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The role of the lateral geniculate nucleus (LGN) in vision has been extensively studied, yet its extraretinal capacities are still being investigated, including its role in arousal from sleep. The β2 nicotinic acetylcholine receptor (nAChR) subunit is involved in the laminal organisation of the LGN with magnocellular (MC) and parvocellular (PC) neurons. Sudden infant death syndrome (SIDS) occurs during a sleep period and, neuropathologically, is associated with increased neuronal cell death and altered nAChRs. A recent qualitative pilot study from our group implicates the possibility of increased neuronal death/apoptosis in the SIDS LGN. The present study used quantitative analysis to report the baseline expression of apoptotic and nAChR subunits α7 and β2 in the PC and MC layers of the LGN, to determine correlations amongst these markers within layers and across layers, and to evaluate changes in the expression of these markers in the LGN of SIDS infants, along with associations with SIDS risk factors, such as age, sex, cigarette smoke exposure, bed-sharing, and presence of an upper respiratory tract infection (URTI). Tissue was immunohistochemically stained for cell death markers of active caspase-3 (Casp-3) and TUNEL, and for the α7 and β2 nAChR subunits. Amongst 43 cases of sudden and unexpected deaths in infancy (SUDI), classifications included explained deaths (eSUDI, n = 9), SIDS I (n = 5) and SIDS II (n = 29). Results indicated a strong correlation of the apoptotic markers and β2 nAChR subunit between the LGN layers, but not across the markers within the layers. Amongst the diagnostic groups, compared to eSUDI, the SIDS II cases had decreased Casp-3 expression while β2 nAChR expression was increased in both PC and MC layers. Amongst the SIDS risk factors, URTI and bed-sharing were associated with changes in neuronal death but not in the α7 and β2 markers. In conclusion, our findings do not support a role for the α7 and β2 nAChRs in apoptotic regulation of the LGN layers during infancy. However, for SIDS victims, an inverse correlation between the changes for markers of apoptosis and the β2 nAChR subunit expression suggests altered LGN function., (© 2023. The Author(s).)

Which neural circuits undergo synaptic changes when an animal learns? Although it is widely accepted that changes in synaptic strength underlie many forms of learning and memory, it remains challenging to connect changes in synaptic strength at specific neural pathways to specific behaviors and memories. Here we introduce SYNPLA (synaptic proximity ligation assay), a synapse-specific, high-throughput, and potentially brain-wide method capable of detecting circuit-specific learning-induced synaptic plasticity., Competing Interests: The authors declare no competing interest.

The basal forebrain provides cholinergic inputs to primary visual cortex (V1) that play a key modulatory role on visual function. While basal forebrain afferents terminate in the infragranular layers of V1, acetylcholine is delivered to more superficial layers through volume transmission. Nevertheless, direct synaptic contact in deep layers 5 and 6 may provide a more immediate effect on V1 modulation. Using helper viruses with cell type specific promoters to target retrograde infection of pseudotyped and genetically modified rabies virus evidence was found for direct synaptic input onto V1 inhibitory neurons. These inputs were similar in number to geniculocortical inputs and, therefore, considered robust. In contrast, while clear evidence for dorsal lateral geniculate nucleus input to V1 excitatory neurons was found, there was no evidence of direct synaptic input from the basal forebrain. These results suggest a direct and more immediate influence of the basal forebrain on local V1 inhibition., (© 2018 Wiley Periodicals, Inc.)

The dorsal striatum has emerged as a key region in sensory-guided, reward-driven decision making. A posterior sub-region of the dorsal striatum, the auditory striatum, receives convergent projections from both auditory thalamus and auditory cortex. How these pathways contribute to auditory striatal activity and function remains largely unknown. Here we show that chemogenetic inhibition of the projections from either the medial geniculate body (MGB) or primary auditory cortex (ACx) to auditory striatum in mice impairs performance in an auditory frequency discrimination task. While recording striatal sound responses, we find that transiently silencing the MGB projection reduced sound responses across a wide-range of frequencies in striatal medium spiny neurons. In contrast, transiently silencing the primary ACx projection diminish sound responses preferentially at the best frequencies in striatal medium spiny neurons. Together, our findings reveal that the MGB projection mainly functions as a gain controller, whereas the primary ACx projection provides tuning information for striatal sound representations.

The ventral lateral geniculate nucleus (LGNv) is a retinorecipient part of the ventral thalamus and in cats, it consists of medial (M), medial intermediate (IM), lateral intermediate (IL), lateral (L), and dorsal (D) subdivisions. These subdivisions can be differentiated not only by their cytoarchitecture, but also by their connectivity and putative functions. The LGNv may play a role in visuomotor gating, in that there is evidence of cerebellar afferent projections to the intermediate subdivisions. The cerebellar posterior interpositus (IP) and lateral (LC) nuclei are known to project to IM and IL, but the specifics of these projections are unclear. We hypothesized that the IP and LC project differentially to IM and IL. To evaluate LGNv innervation by the deep cerebellar nuclei, we injected the tract-tracer wheat germ agglutinin-horseradish peroxidase (WGA-HRP) into several different regions of the LGNv and cerebellar nuclei of adult cats in either sex. Small injections into the middle and posterior LGNv retrogradely labeled cells in the ventral part of the IP. However, injections in the anterior regions of the LGNv, with or without diffusion into the thalamic reticular nucleus (Re), retrogradely labeled cells in the ventral part of both the IP and the LC. Confirmatory injections into the IP and LC produced terminal-like labeling distributed in IM, IL, and Re; injections mostly localized to the LC resulted in labeling mainly in IM and Re. We concluded that the IP projects to IL whereas the LC projects to IM and Re., (© 2017 Wiley Periodicals, Inc.)

To determine whether thalamocortical synaptic circuits differ across cortical areas, we examined the ultrastructure of geniculocortical terminals in the tree shrew striate cortex to compare directly the characteristics of these terminals with those of pulvinocortical terminals (examined previously in the temporal cortex of the same species; Chomsung et al. [] Cereb Cortex 20:997-1011). Tree shrews are considered to represent a prototype of early prosimian primates but are unique in that sublaminae of striate cortex layer IV respond preferentially to light onset (IVa) or offset (IVb). We examined geniculocortical inputs to these two sublayers labeled by tracer or virus injections or an antibody against the type 2 vesicular glutamate antibody (vGLUT2). We found that layer IV geniculocortical terminals, as well as their postsynaptic targets, were significantly larger than pulvinocortical terminals and their postsynaptic targets. In addition, we found that 9-10% of geniculocortical terminals in each sublamina contacted GABAergic interneurons, whereas pulvinocortical terminals were not found to contact any interneurons. Moreover, we found that the majority of geniculocortical terminals in both IVa and IVb contained dendritic protrusions, whereas pulvinocortical terminals do not contain these structures. Finally, we found that synaptopodin, a protein uniquely associated with the spine apparatus, and telencephalin (TLCN, or intercellular adhesion molecule type 5), a protein associated with maturation of dendritic spines, are largely excluded from geniculocortical recipient layers of the striate cortex. Together our results suggest major differences in the synaptic organization of thalamocortical pathways in striate and extrastriate areas., (© 2015 Wiley Periodicals, Inc.)

The primary visual cortex (V1) receives its main thalamic drive from the dorsal lateral geniculate nucleus (dLGN) through synaptic contacts terminating primarily in cortical layer IV. In contrast, the projections from the pulvinar nucleus to the cortex are less clearly defined. The pulvinar projects predominantly to layer I in V1, and layer IV in extrastriate areas. These projection patterns suggest that the pulvinar nucleus most strongly influences (drives) activity in cortical areas beyond V1. Should this hypothesis be true, one would expect the spatiotemporal responses evoked by pulvinar activation to be different in V1 and extrastriate areas, reflecting the different connectivity patterns. We investigated this issue by analyzing the spatiotemporal dynamics of cortical visual areas' activity following thalamic electrical microstimulation in tree shrews, using optical imaging and voltage-sensitive dyes. As expected, electrical stimulation of the dLGN induced fast and local responses in V1, as well as in extrastriate and contralateral cortical areas. In contrast, electrical stimulation of the pulvinar induced fast and local responses in extrastriate areas, followed by weak and diffuse activation in V1 and contralateral cortical areas. This study highlights spatiotemporal cortical activation characteristics induced by stimulation of first (dLGN) and high-order (pulvinar) thalamic nuclei., Significance Statement: The pulvinar nucleus represents the main extrageniculate thalamic visual structure in higher-order mammals, but its exact role remains enigmatic. The pulvinar receive prominent inputs from virtually all visual cortical areas. Cortico-thalamo-cortical pathways through the pulvinar nuclei may then provide a complementary route for corticocortical information flow. One step toward the understanding of the role of transthalamic corticocortical pathways is to determine the nature of the signals transmitted between the cortex and the thalamus. By performing, for the first time, high spatiotemporal mesoscopic imaging on tree shrews (the primate's closest relative) through the combination of voltage-sensitive dye recordings and brain stimulation, we revealed clear evidence of distinct thalamocortical functional connectivity pattern originating from the geniculate nucleus and the pulvinar nuclei., (Copyright © 2015 the authors 0270-6474/15/3511891-06$15.00/0.)

Objective: To explore the changes of lateral geniculate body and visual cortex in monocular strabismus and form deprived amblyopic rat, and visual development plastic stage and visual plasticity in adult rats., Methods: A total of 60 SD rats ages 13 d were randomly divided into A, B, C three groups with 20 in each group, group A was set as the normal control group without any processing, group B was strabismus amblyopic group, using the unilateral extraocular rectus resection to establish the strabismus amblyopia model, group C was monocular form deprivation amblyopia group using unilateral eyelid edge resection + lid suture. At visual developmental early phase (P25), meta phase (P35), late phase (P45) and adult phase (P120), the lateral geniculate body and visual cortex area 17 of five rats in each group were exacted for C-fos Immunocytochemistry. Neuron morphological changes in lateral geniculate body and visual cortex was observed, the positive neurons differences of C-fos expression induced by light stimulation was measured in each group, and the condition of radiation development of P120 amblyopic adult rats was observed., Results: In groups B and C, C-fos positive cells were significantly lower than the control group at P25 (P<0.05), there was no statistical difference of C-fos protein positive cells between group B and group A (P>0.05), C-fos protein positive cells level of group B was significantly lower than that of group A (P<0.05). The binoculus C-fos protein positive cells level of groups B and C were significantly higher than that of control group at P35, P45 and P120 with statistically significant differences (P<0.05)., Conclusions: The increasing of C-fos expression in geniculate body and visual cortex neurons of adult amblyopia suggests the visual cortex neurons exist a certain degree of visual plasticity., (Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.)

We investigated optic nerve and geniculocalcarine tract (GCT) in acquired blindness (AB) using routine cranium magnetic resonance imaging and diffusion tensor imaging. Twenty individuals with AB were compared with 20 normally sighted (NS) individuals. The transverse diameters of optic nerves in NS were significantly bigger than the AB participants in T(1)WI maps. AB participants had higher mean diffusivity and transverse diffusivity and lower fractional anisotropy and primary diffusivity in the optic nerve. This pattern of diffusion change suggests axonal degeneration or atrophy of nerve fibers. No diffusion-index alterations in the GCT were found between AB participants and NS controls. White matter integrity remained normal in the GCT. Thus, the GCT may not rely on visual afferent input to maintain integrity after development., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Endogenous neuropeptides, acting as neurotransmitters or hormones in the brain, carry out important functions including neural plasticity, metabolism and angiogenesis. Previous neuropeptide studies have focused on peptide-rich brain regions such as the striatum or hypothalamus. Here we present an investigation of peptides in the visual system, composed of brain regions that are generally less rich in peptides, with the aim of providing the first broad overview of peptides involved in mammalian visual functions. We target three important parts of the visual system: the primary visual cortex (V1), lateral geniculate nucleus (LGN) and superior colliculus (SC). Our study is performed in the tree shrew, a close relative of primates. Using a combination of data dependent acquisition and targeted LC-MS/MS based neuropeptidomics; we identified a total of 52 peptides from the tree shrew visual system. A total of 26 peptides, for example GAV and neuropeptide K were identified in the visual system for the first time. Out of the total 52 peptides, 27 peptides with high signal-to-noise-ratio (>10) in extracted ion chromatograms (EIC) were subjected to label-free quantitation. We observed generally lower abundance of peptides in the LGN compared to V1 and SC. Consistently, a number of individual peptides showed high abundance in V1 (such as neuropeptide Y or somatostatin 28) and in SC (such as somatostatin 28 AA1-12). This study provides the first in-depth characterization of peptides in the mammalian visual system. These findings now permit the investigation of neuropeptide-regulated mechanisms of visual perception., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The extracellular matrix surrounds different neuronal compartments in the mature nervous system. In a variety of vertebrates, most brain regions are loaded with a distinct type of extracellular matrix around the somatodendritic part of neurons, termed perineuronal nets. The present study reports that chondrotin sulfate proteoglycan-based matrix is structured differently in the human lateral geniculate body. Using various chondrotin sulfate proteoglycan-based extracellular matrix antibodies, we show that perisomatic matrix labeling is rather weak or absent, whereas dendrites are contacted by axonal coats appearing as small, oval structures. Confocal laser scanning microscopy and electron microscopy demonstrated that these typical structures are associated with synaptic loci on dendrites. Using multiple labelings, we show that different chondrotin sulfate proteoglycan components of the extracellular matrix do not associate exclusively with neuronal structures but possibly associate with glial structures as well. Finally, we confirm and extend previous findings in primates that intensity differences of various extracellular matrix markers between magno- and parvocellular layers reflect functional segregation between these layers in the human lateral geniculate body., (Copyright © 2011 Wiley Periodicals, Inc.)

Vesicular glutamate transporters (VGLUTs) reuptake glutamate into synaptic vesicles at excitatory synapses. VGLUT2 is localized in the cortical terminals of neuronal somas located in the main sensory nuclei of the thalamus. Thus, immunolabeling of cortex with antibodies to VGLUT2 can reveal geniculostriate terminal distributions in species in which connectivity cannot be studied with tract-tracing techniques, permitting broader comparative studies of cortical specializations. Here, we used VGLUT2 immunohistochemistry to compare the organization of geniculostriate afferents in primary visual cortex in hominid primates (humans, chimpanzees, and an orangutan), Old World monkeys (rhesus macaques and vervets), and New World monkeys (squirrel monkeys). The New and Old World monkeys had a broad, dense band of terminal-like labeling in cortical layer 4C, a narrow band of labeling in layer 4A, and additional labeling in layers 2/3 and 6, consistent with results from conventional tract-tracing studies in these species. By contrast, although the hominid primates had a prominent layer 4C band, labeling of layer 4A was sparse or absent. Labeling was also present in layers 2/3 and 6, although labeling of layer 6 was weaker in hominids and possibly more individually variable than in Old and New World monkeys. These findings are consistent with previous observations from cytochrome oxidase histochemistry and a very small number of connectivity studies, suggesting that the projections from the parvocellular layers of the lateral geniculate nucleus to layer 4A were strongly reduced or eliminated in humans and apes following their evolutionary divergence from the other anthropoid primates., (Copyright © 2012 S. Karger AG, Basel.)

Cholinergic projections to auditory system are vital for coupling arousal with sound processing. Systematic search with in situ hybridization and immunohistochemistry indicated that the ventral nucleus of the medial geniculate body and the nucleus of the brachium of the inferior colliculus constituted cholinergic synaptic sites in the brainstem auditory system, containing a significant number of cholinergic axon terminals and m2 receptor-expressing cell bodies., ((c) 2010 Wiley-Liss, Inc.)

We studied the distributions of calretinin and calbindin immunoreactivity in subdivisions of the mouse medial geniculate body and the adjacent paralaminar nuclei. We found that the vast majority of labeled cells in the dorsal division of the medial geniculate body were immunoreactive for calbindin-only, whereas most of the remaining labeled cells were double-labeled. Very few calretinin+ only cells were observed. By contrast, we observed significant proportions of calbindin+ only, calretinin+ only and double-labeled cells in the medial division of the medial geniculate body. Further, the distributions of calbindin-only, calretinin-only and double-labeled cells did not differ between the medial division of the medial geniculate body, the suprageniculate nucleus, the peripeduncular nucleus and the posterior intralaminar nucleus. We found essentially no somatic staining for either calbindin or calretinin in the ventral division of the medial geniculate body. These data suggest that there are distinct neurochemical differences between the two non-lemniscal auditory thalamic nuclei. In addition, these data extend previous observations that the medial division of the medial geniculate body shares many properties with the paralaminar group of nuclei.

Purpose: The localization and distribution of neuropeptide expression in the cat visual pathway can provide information about the function of that pathway., Method: Study of optic pathway in eight cats. Following extraction of the brain, slices were prepared using a microkeratome. The slices were examined by indirect immunocytochemistry using anti-metenkephalin as antibody to determine the presence or absence of this pentapeptide in the visual pathway., Results: Met-enkephalin receptors in both cortical and subcortical regions of the brain were detected. This suggests that met-enkephalin could be involved in the visual mechanism., Conclusions: The presence of met-enkephalin receptors in both cortical and subcortical regions of the brain suggests that this pentapeptide could be involved in the visual mechanism.

We investigated the effect of glaucoma (GL) on nerve growth factor (NGF) presence in two brain visual areas. Rats with elevated intraocular pressure (EIOP), induced by hypertonic saline injection in the episcleral vein, were treated with eye topical application of saline or NGF. Rats were subsequently sacrificed, and brain tissues were used for immunohistochemical, biochemical, and molecular analyses. We found that GL alters the basal level of NGF and NGF receptors in brain visual centers and that NGF eye application normalized these deficits. These findings demonstrate that the reduced presence of NGF can arise due to degenerative events in retinal and brain visual areas.

We investigated the distribution pattern of SMI-32-immunopositive cells in the lateral geniculate nucleus (LGN) and in the primary (V1) and middle temporal (MT) cortical visual areas of the adult New World monkey Cebus apella. In the LGN, the reaction for SMI-32 labeled cells in both the magnocellular (M) and parvocellular (P) layers. However, the cellular label was heavier in M layers, which also showed a more intense labeling in the neuropil. In V1, the reaction showed a lamination pattern, with the heaviest labeling occurring in layer 4B and upper layer 6 (layers that project to area MT). Area MT shows a dense band of labeled neuropil and large pyramidal neurons in layer 3, large darkly labeled but less densely packed neurons in layer 5, and a population of small, lightly labeled cells in layer 6. These results resemble those found in other New and Old World monkeys, which suggest that the preferential labeling of projection neurons associated with fast-conducting pathways to the extrastriate dorsal stream is a common characteristic of simian primates. In the superficial layers of V1 in Cebus monkeys, however, SMI-32-labeled neurons are found in both cytochrome oxidase blobs and interblob regions. In this aspect, our results in Cebus are similar to those found in the Old World monkey Macaca and different from those described for squirrel monkey, a smaller New World Monkey. In Cebus, as well as in Macaca, there is no correlation between SMI-32 distribution and the blob pattern.

The visual system encodes and deciphers information using parallel, anatomically segregated, processing streams. To reveal patterns of gene expression in the visual thalamus correlated with physiological processing streams, we designed a custom ferret cDNA microarray. By isolating specific subregions and layers of the thalamus, we identified a set of transcription factors, including Zic2, Islet1, and Six3, the unique distribution profiles of which differentiated the lateral geniculate nucleus (LGN) from the associated perigeniculate nucleus. Within the LGN, odd homeobox1 differentiated the A layers, which contain X cells and Y cells, from the C layers. One neuron-specific protein, Purkinje cell protein 4 (PCP4), was strongly expressed in Y cells in the ferret LGN and in the magnocellular layers of the primate LGN. In the ferret LGN, PCP4 expression began as early as postnatal day 7 (P7), suggesting that Y cells are already specified by P7. These results reveal a rich molecular repertoire that correlates with functional divisions of the LGN.

In most subcortical visual centers in normal mice maintained for a period in the dark, very few neurons express fos-like immunoreactivity (FLI), most likely reflecting c-fos expression, but if an animal is exposed to a flashing light, there is transient increase in the number of FLI-expressing cells. In dark-maintained retinal degeneration (rd) mice, with an inherited photoreceptor defect, numbers of FLI-positive cells, identified immunohistochemically, are anomalously elevated in the superior colliculus (SC) and lateral geniculate nucleus (LGN). Eye removal largely prevents the elevated counts. The difference in number of FLI-positive cells in the SC of rd mice and nondystrophic controls is highly significant (p<0.001). Because we have previously found a similar phenomenon in Royal College of Surgeons (RCS) rats, in which photoreceptor loss is caused by a retinal pigment cell defect, it argues for an effect related to photoreceptor loss rather than its cause.

The pretectal and tectal projections to the dorsal lateral geniculate nucleus (GLd) of two species of turtle (Emys orbicularis and Testudo horsfieldi) were examined under the electron microscope by using axonal tracing techniques (horseradish peroxidase or biotinylated dextran amine) and postembedding gamma-aminobutyric acid (GABA) immunocytochemistry. After injection of tracer into the pretectum, two types of axon terminals were identified as those of pretectogeniculate pathways. Both contained pleomorphic synaptic vesicles and were more numerous in the inner part of the nucleus. They could be distinguished on the bases of size and shape of their synaptic vesicles, type of synaptic contact, and level of GABA immunoreactivity. One type had a higher density of immunolabeling and established symmetric synaptic contacts, whereas the other, less densely immunolabeled, made asymmetric synaptic contacts. In both cases, synaptic contacts were mainly with relay cells and occasionally with interneurons. We suggest that these two types of pretectogeniculate terminals originate in two separate pretectal nuclei. After injection of tracer into the optic tectum, a single population of GABA-immunonegative tracer-labeled terminals was identified as belonging to the tectogeniculate pathway. These were small, had smooth contours, contained very small round synaptic vesicles, and established asymmetric synaptic contacts with long active zones, predominantly with relay cells and less frequently with interneurons, in the inner part of the nucleus. In addition, a population of GABA-negative and occasionally GABA-positive terminals, labeled by tracer injected into either the pretectum or the tectum, was identified as retinal terminals; these were presumably labeled by the retrograde transport of tracer in collateral branches of visual fibers innervating both the GLd and the pretectum or tectum. Comparison of the present ultrastructural findings in turtles with those previously reported in mammals shows that the cytological features, synaptic morphology, and immunochemical properties of the pretectogeniculate and tectogeniculate terminals of both groups share many similarities. Nevertheless, the postsynaptic targets of these two categories of terminals display some pronounced differences between the two groups, which are discussed in terms of their possible functional significance., (Copyright 2004 Wiley-Liss, Inc.)

The pathways ascending through the brain stem to the medial geniculate complex of the thalamus can be distinguished by immunostaining for the calcium binding proteins parvalbumin and calbindin and by the properties of the neurons in the subdivisions of the medial geniculate complex in which they terminate. The parvalbumin pathway, ascending from the central nucleus of the inferior colliculus, is the more direct and terminates in the ventral nucleus. The calbindin pathway is more diffuse in its origins and terminates in the dorsal and medial nuclei. Ventral nucleus neurons are sharply tuned, tonotopically organized and consistent in their responses. They project to core areas of the auditory cortex characterized by high parvalbumin immunoreactivity and by similar neuronal properties. Neurons in the dorsal and medial nuclei are not frequency specific or tonotopic and are labile in their responses. They project more diffusely to belt areas of the auditory cortex in which parvalbumin immunoreactivity is reduced and in which neuronal responses are less specific than in the core. The belt areas are the origins of streams of corticocortical connections leading into the temporal, parietal, and frontal lobes. These routes can be differentially engaged in functional imaging studies of monkeys responding to biologically significant sounds.

A large set of voltage-gated potassium channels is involved in regulating essential aspects of neuronal function in the central nervous system, thus contributing to the ability of neurons to respond to a given input. In the present study, we used immunocytochemical methods to elucidate the regional, cellular and subcellular distribution of the voltage-gated potassium channel subunit Kv1.4, a member of the Shaker subfamily, in the brain. At the light microscopic level, the Kv1.4 subunit showed a unique distribution pattern, being localized in specific neuronal populations of the rat brain. The neuronal regions expressing the highest levels of Kv1.4 protein included the cerebral cortex, the hippocampus, the posterolateral and posteromedial ventral thalamic nuclei, the dorsolateral and medial geniculate nuclei, the substantia nigra and the dorsal cochlear nucleus. The Kv1.4 subunit was also present in other neuronal populations, with different levels of Kv1.4 immunoreactivity. In all immunolabeled regions, the Kv1.4 subunit was mostly diffusely distributed and, to a lesser extent, it stained cell bodies and proximal dendrites. Furthermore, Kv1.4 immunoreactivity was also detected in nerve terminals and axonal terminal fields. At the electron microscopic level, Kv1.4 was located postsynaptically in dendritic spines and shafts at extrasynaptic sites, as well as presynaptically in axon and active zone of axon terminals, in the neocortex and hippocampus. The findings indicate that Kv1.4 channels are widely distributed in the rat brain and suggest that activation of this channel would have different modulatory effects on neuronal excitability.

We compared the ultrastructure and synaptic targets of terminals of cortical or retinal origin in the rat dorsal lateral geniculate nucleus (LGN) and lateral posterior nucleus (LPN). Following injections of biotinylated dextran amine (BDA) into cortical area 17, two types of corticothalamic terminals were labeled by anterograde transport. Type I terminals, found throughout the LGN and LPN, were small, drumstick-shaped terminals that extended from thin axons. At the ultrastructural level in both the LGN and LPN, labeled type I corticothalamic terminals were observed to be small profiles that contained densely packed round vesicles (RS profiles) and contacted small-caliber dendrites. In tissue stained for gamma amino butyric acid (GABA) using postembedding immunocytochemical techniques, most dendrites postsynaptic to type I corticothalamic terminals did not contain GABA (97%). Type II corticothalamic terminals, found only in the LPN, were large terminals that sometimes formed clusters. At the ultrastructural level, type II terminals were large profiles that contained round vesicles (RL profiles) and contacted large-caliber dendrites, most of which did not contain GABA (98%). Retinogeniculate terminals, identified by their distinctive pale mitochondria, were similar to type II corticothalamic terminals except that 26% of their postsynaptic targets were vesicle-containing profiles that contained GABA (F2 profiles). Our results suggest that type I corticothalamic terminals are very similar across nuclei but that the postsynaptic targets of RL profiles vary. Comparison of the responses to retinal inputs in the LGN and to layer V cortical inputs in the LPN may provide a unique opportunity to determine the function of interneurons in the modulation of retinal signals and, in addition, may provide insight into the signals relayed by cortical layer V., (Copyright 2003 Wiley-Liss, Inc.)

We previously established (Datskovskaia et al. [2001] J Comp Neurol 430:85-100) that roughly 40% of Y retinal terminals contact interneurons in the A lamina of the dorsal lateral geniculate nucleus (dLGN) of the cat. However, we did not establish whether the dendritic terminals of interneurons postsynaptic to Y retinal terminals subsequently contact Y thalamocortical cells. To begin to address this issue, we examined the synaptic targets of Y retinal terminals in the magnocellular C lamina of the dLGN, which is populated almost exclusively by Y thalamocortical cells and interneurons. We utilized material generated from our previous work, in which we injected the superior colliculus with biotinylated dextran amine to backfill the geniculate branches of Y retinogeniculate axons in the dLGN. Sections prepared for electron microscopy were stained for gamma aminobutyric acid (GABA) to distinguish interneurons from thalamocortical cells. We found that the majority of profiles postsynaptic to Y retinal axons were the GABA-negative dendrites of thalamocortical cells (116/200, 58%). The remainder were GABA-positive dendrites of interneurons (84/200, 42%), many of which contained vesicles (F2 profiles; 54/200, 27%). In addition, we examined the synaptic targets of F2 profiles and found that almost all contacts of F2 profiles in the magnocellular C lamina were made onto the GABA-negative dendrites of thalamocortical cells (199/200, 99.5%). Thus, Y retinogeniculate axons contact interneurons and interneurons contact Y thalamocortical cells in the magnocellular C lamina of the dLGN. This indicates that interneurons are involved in modulation of the Y pathway., (Copyright 2003 Wiley-Liss, Inc.)

Injections of rhodamine-B into the dorsal raphe nucleus (DRN) and Fluoro-Gold into the lateral geniculate nucleus (LGN) revealed double-labeled retinal ganglion cells (DL RGCs) projecting to both nuclei. The soma-size distribution of DL RGCs was compared with three other distributions: DRN-projecting RGCs, LGN-projecting RGCs, and a large sample of RGCs labeled via the optic nerve with DiI. DL RGC soma diameters fell primarily within the mid-to-upper size range of all three distributions. DL RGCs may provide information to both nuclei concerning comparable aspects of light and visual stimulation via collateralized axons.

The mechanisms of injury- and disease-associated apoptosis of neurons within the CNS are not understood. We used a model of cortical injury in rat and mouse to induce retrograde neuronal apoptosis in thalamus. In this animal model, unilateral ablation of the occipital cortex induces apoptosis of corticopetal projection neurons in the dorsal lateral geniculate nucleus (LGN), by 7 days post-lesion, that is p53 modulated and Bax dependent. We tested the hypothesis that this degenerative process is initiated by oxidative stress and early formation of DNA damage and is accompanied by changes in the levels of pro-apoptotic mediators of cell death. Immunoblotting revealed that the protein profiles of Bax, Bak and Bad were different during the progression of neuronal apoptosis in the LGN. Bax underwent a subcellular redistribution by 1 day post-lesion, while Bak increased later. Bad showed an early sustained increase. Cleaved caspase-3 was elevated maximally at 5 and 6 days. Active caspase-3 underwent a subcellular translocation to the nucleus. A dramatic phosphorylation of p53 was detected at 4 days post-lesion. DNA damage was assessed immunocytochemically as hydroxyl radical adducts (8-hydroxy-2-deoxyguanosine) and single-stranded DNA. Both forms of DNA damage accumulated early in target-deprived LGN neurons. Transgenic overexpression of superoxide dismutase-1 provided significant protection against the apoptosis but antioxidant pharmacotreatments with trolox and ascorbate were ineffective. We conclude that overlapping and sequential signaling pathways are involved in the apoptosis of adult brain neurons and that DNA damage generated by superoxide derivatives is an upstream mechanism for p53-regulated, Bax-dependent apoptosis of target-deprived neurons.

As in other primates, the lateral geniculate nucleus (LGN) of owl monkeys contains three anatomically and physiologically distinct relay cell classes, the magnocellular (M), parvocellular (P), and koniocellular (K) cells. M and P LGN cells send axons to the upper and lower tiers of layer IV, and K cells send axons to the cytochrome oxidase (CO) blobs of layer III and to layer I of primary visual cortex (V1). Our objective was to compare the synaptic arrangements made by these axon classes. M, P, and K axons were labeled in adult owl monkeys by means of injections of wheat germ agglutinin-horseradish peroxidase into the appropriate LGN layers. The neurochemical content of both pre- and postsynaptic profiles were identified by postembedding immunocytochemistry for gamma-aminobutyric acid (GABA) and glutamate. Our key finding is that the synaptic arrangements made by M, P, and K axons in owl monkey exhibit more similarities than differences. They are exclusively presynaptic, contain glutamate and form asymmetric synapses mainly with glutamate-positive dendritic spines. The majority of the remaining axons synapse with glutamatergic dendritic shafts. There are also differences between LGN pathways. M and P terminals are significantly larger and more likely to make multiple synapses than K axons, although M and P axons do not differ from each other in either of these characteristics. Of interest, a larger percentage of M and K axons than P axons make synapses with GABAergic dendritic shafts. Cells directly postsynaptic to M and K axons are known to exhibit orientation selectivity and, in some cases, direction selectivity. Cells postsynaptic to P axons do not show these properties, but instead tend to reflect their LGN inputs more faithfully; therefore, it is possible that these physiologic differences seen in the cortical cells postsynaptic to different LGN pathways reflect the differential involvement of inhibitory circuits., (Copyright 2002 Wiley-Liss, Inc.)

Immediate early gene expression has been used frequently as a marker of activity in the circadian visual system. Recent evidence suggests that the pretectum participates in orchestrating sleep and circadian responses to light. Lesions of the pretectum eliminate dark shift-induced rapid eye movement sleep triggering in albino rats, and compromise circadian phase shifts in hamsters. We hypothesized that regions of the pretectum respond to light with robust and region-specific Fos activation, similar to the suprachiasmatic nucleus and intergeniculate leaflet. We used Fos expression, the protein product of the immediate early gene c-fos, as a functional marker to measure the responses of neurons following acute lighting changes. Rats maintained on a 12:12 light-dark cycle were subjected to a shift from light-to-dark or from dark-to-light at midday (Zeitgeber time 6) or midnight (Zeitgeber time 18). Fos expression was visualized with immunocytochemistry and quantified with an automated scoring system. We found three regions in the pretectum (the olivary pretectal nucleus, posterior limitans, and a region homologous to the hamster commissural pretectal nucleus), and two regions in the lateral geniculate complex (the intergeniculate leaflet and ventral lateral geniculate nucleus) that demonstrated significant Fos activation in response to light. Furthermore, the olivary pretectal nucleus, the posterior limitans, and the ventral lateral geniculate nucleus showed preferential Fos activation after acute light onset rather than following chronic exposure to light at midday, whereas at midnight these nuclei showed Fos activation following both chronic light exposure and acute light onset. Given the extensive anatomical connections between pretectal nuclei and other nuclei in the subcortical visual shell, as well as with centers for sleep and arousal, it is highly plausible that these pretectal nuclei integrate information about changes in illuminance, and aid in the coordination of acute behavioral responses to light., (Copyright 2002 IBRO)

Feedback controlling is an important element in the sensory processing in the auditory system. It has been long recognized that the inferior colliculus (IC) sends direct ascending projections to the medial geniculate body (MGB), but receives feedback regulation from the auditory cortex. In the present study we probed the shorter extracortical projections to the IC, including the direct descending pathway from the MGB. In the rat, the fluorescence retrograde tracers Fluorogold, True Blue or Rhodamine latex microspheres were injected into the IC, and the auditory thalamus and surrounding regions were examined for fluorescent neurones. We did not find any retrograde labelling in the ventral division of the MGB. However, retrogradely labelled neurones were found in the medial and suprageniculate nuclei of the MGB. We also observed densely packed groups of fluorescent neurones in the peripeduncular nucleus and numerous labelled neurones in the nucleus of the brachium of the IC. The existence of a direct descending pathway to the IC from at least some auditory thalamic nuclei challenges the perception of the colliculo-thalamic relationship as one-way traffic and suggests more direct involvement of the auditory thalamus in the feedback regulation of the incoming acoustic signals.

The firing pattern of auditory neurons is determined in part by the type of voltage-sensitive potassium channels expressed. The expression patterns for two high-threshold potassium channels, Kv3.1 and Kv3.3, that differ in inactivation properties were examined in the rat auditory system. The positive activation voltage and rapid deactivation kinetics of these channels provide rapid repolarization of action potentials with little effect on action potential threshold. In situ hybridization experiments showed that Kv3.3 mRNA was highly expressed in most auditory neurons in the rat brainstem, whereas Kv3.1 was expressed in a more limited population of auditory neurons. Notably, Kv3.1 mRNA was not expressed in neurons of the medial and lateral superior olive and a subpopulation of neurons in the ventral nucleus of the lateral lemniscus. These results suggest that Kv3.3 channels may be the dominant Kv3 subfamily member expressed in brainstem auditory neurons and that, in some auditory neurons, Kv3.1 and Kv3.3 may coassemble to form functional channels. The localization of Kv3.1 protein was examined immunohistochemically. The distribution of stained somata and neuropil varied across auditory nuclei and correlated with the distribution of Kv3.1 mRNA-expressing neurons and their terminal arborizations, respectively. The intensity of Kv3.1 immunoreactivity varied across the tonotopic map in the medial nucleus of the trapezoid body with neurons responding best to high-frequency tones most intensely labeled. Thus, auditory neurons may vary the types and amount of K(+) channel expression in response to synaptic input to subtly tune their firing properties., (Copyright 2001 Wiley-Liss, Inc.)

The intergeniculate leaflet (IGL) of the lateral geniculate complex has widespread, bilateral, and reciprocal connections with nuclei in the subcortical visual shell. Its function is poorly understood with respect to its role in visual processing. The most well-known IGL projection, and the only one with a clear function, is the geniculohypothalamic tract (GHT) that terminates in the suprachiasmatic nucleus (SCN), site of the primary circadian clock. The hamster GHT is derived, in part, from IGL neurons containing neuropeptide Y and enkephalin. IGL neurons containing these peptides also project to the pretectal region. The present studies used a combination of immunohistochemical, lesion, and retrograde tracing techniques to study neuron types in the IGL and their projections to hamster SCN and pretectum. Two additional neuromodulators, gamma-aminobutyric acid (GABA) and neurotensin, are shown to be present in IGL neurons. The GABA- and neurotensin-immunoreactive neurons project to the SCN with terminal field patterns very similar to those for neuropeptide Y and enkephalin. IGL neurons of all four types also send projections to the pretectum, but rarely do individual cells project to both the SCN and the pretectum. Nearly all neurotensin is colocalized with neuropeptide Y in IGL neurons, although about half of the neuropeptide Y cells do not contain neurotensin. Otherwise, the extent to which the four neuromodulators are colocalized varies from 6% to 54%. Nearly every SCN neuron appears to contain GABA. In the IGL, the majority of cells studied are not identifiable by GABA immunoreactivity. Putative functions of the various neuromodulator projections from the IGL to pretectum or SCN are discussed., (Copyright 2001 Wiley-Liss, Inc.)

We have examined the patterns of Fos-like immunoreactivity in the ventral thalamus (thalamic reticular nucleus (Rt), zona incerta (ZI) and ventral lateral geniculate nucleus (LGv)) after electrical or chemical stimulation of nuclei in either the brainstem (midbrain reticular nucleus), basal forebrain (substantia innominata) or dorsal thalamus (parafascicular nucleus). Sprague-Dawley rats were anaesthetised with Halothane or Ketamil/Rompun and the above mentioned centres were stimulated either electrically or chemically (using kainic acid). Brains were then processed for Fos-like immunocytochemistry using standard methods. We detected no major differences in the labelling patterns after either electrical or chemical stimulation or after using Halothane or Ketamil/Rompun anaesthesia. After brainstem or dorsal thalamic stimulations, many Fos-like immunoreactive cells were seen within the rostral pole of the Rt, the dorsal sector of the ZI and the parvocellular lamina of the LGv. After basal forebrain stimulations, many Fos-like immunoreactive cells were seen in the rostral pole of the Rt and rostral sector of the ZI, but very few were apparent in the LGv. Overall, our results show that distinct groups of cells in the ventral thalamus show increased levels of Fos-like immunoreactivity after stimulation of different subcortical centres. These activated cells of the ventral thalamus, are in turn, in a position to influence particular thalamocortical pathways through their dorsal thalamic projections.

Marker molecules to visualize specific subsets of neurons are useful for studying the functional organization of the neocortex. One approach to identify such molecular markers is to examine the differences in molecular properties among morphologically and physiologically distinct neuronal cell types. We used differential display to compare mRNA expression in the anatomically and functionally distinct areas of the adult macaque neocortex. We found that a gene, designated occ1, was preferentially transcribed in the posterior region of the neocortex, especially in area 17. Complete sequence analysis revealed that occ1 encodes a macaque homolog of a secretable protein, TSC-36/follistatin-related protein (FRP). In situ hybridization histochemistry confirmed the characteristic neocortical expression pattern of occ1 and showed that occ1 transcription is high in layers II, III, IVA and IVC of area 17. In addition, occ1 transcription was observed selectively in cells of the magnocellular layers in the lateral geniculate nucleus (LGN). Dual labeling immunohistochemistry showed that the occ1-positive neurons in area 17 include both gamma-aminobutyric acid (GABA)-positive aspiny inhibitory cells and the alpha-subunit of type II calcium/calmodulin-dependent protein kinase (CaMKII alpha)-positive spiny excitatory cells. With brief periods of monocular deprivation, the occ1 mRNA level decreased markedly in deprived ocular dominance columns of area 17. From this we conclude that the expression of occ1 mRNA is present in a subset of neurons that are preferentially localized in particular laminae of area 17 and consist of various morphological and physiological neuronal types, and, furthermore, occ1 transcription is subject to visually driven activity-dependent regulation.

Several cortico-cortical and limbic-related circuits are enriched in zinc, which is considered as an important modulator of glutamatergic transmission. While heavy metals have been detected in the thalamus, the specific presence of zinc has not been examined in this region. We have used two highly sensitive variations of the Timm method to study the zinc-rich innervation in the rat thalamus, which was compared to the distribution of acetylcholinesterase activity. The origin of some of these zinc-rich projections was also investigated by means of retrograde transport after intracerebral infusions of sodium selenium (Na2SeO3). The overall zinc staining in the thalamus was much lower than in the neocortex, striatum or basal forebrain; however, densely stained terminal fields were observed in the dorsal tip of the reticular thalamic nucleus, the anterodorsal and lateral dorsal thalamic nuclei and the zona incerta. In addition, moderately stained zinc-rich terminal fields were found in the rostral intralaminar nuclei, nucleus reuniens and lateral habenula. Intracerebral infusions of Na2SeO3 in the lateral dorsal nucleus resulted in retrogradely labeled neurons that were located in the postsubiculum, and also in the pre- and parasubiculum. These results are the first to establish the existence of a zinc-rich subicular-thalamic projection. Similar infusions in either the intralaminar nuclei or the zona incerta resulted in labeling of neurons in several brainstem structures related to the reticular formation. Our results provide morphological evidence for zinc modulation of glutamatergic inputs to highly selective thalamic nuclei, arising differentially from either cortical limbic areas or from brainstem ascending activation systems.

In neurons, neuropeptides and other synaptic components are transported down the axon to the synapse in vesicles using molecular motors of the kinesin family. In the synapse, these neuropeptides are found in dense core vesicles (DCVs), and, following calcium-mediated exocytosis, they interact with receptors on the target cell. We have developed a rapid, large-scale technique for purifying peptide-containing DCVs from specific nuclei in the central nervous system. By using differential velocity gradient and equilibrium gradient centrifugation, neuropeptide-containing DCVs can be separated by size and density from optic nerve (ON) and its termini, the lateral geniculate nuclei and the superior colliculi. Isolated DCVs contain neuropeptides (substance P and brain-derived neurotrophic factor), synaptic vesicle (SV) membrane proteins (SV2, synaptotagmins, synaptophysin, Rab3 and synaptobrevin), SV-associated proteins (alpha-synuclein), secretory markers for DCVs previously isolated (secretogranin II), and beta-amyloid precursor protein. By using electron microscopic techniques, DCV were also visualized and shown to be immunoreactive for neuropeptides, neurotrophins, and SV membrane proteins. Because of the interesting group of physiological and potentially pathophysiological proteins associated with these vesicles; this isolation procedure, applicable to other CNS nuclei, should represent an important research tool., (Copyright 2000 Wiley-Liss, Inc.)

Thalamic relay cells respond in two distinct modes, burst and tonic, that depend on a voltage-dependent, low-threshold, transient Ca(2+) current (I(T)), and these modes relay different forms of information to cortex. I(T) activation evokes a low-threshold spike (LTS), producing a burst of action potentials. Modulatory inputs from cortex and brainstem are known to activate metabotropic receptors on relay cell dendrites at which the T channels underlying I(T) may be concentrated. We thus investigated the influence of activating these receptors on the LTS, using current-clamp intracellular recording in an in vitro slice preparation of the cat's lateral geniculate nucleus. We found a strong correlation between LTS amplitude and the number of action potentials evoked in the burst. We then found that activation of either metabotropic glutamate or muscarinic receptors produced a hyperpolarizing shift in the sigmoid relationship between LTS amplitude and the initial holding potential without affecting the maximum LTS amplitude or slope of the relationship. This hyperpolarizing shift in the voltage dependency of LTS amplitude is best explained by space-clamp limitations and significantly more depolarization of T channels near the dendritic location of activated receptors than at the soma. Thus, nonretinal modulatory inputs may have a stronger influence on I(T) and number of action potentials generated in a burst than previously imagined from somatic recording, because the EPSP amplitudes generated by these inputs at the dendritic location of most T channels are greater than after their electrotonic decay recorded at the soma.

The distribution of mRNA transcripts corresponding to the alpha7 and alpha8 subunits of the nicotinic acetylcholine receptors (nAChRs) was studied in selected structures of the chick visual system with non-radioactive in situ hybridization and immunohistochemical techniques. The results indicated that the alpha7 and alpha8 nAChR transcripts are widely distributed in the brain, exhibiting differential expression in some structures but also some degree of co-localization. The pattern of localization of alpha7 and alpha8 nAChR transcripts was highly correlated with immunohistochemical data, with very few instances of possible mismatches between the distribution of mRNAs and their corresponding proteins.

The distribution of FMRFamide (FMRFa)-like peptides in caiman (Caiman crocodilus) and turtle (Trachemys scripta elegans) brains was studied by immunohistochemistry. In both species, distinct groups of FMRFa-like immunoreactive (ir) perikarya were present in the medial septal nucleus, accumbens nucleus, nucleus of the diagonal band of Broca, suprachiasmatic area, lateral hypothalamic area, and periventricular hypothalamic nucleus. A few FMRFa-ir neurons in the hypothalamic area were located in the neuroepithelial cell lining of the third ventricle. FMRFa-ir fibers were scattered in all major areas of the brain, from the olfactory bulbs to the rhombencephalon. They formed dense aggregates in the medial septal area, basal telencephalon, median eminence, and infundibulum, and adjacent to the fourth ventricle. The most obvious difference between the FMRFa-ir systems in caimans and turtles concerned the number of nuclei that contained neurons with this immunoreactivity. Eight such clusters were present in the caiman brain, whereas thirteen clusters were found in the turtle brain. The turtle also displayed scattered FMRFa-ir somata in the anterior olfactory nucleus, striatum, lateral septal nucleus, medial and lateral cortex, medial forebrain bundle, lateral preoptic area, and lateral geniculate nucleus. In the caiman brain, a few FMRFa-ir neurons were noted in the ventrolateral area of the pallial commissure and an even smaller number of ir neurons was found dispersed in the optic tracts. Neither formed nuclear aggregates. The results are compared with those described for other vertebrates.

Fluorescence-coupled peptides allow a non-radioactive receptor binding study whereby single cells can be examined under a fluorescence microscope. By the combination of such a method with immunohistochemistry, using an HRP-coupled anti-fluorescein antibody, a permanent labeling can be achieved. By using this method the distribution of angiotensin II binding sites has been examined in the mouse thalamus. The results show that a moderate staining was obvious within the thalamus and that the distribution of binding sites in the thalamus is very homogeneous in the mouse brain. In detail, angiotensin II binding sites were found in the anterodorsal nucleus, in the laterodorsal and posterior nucleus of the thalamus, as well as in the lateral geniculate nucleus, the reticular thalamic nucleus and in the zona incerta., (Copyright 1999 Harcourt Publishers Ltd.)

The parabrachial region of the brainstem reticular formation projects to the dorsal lateral geniculate nucleus of the thalamus and to the intermediate gray layer of the superior colliculus. We used the retrograde axonal transport of two fluorescent labels to demonstrate that individual parabrachial cells project to both structures. The results suggest that cholinergic cells of the parabrachial region may coordinate the relay of visuosensory information to the cortex with the onset of orienting movements.

The aim of this study was to describe the distribution patterns of neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP)-immunoreactive (ir) neuronal elements in subcortical visual centers of the cat. Numerous NPY-ir neurons were present in the feline nucleus of the optic tract and in the anterior pretectal nucleus. Only a few NPY-ir neurons were found in the posterior, medial and olivary pretectal nuclei and in the accessory optic nuclei. Diffuse and heavily beaded NPY-ir fiber plexuses were observed throughout the superior colliculus, pretectum, and accessory optic system. Extensively arborising NPY-ir fibers were present in the mesencephalon and ventral lateral geniculate nucleus, while the dorsal visual thalamic nuclei contained only a few NPY-ir fibers. VIP-ir cells were present mainly in the accessory optic nuclei, and they were absent in the dorsal visual thalamus. Both NPY- and VIP-ir neurons were multipolar and fusiform in shape in the regions studied. Enucleation did not alter the appearance of NPY- and VIP-containing neuronal elements in the superior colliculus and pretectum while in the thalamus a subset of NPY-ir fiber population disappeared, indicating their retinal origin. Although there is a partial overlap in the topographical localization of the NPY- and VIP-ergic neurons in the pretectum, the colocalization of the two peptides could not be demonstrated. The present observations demonstrate the existence of two different and separate peptidergic (NPY and VIP) neuronal populations in the pretectum.

The present study was undertaken to explore the possibility that the integration of hormonal cues in the regulation of neuroendocrine mechanisms may occur outside of the hypothalamus at the level of the lateral geniculate body. In situ hybridization for mRNA encoding estrogen receptor beta and progesterone receptor was carried out on sections containing the lateral geniculate body using [35S]-labeled antisense riboprobes. Labeled cells were present in different limbic and hypothalamic sites as described previously. Populations of cells distributed homogeneously in the ventral lateral geniculate nucleus and intergeniculate leaflet were also found to express mRNA for estrogen receptor beta and progesterone receptor. The dorsal lateral geniculate nucleus lacked specific labeling for either type of gonadal steroid hormone receptor mRNA. The present observation together with the recent demonstration of a direct pathway between the intergeniculate leaflet and hypothalamic neuroendocrine cells indicate that integration of hormonal and photic stimuli in the central regulation of endocrine mechanisms occurs outside of the hypothalamus in the lateral geniculate body.

GABA(B) receptors are believed to be associated with the efferents of the nucleus reticularis thalami, which is implicated in the regulation of activity in the thalamocortical-corticothalamic circuit and plays a role in absence seizures. Yet, the distribution of GABA(B) receptors in the thalamus has only been studied in the rat, and there is no comparable information in primates. The potent GABA(B) receptor antagonist [3H]CGP 62349 was used to study the distribution and binding properties of the receptor in control monkeys and those with small ibotenic acid lesions in the anterodorsal segment of the nucleus reticularis thalami. Eight-micrometer-thick cryostat sections of the fresh frozen brains were incubated in the presence of varying concentrations of the ligand. Autoradiographs were analysed using a quantitative image analysis technique, and binding parameters were calculated for select thalamic nuclei as well as basal ganglia structures present in the same sections. The overall number of GABA(B) binding sites in the monkey thalamus and basal ganglia was several-fold higher than previously reported values for the rat. In the thalamus, the receptors were distributed rather uniformly and the binding densities and affinities were high (Bmax range of 245.5-437.9 fmol/ mg of tissue, Kd range of 0.136-0.604 nM). In the basal ganglia, the number of binding sites and the affinities were lower (Bmax range of 51.1-244.2 fmol/mg of tissue; K(d) range of 0.416-1.394 nM), and the differences between nuclei were more pronounced, with striatum and substantia nigra pars compacta displaying the highest binding densities. Seven days post-lesion, a 20-30% decrease in Bmax values (P < 0.05) was found in the nuclei receiving input from the lesioned nucleus reticularis thalami sector (the mediodorsal nucleus and densicellular and magnocellular parts of the ventral anterior nucleus) without changes in affinity. No significant changes were detected in any other structures. The results of the lesioning experiments suggest that a portion of thalamic GABA(B) receptors is in a presynaptic location on the nucleus reticularis thalami efferents. The overall distribution pattern in the thalamus also suggests a partial association of GABA(B) receptors with corticothalamic terminals presynaptically.

Studies carried out within the primary visual system have shown that neural responses to light stimuli transmitted via the retinogeniculate pathway are significantly altered when these stimuli are made aversive through conditioning. The effect of such aversive conditioning on neural responses to light transmitted within the circadian visual system has not been investigated. In mammals, the principal projection of the circadian visual system, the retinohypothalamic tract, is functionally and anatomically distinct from the primary visual pathway allowing for direct transmission of light from the retina to the suprachiasmatic nucleus of the hypothalamus, the circadian clock. Light transmitted within this pathway is essential for entrainment of circadian rhythms providing the critical stimulus for resetting the circadian clock. We asked whether the response of neural elements within the suprachiasmatic nucleus to a resetting light stimulus would be altered if that stimulus had acquired aversive properties through conditioning. To study this we assessed the effect of a light stimulus made aversive through pairings with footshock on a cellular correlate of clock resetting, the expression of the transcription factor Fos in neurons of the suprachiasmatic nucleus. We show that Fos expression in the suprachiasmatic nucleus in response to light previously paired with footshock is significantly suppressed. This finding provides the first evidence that the effectiveness of a light as a resetting stimulus can be modulated by its conditioned aversive properties.

The serotonin transporter MRNA has been found throughout the trigeminal sensory system late in gestation and during early postnatal development, a period known to be critical for maturation of the sensory circuitry. The purpose of the present study was to determine whether sensory denervation in newborn rat pups would alter either the density or pattern of expression of the 5-HT transporter (5-HTT) within the trigeminal system. We combined autoradiographic localization of 5-HT transporters and in situ hybridization techniques to visualize both the transporter protein and mRNA in thalamic sensory neurons and in the somatosensory cortex following unilateral infraorbital nerve transection at postnatal day 1. For comparative purposes, similar measurements were conducted in thalamic visual neurons as well as in the visual cortex. Lesion of the infraorbital nerve decreased the [3H]citalopram labelling of 5-HT transporters in the ventral basal and ventral medial areas of the thalamus contralateral to the lesion, while labelling of 5-HT transporters was decreased in both contralateral and ipsilateral sides of the lateral genicuate (visual thalamus). Citalopram labelling of 5-HT transporters was not significantly altered in somatosensory or in cingulate cortex, however a significant decrease was observed in the visual cortex. In contrast, there were no obvious changes in the intensity of the 5-HT mRNA hybridization signal in sensory or visual thalamic areas. Given that the serotonin transporter regulates extracellular concentrations of 5-HT, the present data suggest that altered peripheral innervation and thereby altered sensory inputs to the thalamus during fetal development can potentially influence 5-HT transporter densities and thus, may influence extracellular levels of 5-HT in thalamus and cortex during a critical period of synapse formation. In turn, modulation of 5-HT transporter levels may influence extracellular concentrations of 5-HT in thalamus and cortex during a critical period of synapse formation., (Copyright 1998 Elsevier Science B.V.)

Thalamocortical and perigeniculate (PGN) neurons can generate action potentials either as Ca2+ spike-mediated high-frequency bursts or as tonic trains. Using dual intracellular recordings in vitro in monosynaptically connected pairs of PGN and dorsal lateral geniculate nucleus (LGNd) neurons, we found that the functional effect of synaptic transmission between these cell types was strongly influenced by the membrane potential and hence the firing mode of both the pre- and postsynaptic neurons. Activation of single action potentials or low-frequency spike trains in PGN or thalamocortical neurons resulted in the generation of PSPs that were 0.5-2.0 mV in amplitude. In contrast, the generation of Ca2+ spike-mediated bursts of action potentials in the presynaptic cell increased these PSPs to an average of 4.4 mV for the IPSP and 3.0 mV for the EPSP barrage, because of temporal summation and/or facilitation. If the postsynaptic neuron was at a resting membrane potential (e.g., -65 mV), these PSP barrages could result in the activation of a low-threshold Ca2+ spike and burst of action potentials. These results demonstrate that the burst firing mode of action potential generation is a particularly effective means by which perigeniculate and thalamocortical neurons may influence one another. We propose that the activation of burst discharges in these cell types is essential for the generation of some forms of synchronized rhythmic oscillations of sleep and of epileptic seizures.

We combined fluorescent tracing with immunohistochemistry to examine nitric oxide synthase (NOS) containing neurons in the rat ventral lateral geniculate nucleus (vLGN). NOS immunopositive neurons in vLGN had a similar appearance to previously described NADPH-d positive neurons. The majority (96%) of the NOS immunopositive neurons in vLGN projected to the pretectal nuclei and these represented 16% of all the vLGN neurons that project to the pretectal nuclei. No NOS immunopositive neurons projected to the superior colliculus. Co-localization of NADPH-d and gamma-aminobutyric acid (GABA) has been reported in vLGN neurons but we found few cells containing both NOS and GABA.

Recent studies of the rabbit auditory forebrain have shown that antibodies directed against the calcium-binding protein parvalbumin (PV) specifically demarcate auditory neocortex and the ventral division of the medial geniculate body (MGV). The auditory cortex is characterized by two PV- immunoreactive bands: dense terminal-like labeling within layer III/IV and a prominent band of PV+ somata in the upper half of layer VI. In some cases, there are distinct patches of PV immunoreactivity within layers III/IV of auditory cortex that appear similar to the patchy termination of thalamocortical axons labeled by the injection of anterograde tracers into MGV. The presence of PV+ patches in III/IV, PV+ somata in layer VI, and the high density of PV+ neurons and terminals in the MGV suggest the existence of a reciprocal PV+ circuit linking primary auditory cortex (AI) and the MGV. In the present study, double-labeling experiments in adult rabbits were carried out to provide evidence for this circuit. Focal injections of the tracers biocytin or biotinylated dextran amine (BDA) into the MGV labeled thalamocortical afferent patches within layer III/IV and retrogradely labeled corticothalamic neurons in layer VIa of the ipsilateral auditory cortex. Adjacent sections stained with antibodies against PV revealed terminal-like PV-immunoreactive patches in III/IV and PV+ somata in VIa that were in register with those labeled by BDA injections into the MGV. Serial section reconstruction of BDA-labeled corticothalamic neurons in VIa revealed pyramidal cells with tangentially oriented basal dendrites and sparsely branched apical dendrites that ascended to layer I. Fluorescent double-labeling studies demonstrated that a subpopulation of corticothalamic neurons also express PV. PV-negative corticothalamic neurons were also found. Discrete injections of BDA into auditory cortex labeled bands of neurons in the ipsilateral MGV, whose orientation paralleled the fibrodendritic laminae characteristic of this subdivision. Retrograde double-labeling experiments showed that most MGV relay neurons also express PV. Small numbers of PV-negative relay neurons were also found. These studies provide evidence for the existence of multiple, chemically coded pathways linking primary auditory cortex and the MGV.

The effects of excitatory classical conditioning on cytochrome oxidase activity in the central auditory system were investigated using quantitative histochemistry. Rats in the conditioned group were trained with consistent pairings of a compound conditional stimulus (a tone and a light) with a mild footshock, to elicit conditioned suppression of drinking. Rats in the pseudorandom group were exposed to pseudorandom presentations of the same tone, light and shock stimuli without consistent pairings. Untrained rats in a naive group did not receive presentations of the experimental stimuli. The findings demonstrated that auditory fear conditioning modifies the metabolic neuronal responses of the auditory system, supporting the hypothesis that sensory neurons are responsive to behavioural stimulus properties acquired by learning. There was a clear distinction between thalamocortical and lower divisions of the auditory system based on the differences in metabolic activity evoked by classical conditioning, which lead to an overt learned behavioural response versus pseudorandom stimulus presentations, which lead to behavioural habituation. Increases in cytochrome oxidase activity indicated that tone processing is enhanced during associative conditioning at upper auditory structures (medial geniculate nucleus and secondary auditory cortices). In contrast, metabolic activation of lower auditory structures (cochlear nuclei and inferior colliculus) in response to the pseudorandom presentation of the experimental stimuli suggest that these areas may be activated during habituation to tone stimuli. Together these findings show that mapping the metabolic activity of cytochrome oxidase with quantitative histochemistry can be successfully used to map regional long-lasting effects of learning on brain systems.

The rat suprachiasmatic nucleus (SCN) contains a dense plexus of low-affinity p75 neurotrophin receptor (p75NTR)-immunoreactivity. Scattered patches of p75NTR immunoreactivity are present in the intergeniculate leaflet (IGL). Both SCN and IGL receive a direct retinal input. After binocular enucleation, there is a delayed loss of p75NTR-immunoreactivity in the SCN and IGL beginning at, respectively, 4 and 8 weeks post-enucleation, with complete loss occurring in both nuclei by week 12. This delayed loss may be due to an up-regulation of growth factor secretion by local cells in response to retinal axon degeneration.