Medical Neurobiology Commons^™

Direct Demonstration Of Retroviral Recombination In A Rhesus Monkey, Dawn P. Wooley, Randall A. Smith, Susan Czajak, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

Recombination may be an important mechanism for increasing variation in retroviral populations. Retroviral recombination has been demonstrated in tissue culture systems by artificially creating doubly infected cells. Evidence for retroviral recombination in vivo is indirect and is based principally on the identification of apparently mosaic human immunodeficiency virus type 1 genomes from phylogenetic analyses of viral sequences. We infected a rhesus monkey with two different molecularly cloned strains of simian immunodeficiency virus. One strain of virus had a deletion in vpx and vpr, and the other strain had a deletion in nef. Each strain on its own induced low virus …

The Type 2 Iodothyronine Deiodinase Is Expressed Primarily In Glial Cells In The Neonatal Rat Brain, Ana Guadaño-Ferraz, Maria Jesus Obregón, Donald L. St. Germain, Juan Bernal

Dartmouth Scholarship

Thyroid hormone plays an essential role in mammalian brain maturation and function, in large part by regulating the expression of specific neuronal genes. In this tissue, the type 2 deiodinase (D2) appears to be essential for providing adequate levels of the active thyroid hormone 3,5,3′-triiodothyronine (T3) during the developmental period. We have studied the regional and cellular localization of D2 mRNA in the brain of 15-day-old neonatal rats. D2 is expressed in the cerebral cortex, olfactory bulb, hippocampus, caudate, thalamus, hypothalamus, and cerebellum and was absent from the white matter. At the cellular level, D2 is expressed predominantly, if not …

Transgenic Mice Which Overexpress Nerve Growth Factor, Kathryn M. Albers, Brian M. Davis

Neuroscience Faculty Patents

Transgenic mice that express increased levels of nerve growth factor (NGF) in the epidermis and other stratified, keratinized epithelium. The nerve growth factor expressing transgenic mice are useful in the study of neurodegenerative disorders of the brain such as Parkinson's syndrome and Alzheimer's disease and for testing for drug candidates for the treatment of these diseases.

A Recombinant Inwardly Rectifying Potassium Channel Coupled To Gtp-Binding Proteins, Kim W. Chan, M. Noelle Langan, Jin Liang Sui, J. Ashot Kozak, Amanda Pabon, John A. A. Ladias, Diomedes E. Logothetis

Neuroscience, Cell Biology & Physiology Faculty Publications

GTP-binding (G) proteins have been shown to mediate activation of inwardly rectifying potassium (K+) channels in cardiac, neuronal and neuroendocrine cells. Here, we report functional expression of a recombinant inwardly rectifying channel which we call KGP (or hpKir3.4), to signify that it is K+ selective, G-protein-gated and isolated from human pancreas. KGP expression in Xenopus oocytes resulted in sizeable basal (or agonist-independent) currents while coexpression with a G-protein-linked receptor, yielded additional agonist-induced currents. Coexpression of KGP and hGIRK1 (a human brain homolog of GIRK1/Kir3.1) produced much larger basal currents than those observed with KGP or hGIRK1 alone, and upon coexpression …

Gtpase-Deficient G Alpha 16 And G Alpha Q Induce Pc12 Cell Differentiation And Persistent Activation Of Cjun Nh2-Terminal Kinases., Lynn E. Heasley, Brooke Storey, Gary R. Fanger, Laura Butterfield, J Zamarripa, D Blumberg, R A. Maue

Dartmouth Scholarship

Persistent stimulation of specific protein kinase pathways has been proposed as a key feature of receptor tyrosine kinases and intracellular oncoproteins that signal neuronal differentiation of rat pheochromocytoma (PC12) cells. Among the protein serine/threonine kinases identified to date, the p42/44 mitogen-activated protein (MAP) kinases have been highlighted for their potential role in signalling PC12 cell differentiation. We report here that retrovirus-mediated expression of GTPase-deficient, constitutively active forms of the heterotrimeric Gq family members, G alpha qQ209L and G alpha 16Q212L, in PC12 cells induces neuronal differentiation as indicated by neurite outgrowth and the increased expression of voltage-dependent sodium channels. Differentiation …

Excitatory Amino Acids And Seizures, Michael A. Rogawski

Michael A. Rogawski

No abstract provided.

Effects Of Natural Sequence Variation On Recognition By Monoclonal Antibodies Neutralize Simian Immunodeficiency Virus Infectivity, Weon Sang Choi, Catherine Collignon, Clotilde Thiriart, Dawn P. Wooley, E. J. Scott, Karen A. Kent, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

The determinants of immune recognition by five monoclonal antibodies (KK5, KK9, KK17, Senv7.1, and Senv101.1) that neutralize simian immunodeficiency virus infectivity were analyzed. These five neutralizing monoclonal antibodies were generated to native SIVmac251 envelope glycoprotein expressed by a vaccinia virus recombinant vector. All five recognize conformational or discontinuous epitopes and require native antigen for optimal recognition. These monoclonal antibodies also recognize SIVmac239 gp120, but they do not recognize gp120 of two natural variants of SIVmac239, 1-12 and 8-22, which evolved during the course of persistent infection in vivo (D.P.W. Burns and R.C. Desrosiers, J. Virol. 65:1843-1854, 1991). Recombinant viruses which …

High Rates Of Frameshift Mutations Within Homo-Oligomeric Runs During A Single Cycle Of Retroviral Replication, Dawn P. Wooley, H. M. Temin

Neuroscience, Cell Biology & Physiology Faculty Publications

Homo-oligomeric runs were inserted into a spleen necrosis virus-based retrovirus vector to determine the nature and rate of mutations within runs of 10 to 12 identical nucleotides during a single replication cycle. Clones of helper cells containing integrated copies of retroviral vectors were used to produce virus for infection of target (nonhelper) cells. Proviral sequences from target cell clones were compared with proviral sequences from helper cell clones to study mutations that occurred during a single cycle of replication. In addition to the internal region spanning the homo-oligomeric inserts, a naturally occurring run of 10 T's in the long terminal …

Simian Immunodeficiency Virus Mutants Resistant To Serum Neutralization Arise During Persistent Infection Of Rhesus Monkeys, Dawn P. Wooley, Catherine Collignon, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

We previously described the pattern of sequence variation in gp120 following persistent infection of rhesus monkeys with the pathogenic simian immunodeficiency virus SIVmac239 molecular clone (D.P.W. Burns and R.C. Desrosiers, J. Virol. 65:1843, 1991). Sequence changes were confined largely to five variable regions (V1 to V5), four of which correspond to human immunodeficiency virus type 1 (HIV-1) gp120 variable regions. Remarkably, 182 of 186 nucleotide substitutions that were documented in these variable regions resulted in amino acid changes. This is an extremely nonrandom pattern, which suggests selective pressure driving amino acid changes in discrete variable domains. In the present study, …

Gelatinase Activity In The Mouse Uterus During Early Pregnancy, Carol A. Brenner, Richard R. Adler, Gary L. Nieder

Gary L. Nieder

No abstract provided.

Gelatinase Activity In The Mouse Uterus During Early Pregnancy, Carol A. Brenner, Richard R. Adler, Gary L. Nieder

Neuroscience, Cell Biology & Physiology Faculty Publications

No abstract provided.

Anion Permeation In An Apical Membrane Chloride Channel Of A Secretory Epithelial Cell, Dan R. Halm, Raymond A. Frizzell

Neuroscience, Cell Biology & Physiology Faculty Publications

Single channel currents though apical membrane Cl channels of the secretory epithelial cell line T84 were measured to determine the anionic selectivity and concentration dependence of permeation. The current-voltage relation was rectified with single channel conductance increasing at positive potentials. At 0 mV the single channel conductance was 41 ± 2 pS. Permeability, determined from reversal potentials, was optimal for anions with diameters between 0.4 and 0.5 nm. Anions of larger diameter had low permeability, consistent with a minimum pore diameter of 0.55 nm. Permeability for anions of similar size was largest for those ions with a more symmetrical charge …

Strain-Specific Neutralizing Determinant In The Transmembrane Protein Of Simian Immunodeficiency Virus, Toshiaki Kodama, Dawn P. Wooley, Daniel P. Silva, Fulvia Dimarzo Veronese, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

Monoclonal antibody SF8/5E11, which recognizes the transmembrane protein (TMP) of simian immunodeficiency virus of macaque monkeys (SIVmac), displayed strict strain specificity. It reacted with cloned and uncloned SIVmac251 but not with cloned SIVmac142 and SIVmac239 on immunoblots. This monoclonal antibody neutralized infection by cloned, cell-free SIVmac251 and inhibited formation of syncytia by cloned SIVmac251-infected cells; these activities were specific to cloned SIVmac251 and did not occur with the other viruses. Site-specific mutagenesis was used to show that TMP amino acids 106 to 110 (Asp-Trp-Asn-Asn-Asp) determined the strain specificity of the monoclonal antibody. This strain-specific neutralizing determinant is located within a …

Selection Of Genetic Variants Of Simian Immunodeficiency Virus In Persistently Infected Rhesus Monkeys, Dawn P. Wooley, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

Genetic and antigenic variation may be one means by which lentiviruses that cause AIDS avoid elimination by host immune responses. Genetic variation in the envelope gene (env) was studied by comparing the nucleotide sequences of 27 clones obtained from two rhesus monkeys infected with molecularly cloned simian immunodeficiency virus. All 27 clones differed from each other and differed from the input clone in the gp120 (SU) portion of the envelope gene. Nucleotide substitutions were shown to accumulate with time at an average rate of 8.5 per 1,000 per year in SU. Surprisingly, the majority of nucleotide substitutions (81%) resulted in …

Antiepileptic Drugs: Pharmacological Mechanisms And Clinical Efficacy With Consideration Of Promising Developmental Stage Compounds, Michael A. Rogawski, Roger J. Porter

Michael A. Rogawski

No abstract provided.

Significance Of Premature Stop Codons In Env Of Simian Immunodeficiency Virus, Toshiaki Kodama, Dawn P. Wooley, Yathirajulu M. Naidu, Harry W. Kestler Iii, Muthiah D. Daniel, Yen Li, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

The location of the translational termination codon for the transmembrane protein (TMP) varies in three infectious molecular clones of simian immunodeficiency virus from macaques (SIV_mac). The SIV_mac251 and SIV_mac142 infectious clones have premature stop signals that differ in location by one codon; transfection of these DNAs into human HUT-78 cells yielded virus with a truncated TMP (28 to 30 kilodaltons [kDa]). The SIV_mac239 infectious clone does not have a premature stop codon in its TMP-coding region. Transfection of HUT-78 cells with this clone initially yielded virus with a full-length TMP (41 kDa). …

Aldosterone Stimulates K Secretion Across Mammalian Colon Independent Of Na Absorption, Gerhard Rechkemmer, Dan R. Halm

Neuroscience, Cell Biology & Physiology Faculty Publications

K transport across guinea pig (Cavia porcellus) distal colon was measured in vitro using isotopically determined unidirectional fluxes. Aldosterone stimulated electrogenic Na absorption, as measured by amiloride-sensitive short-circuit current (I_sc), and reduced net K absorption from +2.5 ± 0.2 µEq/cm² per hr to +0.8 ± 0.3 µEq/cm² per hr (mean ± SEM). Amiloride addition to the mucosal solution did not enhance net K absorption, as expected if inhibiting active Na absorption would reduce active K secretion as in the distal nephron. The amiloride-insensitive I_sc was -1.0 ± 0.2 µEq/cm² per hr (mean ± SEM) …

Hyperosmotic Modification Of Brain Extracellular Space And Blood-Brain Barrier In The Rat As Observed By Electron Microscopy, Susan A. Meacham

Loma Linda University Electronic Theses, Dissertations & Projects

The highly selective permeability of cerebral blood vessels, termed the blood-brain barrier, is due to endothelial tight junctions (zonulae occludentes). Hyperosmotic solutions reversibly open the blood-brain barrier. The purpose of this study is to determine the immediate effect of hyperosmotic arabinose perfusion upon the cerebrovascular endothelium, the morphology of the cerebral cortex, and the cortical extracellular space.

Twelve adult male Sprague-Dawley rats were separated into three groups of four animals each: control, intermediate, and extreme. Controls were perfused with a Tyrode's solution prewash followed by trialdehyde fixative. The intermediate group received a prewash of 740 mmol/kg arabinose followed by the …

A Semiquantitative Analysis Of The Chorda Tympani Taste Pathway In The Rat Brain With The 2-Deoxyglucose Method, Charles L. Hardison

Loma Linda University Electronic Theses, Dissertations & Projects

This thesis is based on a semiquantitative analysis of four nuclear relay centers [nucleus tractus solitarius (NTS), parabrachial nucleus (PBN), ventroposteromedial nucleus (VPM), gustatory neocortex (GNC)] in the chorda tympani nerve pathway. Metabolic changes were examined by the 2-deoxyglucose method in which one chorda tympani nerve was chemically stimulated and its contralateral side lesioned. Coronal sections of four taste centers were prepared for autoradiography and Nissl staining. Densities of the autoradiograms were determined by means of a video based image processor. Relative differences in the optical density ratios of the stimulated versus lesioned nuclear grey matter areas to white matter …

Electrophysiology Of Flounder Intestinal Mucosa. I. Conductance Properties Ofthe Cellular And Paracellular Pathways, Dan R. Halm, Edward J. Krasny Jr., Raymond A. Frizzell

Neuroscience, Cell Biology & Physiology Faculty Publications

We evaluated the conductances for ion flow across the cellular and paracellular pathways of flounder intestine using microelectrode techniques and ion-replacement studies. Apical membrane conductance properties are dominated by the presence of Ba-sensitive K channels. An elevated mucosal solution K concentration, [K]_m, depolarized the apical membrane potential (ψ_a) and, at [K]_m less than 40 mM, the K dependence of ψ_a was abolished by 1-2 mM mucosal Ba. The basolateral membrane displayed Cl conductance behavior, as evidenced by depolarization of the basolateral membrane potential (ψ_b) with reduced serosal Cl concentrations, [Cl]_s. …

Electrophysiology Of Flounder Intestinal Mucosa. Ii. Relation Of The Electrical Potential Profile To Coupled Nacl Absorption, Dan R. Halm, Edward J. Krasny Jr., Raymond A. Frizzell

Neuroscience, Cell Biology & Physiology Faculty Publications

We characterized the hyperpolarization of the electrical potential profile of flounder intestinal cells that accompanies inhibition of NaCl cotransport. Several observations indicate that hyperpolarization of ψ_a and ψ_b (Δψ_a,b) results from inhibition of NaCl entry across the apical membrane: (a) the response was elicited by replacement of mucosal solution Cl or Na by nontransported ions, and (b) mucosal bumetanide or serosal cGMP, inhibitors of NaCl influx, elicited Δψ_a,b and decreased the transepithelial potential (ψ_t) in parallel. Regardless of initial values, ψ_a and ψ_b approached the equilibrium potential for K (E^{K …}

Recognition And Characterization Of Stage-Specific Oocyst/Sporozoite Antigens Of Toxoplasma Gondii By Human Antisera., Lloyd H. Kasper, Patricia L. Ware

Dartmouth Scholarship

Human infection with Toxoplasma gondii is presumed due to the ingestion of either tissue cysts containing bradyzoites or oocyst/sporozoites that are excreted in the feces of infected cats. The incidence of human infection in the general population by either of these routes is unknown. We have previously described unique stage-specific oocyst/sporozoite antigens identified by murine hybridoma monoclonal antibodies. We obtained acute and convalescent antitoxoplasma antisera from patients in an epidemiologically well-documented outbreak of oocyst-transmitted infection associated with the ingestion of contaminated water. An enzyme-linked immunosorbent assay comparing equal numbers of tachyzoites (invasive stage) and oocyst/sporozoite (excreted stage) indicated that these …

Metabolism In Preimplantation Mouse Embryos, Harry M. Weitlauf, Gary L. Nieder

Neuroscience, Cell Biology & Physiology Faculty Publications

The ability of preimplantation mouse embryos to utilize glucose oxidatively is controlled, in part at least, at the level of glycolysis. Various experimental observations are reviewed that indicate the regulatory mechanism in delayed implanting blastocysts involves the classic negative allosteric feedback of high levels of ATP on phosphofructokinase while the situation in 2-cell embryos appears to be more complicated. That is, in addition to the usual negative effect of ATP and citrate on phosphofructokinase, there appears to be a modification of hexokinase that prevents phosphorylation of adequate amounts of glucose and results in low levels of fructose-6-phosphate at the 2-cell …

Activation Of Long Descending Propriospinal Neurons In Cat Spinal Cord, Robert J. Adams, Robert D. Skinner, Ronald S. Remmel

Journal of the Arkansas Academy of Science

Isolated mammalian spinal cord has been shown capable of generating locomotor activity. Propriospinal systems assumed to coordinate fore- and hindlimb activity are poorly understood. This study characterizes the long descending propriospinal (LDP) neurons in terms of the location of the somas and their peripheral inputs by direct neuronal recording. Anatomical studies using axonal retrograde transport of horseradish peroxidase from the lumbar to the cervical spinal cord as a tracer first described these neurons. Two hundred and thirty-one LDP neurons were identified in electrophysiological experiments. Of these, 123 responded to natural stimulation, and about 50% of the others were activated only …

Connections Of The Mesencephalic Locomotor Region (Mlr) In The Cat, E. Garcia-Rill, Robert D. Skinner, S. A. Gilmore

Journal of the Arkansas Academy of Science

The cat entopeduncular nucleus (EN), which is the main output of the basal ganglia, is known to project to the mesencephalic tegmentum. We have been able to elicit antidromic responses in single EN neurons from the region of the mesencephalic locomotor region (MLR), then transect (precollicular-postmamillary) the brainstem and elicit rhythmic movements of the limbs by stimulation of the same site in the same animal. Injections of the fluorescent dye 2,4 diamidino phenylindole 2 HCL (DAPI) into this area induces retrograde labeling of cell bodies in EN and motor cortex. Injections of a tritiated amino acid (leucine) into the motor …

Interplanetary Magnetic Field Sector Polarity And Neuropsychiatric Adaption: A Reanalysis, Hans W. Wendt

Journal of the Minnesota Academy of Science

Recent Russian studies reported associations between adaptation syndromes and (solar activity-related) interplanetary magnetic field (IMF) sector polarity. Certain ambiguities in the original work suggested some reanalyses, based on the graphs and data made available. Syndrome status exhibited a biphasic (semi annual) trend. It also correlated with specific geomagnetic changes during magnetic storms and possibly after sudden impulses. Moreover, lag effects and higher order interactions were noted among all parameters investigated. The restudy tends to support some of the original findings but also suggests new conclusions regarding potential relationships between geophysical/solar and psychophysiological processes.

Taste-Masking: Function Of Exaggerated Prandial Drinking In Desalivate Mice, Richard C. Lewis

Journal of the Arkansas Academy of Science

Taste thresholds for the bitter compound sucrose octaacetate (SOA) were elevated by desalivation in mice. Thresholds were determined for control and experimental animals both before and after ligation of all salivary ducts. There was a significant increase in SOA thresholds in the desalivate mice, and the pre- to post-operative differences in threshold between the control and experimental groups were significant. The altered response to SOA by desalivate mice is shown to be due to the assumption of a prandial pattern of drinking as a result of desalivation. This conclusion is based on experiments with wet mash which failed to show …

Stereospecific Opiate Binding In Human Erythrocyte Membranes And Changes In Heroin Addicts, Leo G. Abood, Holly G. Atkinson, Mary Macneil

Publications and Research

Stereospecific opiate binding has been demonstrated in human erythrocyte membranes, having a Kd of 9-10(-9) M. In most respects the binding characteristics resemble those of synaptic membranes. These included the correlation of binding affinity and pharmacological potency of opiates; competition by naloxone; inhibition by Ca2+ and Na+; and sensitivity to phospholipases and trypsin. A comparison of stereospecific opiate binding in control human subjects and heroin addicts revealed a 43% increase in the addict group.

A Study Of Regeneration Of Peripheral Nerve In The Rat, James C. Jay

Loma Linda University Electronic Theses, Dissertations & Projects

Due to improvements in fixation and embedding techniques a restudy of regenerating peripheral nerve was thought advisable in order to better understand and describe the processes which take place.

Femoral nerves of white Wistar rats were used. The lesions were made by either crushing or severing the nerve. After a period of regeneration, varying from 12 hours to six days, the wound was reopened and the injured portion fixed in situ with 1% osmium tetroxide. The tissues were then removed, dehydrated, embedded and sectioned. The sections were stained and studied with a modified RCA EMU2A electron microscope.

Observations revealed that …

Cerebral Anoxia And Its Residuals: Pt. Iii. The Structural Changes, Cyril B. Courville

Medical Arts and Sciences: A Scientific Journal of the College of Medical Evangelists

No abstract provided.