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PubMed2001

Thalamo-cortical connections of areas 3a and M1 in marmoset monkeys.

The present investigation is part of a broader effort to examine cortical areas that contribute to manual dexterity, reaching, and grasping. In this study we examine the thalamic connections of electrophysiologically defined regions in area 3a and architectonically defined primary motor cortex (M1). Our studies demonstrate that area 3a receives input from nuclei associated with the somatosensory system: the superior, inferior, and lateral divisions of the ventral posterior complex (VPs, VPi, and VPl, respectively). Surprisingly, area 3a receives the majority of its input from thalamic nuclei associated with the motor system, posterior division of the ventral lateral nucleus of the thalamus (VL), the mediodorsal nucleus (MD), and intralaminar nuclei including the central lateral nucleus (CL) and the centre median nucleus (CM). In addition, sparse but consistent projections to area 3a are from the anterior pulvinar (Pla). Projections from the thalamus to the cortex immediately rostral to area 3a, in the architectonically defined M1, are predominantly from VL, VA, CL, and MD. There is a conspicuous absence of inputs from the nuclei associated with processing somatic inputs (VP complex). Our results indicate that area 3a is much like a motor area, in part because of its substantial connections with motor nuclei of the thalamus and motor areas of the neocortex (Huffman et al. [2000] Soc. Neurosci. Abstr. 25:1116). The indirect input from the cerebellum and basal ganglia via the ventral lateral nucleus of the thalamus supports its role in proprioception. Furthermore, the presence of input from somatosensory thalamic nuclei suggests that it plays an important role in somatosensory and motor integration.

Action Potentials↗
PubMed2002

Few intrinsic connections cross the hand-face border of area 3b of New World monkeys.

After long-standing loss of afferents from the hand, the hand representation in area 3b of the somatosensory cortex of monkeys becomes responsive to touch on the face. Because the reactivation of deprived hand cortex by the face inputs could depend on axonal connections across the hand-face border, we determined the extent of such connections in New World marmosets, owl monkeys, and squirrel monkeys. Small injections of anatomic tracers were placed in the hand or the face representations after these representations were identified by microelectrode recordings. The positions of retrogradely labeled neurons were plotted in processed brain sections cut parallel to the brain surface, and their locations were related to anatomic isomorphs of the hand and face representations revealed in adjacent brain sections stained for myelin. In these sections, the hand-face border was clearly visualized as a myelin-poor septum. The intrinsic connections of area 3b labeled by injections in either the hand or face representations were almost completely confined to their respective representation, and very few neurons projected across the border. In addition, neurons in the somatosensory thalamus labeled by injections in either face or hand representations were confined to either VPM, representing the face, or the hand subnucleus of VPL. Thus the reactivation of hand cortex by face stimulation does not depend on a previously existing network of intrinsic cortical connections across the hand-face border, or mismatched thalamocortical projections.

Afferent Pathways↗
PubMed2001

Parkin immunoreactivity in the brain of human and non-human primates: an immunohistochemical analysis in normal conditions and in Parkinsonian syndromes.

The etiology of Parkinson's disease is unknown, but the gene involved in an autosomic recessive form of the disease with early onset has recently been identified. It codes for a protein with an unknown function called parkin. In the present study we produced a specific polyclonal antiserum against human parkin. Immunohistochemical analysis showed that parkin is expressed in neuronal perikarya and processes but also in glial and blood vessels in the primate brain (human and monkey). Electron microscopy indicated that parkin immunoreactivity is mostly located in large cytoplasmic vesicles and at the level of the endoplasmic reticulum. Parkin was expressed heterogeneously in various structures of the brain. It was detectable in the dopaminergic systems at the level of the perikarya in the mesencephalon but also in the striatum. However, parkin was also expressed by numerous nondopaminergic neurons. The staining intensity of parkin was particularly high in the hippocampal formation, the pallidal complex, the red nucleus, and the cerebellum. Comparison of control subjects with patients with Parkinson's disease and control animals with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-intoxicated animals revealed a loss of parkin-immunoreactive neurons only in the substantia nigra pars compacta. Furthermore, the surviving dopaminergic neurons in the parkinsonian state continued to express parkin at a level similar to that observed in the control situation. These data indicate that parkin is a widely expressed protein. Thus, the degeneration of dopaminergic neurons in familial cases of Parkinson's disease with autosomal recessive transmission cannot be explained solely in terms of an alteration of this protein.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed2004

Synaptic connectivity of the diffuse bipolar cell type DB6 in the inner plexiform layer of primate retina.

Diffuse bipolar cells in primate retina receive synaptic input from multiple cones and provide output to ganglion cells. Diffuse bipolar cells can be subdivided into six types (DB1-DB6) according to the stratification of their axon terminals in the inner plexiform layer, but their synaptic connectivity in the inner plexiform layer is not well understood. Here the stratification and synaptic connectivity of DB6 axon terminals were studied in the retinae of New World (marmoset) and Old World (macaque) monkeys. Immunohistochemical markers were applied to retinal sections. The sections were analyzed by confocal and deconvolution light microscopy as well as electron microscopy. The DB6 cells were identified with antibodies against CD15; rod bipolar cells were identified with antibodies against protein kinase Calpha (PKCalpha); and AII amacrine cells were identified with antibodies against calretinin. The axons of DB6 and rod bipolar cells occupy distinct regions in stratum 5 of the inner plexiform layer. The distal processes of calretinin-labeled AII cells are usually closely associated with rod bipolar axons but sometimes also with DB6 axons. Pre-embedding immunoelectron microscopy showed that the vast majority (over 86%) of the synaptic output of DB6 cells is onto amacrine cell processes, whereas less than 14% goes to ganglion cell processes. In double-labeled preparations DB6 axons occasionally made output onto calretinin-labeled amacrine processes. Thus it is possible that AII cells receive some input from DB6 cells.

Amacrine Cells↗
PubMed2005

Species differences in tachykinin receptor distribution: further evidence that the substance P (NK1) receptor predominates in human brain.

Marked species differences in the distribution of central tachykinin receptors are reported but uncertainty remains about the ability of available ligands to detect NK2 and NK3 receptors in human brain. We compared the distribution of NK1, NK2, and NK3 receptors in sections from rodent, primate, and human brain using the 125I-labeled ligands substance P (SP) for the NK1 receptor, neurokinin A (NKA) for the NK2 receptor, and neurokinin B (NKB) and eledoisin for NK3 receptors. Duration of exposure to autoradiographic film was from 7 days for [125I]SP up to 90 days for the other ligands. High levels of specific [125I]SP binding were seen throughout the brains of all species studied. Specific [125I]NKA binding was detected in brains from neonatal rat, and to a lesser level in adult rat, gerbil, and guinea pig; it was not detected in monkey or human brain, but was present in circular muscle of human duodenum, confirming that this ligand binds to human NK2 receptors under our experimental conditions. Specific [125I]NKB and [125I]eledoisin binding was widespread in brain sections from rats, gerbils, and guinea pigs, and very low levels were also detected in marmoset, squirrel monkey, and rhesus monkey brain after prolonged (up to 90 days) exposure. We failed to identify specific eledoisin binding in human brain, even after prolonged exposures. These findings demonstrate that the NK1 receptor is the predominant tachykinin receptor expressed in primate and human brain, but that low levels of NK3 receptor are present in nonhuman, primate brain.

Animals↗
PubMed2006

Cortical connections of the auditory cortex in marmoset monkeys: core and medial belt regions.

The auditory cortex of primates contains a core region of three primary areas surrounded by a belt region of secondary areas. Recent neurophysiological studies suggest that the belt areas medial to the core have unique functional roles, including multisensory properties, but little is known about their connections. In this study and its companion, the cortical and subcortical connections of the core and medial belt regions of marmoset monkeys were compared to account for functional differences between areas and refine our working model of the primate auditory cortex. Anatomical tracer injections targeted two core areas (A1 and R) and two medial belt areas (rostromedial [RM] and caudomedial [CM]). RM and CM had topographically weighted connections with all other areas of the auditory cortex ipsilaterally, but these were less widespread contralaterally. CM was densely connected with caudal auditory fields, the retroinsular (Ri) area of the somatosensory cortex, the superior temporal sulcus (STS), and the posterior parietal and entorhinal cortex. The connections of RM favored rostral auditory areas, with no clear somatosensory inputs. RM also projected to the lateral nucleus of the amygdala and tail of the caudate nucleus. A1 and R had topographically weighted connections with medial and lateral belt regions, infragranular inputs from the parabelt, and weak connections with fields outside the auditory cortex. The results indicated that RM and CM are distinct areas of the medial belt region with direct inputs from the core. CM also has somatosensory input and may correspond to an area on the posteromedial transverse gyrus of humans and the anterior auditory field of other mammals.

Acoustic Stimulation↗
PubMed2006

Thalamic connections of the auditory cortex in marmoset monkeys: core and medial belt regions.

In this study and its companion, the cortical and subcortical connections of the medial belt region of the marmoset monkey auditory cortex were compared with the core region. The main objective was to document anatomical features that account for functional differences observed between areas. Injections of retrograde and bi-directional anatomical tracers targeted two core areas (A1 and R), and two medial belt areas (rostromedial [RM] and caudomedial [CM]). Topographically distinct patterns of connections were revealed among subdivisions of the medial geniculate complex (MGC) and multisensory thalamic nuclei, including the suprageniculate (Sg), limitans (Lim), medial pulvinar (PM), and posterior nucleus (Po). The dominant thalamic projection to the CM was the anterior dorsal division (MGad) of the MGC, whereas the posterior dorsal division (MGpd) targeted RM. CM also had substantial input from multisensory nuclei, especially the magnocellular division (MGm) of the MGC. RM had weak multisensory connections. Corticotectal projections of both RM and CM targeted the dorsomedial quadrant of the inferior colliculus, whereas the CM projection also included a pericentral extension around the ventromedial and lateral portion of the central nucleus. Areas A1 and R were characterized by focal topographic connections within the ventral division (MGv) of the MGC, reflecting the tonotopic organization of both core areas. The results indicate that parallel subcortical pathways target the core and medial belt regions and that RM and CM represent functionally distinct areas within the medial belt auditory cortex.

Animals↗
PubMed2006

Transitory uptake of serotonin in the developing sensory pathways of the common marmoset.

Serotonin (5-HT) affects brain development during sensitive developmental periods. In rodents, transient sites of high affinity capture of 5-HT were demonstrated in the primary sensory neurons and in the sensory thalamocortical afferents. This uptake is required to adjust 5-HT receptor stimulation during the formation of sensory maps. To determine whether similar mechanisms exist in primates, we analyzed staged embryos and postnatal pups in the common marmoset (total gestation time, 142 days). Immunocytochemical analyses were performed using antisera to 5-HT, to the serotonin transporter (SERT), and to the vesicular monoamine transporter (VMAT2). 5-HT, SERT, and VMAT2 labeled the raphe neurons and their terminal network from embryonic day (E)70 to adulthood. In addition, from E70-130 VMAT2 and SERT were observed in all the sensory cranial nerves, the olfactory nerve, the gustatory, the trigeminal, the auditory fibers, in the retinal ganglion cells, and the optic tract up to the lateral geniculate nucleus and the superior colliculus. All the spinal sensory ganglia and their peripheral sensory branches were labeled. Accumulation of 5-HT was observed in all the sensory neurons expressing SERT and the corresponding axon tracts. Since these neurons were missing tryptophan hydroxylase (TPH), the synthesizing enzyme for 5-HT, they most likely accumulated 5-HT through the action of the amine transporters, as has been shown in rodents. No transient expression of 5-HT markers was detectable in the sensory thalamocortical axons at any of the ages examined. Thus, the existence of 5-HT uptake in nonserotoninergic neurons appears to be a conserved feature in primates, although the topographic extent of this transient expression is more restricted than that previously demonstrated in rodents.

Afferent Pathways↗
PubMed1985

Cortical and subcortical afferent connections of the primate's temporal pole: a study of rhesus monkeys, squirrel monkeys, and marmosets.

The afferent connections of the primate's temporopolar cortex were investigated with the retrograde horseradish peroxidase technique. Old World and New World monkeys received small unilateral injections of horseradish peroxidase. These labeled cells in a number of cortical, thalamic, and brainstem regions and in a few further telencephalic and diencephalic regions. Cortically, the neighboring areas of the inferior and superior temporal gyrus and the insula contained a considerable number of labeled cells. Furthermore, a substantial projection arose from the orbitofrontal and the frontopolar cortex. The cingulate gyrus contained only very few labeled cells. Interhemispherically, corticocortical connections arose mainly from temporal lobe areas. Labeled cells were seen in various regions of the basal forebrain and cells labeled only faintly in the lateral and basal accessory nuclei of the amygdala. The claustrum contained labeled neurons only in one rhesus monkey. On the diencephalic level, the caudal medial portion of the medial pulvinar was the principal thalamic source of afferents to the temporopolar cortex. Furthermore, labeled cells were found in the neighboring, caudal part of the mediodorsal nucleus, within and along the nucleus limitans, in the medial geniculate nucleus, and in several nuclei of the nonspecific system. The fields of Forel, the zona incerta, and lateral and dorsomedial hypothalamic areas contained a few labeled cells. Within the brainstem of the rhesus monkeys those regions projecting diffusely to the cortex contained a few labeled neurons. Furthermore, these brains had some labeled cells in the regions of the nuclei medialis annuli aqueductus, tractus mesencephalicus nervi trigemini, and trochlearis. Although among the three species differences in the cortical and thalamic projection patterns were observed, the regions projecting most densely to the temporal pole were similar in principle. This statement holds in particular for cortical and thalamic sites. However, the greatest number of labeled cells was found in the rhesus monkey, a fact that cannot be attributed solely to the size of the horseradish peroxidase injections and the size of the brain, but that appears rather to represent a true species difference. From our results we conclude that the temporopolar cortex constitutes a cortical area necessary for effective affectional-sensory integration.

Afferent Pathways↗
PubMed1992

Dendritic sampling across processing streams in monkey striate cortex.

Cytochrome oxidase (CO) dense blobs in primate striate cortex provide a striking example of parallel processing of visual information. The level of isolation of the blobs from the surrounding interblob tissue was investigated in the present study by combining CO staining with Golgi impregnation of dendritic arbors in the same tissue sections. The data are based on material from two marmoset and three squirrel monkeys. The analysis was conducted on two types of Golgi preparations. In the first preparation, dense networks of overlapping dendrites were impregnated over blob margins. The results of analyzing these networks with transmission and confocal microscopy revealed that dendritic arbors penetrate freely through blob margins. Statistical analysis revealed that the density of dendritic crossings at blob margins was similar to that found at blob and interblob centers. In the second type of Golgi preparation, single, isolated neurons were impregnated. Studies of such neurons revealed occasional examples of dendritic arbors that appeared to reflect back from blob margins, but counter examples were equally abundant. Bias index analysis indicated that dendritic arbors were generally unaffected by the presence of a nearby blob margin. Scanning a large number of impregnated arbors indicated that at least half of the population of blob-related neurons had dendrites in both blob and interblob territory. Under the conditions of free dendritic penetration of blob margins, the sole factor that determines the level of blob/interblob mixing appears to be the relationship between blob size and the dendritic spread of blob neurons. Interestingly, in both the marmoset and squirrel monkeys this size ratio is similar despite a large difference in their cortical surface area. Thus, it is hypothesized that blob size is optimally matched to the dendritic span so as to create a smooth transition of dendritic sampling from blob to interblob-related processing streams.

Animals↗
PubMed1992

The somatosensory thalamus of monkeys: cortical connections and a redefinition of nuclei in marmosets.

Thalamic connections of three subdivisions of somatosensory cortex in marmosets were determined by placing wheatgerm agglutinin conjugated with horseradish peroxidase and fluorescent dyes as tracers into electrophysiologically identified sites in S-I (area 3b), S-II, and the parietal ventral area, PV. The relation of the resulting patterns of transported label to the cytoarchitecture and cytochrome oxidase architecture of the thalamus lead to three major conclusions. 1) The region traditionally described as the ventroposterior nucleus (VP) is a composite of VP proper and parts of the ventroposterior inferior nucleus (VPi). Much of the VP region consists of groups of densely stained, closely packed neurons that project to S-I. VPi includes a ventral oval of pale, less densely packed neurons and finger-like protrusions that extend into VP proper and separate clusters of VP neurons related to different body parts. Neurons in both parts of VPi project to S-II rather than S-I. Connection patterns indicate that the proper and the embedded parts of VPi combine to form a body representation paralleling that in VP. 2) VPi also provides the major thalamic input into PV. 3) In architecture, location, and cortical connections, the region traditionally described as the anterior pulvinar (AP) of monkeys resembles the medial posterior nucleus, Pom, of other mammals and we propose that all or most of AP is homologous to Pom. AP caps VP dorsomedially, has neurons that are moderately dense in Nissl staining, and reacts moderately in CO preparations. AP neurons project to S-I, S-II, and PV in somatotopic patterns.

Animals↗
PubMed1992

The selective innervation by serotoninergic axons of calbindin-containing interneurons in the neocortex and hippocampus of the marmoset.

The serotoninergic input to the mammalian cerebral cortex originates in the median and the dorsal raphe nuclei. Median raphe neurons have been previously shown to give rise to beaded varicose axons which form dense pericellular arrays (baskets) surrounding the soma and the proximal dendrites of certain cortical neurons. In the present study, we have searched for specific markers characterizing the neurons of the marmoset neocortex and hippocampus surrounded by these thick varicose serotonin-containing fibers. The non-pyramidal nature of these neurons, suggested by their dendritic arborization, was correlated, in immunocytochemical experiments with double-labelling to demonstrate their surrounding serotonin-containing basket and their content of glutamic acid decarboxylase (GAD) or of the calcium-binding protein calbindin. Another calcium-binding protein common in numerous non-pyramidal cortical neurons, parvalbumin, was never found in neurons surrounded by serotonin-containing baskets. This organization was found in all areas of the neocortex and of the hippocampus where serotonin-containing baskets were present. One of the serotoninergic cortical inputs which originates from the brainstem tegmentum, traditionally described as "diffuse," proves to be highly selective in that a subset of its axons terminates preferentially on a subpopulation of inhibitory interneurons of the cerebral cortex. It may be emphasized that this subset of cortical interneurons has now been shown to be characterized not only by its axonal and dendritic arborization and its neurotransmitter, but also by a specific type of input which can modulate cortical function in a unique manner.

Animals↗
PubMed1995

Visual areas in the dorsal and medial extrastriate cortices of the marmoset.

To define the number and limits of the visual areas in the primate extrastriate cortex, the visuotopy of the dorsal convexity and medial wall was studied by electrophysiological recordings in five marmosets anaesthetised with sufentanil and nitrous oxide and paralysed with pancuronium bromide. We identified five visuotopic representations in and around the densely myelinated zone between visual area 2 (V2) and the posterior parietal cortex. Most of the densely myelinated zone is formed by the homologue of the owl monkey's dorsomedial area (DM); thus, we also termed this area DM in the marmoset. Within DM, the lower quadrant representation is continuous, with central vision represented laterally, peripheral vision medially, the horizontal meridian caudally, and the vertical meridian rostrally. In contrast, the upper quadrant representation is split, with the central portion represented at the lateral edge of DM on the dorsal surface, and the periphery along the midline. Two other visual field representations, corresponding to the dorsointermediate area (DI) and to a new subdivision termed the dorsoanterior area (DA), are also densely myelinated but can be distinguished from DM based on the separation of the bands of Baillarger and visual topography. In addition, a homologue of the medial visual area (M) was identified. Our results reveal a highly complex visuotopy in primate cortex, with local discontinuities in representation and borders between areas that are often not coincident with either the horizontal or the vertical meridian. The topography of the dorsal extrastriate cortex in the marmoset strongly suggests that both visual area 3 (V3) and the parietooccipital area (PO) of other primates are portions of a single visuotopic representation, DM, and calls into question the existence of visual areas with partial or quadrantic representations of the visual field.

Animals↗
PubMed1993

Human immunoglobulin class and IgG subclass regulation: dual action of interleukin-4.

Epstein-Barr virus (EBV) was used as a polyclonal human B cell mitogen to investigate the regulation of immunoglobulin class and IgG subclass responses by interleukin-4 (IL-4). Activation of tonsillar B cells with EBV resulted in an early peak of polyclonal immunoglobulin secretion between days 13 and 14 consisting of IgM, IgA, and IgG1, IgG2, IgG3 and IgG4, but not IgE. Addition of IL-4 to EBV-activated B cells at concentrations of 100 U/ml or greater induced the production of IgE and enhanced IgG4 secretion, but had no effect, or more often inhibited the other isotypes. In contrast, low concentrations of IL-4 (1-5 U/ml) significantly increased the production of IgM, IgA, IgG1, IgG2 and IgG3, but had no effect on IgG4 or IgE. The increase in immunoglobulin secretion obtained with low concentrations of IL-4 was found to occur only with high-density (resting) B cells, suggesting that IL-4 was not functioning simply as a late-acting differentiation factor. Low concentrations of IL-4 significantly increased IgG1, IgG2, IgG3, and IgA production by surface (s) IgM+ (sIgG-/sIgA-) B cells which is consistent with heavy chain switching. In some experiments, however, IL-4 enhanced IgM secretion by sIgM+ B cells, and IgA, IgG1, IgG2, IgG3 by sIgM- B cells, suggesting that it may have an additional B cell differentiation factor activity which was not isotype specific. The different effect of IL-4 at high and low concentrations were similar to those observed in B cell activation experiments, and may be due to the existence of high- and low-affinity IL-4 receptors.

Animals↗
PubMed2004

Frontline: Epitope recognition on the myelin/oligodendrocyte glycoprotein differentially influences disease phenotype and antibody effector functions in autoimmune demyelination.

Preliminary observations of humoral immunity against the myelin oligodendrocyte glycoprotein (MOG) in experimental allergic encephalomyelitis (EAE) and human multiple sclerosis (MS) suggest that a subset of anti-MOG autoantibodies directed against conformational epitopes is of pathogenic predominance. Here, we provide proof that in marmoset EAE, autoantibodies reactive against conformational epitopes of MOG are not only responsible for aggravating demyelination, but also an essential factor for disease dissemination in space within the central nervous system, a hallmark for typical forms of human MS. In terms of effector mechanisms, IgG deposition and complement activation occur exclusively in association with presence of these conformational antibodies, while microglial/macrophage activation appears to be a common immunopathological finding regardless of the fine determinant specificity of anti-MOG antibodies. These findings highlight for the first time the complex heterogeneity of function and pathogenicity in the polyclonal anti-MOG antibody repertoire of outbred species. Because the linear and conformational antibody determinants of MOG are shared between marmosets and humans, these results are directly relevant to understanding effector mechanisms of organ damage in MS.

Animals↗
PubMed2004

Commentary: Sorting the wheat from the chaff: identifying demyelinating components of the myelin oligodendrocyte glycoprotein (MOG)-specific autoantibody repertoire.

Myelin oligodendrocyte glycoprotein (MOG) is the only myelin protein known to initiate a demyelinating autoantibody response in EAE, an animal model for multiple sclerosis (MS). The pathophysiological significance of MOG-specific autoantibodies in MS is, however, controversial, as high titer antibody responses to MOG are also found in many patients with non-demyelinating neurological diseases. In this issue of the European Journal of Immunology, von Büdingen et al. demonstrate that demyelination in a primate model of MOG-induced EAE is mediated by MOG-specific antibodies directed against discontinuous, rather than linear, MOG epitopes. This functional segregation of pathogenic vs. non-pathogenic autoantibodies in terms of epitope specificity may be crucial to understand the relevance of MOG-specific responses in human disease. This commentary discusses these findings in the context of the structure and immunobiology of MOG, and their implications with respect to antibody-mediated demyelination in MS.

Animals↗
PubMed2005

Glial glutamate transporter expression patterns in brains from multiple mammalian species.

It is generally assumed that rodent brains can be used as representative models of neurochemical function in other species, such as humans. We have compared the distributions of the predominant glial glutamate transporters in rodents, rabbits, cats, pigs, monkeys, and humans. We identify similarities but also significant differences between species. GLT-1v, which is abundantly expressed by rodent astrocytes, is expressed only in a rare subset of astrocytes of cats and humans, and appears to be absent from brains of rabbits and monkeys. Conversely, in the pig brain GLT-1v is expressed only by oligodendrocytes. GLAST and GLT-1alpha expression differed significantly between species; while rodents and rabbits exhibited uniform expression patterns in cortex, higher species, including cats, pigs, monkeys, and humans, exhibited heterogeneities in cortical and hippocampal expression. Patches devoid of labeling intermingling with patches of strong labeling were evident in areas such as temporal cortex and frontal cortex. In addition, we noted that in human motor cortex, there were inconsistencies in labeling for the C-terminal of GLT-1alpha and common domains of GLT-1, suggesting that the C-terminal region may be missing or that an unidentified splicing is present in many human astrocytes. Collectively our data suggest that assumptions as to the roles of glutamate transporters in any species may need to be tested empirically.

Aged↗
PubMed1980

Radioimmunoassay for infectious primate retrovirus reverse transcriptase: characterization, comparison with conventional immunologic assays and applicability to cellular extracts.

SSV reverse transcriptase (RT) was purified to homogeneity and used in a radioimmunoassay. Following iodination, the homogeneity of the protein and its identity with RT were confirmed by several criteria: (1) its molecular weight on an SDS-polyacrylamide gel; (2) its precipitation by anti-SSV RT but not by antisera to other SSV proteins; (3) its cross-reactivity in RIA with antisera to other retroviral polymerases; (4) its competition in RIA by active homogeneous SSV RT but not by other purified SSV proteins; and (5) its competition in RIA by only those fractions from a poly(U)-Sepharose column possessing SSV RT activity. Competition of the labelled probe with disrupted retroviruses of the infectious primate group showed that, while a homologous RIA detected only type-specific enzyme determinants, it did not distinguish the various woolly-gibbon retroviral DNA polymerases. A more broadly reactive heterologous assay utilizing an antiserum to R-MuLV RT detected group- but not interspecies-specific enzyme determinants. A comparison of immunologic assays for RT showed that: (1) highly purified RT is not essential for reliable results in enzyme neutralization or enzyme binding assays; (2) the greater sensitivity of enzyme binding compared to enzyme neutralization assays is a function of the antibody, not of the antigen. Competition RIAs using extracts of virus-infected cells showed that infectious primate retrovirus RT could be measured in a crude system and that cellular DNA polymerases alpha, beta and gamma did not compete with the labelled probe.

Animals↗