergic terminals in cytochrome oxidase-rich and -poor regions of the visual cortex. The NMDA type of glutamate receptors have been proposed to be involved in the activation of nitric oxide synthase to produce nitric oxide, the neurotransmitter. In the present study, we hypothesized that the expressions of glutamate receptor, NMDA receptors (NMDAR1) and neuronal nitric oxide synthase (nNOS) were colocalized and were also correlated with that of cytochrome oxidase (CO) in a subset of neurons. We used primary cultures of postnatal rat visual cortical neurons as a model system, so that we could examine both the somatic and dendritic expressions of these neurochemicals in individual neurons. We found a difference in the sequence of developmental expressions of NMDAR1, nNOS, CO, and Na(+)/K(+) ATPase. Triple labeling showed that all nNOS-positive neurons were immunoreactive for NMDAR1, and a subpopulation of them had high CO activity. The expression of NMDAR1 was positively correlated with CO activity. This is consistent with our previous finding that CO activity is strongly governed by excitatory glutamatergic synapses. After 40 hours of depolarizing potassium chloride treatment, CO activity was increased, and NMDAR1and nNOS levels were up-regulated in parallel. One week of tetrodotoxin significantly decreased the expression of NMDAR1, nNOS, and CO activity. Our results demonstrate that NMDA receptors and nNOS do co-exist in a subset of neurons that have high CO activity and their expressions are under the control of neuronal activity.

Plasticity of metabolic whisker maps in somatosensory brainstem and thalamus of mice with neonatal lesions of whisker follicles.

We employed the autoradiographic deoxyglucose method to study metabolic whisker maps of the adult mouse somatosensory brainstem and thalamus after the neonatal removal of left whisker follicles C1, C2 and C3. Left whiskers B1-3 and D1-3 were deflected to metabolically activate the somatosensory pathway. Unoperated mice that were stimulated in the same fashion served as controls. Whisker stimulation resulted in an ipsilateral increase in metabolic activity in the three trigeminal brainstem structures in which the whiskers are represented topologically by segments of high cytochrome oxidase activity, i.e. subnucleus caudalis, subnucleus interpolaris and nucleus principalis. In the two subnuclei of mice with lesions and of controls, there was an increase in metabolic activity of the representations of the deflected whiskers, whereas the metabolic activity of representations A1-3 and E1-3 was low. Apart from these similarities, the metabolic activation of the representations originally representing whiskers C1-3 was remarkably greater in mice with lesions than in controls. This increase reached statistical significance in subnucleus caudalis and approached statistical significance in subnucleus interpolaris. In nucleus principalis the deprived territory was only partially activated and the degree of metabolic activation was less than in the subnuclei. In the thalamic ventrobasal complex of mice with lesions metabolic activity was unpatterned whereas two areas of metabolic activation were distinct in controls. Hence, the removal of whisker follicles in newborn mice resulted in the suppression of localized metabolic responses to whisker stimulation in the thalamus, whereas in the brainstem stimulus-related activity was prominent and the deprived territory became responsive to the stimulation of whisker follicles adjacent to the lesion. Apparently, the modification of the whisker representation at the first synapse of the pathway induces a diminution of localized responsivity in the thalamus.

Cytochrome oxidase 'blobs' and other characteristics of primary visual cortex in a lemuroid primate, Cheirogaleus medius.

We recently obtained the brain of a rare lemuroid primate, Cheirogaleus medius. The brain was not perfused before death, but rather fixed by immersion shortly thereafter. In both flat-mounted and transversely sectioned tissue, we were able to clearly demonstrate periodic zones of high cytochrome oxidase (CO) activity in the primary visual cortex, resembling the so-called 'blobs' described in many other primate species. Our results contrast with a previous report indicating that blobs are absent in Cheirogaleus medius and provide support for the view that blobs are an evolutionary specialization of primate visual cortex that evolved only once, early in primate history. In other aspects of architectonic organization, area V1 of this Cheirogaleus individual closely resembles that of other strepsirhine primates, such as Galago. We were able to identify additional divisions of cortex in this individual, including the middle temporal visual area (MT), auditory cortex, and the primary somatosensory area (S1 or area 3b). These observations indicate that valuable neuroanatomical information can, in favorable cases, be obtained from rare mammalian species that die of natural causes in captivity or which must be euthanized, even though the animals have not been perfused.