Thimet oligopeptidase (THOP 1) distribution in cane toad (Bufo Marinus, Linnaeus, 1758) brain.

Thimet oligopeptides (THOP 1) is a metal-dependent peptidase involved in the metabolism of neuropeptides and the presentation of peptides via MHC-1. It has been shown to play a role in the regulation of protein-protein interactions and the metabolism of intracellular peptides. THOP 1 is associated with important biological processes such as metabolism and neurodegenerative diseases, among others. The objective of this study is to elucidate the distribution of THOP 1 in the Bufo marinus brain. The analysis of THOP 1 amino acid sequences indicates that they have been conserved throughout evolution, with significant homology observed across various phyla. When comparing amphibians with other species, more than 70% identity can be identified. Immunohistochemistry analysis of the toad's brain has demonstrated that the enzyme has a ubiquitous distribution, consistent with previous findings in mammals. THOP 1 can be found in important areas of the brain, such as bulb, thalamic nuclei, striatum, hypothalamus, and among others. Nonetheless, THOP 1 is consistently localized within the nucleus, a pattern also observed in the rat brain. Therefore, based on these results, the toad appears to be an excellent model for studying the general biology of THOP 1, given the substantial homology of this enzyme with mammals and its similarity in distribution within the brain.

Thalamic reticular nucleus in Caiman crocodilus: Relationship with the dorsal thalamus.

The thalamic reticular nucleus was investigated in one group of crocodilians, Caiman crocodilus. This neuronal aggregate is composed of two parts: a compact portion and a diffuse region made up of scattered cells within the forebrain bundles. In Caiman, both the lateral and medial forebrain bundles project to the telencephalon and the thalamic reticular nucleus is associated with each fiber tract. In the lateral forebrain bundle, the compact area is termed the nucleus of the dorsal peduncle (dorsal peduncular nucleus) while the diffuse part is called the perireticular area. In the medial forebrain bundle, the interstitial nucleus comprises one part of the compact area while another region without a specific neuronal label is also present. Similar to the perireticular cells of the lateral forebrain bundle, scattered cells are also present in the medial forebrain bundle. Morphological features of the thalamic reticular nucleus are revealed with stains for the following: fibers; cells; succinic acid dehydrogenase; and acetylcholinesterase. Regardless of which dorsal thalamic nucleus was injected, a localized region of the thalamic reticular nucleus contained retrogradely labeled cells and anterogradely labeled axons and terminals. This grouping was termed clusters and was felt to represent the densest interconnection between the dorsal thalamus and the reticular nucleus. Using clusters as an index of interconnections, the reticular nucleus was divided into sectors, each of which was associated with a specific dorsal thalamic nucleus. An organization similar to that found in Caiman is present in other sauropsids as well as in mammals. These data suggest that a thalamic reticular nucleus is present in all amniotes and has morphological properties similar to those described in this analysis. Lastly, a hypothesis is presented to explain how the external shape of the reticular nucleus in Caiman might be transformed into the homologous area in a representative bird and mammal.

[Activity of cytochrome oxidase in centers of tectofugal and thalamofugal channels of the visual system of pigeon Columba 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.

Effects of genetic deficiency of cyclooxygenase-1 or cyclooxygenase-2 on functional and histological outcomes following traumatic brain injury in mice.

BACKGROUND: Neuroinflammation contributes to the pathophysiology of acute CNS injury, including traumatic brain injury (TBI). Although prostaglandin lipid mediators of inflammation contribute to a variety of inflammatory responses, their importance in neuroinflammation is not clear. There are conflicting reports as to the efficacy of inhibiting the enzymes required for prostaglandin formation, cyclooxygenase (COX) -1 and COX-2, for improving outcomes following TBI. The purpose of the current study was to determine the role of the COX isoforms in contributing to pathological processes resulting from TBI by utilizing mice deficient in COX-1 or COX-2. RESULTS: Following a mild controlled cortical impact injury, the amount of cortical tissue loss, the level of microglial activation, and the capacity for functional recovery was compared between COX-1-deficient mice or COX-2-deficient mice, and their matching wild-type controls. The deficiency of COX-2 resulted in a minor (6%), although statistically significant, increase in the sparing of cortical tissue following TBI. The deficiency of COX-1 resulted in no detectable effect on cortical tissue loss following TBI. As determined by 3[H]-PK11195 autoradiography, TBI produced a similar increase in microglial activation in multiple brain regions of both COX-1 wild-type and COX-1-deficient mice. In COX-2 wild-type and COX-2-deficient mice, TBI increased 3[H]-PK11195 binding in all brain regions that were analyzed. Following injury, 3[H]-PK11195 binding in the dentate gyrus and CA1 region of the hippocampus was greater in COX-2-deficient mice, as compared to COX-2 wild-type mice. Cognitive assessment was performed in the wild-type, COX-1-deficient and COX-2-deficient mice following 4 days of recovery from TBI. There was no significant cognitive effect that resulted from the deficiency of either COX-1 or COX-2, as determined by acquisition and spatial memory retention testing in a Morris water maze. CONCLUSION: These findings suggest that the deficiency of neither COX-1 nor COX-2 is sufficient to alter cognitive outcomes following TBI in mice.

[Chemical anatomy of the thalamus in the schizophrenia]. / Anatomía química del tálamo en la esquizofrenia.

The thalamus is considered nowadays as a key structure in the whole organization of the cortico-subcortical relationships. In the last few years, there has been a growing interest in analyzing the potential alterations that could occur in this diencephalic structure in certain psychiatric disorders, prominently in the schizophrenia. In this contribution we describe some of the results obtained in various studies focused on the synaptic modifications of the thalamus observed in schizophrenic patients.

Connections of thalamic modulatory centers to the vocal control system of the zebra finch.

The vocal control system of zebra finches shows auditory gating in which neuronal responses to the individual bird's own song vary with behavioral states such as sleep and wakefulness. However, we know neither the source of gating signals nor the anatomical connections that could link the modulatory centers of the brain with the song system. Two of the song-control nuclei in the forebrain, the HVC (used as the proper name) and the interfacial nucleus of the nidopallium, both show auditory gating, and they receive input from the uvaeform nucleus (Uva) in the thalamus. We used a combination of anterograde and retrograde tracing methods to show that the dorsal part of the reticular formation and the medial habenula (MHb) project to the Uva. We also show by choline acetyl transferase immunohistochemistry that the MHb is cholinergic and sends cholinergic fibers to the Uva. Our findings suggest that the Uva might serve as a hub to coordinate neuromodulatory input into the song system.

NADPH-diaphorase-positive cells in the thalamic nuclei and internal capsule in humans.

The nuclei of the dorsal thalamus and reticular nucleus in humans were found to contain separated NADPH-diaphorase (NADPH-d)-positive neurons. Staining of NADPH-d-positive neurons and all their processes, along with previous studies of neurons in the nuclei of the dorsal thalamus based on the Golgi method, allowed the type of these cells to be identified as sparsely branched. The main, densely branched, efferent neurons did not contain NADPH-d. NADPH-d-positive neurons included reticular cells and cells of one of the types of short-axon interneurons. The internal capsule contained large numbers of NADPH-d-positive reticular neurons. NADPH-d-positive neurons were found in contact with vessels. Thus, NADPH-d-positive cells of the dorsal thalamus, reticular nucleus, and internal capsule were evolutionarily more ancient and less structurally complex cells.

[NADPH-diaphorase positive cells of the human thalamic nuclei and internal capsule]. / NADPH-diaforazo-pozitivnye kletki iader talamusa i vnytrennei kapsuly.

Isolated NADPH-diaphorase (NADPH-d)-positive neurons were demonstrated in the nuclei of human dorsal thalamus and nucleus reticularis. Staining of NADPH-d-positive neurons with all their processes and preceding study of neurons of dorsal thalamus using Golgi method enabled the identification of their types and their determination as sparsely-branched cells. Main types of efferent densely-branched neurons had no demonstrable NADPH-d activity. NADPH-d-positive neurons were represented by reticular neurons and by one type of short-axon interneurons. Capsula interna contains numerous NADPH-d-positive reticular neurons. NADPH-d-positive cells forming contacts with blood vessels were found. Thus, NADPH-d-positive cells of dorsal thalamus, reticular nucleus and capsula interna appear to be evolutionally more ancient and structurally less complex.

Region-specific and epileptogenic-dependent expression of six subtypes of alpha2,3-sialyltransferase in the adult mouse brain.

Sialylated glycoconjugates play important roles in various biological functions. The structures are also observed in brains and it has been proposed that sialylation may affect neural plasticity. To clarify the effects of sialylation in the brain, particular neurons that exhibit sialylation should first be determined. Using in situ hybridization, we performed systematic surveys of the localization of mRNAs encoding the six alpha2,3-sialyltransferases (ST3Gal I-VI) in the adult mouse brain with or without physiological stimulation. First, striking region-specific patterns of expression were observed: While ST3Gal II, III, and V mRNAs were in neuronal cells throughout the brain, ST3Gal I, IV, and VI mRNAs were in restricted brain regions. Next, to assess whether the expression of the six mRNAs can be regulated, we examined the effect of kindling epileptogenesis on the six mRNA levels. Of the six subtypes, upregulation in the ST3Gal IV level in the thalamus was most pronounced; the number of ST3Gal IV-expressing neurons in the anterior thalamic nuclei increased from 2% to 21% in a time-dependent manner during epileptogenesis. Western blot analysis evaluated the increase of the end-products in the thalamus. These findings provide a molecular basis to clarify when and where sialylated glycoconjugates function accompanied by neural plasticity.

Deficit of endogenous kynurenic acid in the frontal cortex of rats with a genetic form of absence epilepsy.

The present studies sought to determine the concentrations of endogenous kynurenic acid (KYNA) and to measure the activity of kynurenine aminotransferases (KAT) I and II in the discrete brain regions of 3- and 6-month old WAG/Rij rats, a genetic model of absence epilepsy. Analogues experiments were performed using age-matched ACI rats, which served as a non-epileptic control. The age-dependent increase in KYNA concentration in the frontal cortex of WAG/Rij rats was considerably reduced in comparison to what was found in ACI rats. Consequently, the concentration of KYNA in the frontal cortex of epileptic rats was significantly lower than in non-epileptic controls. There were no such strain differences in other brain regions. The activities of KAT I and II also showed age-dependent increase with an exception for KAT II in the frontal cortex. Our data suggest that selective deficits of endogenous KYNA may account for increased excitability in the frontal cortex, which in turn may lead to the development of spontaneous spike-wave discharges in WAG/Rij rats.

A quantitative histochemistry technique for measuring regional distribution of acetylcholinesterase in the brain using digital scanning densitometry.

Studies of brain acetylcholinesterase (AChE) are traditionally based on biochemical assays, immunoreactivity, and histochemistry. Conventional histochemistry yields rich morphological data from tissue sections but yields quantitative results only with great difficulty. Several histochemical methods developed in recent years, including microdensitometry, microphotometry, and video-based histochemistry, are effective in quantitative and detailed study of AChE in tissue sections. However, they are usually time-consuming. As we report here, we adapted digital scanning densitometry to quantitate AChE histochemical staining in brain sections. The AChE and butyrylcholinesterase (BuChE), as measured by the method, were heterogeneously distributed throughout the brain, results that are consistent with those obtained by biochemical methods. The staining intensity is dependent on section thickness, substrate concentration, and reaction time. The cholinesterase inhibitor methyl paraoxon significantly decreased AChE staining intensity. Furthermore, data acquired from densitometry are similar to those obtained by video-based microscopy or by spectrophotometry. The advantage of the densitometric measurements compared to other quantitative histochemical methods is that it is very rapid while collecting data that are equivalent in quality. Because the digital scanning densitometers provide high quality and sensitive imaging, wide dynamic ranges, and convenient image analysis software, they are very useful tools in quantitative histochemistry.

Size gradients of barreloids in the rat thalamus.

The spatial organization of the anatomical structures along the trigeminal afferent pathway of the rat conserves the topographical order of the receptor sheath: The brainstem barrelettes, thalamic barreloids, and cortical barrels all reflect the arrangement of whiskers across the mystacial pad. Although both the amount of innervation in the mystacial pad and the size of cortical barrels were shown previously to exhibit increasing gradients toward the ventral and caudal whiskers, whether similar gradients existed in the brainstem and thalamus was not known. Here, the authors investigated the size gradients of the barreloids in the ventral posteromedial nucleus of the rat thalamus. Because the angles used to cut the brain were crucial to this study, the optimal cutting angles were determined first for visualization of individual barreloids and of the entire barreloid field. Individual barreloids, arcs, and rows as well as entire barreloid fields were clearly visualized using cytochrome oxidase staining of brain slices that were cut with the optimal cutting angles. For the first five arcs (including straddlers), the length of barreloids increased in the direction of dorsal-to-ventral whiskers and of caudal-to-rostral whiskers. These gradients reveal an inverse relationship between the size of barreloids and whiskers (length and follicle diameter) along arcs and rows. The largest barreloids in the ventral posteromedial nucleus were those that represent whiskers C2-C4, D2-D4, and E2-E4, which are neither the largest nor the most innervated whiskers in the mystacial pad. This implies that the extended representation is not merely a reflection of peripheral innervation biases and probably serves an as yet unknown processing function.

Nitric oxide synthase and glutamate receptor immunoreactivity in the rat spinal trigeminal neurons expressing Fos protein after formalin injection.

Although recent studies implicated glutamate receptors and nitric oxide in nociception, much still needs to be known about their localisation in neurons involved in nociceptive transmission from the orofacial region. In this study, c-fos expression indicated by Fos immunohistochemistry in the caudal spinal trigeminal nucleus induced by subcutaneous injection of formalin into the lateral face of the rat was used as a marker for nociceptive neurons. The study sought to determine whether Fos-positive neurons express nitric oxide synthase, glutamate N-methyl-D-aspartate type receptor subunit 1, and glutamate alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid type receptor subunit 2/3; and whether they project to the thalamus. After formalin injection, many Fos-positive nuclei appeared in the superficial laminae of the ipsilateral trigeminal nucleus. Confocal laser scanning microscope revealed that almost all neurons with Fos immunofluorescent nuclei were colocalised with N-methyl-D-aspartate receptor 1, 94% with glutamate receptor 2/3 and 14% with nitric oxide synthase. Some of them were closely related to neurons labelled by nitric oxide synthase. Lastly, some of the Fos-positive neurons were labelled by tetramethylrhodamine-dextran injected into the trigeminothalamic tract or the thalamic region. The results suggested that activation of N-methyl-D-aspartate receptor 1 and glutamate receptor 2/3 upon glutamate release in response to noxious stimulation to the orofacial region might mediate c-fos expression in neurons involved in nociception. The expression of Fos in the neurons could also be mediated by nitric oxide produced from the same, as well as neighbouring neurons, when nociceptive stimulation persisted. Fos-positive neurons in the spinal trigeminal nucleus may project to the thalamus, relaying orofacial nociception to the higher sensory centre.

Cutaneous receptive field organization in the ventral posterior nucleus of the thalamus in the common marmoset.

The organization of cutaneous receptive fields in the ventroposterior (VP) thalamus of the common marmosets (Callithrix jacchus) was determined from single-unit recordings, and these data were correlated with the cytochrome oxidase (CO) histochemistry of the thalamus in the same animals. Under continuously maintained ketamine anesthesia, the receptive fields of a total of 192 single units were recorded from the right VP thalamus using 2 MOhms glass electrodes. After the receptive fields were mapped, the brains were reacted for CO histochemistry on 50-microm coronal frozen sections through the entire VP thalamus. The majority of units were localized to the CO-reactive regions that define the medial and lateral divisions of VP (VPm and VPl). Apart from the expected finding of the face being represented in VPm and the body in VPl, reconstructing the electrode tracks and unit locations in the histological sections revealed a general association between discrete regions of CO reactivity and the representation of specific body regions. Some low-threshold cutaneous units were apparently localized to VPi (the CO weak regions dorsal, ventral, and interdigitating with, the CO regions of VP). These VPi units were clearly part of the same representational map as the VPl and VPm units. We conclude that the low-threshold cutaneous receptive fields of the marmoset are organized in a single continuous representation of the contralateral body surface, and that this representation can most simply be interpreted as being folded or crumpled into the three-dimensional space of VP thalamus. The folded nature of the body map in VP may be related to the folded nature of VP as revealed by CO histochemistry.

Tyrosine hydroxylase-immunoreactive elements in the human globus pallidus and subthalamic nucleus.

In contrast to the well-established dopaminergic innervation of the neostriatum, the existence of dopaminergic innervation of the subthalamic nucleus and globus pallidus is controversial. In the present study, tyrosine hydroxylase (TH)-immunoreactive elements were observed by light microscopy after antigen retrieval in the subthalamic nucleus and in the internal and external segments of the globus pallidus in postmortem human brain. Small islands of apparent neostriatal tissue with abundant arborization of fine, TH-immunoreactive axons in the vicinity of calbindin-positive small neurons resembling neostriatal medium spiny neurons were present in the external segment of the globus pallidus. Large numbers of medium-large, TH-immunoreactive axons were observed passing above and through the subthalamic nucleus and through both pallidal segments; these are presumed to be axons of passage on their way to the neostriatum. In addition, fine, TH-immunoreactive axons with meandering courses, occasional branches, and irregular outlines, morphologically suggestive of terminal axon arborizations with varicosities, were seen in both pallidal segments, including the ventral pallidum, and the subthalamic nucleus, consistent with a catecholaminergic (probably dopaminergic) innervation of these nuclei. This finding suggests that, in Parkinson's disease and in animal models of this disorder, loss of dopaminergic innervation might contribute to abnormal neuronal activation in these three nuclei.

Neurochemical organization of inferior pulvinar complex in squirrel monkeys and macaques revealed by acetylcholinesterase histochemistry, calbindin and Cat-301 immunostaining, and Wisteria floribunda agglutinin binding.

To investigate whether the inferior pulvinar complex has a common organization in different primates, the chemoarchitecture of the visual thalamus was re-examined in squirrel monkeys (Saimiri sciureus) and macaques (Macaca mulatta). The inferior pulvinar (PI) complex consisted of multiple subdivisions and encompassed the classic PI, and adjacent ventral parts of the lateral and medial pulvinar (PL and PM, respectively). In keeping with nomenclature suggested previously for macaques, the PI subdivisions were termed the posterior, medial, central, lateral, and lateral-shell (PI(P), PI(M), PI(C), PI(L), and PI(L-S)). In both species, PI(P) was intense for calbindin, light for acetylcholinesterase (AChE), and very light for Wisteria floribunda agglutinin (WFA) histochemistry. The PI(M) was calbindin poor, AChE rich, and moderate for WFA. The PI(C) was calbindin intense, lighter for AChE, and exhibited little WFA binding. PI(L) and PI(L-S) contained populations of large calbindin or WFA cells that were more numerous in PI(L-S). Although staining with the monoclonal antibody Cat-301 differed between macaques and squirrel monkeys, the same subdivisions were displayed. Moderately dense, patchy Cat-301 stain was found in PI(M) of macaques, whereas in squirrel monkeys PI(M) was light. Connections of the rostral dorsolateral (DLr) and middle temporal (MT) areas of visual cortex in squirrel monkeys were compared with PI subdivisions revealed by the newer histochemical methods in the same cases. The major connections of DLr were with PI(C) and of MT were with PI(M).

Thalamic and basal forebrain cholinergic connections of the rat posterior parietal cortex.

The thalamic connectivity and basal forebrain cholinergic input to the posterior parietal cortex (PPC) of Long-Evans rats was examined using combined retrograde tracing and immunocytochemical methods. As in previous studies, the PPC could be distinguished by its input from the lateral posterior, lateral dorsal, and posterior nuclei of the thalamus, but not the lateral geniculate nucleus or ventrobasal complex. These nuclei were also observed to receive reciprocal projections from the ipsilateral PPC. Cholinergic neurons innervating the PPC were primarily localized to the substantia innominata/nucleus basalis region. The implications of these data for possible functions of the cholinergic input to PPC are discussed.

Long-range GABAergic projection in a circuit essential for vocal learning.

The anterior forebrain pathway (AFP) in the passerine song system is essential for song learning but not for song production. Several lines of evidence suggest that area X, a major nucleus in the AFP, forms part of the avian striatum. A key feature of striatal projection neurons is that they use the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). Some area X neurons express GABA-like immunoreactivity, but the neurotransmitter phenotype of the projection neurons is largely unknown. To determine whether area X projection neurons are GABAergic, we used immunocytochemistry and confocal microscopy to examine whether these neurons in adult male zebra finches express the GABA synthetic enzyme glutamic acid decarboxylase (GAD). We observed numerous large and small GAD+ somata in area X, and dense GAD+ terminals, but no GAD+ somata in the target of area X, the medial nucleus of the dorsolateral thalamus (DLM). The density of GAD+ terminals in DLM was strongly reduced by ibotenic acid lesions of area X. After tracer injection into the DLM, all of the retrogradely labeled neurons in area X were GAD+. After tracer injection into area X, the vast majority of anterogradely labeled terminals in DLM were GAD+. We conclude that area X neurons projecting to DLM express GAD and are thus likely GABAergic. If this projection is indeed inhibitory, information processing in the AFP is substantially more complicated than previously realized. Moreover, because a GABAergic projection to a thalamic target is reminiscent of pallidal rather than of striatal circuitry, area X may contain both striatal and pallidal components.

Changes in the mitochondrial enzyme activity in striatal projection areas after unilateral excitotoxic striatal lesions: partial restoration by embryonic striatal transplants.

It is well established that the activity of cytochrome oxidase (CO), a mitochondrial enzyme, reflects the long-term, steady-state levels of neuronal activity. The present study investigated the long-term effects of unilateral striatal lesions induced by quinolinic acid on CO activity in primary striatal targets, including the globus pallidus (GP), entopeduncular nucleus (EP), and substantia nigra pars reticulata (SNR) and a secondary striatal projection area, such as subthalamic nucleus (STN), in rats. The activity of CO was determined by measuring staining intensity on brain sections processed for CO histochemistry. We also examined whether intrastriatal transplants of embryonic striatal tissue could affect the lesion-induced changes in the CO activity of those brain structures. Unilateral striatal lesions were found to lead to increases in the CO activity of the GP, EP, and SNR ipsilateral to the lesions. By contrast, the activity of the ipsilateral STN was decreased following striatal lesions, probably due to the increased inhibitory effect of the GP on the STN. Intrastriatal implantation of the lateral ganglionic eminence (LGN), but not the medial ganglionic eminence (MGE), reversed the lesion-induced changes in the CO activity of the GP and STN with concomitant attenuation of apomorphine-induced rotational asymmetry. The grafts failed to affect the activity of either the EP or SNR. The present results indicate that striatal lesions induce changes in the functional activity of basal ganglia nuclei and that the LGE grafts placed in the damaged striatum partly reverse the alterations in the functional state of the basal ganglia circuitry.

Afferent projections to the mediodorsal and anterior thalamic nuclei in the cat. Anatomical-clinical correlations.

Afferent projections to the mediodorsal and anterior thalamic nuclei in the cat were studied by means of stereotaxic injection of neuronal tracers (horseradish peroxidase and fluorochromes). Acetylcholinesterase reaction was studied, as well as horseradish peroxidase and NADPH-diaphorase colocation in neuronal bodies which send and receive projections to and from the mediodorsal thalamic nucleus. Based on the connectivity and histochemistry findings, the possibility that prion agents responsible for fatal familial insomnia spread from the mediodorsal and anterior thalamic nuclei through a retrograde pathway is discussed. The possible pathophysiological implication of nitrergic systems in fatal familial insomnia is also considered.