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J C Roder

Publications and source records attributed to J C Roder.

At least 37 records · Page 2Linked to original sources
PubMed1997

Mice lacking metabotropic glutamate receptor 5 show impaired learning and reduced CA1 long-term potentiation (LTP) but normal CA3 LTP.

Class I metabotropic glutamate receptors (mGluRs) have been postulated to play a role in synaptic plasticity. To test the involvement of one member of this class, we have recently generated mutant mice that express no mGluR5 but normal levels of other glutamate receptors. The CNS revealed normal development of gross anatomical features. To examine synaptic functions we measured evoked field EPSPs in the hippocampal slice. Measures of presynaptic function, such as paired pulse facilitation in mutant CA1 neurons, were normal. The response of mutant CA1 neurons to low concentrations of (1S,3R)-1-amino-cyclopentane-1,3-dicarboxylic acid (ACPD) was missing, which suggests that mGluR5 may be the primary high affinity ACPD receptor in these neurons. Long-term potentiation (LTP) in mGluR5 mutants was significantly reduced in the NMDA receptor (NMDAR)-dependent pathways such as the CA1 region and dentate gyrus of the hippocampus, whereas LTP remained intact in the mossy fiber synapses on the CA3 region, an NMDAR-independent pathway. Some of the difference in CA1 LTP could lie at the level of expression, because the reduction of LTP in the mutants was no longer observed 20 min after tetanus in the presence of 2-amino-5-phosphonopentanoate. We propose that mGluR5 plays a key regulatory role in NMDAR-dependent LTP. These mutant mice were also impaired in the acquisition and use of spatial information in both the Morris water maze and contextual information in the fear-conditioning test. This is consistent with the hypothesis that LTP in the CA1 region may underlie spatial learning and memory.

Animals↗
PubMed1997

Cellular distribution of a natural killer cell tumour recognition-related surface antigen in purified human lymphocytes.

Natural killer (NK) cells are large granular lymphocytes capable of human leucocyte antigen (HLA) unrestricted killing of tumour cells. A putative NK cell tumour-recognition molecule (NK-TR) was previously isolated and cloned. The predicted primary structure of the NK-TR revealed that the amino terminus of the protein shared high homology with cyclophilin proteins. In this study, we used rabbit antibodies directed against synthetic peptides corresponding to amino acids 476-497 of the NK-TR protein, to examine the expression of the NK-TR antigen in freshly purified human lymphocytes. Cell-surface staining experiments using these peptide antibodies indicated the presence of the NK-TR protein on the surface of human CD3+ T-cell populations purified from peripheral blood. There were individual donor differences in the levels of cell-surface expression of this antigen ranging from 35 to 90% in T lymphocytes and, NK cells purified from different healthy volunteers. The immunoreactivity of our peptide antibodies in immunoprecipitation showed that the NK-TR-related protein expressed in purified T cells is similar to that expressed in NK cells in terms of its electrophoretic mobility. Cell-surface staining experiments using the peptide antibodies revealed that the NK-TR-related protein is more abundantly expressed on the surface of purified T cells compared with NK cells. Northern blot analysis of the mRNA species transcribed in human lymphocytes revealed abundant expression of NK-TR-specific mRNA species in purified T cells. Furthermore, another mRNA species smaller than 7 kb was detected in both NK and T-cell populations of lymphocytes freshly isolated from peripheral blood. Expression at the cell surface of a cyclophilin-homologous protein in purified human T lymphocytes may indicate another function for the reported NK-TR protein, that is, distinct from tumour-cell recognition and cytosis.

Antigens, Surface↗
PubMed1997

Regulation of myelin-associated glycoprotein binding by sialylated cis-ligands.

Myelin-associated glycoprotein (MAG) and Schwann cell myelin protein (SMP) are highly glycosylated members of a newly defined family of cell adhesion molecules belonging to the immunoglobulin superfamily that recognize terminal sialic acid residues on N- and O-linked oligosaccharides. The importance of the N-linked oligosaccharides on MAG were determined by removal of the eight predicted carbohydrate addition sites by site-directed mutagenesis. The results suggest that all eight N-linked glycosylation sites are utilized in COS cells. N-linked glycosylation does not appear to be required for sialic acid-dependent MAG binding to erythrocytes. However, N-linked glycosylation of MAG does play a role in the proper folding of MAG. It was also shown that sialylation in the host cell expressing MAG and SMP could inhibit binding to erythrocytes. The degree to which SMP and MAG erythrocyte binding was affected by sialylation in the host cell was dependent on (a) the level at which MAG was expressed on the surface of the host cell and (b) the presence of MAG ligands on the host cell. The data suggest that cis-ligands on the host cell compete with trans-ligands on the target cell for the binding site(s) on MAG.

Animals↗
PubMed1996

Reduction of lower motor neuron degeneration in wobbler mice by N-acetyl-L-cysteine.

The murine mutant wobbler is a model of lower motoneuron degeneration with associated skeletal muscle atrophy. This mutation most closely resembles Werdnig-Hofmann disease in humans and shares some of the clinical features of amyotrophic lateral sclerosis (ALS). It has been suggested that reactive oxygen species (ROS) may play a role in the pathogenesis of disorders such as ALS. To examine the relationship between ROS and neural degeneration, we have studied the effects of agents such as N-acetyl-L-cysteine (NAC), which reduce free radical damage. Litters of wobbler mice were given a 1% solution of the glutathione precursor NAC in their drinking water for a period of 9 weeks. Functional and neuroanatomical examination of these animals revealed that wobbler mice treated with NAC exhibited (1) a significant reduction in motor neuron loss and elevated glutathione peroxidase levels within the cervical spinal cord, (2) increased axon caliber in the medial facial nerve, (3) increased muscle mass and muscle fiber area in the triceps and flexor carpi ulnaris muscles, and (4) increased functional efficiency of the forelimbs, as compared with untreated wobbler littermates. These data suggest that reactive oxygen species may be involved in the degeneration of motor neurons in wobbler mice and demonstrate that oral administration of NAC effectively reduces the degree of motor degeneration in wobbler mice. This treatment thus may be applicable in the treatment of other lower motor neuropathies.

Acetylcysteine↗
PubMed1996

Impaired cerebellar synaptic plasticity and motor performance in mice lacking the mGluR4 subtype of metabotropic glutamate receptor.

The application of the glutamate analog L-2-amino-4-phosphonobutyric acid (L-AP4) to neurons produces a suppression of synaptic transmission. Although L-AP4 is a selective ligand at a subset of metabotropic glutamate receptors (mGluRs), the precise physiological role of the L-AP4-activated mGluRs remains primarily unknown. To provide a better understanding of the function of L-AP4 receptors, we have generated and studied knockout (KO) mice lacking the mGluR4 subtype of mGluR that displays high affinity for L-AP4. The mGluR4 mutant mice displayed normal spontaneous motor activity and were unimpaired on the bar cross test, indicating that disruption of the mGluR4 gene did not cause gross motor abnormalities, impairments of novelty-induced exploratory behaviors, or alterations in fine motor coordination. However, the mutant mice were deficient on the rotating rod motor-learning test, suggesting that mGluR4 KO mice may have an impaired ability to learn complex motor tasks. Patch-clamp and extracellular field recordings from Purkinje cells in cerebellar slices demonstrated that L-AP4 had no effect on synaptic responses in the mutant mice, whereas in the wild-type mice 100 microM L-AP4 produced a 23% depression of synaptic responses with an EC50 of 2.5 microM. An analysis of presynaptic short-term synaptic plasticity at the parallel fiber-->Purkinje cell synapse demonstrated that paired-pulse facilitation and post-tetanic potentiation were impaired in the mutant mice. In contrast, long-term depression (LTD) was not impaired. These results indicate that an important function of mGluR4 is to provide a presynaptic mechanism for maintaining synaptic efficacy during repetitive activation. The data also suggest that the presence of mGluR4 at the parallel fiber-->Purkinje cell synapse is required for maintaining normal motor function.

Animals↗
PubMed1996

Physiological effects of CNTF-induced wasting.

The authors have shown previously that in addition to its survival effects on neurons and glia, ciliary neurotrophic factor (CNTF) induced potent cachectic effects and acute phase proteins when present in the peripheral circulation at concentrations of < or = 10 ng/ml. These effects did not depend upon the induction of other cytokine family members. Described here are the specific physiological effects which systemic administration of CNTF can induce in somatic tissue. Mice implanted with C6 glioma cells, genetically modified to secrete CNTF, exhibited rapid catabolism of adipose tissue and skeletal muscle, depressed steady-state levels of glucose and triglycerides, elevations in red blood cell content, gall bladder hypertrophy and thymic atrophy, with a disproportionate loss of CD4+/CD8+ T cells. This cachectic wasting resulted in death over a period of 7-10 days. Implantation of the parental C6 line, or C6 cells which express a non-secreted form of CNTF, did not result in overt effects over this time period. These findings have implications both for the biology of CNTF family members, and the therapeutic use of factors such as CNTF in vivo.

Adipose Tissue↗
PubMed1996

Conspecific exploration in the T-maze: abnormalities in S100 beta transgenic mice.

S100 beta, a calcium binding brain protein expressed by astrocytes, has been shown to be involved in higher neural processes, including hippocampal-dependent behavioral traits and hippocampal neuronal long-term potentiation (LTP) and depression (LTD), neurophysiological phenomena that may be involved in exploring, learning and remembering novel stimuli. In the present study, the exploratory behavior of previously generated transgenic mice overexpressing the protein are compared to that of normal control mice of identical genetic background and age in a T-maze. The test mice encountered a normal control and an S100 beta transgenic mouse (the choice mice) in the goal arms of the T-maze. We show that no test mice exhibited any preference for either genotype of choice mouse. However, there was a significant difference in the spatial and temporal exploratory pattern between control and S100 beta test mice, demonstrating that S100 beta overexpression significantly altered the behavior of the transgenic mice. We suggest that one probable factor underlying the abnormalities observed is impaired short-term memory.

Animals↗
PubMed1996

Memory and the effect of cold shock in the water maze in S100 beta transgenic mice.

S100 beta, a calcium binding astrocytic brain protein, influences hippocampal long-term potentiation (LTP) and depression (LTD), synaptic processes suggested to play role in spatial (contextual) learning and memory. In the present study we trained S100 beta transgenic and wild-type control mice in a nonspatial version of the Morris water maze, the visible platform task, and analyzed retention of memory over periods of 18 h, several days, and weeks. The results show that acquisition and retention were not altered in the S100 beta transgenic mice compared to control. However, a single alteration of an environmental stimulus, water temperature, significantly worsened the performance of transgenic mice. This impairment lasted for two consecutive trials separated by a 2-week intertrial interval, suggesting a temporary disturbance associated with memory processes. We discuss the possibility that these results are compatible with normal cortical but abnormal hippocampal functioning in the S100 beta transgenic mice.

Animals↗
PubMed1995

Ciliary neurotrophic factor promotes the terminal differentiation of v-myc immortalized sympathoadrenal progenitor cells in vivo.

Survival and differentiation of a sympathoadrenal progenitor cell line (termed MAH), transduced with a v-myc oncogene, was studied subsequent to transplantation in the peripheral and central nervous system of adult rats. In the brain, MAH cell survival depended on the secretion of ciliary neurotrophic factor (CNTF) by co-grafts of genetically modified glioma cells. No trophic factor supplement was required for development of the MAH cells in the peripheral nerve environment. Transplanted progenitor cells withdrew from the cell cycle within 48 h and differentiated into a prominent population of large sympathetic-like neurons. The neurons expressed the alpha subunit of the CNTF receptor and appropriate spatial distributions of cytoskeletal proteins and catecholamine related enzymes. The results identify a role for CNTF in the development of the sympathoadrenal cell lineage and support the concept of immortalized progenitor cells as alternatives to primary cells for cell replacement strategies in the nervous system.

Adrenal Medulla↗
PubMed1995

Neurons express ciliary neurotrophic factor mRNA in the early postnatal and adult rat brain.

The regional and subcellular localization in the central nervous system (CNS) of postnatal day 5, day 15, and adult rats of ciliary neurotrophic factor (CNTF) mRNA was examined by in situ hybridization with biotinylated riboprobes. Probe specificity was determined by Northern blot analysis of poly(A)+ RNA extracted from adult rat brain using digoxigenin labeled riboprobes and chemiluminescent detection. Both a 4 kb and a 1.2 kb transcript were detected in the cortex and brainstem. In situ hybridization revealed that CNTF mRNA was widely distributed in neurons and glia throughout the CNS at each of the developmental time points. The density of the neuronal hybridization signal was found to be greater in neuronal nuclei than in their cytoplasm. In the nucleus of most neurons, CNTF mRNA distribution was concentrated in a perinucleolar fashion. Alternate sections from the same animals, which were incubated with a specific polyclonal antibody against a CNTF peptide fragment, revealed that both neurons and glia in postnatal day 5, day 15, and adult rat brain were immunoreactive for CNTF.

Aging↗
PubMed1995

Functional topography of myelin-associated glycoprotein. II. Mapping of domains on molecular fragments.

The myelin-associated glycoprotein (MAG), an adhesion molecule of the immunoglobulin (Ig) superfamily with five Ig-like domains, was investigated with regard to its binding site(s) for the neuronal cell surface, collagen I, and heparin, using a panel of new monoclonal antibodies and cyanogen bromide cleavage fragments of MAG. All antibodies generated competed with each other for binding to MAG, indicating that they reacted with identical or closely related epitopes. Mapping of the reactive epitopes on recombinant deletion fragments of MAG expressed by Chinese hamster ovary (CHO) fibroblasts showed reactivity of monoclonal antibody 513 with domains I, II, and III, comprising the amino-terminal end of the extracellular domain. Monoclonal antibody 15 recognized domain III only. Binding of MAG-containing liposomes to neurons was blocked by antibodies 15 and 513. Cyanogen bromide (CNBr) fragments of domains I, II, and III bound to collagen type I under isotonic buffer conditions. CNBr fragments containing domains I and II were involved in binding to heparin. These observations suggest that domain III may be important for binding to the neuronal cell surface receptor for MAG, while domains I, II, and III interact with collagen type I and domains II and III with heparin.

3T3 Cells↗
PubMed1995

Conservation of functionally important epitopes on myelin associated glycoprotein (MAG).

Phylogenetic conservation of protein domains often points to functionally important regions. As a step toward mapping these sites on myelin associated glycoprotein (MAG) we have determined the species distribution of epitopes recognized by a panel of anti-MAG antibodies (Ab). Monoclonal antibodies (mAb) B11F7, GenS3 and 28 recognized MAG only in mammalian species. However, the mAb 513 which inhibits MAG binding recognized a conformational epitope in a wider distribution of species including, human (Homo sapiens), bovine (Bos taurus), rat (Rattus norvegicus), chicken (Gallus gallus), quail (Coturnix coturnix japonica), lizard (Iguana iguana), snake (Thamnophis sirtalis), frog (Xenopus laevis) and turtle (all tetrapods) but not in goldfish (Crassius aurata) (a teleost). However, only MAG from mammals was shown to bind rat dorsal ganglion neurons (DRGs) suggesting that structures additional to those recognized by mAb 513 must be involved in function. Antibody 28, on the other hand, recognized only MAG species which bound to neurons, suggesting that this epitope, in comparison with mAb 513, more closely represented the functionally important region of MAG. Observed species differences in glycosylation of MAG may be functionally significant. A newly developed polyclonal Ab against MAG recognized the protein in tetrapods and teleosts, but not chondricthyes. The results show that MAG is present in a wide spectrum of species.

Animals↗
PubMed1995

Over-expression of the DM-20 myelin proteolipid causes central nervous system demyelination in transgenic mice.

We have created transgenic mice bearing varying copy numbers of a transgene coding for normal DM-20, the alternatively spliced quantitatively minor isoform of myelin proteolipid protein. Demyelination of the CNS occurs as a consequence of 70 copies of this transgene. Overt symptoms begin at approximately 3 months with a wobbling gait. Occasional seizures lasting a few seconds begin at 3-4 months. These symptoms progress in severity with age. Death occurs by 8-10 months. Myelination in 2-month-old animals, before the onset of any overt symptoms, appears morphologically normal at the electron microscopic level. However, the myelin in these 2-month-old animals has a reduced amount of the major myelin proteolipid protein and about three times as much DM-20 as normal animals. In 7-month-old animals that appear to be undergoing demyelination in the CNS, both the major myelin proteolipid protein and DM-20 are greatly reduced relative to the 2-month-old animal. Mice with 17 copies of the transgene also have a reduced amount of the major myelin proteolipid protein but appear to be otherwise normal and have normal life spans (> 2 yr). Mice with low copy numbers of the transgene (2-4 copies) appear to be unaffected and have normal life spans.

Alternative Splicing↗
PubMed1994

High level functional engraftment of severe combined immunodeficient mice with human peripheral blood lymphocytes following pretreatment with radiation and anti-asialo GM1.

The severe combined immunodeficient (SCID) mouse engrafted with human peripheral blood lymphocytes (PBLs) is a potentially useful model for the study of cancer immunotherapy. For this application, rapid, consistent, and high level engraftment of SCID mice with functional human cytotoxic effector cells is necessary. To date, short term human lymphoid cell engraftment in SCID mice has generally been low and variable. Further, most of the human cells detected within the first 30 days are found in the peritoneal cavity. The purpose of the present study was to improve short term reconstitution of human PBLs in the SCID mouse. When untreated SCID mice were injected with human PBLs, the mean level of CD3+ cells in the spleens was < 5% on days 6-32 after injection, as determined by flow cytometry (FCM). Depletion of SCID mouse natural killer (NK) cells with anti-asialo GM1 only marginally improved short term reconstitution with human CD3+ cells. Preirradiation of SCID mice with 3 Gy improved reconstitution to over 16% CD3+ cells on days 12-14 following engraftment. However, the combination of pretreatment with anti-asialo GM1 plus radiation, significantly increased the mean percentage of human CD3+ cells in the spleen to 40% within 2 weeks following injection of PBLs. Human T cells positive for CD4, CD8, TcR alpha beta, and TcR gamma delta, and human NK and B cells were detected in the spleens of irradiated plus anti-asialo GM1 pretreated SCID mice. The presence of human lymphoid cells was confirmed by immunohistologic staining. The human immune cells in these mice were shown to be functional by the in vivo demonstration of an appropriate secondary immune response to the injection of tetanus toxoid and by an in vivo proliferative response to phytohemagglutinin. Human NK cells could be found in the spleens and peripheral blood of irradiated plus anti-asialo GM1 pretreated mice. These cells were also shown to be competent by their ability to lyse the human NK sensitive tumor targets K562 and MOLT-4 in 51Cr release assays. Thus, pretreatment of SCID mice with radiation plus anti-asialo GM1 significantly improves short term human PBL engraftment and provides a potentially useful model for the study of cancer immunotherapy.

Animals↗
PubMed1994

Increased CNTF gene expression in process-bearing astrocytes following injury is augmented by R(-)-deprenyl.

R(-)-deprenyl has been shown to rescue axotomized immature facial motoneurons with an efficacy comparable to that of the neurotrophic factors CNTF and BDNF (Salo and Tatton, J Neurosci Res 31:394-400, 1992; Ansari et al., J Neurosci 13:4042-4053, 1993). Recent work has suggested that some of the actions of (-)-deprenyl may be mediated through reactive astrocytes (Biagini et al., NeuroReport 4:955-958, 1993). To test this proposal we have developed an in vitro model of reactive gliosis consisting of a mixed astrocyte population of flat and process-bearing (PB) astroglia taken from postnatal day (PD) 2 or PD5 rat cerebral cortex. After mechanical wounding, PB astrocytes preferentially migrate into the wound zone while flat astrocytes maintain their position at the wound edge. CNTF mRNA was localized to PB astrocytes, but not flat astrocytes, as determined by in situ hybridization using biotin-labelled riboprobes. Following "wounding," there was an increase in CNTF mRNA in PB astrocytes only, which could be further enhanced by a single pulse of (-)-deprenyl (10(-8)-10(-11) M) 48 hr after injury. (-)-Deprenyl also increased the total process length of PB astrocytes after wounding by an average of 50%. The stereoisomer (+)-deprenyl (10(-9) M) had no effect on either astrocyte process length or CNTF mRNA content. This is the first report to our knowledge of an agent which can upregulate CNTF gene expression in astroglial cell culture as well as influence glial cell process length.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Localization of CNTF immunoreactivity to neurons and astroglia in the CNS.

Species specific antibodies were raised to a peptide of rat ciliary neurotrophic factor (CNTF-amino acids number 131-147). Following affinity purification, these antibodies were used to determine the pattern of CNTF immunoreactivity in adult rat and mouse brain, spinal cord, and sciatic nerve. Alternate sections stained using neurofilament and the affinity purified anti-CNTF antibody (HARC-1) demonstrate that CNTF immunoreactive neurons are present within the facial nucleus, dentate gyrus, olfactory bulb, basal forebrain, locus coeruleus, cortex and substantia nigra. In addition, neurons throughout the hippocampus, and Purkinje cells within the cerebellum also exhibit CNTF immunoreactivity. CNTF immunopositive neurons demonstrate a preponderance of nuclear staining, with some staining present in the cytoplasm. Alternate sections incubated with glial fibrillary acidic protein (GFAP) antibody also demonstrate glia which are positive for CNTF. In the peripheral nervous system, Schwann cells of the sciatic nerve exhibit strong immunoreactivity for CNTF, however staining is confined to the cytoplasm and is absent from the cell nucleus. These data demonstrate that CNTF immunoreactivity is broadly distributed throughout neurons and glia of the adult rodent nervous system.

Amino Acid Sequence↗
PubMed1994

High-resolution mapping of GenS3 and B11F7 epitopes on myelin-associated glycoprotein by expression PCR.

The GenS3 and B11F7 monoclonal antibodies (MAbs) have been widely used for biochemical and immunocytochemical experiments on myelin-associated glycoprotein (MAG), a cell adhesion molecule mediating the interaction between myelinating glia and axons. We have mapped the epitopes to within several amino acids on Ig domain 2 (D2) (amino acids 167-77) and domain 4 (D4) (amino acids 375-388) for GenS3 and B11F7, respectively. Domain deletion and substitution mutants of the MAG cDNA were first used to map the epitopes to a given domain. In the cases of GenS3, insertion mutants were used to resolve the epitope to a small region of D2. For the B11F7 epitope, a novel technique combining PCR and in vitro transcription and translation was used to generate small C-terminal deletions and map the epitope to 13 amino acids. Then, inhibition by peptides corresponding to the GenS3 (ELRPELSWLGHE; amino acids 167-177) and B11F7 (QLELPAVTPEDDGE; amino acids 375-388) epitopes was used to confirm the position of the epitopes based on the mutant data. Interestingly, the GenS3 epitope maps to a region predicted to be sequestered within the hydrophobic core of D2. This is consistent with the inability of GenS3 to recognize the epitope in native MAG; GenS3 epitope recognition occurs only in denatured MAG, where the epitope is more accessible. With the definition of the GenS3 and B11F7 epitopes, these antibodies will be useful for further structure-function studies on MAG.

Animals↗