Effect of acute hypoxia on the intensity of free radical processes in the basal nuclei of the brain, and the rat behaviour in the open field test under conditions of altered photoperiod

The effect of acute hypoxia on the intensity of free radical processes in the basal nuclei (the nucleus caudatus, globus pallidus. nucleus accumbens. amygdaloid complex) of the brain, and the rat behaviour in the open field test has been studied under conditions of altered photoperiod. It has been shown that constant darkness levels the effect of acute hypoxia on the intensity of lipid peroxidation, preserves the activity of superoxide dismutase and catalase at a higher level, lowers the activity of glutathione peroxidase. Under light, the sensitivity of basal nuclei neurons to acute hypoxia is enhanced, the latter being reflected in intensification of lipid peroxidation at the expense of increased formation of dien conjugates. The activity of catalase at that considerably exceeds the level of even intact rats in all the structures. It has been established that an altered photoperiod modulates the effect of acute hypoxia on the parameters of rat's activity in the open field, the character of their change depending on the nature of a photophase change.     

Structural Connectivity of the Developing Human Amygdala

A large corpus of research suggests that there are changes in the manner and degree to which the amygdala supports cognitive and emotional function across development. One possible basis for these developmental differences could be the maturation of amygdalar connections with the rest of the brain. Recent functional connectivity studies support this conclusion, but the structural connectivity of the developing amygdala and its different nuclei remains largely unstudied. We examined age related changes in the DWI connectivity fingerprints of the amygdala to the rest of the brain in 166 individuals of ages 5-30. We also developed a model to predict age based on individual-subject amygdala connectivity, and identified the connections that were most predictive of age. Finally, we segmented the amygdala into its four main nucleus groups, and examined the developmental changes in connectivity for each nucleus. We observed that with age, amygdalar connectivity becomes increasingly sparse and localized. Age related changes were largely localized to the subregions of the amygdala that are implicated in social inference and contextual memory (the basal and lateral nuclei). The central nucleus’ connectivity also showed differences with age but these differences affected fewer target regions than the basal and lateral nuclei. The medial nucleus did not exhibit any age related changes. These findings demonstrate increasing specificity in the connectivity patterns of amygdalar nuclei across age.     

New data on the immunocytochemical localization of thyrotropin-releasing hormone in the rat central nervous system

An antiserum raised against the synthetic tripeptide pyroglutamyl-histidyl-proline (free acid) was used to localize thyrotropin-releasing hormone (TRH) in the rat central nervous system (CNS) by immunocytochemistry. The distribution of TRH-immunoreactive structures was similar to that reported earlier; i.e., most of the TRH-containing perikarya were located in the parvicellular part of the hypothalamic paraventricular nucleus, the suprachiasmatic portion of the preoptic nucleus, the dorsomedial nucleus, the lateral basal hypothalamus, and the raphe nuclei. Several new locations for TRH-immunoreactive neurons were also observed, including the glomerular layer of the olfactory bulb, the anterior olfactory nuclei, the diagonal band of Broca, the septal nuclei, the sexually dimorphic nucleus of the preoptic area, the reticular thalamic nucleus, the lateral reticular nucleus of the medulla oblongata, and the central gray matter of the mesencephalon. Immunoreactive fibers were seen in the median eminence, the organum vasculosum of the lamina terminalis, the lateral septal nucleus, the medial habenula, the dorsal and ventral parabrachial nuclei, the nucleus of the solitary tract, around the motor nuclei of the cranial nerves, the dorsal vagal complex, and in the reticular formation of the brainstem. In the spinal cord, no immunoreactive perikarya were observed. Immunoreactive processes were present in the lateral funiculus of the white matter and in laminae V-X in the gray matter. Dense terminal-like structures were seen around spinal motor neurons. The distribution of TRH-immunoreactive structures in the CNS suggests that TRH functions both as a neuroendocrine regulator in the hypothalamus and as a neurotransmitter or neuromodulator throughout the CNS.     

Proteinase activity of nuclei from various tissues towards endogenous histones]

The proteinase activities of nuclei isolated from tissues differing in their mitotic activities (brain, thymus, liver, ascite lymphoma) towards the histones and non-histone acid -- extractable proteins were studied. The sensitivity of different histone fractions to nuclear proteinase depends on temperature and time of nuclei incubation under conditions providing for complete dissociation of chromatin proteins from DNA (2 M NaCl--5 M urea). The proteinase activity in the brain and thymus nuclei is revealed only under prolonged (43 hrs) incubation of the nuclei at 25 degrees C, which is accompanied by partial proteolysis of histone H1. Histone H4 from brain nuclei and histone H2a from thymus nuclei are preferably degraded. In the nuclei isolated from the mice ascite cell lymphoma NK/ly and from rat liver the enzyme activity is revealed mainly towards the arginine-enriched histones H3 and H4. The proteolysis of the arginine-enriched histones in tumour cell nuclei is more complete. A high sensitivity to proteolysis was observed for non-histone acid-extractable proteins with low electrophoretic mobility, which were found in brain and tumour cell nuclei.     

A light and electron microscopic study of the GABA transporter GAT-3 in the monkey basal ganglia and brainstem

The present study aimed to elucidate the distribution of the GABA transporter GAT-3 in the monkey basal ganglia and brainstem. Very dense GAT-3 immunoreactivity was observed in the medial septum, diagonal band, basal nucleus of Meynert, thalamus, globus pallidus, and substantia nigra. Moderate levels were observed in the subthalamic nucleus, periaqueductal grey, spinal trigeminal and vestibular nuclei. A general light level of staining was observed in the remainder of the brainstem regions, and very light staining was observed in the caudate nucleus and putamen. Electron microscopy showed that GAT-3 immunoreactivity was present in cell bodies with light cytoplasm and dense bundles of glial filaments, and features of astrocytes. Large numbers of astrocytic processes were also labeled in the neuropil. The cell bodies and processes of neurons were unlabeled. Further study is necessary to elucidate GAT-3 expression in neurological conditions, including hyperalgesia and Parkinson's disease.     

In vivo release of transmitters in the cat basal ganglia

The release of transmitters was studied in various structures of the basal ganglia in cats implanted with several push-pull cannulas. Local depolarization enhanced Met-enkephalin release in the globus pallidus. Activation of striatonigral substance P(SP) neutrons stimulated the transmitter release from terminals. Unilateral electrical stimulation of the caudate nucleus evoked GABA release in both substantia nigrae and pallidoentopeduncular nuclei. The unilateral facilitation or interruption of nigral SP transmission modified dopamine (DA) release in the ipsilateral caudate nucleus in contrast, modifications of GABAergic or glycinergic nigral transmissions induced bilateral symmetrical effects, whereas bilateral asymmetrical changes in DA release in the two caudate nuclei were seen during the unilateral modification of nigral DA transmission. Changes in the dendritic release of DA induced changes in serotonin release both in the substantia nigra and in the ipsilateral caudate nucleus. Finally, it will be shown that acetylcholinesterase can be released from the substantia nigra and the caudate nucleus through processes dependent on nerve activity.     

Neuropeptide Y localization in telencephalic and diencephalic structures of the ground squirrel brain

The distribution of neuropeptide Y-immunoreactive (NPY-IR) perikarya, fibers, and terminals was investigated in the brain of two species of hibernatory ground squirrels, Spermophilus tridecemlineatus and S. richardsonii, by means of immunohistochemistry. In the telencephalic and diencephalic structures studied, distinct patterns of NPY-IR were observed which were essentially identical in male and female animals of both species. No differences in amount or distribution of NPY-IR structures were observed between animals which had been in induced hibernation for several months before sacrifice in March/April and those sacrificed one week after their capture in May. In some brain structures (e.g., the hypothalamic arcuate nucleus), IR cell bodies were observed only after pretreatment with colchicine. NPY-IR perikarya and fibers were found in the cerebral cortex, caudate nucleus-putamen, and dorsal part of the lateral septal nucleus. Dense fiber plexuses were seen in the lateral and medial parts of the bed nucleus of the stria terminalis. The numbers of IR perikarya observed in the medial part of the nucleus increased following intraventricular colchicine injections. The accumbens nucleus exhibited few IR cells and many fibers. Claustrum and endopiriform nuclei showed a considerable number of stained cells and fibers that increased in number and staining intensity in colchicine-treated ground squirrels. The induseum griseum showed a small band of IR cell bodies and varicose fibers. Bipolar of multipolar IR cells and varicose fibers were found in the basal nucleus of the amygdala. Dense fiber plexuses as well as IR terminals were seen in the median, medial, and lateral preoptic areas of the hypothalamus. Terminals and relatively few fibers were located in the periventricular, paraventricular, and supraoptic nuclei. The anterior, lateral, dorsomedial, and ventromedial hypothalamic nuclei contained relatively large numbers of terminals and fibers. In the suprachiasmatic nuclei, dense terminals were distributed mainly in the ventromedial subdivision. In the median eminence, immunoreactive terminals were concentrated in the external layer, with fibers predominant in the internal layer. NPY-IR perikarya were observed only in the arcuate nucleus of the hypothalamus and only following colchicine treatment. In the epithalamus (superficial part of the pineal gland and habenular nuclei), varicose fibers appeared mainly in perivascular locations (pineal) or as a dense plexus (habenular nuclei). These results from ground squirrels are discussed in comparison to those obtained in other species and with regard to considerations of the physiological role of NPY.     

Configuration of the magnocellular nuclei in the basal forebrain of the human adult

With the use of gapless series of thick sections (800 microns) stained for lipofuscin pigment as a stable intraneuronal characteristic, it was found in the human brain that the three magnocellular nuclei in the basal forebrain-the medial septal nucleus, the nucleus of the diagonal band (vertical and horizontal limb) and the basal nucleus of Meynert located within the substantia innominata-were tightly connected with each other. A band-like anteromedial subnucleus and a semilunar posterolateral subnucleus could be delineated in the basal nucleus and were found to be embedded in a zone of low cell density, the substantia innominata, which medially blends into the horizontal limb of the diagonal band nucleus. A three-dimensional reconstruction was made to demonstrate the complex shape of the magnocellular basal forebrain nuclei.     

Thiamine-like fibers in the monkey brain: an immunocytochemical study

The distribution of thiamine-immunoreactive structures was studied in the brain of the monkey using an indirect immunoperoxidase technique. Fibers containing thiamine, but no thiamine-immunoreactive cell bodies, were found. The highest density of fibers containing thiamine was observed in the pulvinar nucleus and in the region extending from the pulvinar nucleus to the caudate nucleus. In the mesencephalon, immunoreactive fibers containing thiamine were only found at rostral level close to the medial lemniscus (at the mesencephalic-diencephalic junction). In the thalamus, the distribution of thiamine-immunoreactive structures was more widespread. Thus, immunoreactive fibers were found in nuclei close to the midline (centrum medianum/parafascicular complex), in the ventrolateral thalamus (medial geniculate nucleus, inferior pulvinar nucleus), and in the dorsolateral thalamus (lateral posterior nucleus, pulvinar nucleus). Finally, in the anterior commissure and in the cerebral cortex a low density immunoreactive fibers was visualized. Thus, in the brainstem, no immunoreactive structures were visualized in the medulla oblongata, pons, or in the medial-caudal mesencephalon, and no immunoreactive fibers were observed in the cerebellum, hypothalamus and in the basal ganglia. The present report describes the first visualization and the morphological characteristics (thick, smooth and short, medium or long in length) of the thiamine-immunoreactive fibers in the primate central nervous system using an antiserum directed against this vitamin. The distribution of thiamine-immunoreactive structures in the monkey brain suggests that this vitamin could be involved in several physiological mechanisms.     

Light-induced tyrosine phosphorylation of BIT in the rat suprachiasmatic nucleus

Circadian changes of protein tyrosine phosphorylation in the hypothalamic suprachiasmatic nucleus have been studied using rats maintained under 12-h light/ 12-h dark cycles as well as constant dark conditions. We found that tyrosine phosphorylation of BIT (brain immunoglobulin-like molecule with tyrosine-based activation motifs), a transmembrane glycoprotein of 90-95 kDa, was higher in the light period than in the dark period and was increased after light exposure in the dark period. Similar changes in tyrosine phosphorylation were observed under constant dark conditions, but its amplitude was weaker than that in 12-h light/12-h dark cycles. As the tyrosine-phosphorylated form of BIT is able to bind to the Src homology 2 domain of a protein tyrosine phosphatase, SHP-2, we examined association of these proteins in suprachiasmatic nucleus extracts and found that SHP-2 was coprecipitated with BIT in parallel with its tyrosine phosphorylation. These results suggest that tyrosine phosphorylation of BIT might be involved in light-induced entrainment of the circadian clock.     

Structural basis of frontal cortex control of thalamic and basal nuclei in dogs

Fibrillar degeneration after extirpation of the frontal cortical field F2, central part, was observed in dogs by Nauta--Gygax and Fink--Heimer's methods. Degenerated fibrillae were stated to form two bundles and terminate near the neurons of nonspecific thalamic nuclei: n. ret, MD, pf, sprf, as well as in the neurons of specific thalamic nuclei: n. Vna, cgl, cgm, pul. Degenerated processes of the frontal cortex cells projected to the neurons of basal nuclei: the head of nucleus caudatus, putamen, globus pallidus. In the claustrum degenerative changes were not observed.     

Neuronal pathways involved in the optokinetic head nystagmus of the frog

The morphology of projection neurons in the basal optic nucleus and the pretectal area and the interconnections of these brain regions were studied with the aid of the cobalt-filling technique. It was found that the nucleus-sends long descending axons to the medullary reticular formation. The two basal optic nuclei are reciprocally interconnected and do not give a direct descending pathway. The pretectal nuclei and the basal optic nucleus are also reciprocally coupled. It is supposed that the described pathways mediate commands for horizontal and vertical nystagmic head movements.     

Immunocytochemical localization of growth hormone releasing factor (GHRF)-containing structures in the rat brain using anti-rat GHRF serum

The distribution of growth hormone releasing factor (GHRF) immunoreactive structures in the rat hypothalmus was studied after colchicine treatment with PAP immunocytochemistry in vibratome sections using an antiserum directed to rat hypothalamic GHRF. The majority of the GHRF-immunoreactive cell bodies were found in the arcuate nucleus, the medial perifornical region, and the ventral premammillary nuclei of the hypothalamus. Scattered cells were seen in the lateral basal hypothalamus, the medial and lateral portions of the ventromedial nucleus, and the dorsomedial and paraventricular nuclei. Immunoreactive fibers were observed in all the regions mentioned above. GHRF terminals were located in the central region of the median eminence. In addition, GHRF-immunoreactive neuronal processes were seen in the ventral region of the dorsomedial nucleus, the medial preoptic and suprachiasmatic regions, dorsal portion of the suprachiasmatic nucleus, bed nucleus of the stria terminals and the hypothalamic portion of the stria terminals. The localization of GHRF-immunoreactive terminals in the median eminence reinforces the view that GHRF plays a physiological role in the regulation of pituitary function. In addition, the localization of GHRF-immunoreactive structures in areas not usually considered to project to the median eminence suggest that GHRF may act as a neuromodulator or neurotransmitter.     

Fractionation of nuclei from brain by zonal centrifugation and a study of the ribonucleic acid polymerase activity in the various classes of nuclei

1. The nuclei of the cells of the whole rat brain have been fractionated in a B-XIV zonal rotor with a discontinuous gradient of sucrose. Five fractions were obtained. Zone (I) contained neuronal nuclei (70%) and astrocytic nuclei (23%). Zone (II) contained astrocytic nuclei (81%) and neuronal nuclei (15%). Zone (III) contained astrocytic nuclei (84%) and oligodendrocytic nuclei (15%). Zone (IV) contained oligodendrocytic nuclei (92%) and zone (V) contained only oligodendrocytic nuclei. 2. The content of DNA, RNA and protein per nucleus was determined for each zone. Although the amount of DNA per nucleus is constant (7pg) the RNA varies from 4.5 to 2.5pg/nucleus and the protein from 38 to 17.6pg/nucleus. The neuronal nuclei have the greatest amounts of protein. The oligodendrocytic nuclei have the least content of RNA and protein. 3. The effects of pH, ionic strength, and Mg(2+) and Mn(2+) concentration on the activity of the nuclear system for synthesis in vitro of RNA have been investigated for unfractionated nuclei. From these studies a standard set of conditions for the assay of nuclear RNA polymerase has been established. 4. The activity of the RNA polymerase in each of the zonal fractions has been determined in the presence and in the absence of alpha-amanitin. Zone (II) is the most active, followed by zone (I). The nuclei of zones (IV) and (V) have comparable activity, which is 40% of that of zone (II). 5. The extent of incorporation of each of the four labelled nucleoside triphosphates by the nuclei from each zone has been measured. These values have been used to calculate the base composition of the RNA synthesized in vitro in each class of nucleus. 6. The effect of changes in the condition of assay of RNA polymerase in the different classes of nuclei has been investigated. Significant differences in the response to concentrations of metal ions and ammonium sulphate have been observed. 7. Homopolymer formation in each zone of brain nuclei has been determined. The extent of formation of the four homopolymers roughly parallels the RNA polymerase activity.     

A comparative volumetric analysis of the amygdaloid complex and basolateral division in the human and ape brain

The amygdaloid complex functions to facilitate effective appraisal of the social environment and is an essential component of the neural systems subserving social behavior. Despite its critical role in mediating social interaction, the amygdaloid complex has not attracted the same attention as the isocortex in most evolutionary analyses. We performed a comparative analysis of the amygdaloid complex in the hominoids to address the lack of comparative information available for this structure in the hominoid brain. We demarcated the amygdaloid complex and the three nuclei constituting its basolateral division, the lateral, basal, and accessory basal nuclei, in 12 histological series representing all six hominoid species. The volumes obtained for these areas were subjected to allometric analyses to determine whether any species deviated from expected values based on the other hominoids. Differences between groups were addressed using nonparametric comparisons of means. The human lateral nucleus was larger than predicted for an ape of human brain size and occupied the majority of the basolateral division, whereas the basal nucleus was the largest of the basolateral nuclei in all ape species. In orangutans the amygdala and basolateral division were smaller than in the African apes. While the gorilla had a smaller than predicted lateral nucleus, its basal and accessory basal nuclei were larger than predicted. These differences may reflect volumetric changes occurring in interconnected cortical areas, specifically the temporal lobe and orbitofrontal cortex, which also subserve social behavior and cognition, suggesting that this system may be acted upon in hominoid and hominid evolution.     

The distribution of tyrosine hydroxylase immunoreactivity in the brains of male and female zebra finches

A system of brain nuclei controls song learning and behavior in zebra finches (Poephila guttata). The size of song-control nuclei are much larger in males, which sing, than in females, which do not sing. This study examined the distribution of fibers, terminals, and cell bodies that are immunoreactive for tyrosine hydroxylase (TH) (the rate-limiting enzyme in the synthesis of catecholamines) in song-control nuclei of adult males and females and juvenile males. In addition, the broad pattern of TH staining throughout the brain was described. There was a sex difference in TH immunoreactivity within song-control nuclei: males had light to moderate staining in all three cortical nuclei examined, whereas females had little or no label in corresponding areas [lateral magnocellular nucleus of the anterior neostriatum (IMAN), higher vocal center (HVC), and robust nucleus of the archistriatum (RA)]. The song-control nucleus area X (X), located in the striatum of avian basal ganglia, was more darkly stained than the surrounding striatum only in males; X was not defined by more intense immunoreactivity in females and hence could not be visualized. There were no apparent differences in TH staining in males ranging in age from 50 days to adulthood (>90 days). Outside of the song-control system there were no substantive differences as a function of sex or age in the pattern or intensity of TH labeling. Major areas of telencephalic staining included the striatal region of basal ganglia, which was covered with dense, fine-grained label, and the septum, where cell bodies were encircled by extremely well-labeled thick processes. In the diencephalon, the preoptic area and hypothalamus included a complex pattern of darkly stained somata and fiber and terminal labeling. Darkly stained somata surrounded the pretectal nucleus, and labeled processes ramified throughout the superficial layers of the optic tectum. The midbrain and hindbrain contained a dense plexus of extremely dark cell bodies corresponding to mammalian substantia nigra, adjacent tegmental areas, and locus ceruleus. Labeled hindbrain cells were also seen in the pontine region, around nucleus solitarius, and in the ventrolateral medulla. © 1993 John Wiley & Sons, Inc.     

Neuroglobin, a novel member of the globin family, is expressed in focal regions of the brain.

Hemoproteins are widely distributed among unicellular eukaryotes, plants, and animals. In addition to myoglobin and hemoglobin, a third hemoprotein, neuroglobin, has recently been isolated from vertebrate brain. Although the functional role of this novel member of the globin family remains unclear, neuroglobin contains a heme-binding domain and may participate in diverse processes such as oxygen transport, oxygen storage, nitric oxide detoxification, or modulation of terminal oxidase activity. In this study we utilized in situ hybridization (ISH) and RT-PCR analyses to examine the expression of neuroglobin in the normoxic and hypoxic murine brain. In the normoxic adult mouse, neuroglobin expression was observed in focal regions of the brain, including the lateral tegmental nuclei, the preoptic nucleus, amygdala, locus coeruleus, and nucleus of the solitary tract. Using ISH and RT-PCR techniques, no significant changes in neuroglobin expression in the adult murine brain was observed in response to chronic 10% oxygen. These results support the hypothesis that neuroglobin is a hemoprotein that is expressed in the brain and may have diverse functional roles.     

Basal ganglia influences on the cerebellum of the cat

The changes in firing rate of intracerebellar nuclear neurons following electrical stimulation of the contralateral basal ganglia were investigated in adult cats, in which antidromic activation of cortico-pontine and/or cortico-olivar fibers arising in the area 6 had been excluded by chronic ablation of the motor cortex. Activation of putamen and caudate nucleus induced discharge changes in a low percentage (below 12.5%) of both medial and lateral cerebellar nuclei neurons, while stimulation of globus pallidus and especially of entopeduncular nucleus modified the spontaneous discharge of a greater percent of cells (up to 29%), mainly in the most lateral cerebellar portions. The basal ganglia-induced effects were abolished upon section of the brachium pontis but not of the restiform body. Latency values of the responses, which were predominantly excitatory in nature, suggest the involvement of structures interposed between basal ganglia and precerebellar systems. We postulated that impulses issued by the basal ganglia could reach the cerebellum through a pathway that involves the pedunculopontine nucleus and the nucleus reticularis tegmenti pontis.     

Localization of corticoliberin receptors in the rat brain

Using the in situ hybridization, a distribution of corticotropin-releasing hormone (CRH) receptors CRH-1 and CRH-2 in the rat brain has been studied. Brain levels of CRH-1 mRNA were higher than the those of CRH-2 mRNA. Most intensive CRH-1 gene expression was observed in the forebrain, including neocortex, archicortex, paleocortex and cerebellar cortex. In addition, significant CRH-1 mRNA expression was detected in the red nucleus, pontine nucleus, cochlear nucleus and reticular tegmental nucleus. CRH-2 mRNA was intensively expressed in the olfactory structures, corticomedial amygdala, CA1-CA4 of hippocampus, ventramedial hypothalamus and several modullar nuclei. Moderate CRH-2 mRNA level were seen in the dorsomedial neostriatum. The findings indicate that within the brain two types of CRH receptors are widely expressed. A distinct pattern of CRH-1 and CRH-2 expression in the brain appears to underline the functional specifics of the CRH actions in brain structures.     

Food entrainment modifies the c-Fos expression pattern in brain stem nuclei of rats

When food is restricted to a few hours daily, animals increase their locomotor activity 2-3 h before food access, which has been termed food anticipatory activity. Food entrainment has been linked to the expression of a circadian food-entrained oscillator (FEO) and the anatomic substrate of this oscillator seems to depend on diverse neural systems and peripheral organs. Previously, we have described a differential involvement of hypothalamic nuclei in the food-entrained process. For the food entrainment pathway, the communication between the gastrointestinal system and central nervous system is essential. The visceral synaptic input to the brain stem arrives at the dorsal vagal complex and is transmitted directly from the nucleus of the solitary tract (NST) or via the parabrachial nucleus (PBN) to hypothalamic nuclei and other areas of the forebrain. The present study aims to characterize the response of brain stem structures in food entrainment. The expression of c-Fos immunoreactivity (c-Fos-IR) was used to identify neuronal activation. Present data show an increased c-Fos-IR following meal time in all brain stem nuclei studied. Food-entrained temporal patterns did not persist under fasting conditions, indicating a direct dependence on feeding-elicited signals for this activation. Because NST and PBN exhibited a different and increased response from that expected after a regular meal, we suggest that food entrainment promotes ingestive adaptations that lead to a modified activation in these brain stem nuclei, e.g., stomach distension. Neural information provided by these nuclei to the brain may provide the essential entraining signal for FEO.