bluuebunny/crossref_metadata_2025 huggingface dataset:
Synaptic plasticity in the basolateral amygdala induced by hippocampal formation stimulation in vivo
Authors: Maren S , Fanselow MS
Published: April 2018

Several studies suggest that axonal projections from the hippocampal formation (HF) to the basolateral amygdala (BLA) play a role in Pavlovian fear conditioning to contextual conditional stimuli. We have used electrophysiological techniques to characterize neuronal transmission in these projections in urethane-anesthetized rats. Single- pulse electrical stimulation of the ventral angular bundle (VAB), which carries projections from the HF to the BLA, reliably evoked a biphasic extracellular field potential in the BLA that consisted of an early, negative and a late, positive component. The negative component of the field potential occurred at a short latency (3–8 msec), was both temporally and spatially correlated with VAB-evoked multiple-unit discharges in the BLA, and exhibited properties typical of a monosynaptic response. Infusion of lidocaine or glutamate receptor antagonists into the BLA attenuated VAB-evoked field potentials, indicating that they are generated by local synaptic glutamatergic transmission. Both paired-pulse stimulation and brief trains of high- frequency stimulation (HFS) induced a short-lasting facilitation of BLA field potentials, whereas longer and more numerous trains of HFS produced an enduring, NMDA receptor-dependent long-term potentiation (LTP) of the potentials. The induction of LTP was accompanied by a decrease in paired-pulse facilitation (PPF), suggesting a presynpatic modification underlying its expression. Electrolytic lesions placed in regions of the HF that project to the BLA or excitotoxic lesions placed in the BLA eliminated Pavlovian fear conditioning to a contextual conditional stimulus. The critical role of both structures in context conditioning implicates plasticity at HF-BLA synapses in this form of learning.
The behavior of APO-SUS rats in animal models with construct validity for schizophrenia
Authors: Ellenbroek BA , Geyer MA , Cools AR
Published: April 2018

Schizophrenic patients are known to suffer from a number of information processing disturbances, including deficits in both prepulse inhibition of startle and latent inhibition. Since these behavioral phenomena can also be observed in animals, they represent an ideal starting point for developing animal models having construct validity for specific deficits observed in schizophrenia. The principal question is how to induce a condition in animals most similar to the schizophrenic deficit. In the present study, we have selected rats on the basis of their response to an open filed or to the dopaminergic agonist apomorphine, and evaluated their prepulse inhibition and latent inhibition. We used three different selection procedures (open field selection for novelty response, gnawing cage selection for apomorphine response, and pharmacogenetic selection for apomorphine response). The results show that, irrespective of the selection procedure used, rats with a high response to novelty or apomorphine susceptible (collectively called APO-SUS rats) show diminished prepulse inhibition of the acoustic startle response as compared to rats with a low response to novelty or apomorphine unsusceptible (collectively called APO-UNSUS rats). This difference was apparent only at low prepulse intensities. Moreover, these APO-SUS rats show diminished latent inhibition in a conditioned taste aversion paradigm as compared to APO- UNSUS rats. Given the fact that the pharmacogenetically bred APO-SUS rats show several central nervous, endocrinological, and immunological similarities to schizophrenic patients, they are hypothesised to represent an interesting nonpharmacological animal model for schizophrenia-prone patients.
Effects of delayed visual information on the rate and amount of prism adaptation in the human
Authors: Kitazawa S , Kohno T , Uka T
Published: April 2018

Accurate reaching towards a visual target is initially disturbed when the visual field is displaced by prisms, but recovers with successive trials. To determine how the improvement depends on the visual error signals associated with the motor output, the time course of prism adaptation was studied with delayed visual information on the error. Subjects were trained to reach rapidly at a target on a tangent screen. Vision was always blocked during the movement, and allowed again only after the index finger touched the screen. One experiment consisted of three sets of 30 trials. In the first set, the subject wore no prisms and vision was allowed without delay. In the second, the visual field was displaced by prisms, and vision was available only after a delay period of 0–10,000 msec while the subjects maintained their final pointing position. Initially, the subject misreached the target by about the amount of visual displacement (60 mm). Errors decreased with trials by an amount proportional to the error in the preceding trial. The rate of decrease of error was generally largest when the delay was 0 msec, became significantly smaller when the delay was 50 msec, and showed only gradual change with longer delays. In the third set, the subject wore no prisms and vision was allowed without delay. Initial misreaching in the direction opposite to the visual displacement, reflecting the amount of adaptation in the second set, was generally largest with no delay (median of 46 mm) and significantly smaller with 50 msec and longer delays (17–33 mm).(ABSTRACT TRUNCATED AT 250 WORDS)
The chronology of oligodendrocyte differentiation in the rat optic nerve: evidence for a signaling step initiating myelination in the CNS
Authors: Colello RJ , Devey LR , Imperato E , Pott U
Published: April 2018

In order to determine the signals that initiate axon myelination in the CNS, we have chronicled the differentiation of oligodendrocytes in the rat optic nerve and related this to the time course and spatial gradient seen for optic axon myelination. By using markers specific to the varying stages of oligodendrocyte differentiation we found that oligodendrocyte progenitor cells, present throughout the length of the nerve at postnatal day 2, mature into GC+ oligodendrocytes in a chiasm to eye progression. This gradient along the nerve of oligodendrocyte differentiation continues with oligodendrocytes near the chiasm expressing the genes and encoded proteins to MBP and PLP 3 d before oligodendrocytes near the eye. Although oligodendrocyte differentiation and maturation occurs in a chiasm to eye gradient along the nerve, optic axon segments near the eye are ensheathed with myelin before segments near the chiasm. This suggests that the myelination of optic axons is initiated by a signaling step that is independent of oligodendrocyte differentiation and is stronger near the eye than the chiasm region of the nerve. By examining proposed axonal signals, we found that the onset of myelination is independent of the electrical activity of an axon but can be correlated to the size of an axon.
A single histidine residue is essential for zinc inhibition of GABA rho 1 receptors
Authors: Wang TL , Hackam A , Guggino WB , Cutting GR
Published: April 2018

The GABA rho 1 subunit, cloned from a human retina library, can form homooligomeric receptors with properties similar to GABAc receptors characterized in retinal cells. The divalent cation Zn2+, abundant in the CNS and retina, was found to inhibit GABA rho 1 receptors in a voltage-independent manner. Varying the extracellular pH from 7.4 to 5.6 significantly reduced this inhibitory effect. This pH profile suggested that one or more histidine residues might play a role in the interaction between Zn2+ and the GABA rho 1 receptor. Site-directed mutagenesis revealed that a single histidine residue (His 156) in the putative extracellular domain of rho 1 was critical for Zn2+ sensitivity. Substitution of this amino acid with tyrosine (H156Y) created a functional GABA receptor with agonist and channel properties indistinguishable from wildtype. However, the H156Y mutant was insensitive to Zn2+, even at concentrations as high as 1 mM. Mutation to aspartic acid, an amino acid that can interact with Zn2+ in other proteins, preserved sensitivity to Zn2+ but abolished the pH-dependent effect. This histidine residue is also involved in Ni2+ and Cd2+ interaction since the H156Y mutation completely suppressed the inhibition effects of these two cations. These data demonstrate that an extracellular histidine residue is critical for transition metal cation sensitivity of GABA rho 1 receptors.
Cell surface morphology identifies microglia as a distinct class of mononuclear phagocyte
Authors: Giulian D , Li J , Bartel S , Broker J , Li X , Kirkpatrick JB
Published: April 2018

To investigate differences among brain-derived microglia and other classes of immune cells, we compared the morphologies and growth properties of mononuclear phagocytes isolated from tissues of the newborn rat. Scanning EM shows that microglia from postnatal rat brain are covered with spines (typically > 20 per cell body) in a distinctive manner which contrasts the smooth surfaces of bone marrow cells and the ruffled surfaces of tissue macrophages from spleen, liver, and peritoneum. The spine-bearing surface of microglia is a specific cell marker, for it does not change with age or after exposure to cytokines or other immunostimulants. Approximately 99% of mononuclear phagocytes cultured from normal adult rat brain are spinous microglia. Five days after injury to rat brain, cells at sites of Wallerian degeneration are essentially all spinous ones while nearly 30% of cells found within areas of infarction or penetrating trauma are invading macrophages. In a similar way, nearly all cells isolated from normal, postmortem adult human brain are spine-bearing microglia (> 99% homogeneity). Brains from patients with amyotrophic lateral sclerosis contain only spinous microglia whereas cells from HIV-1 infected brains include significant numbers of invading ruffled macrophages. Cultured microglia, unlike cultured bone marrow precursors, monocytes, or tissue macrophages, spontaneously develop long, thin processes that extend hundreds of microns in length. Microglia retract these processes after exposure to fetal bovine serum, laminin, or such immunostimulants as recombinant murine interferon gamma (rmIFN gamma) and lipopolysaccharide. Of all types of mononuclear phagocytes tested, only microglia differentiate into quiescent ramified cells when in contact with astrocytes. Thus, microglia represent a unique class of cell maintained, in part, by astroglia as dormant, ramified mononuclear phagocytes in mature CNS. Application of cell surface criteria described here will allow study of distinct populations of mononuclear phagocytes associated with neurologic disorders.
Observation and quantitation of exocytosis from the cell body of a fully developed neuron in Planorbis corneus
Authors: Chen G , Gavin PF , Luo G , Ewing AG
Published: April 2018

We have discovered a neuronal system that releases neurotransmitter via exocytosis from the cell body. In the large dopamine cell of the pond snail Planorbis corneus, depolarization induces rhythmic release of dopamine from the cell body. When a stimulant is applied extracellularly or intracellularly in situ to the cell body, transient dopamine concentration packets that appear in a bursting pattern are observed. Dopamine release is calcium dependent and release is on the time scale expected for exocytosis (2 to 4 msec rise times). Quantitation of individual events reveals an average of 818,000 molecules per exocytotic event. As many as 89,000 individual exocytotic events have been observed following a single stimulation of one cell. Neurotransmitter exocytosis from the neuronal cell body appears to represent an alternative form of neurocommunication to synaptic transmission.
The migration of luteinizing hormone-releasing hormone neurons in the developing rat is associated with a transient, caudal projection of the vomeronasal nerve
Authors: Yoshida K , Tobet SA , Crandall JE , Jimenez TP , Schwarting GA
Published: April 2018

Luteinizing hormone-releasing hormone (LHRH) neurons originate in the olfactory placode and vomeronasal organ and migrate to the brain from embryonic day 14 (E14) in the rat. We investigated the development of the vomeronasal nerve and its role as a guide for migrating LHRH neurons. Using fluorescent, lipophilic dye tracing methods, we observed axons that emerge from the vomeronasal organ and cross the nasal septum as several large fascicles. At E14–15, these fascicles converge as they enter the region of the cribriform plate and subsequently disperse, projecting dorsally and caudally across the olfactory bulb and rostral forebrain. At E16, the dorsal branch of the vomeronasal nerve forms a more tightly fasciculated projection; the caudal fibers remain dispersed, extending along the medial forebrain. The number of caudally directed axons decreases during development, leaving four or five present at postnatal day 4 (P4). Immunohistochemical studies indicate that the vomeronasal nerve can be divided into four spatially distinct subpopulations of fibers. One subset, composed of caudal fibers that terminate in the lamina terminalis, selectively expresses TAG-1, a transient axonal surface glycoprotein and PSA-N-CAM, a highly polysialated form of neural cell adhesion molecule. The extension and subsequent retraction of this branch of the vomeronasal nerve corresponds spatially and temporally with the migration of LHRH neurons from the nasal cavity to the brain. Our studies show that between E14 and E18, LHRH neurons migrate in contact with the TAG-1+, PSA-N-CAM+ caudal branch of the vomeronasal nerve.
Perisynaptic surface distribution of multiple classes of nicotinic acetylcholine receptors on neurons in the chicken ciliary ganglion
Authors: Horch HL , Sargent PB
Published: April 2018

Most nicotinic ACh receptors (AChRs) studied to date in the embryonic chicken ciliary ganglion are recognized either by monoclonal antibody (mAb) 35 or by alpha-bungarotoxin (alpha-Bgt). Previous studies found that mAb 35-AChRs are found at both synaptic and extrasynaptic sites, while alpha-Bgt-AChRs are found exclusively at extrasynaptic sites. To gain a three-dimensional understanding of the distribution of these two AChR classes and their spatial relationship to synaptic sites, we have visualized mAb 35-AChRs and alpha-Bgt-AChRs immunofluorescently using laser scanning confocal microscopy and have compared their distribution with that of the synaptic vesicle antigen SV2. Both mAb 35-AChR and alpha-Bgt-AChRs are found in clusters that are widely distributed over the surface of embryonic chicken ciliary ganglion neurons. Some mAb 35- AChRs are located at synaptic sites, but the bulk of them are located extrasynaptically in well-defined patches measuring 1–4 micron in diameter. alpha-Bgt-AChRs are found almost exclusively in these extrasynaptic sites, which thus contain both AChR types. These sites are often surrounded by elements of the synaptic calyx but are themselves largely free of SV2 antigen. In 14 week chickens the relationship between mAb 35-AChRs, alpha-Bgt-AChRs, and synaptic sites is similar to that in embryos except that in this instance individual synaptic boutons are often surrounded by AChR-containing patches. These results suggest that most surface AChRs in both embryonic and mature chicken ciliary neurons are perisynaptic, which raises questions about the function of these AChRs.
Intrastriatal implantation of fibroblasts genetically engineered to produce brain-derived neurotrophic factor prevents degeneration of dopaminergic neurons in a rat model of Parkinson's disease
Authors: Levivier M , Przedborski S , Bencsics C , Kang UJ
Published: April 2018

Parkinson's disease (PD) is a neurodegenerative disorder characterized by a progressive loss of the dopaminergic neurons of the substantia nigra pars compacta (SNpc). Although various treatments are successfully used to alleviate the symptoms of PD, none of them prevents or halts the neurodegenerative process of the disease. Brain- derived neurotrophic factor (BDNF), a member of the neurotrophin family of proteins, supports the survival and the differentiation of dopaminergic neurons. BDNF also prevents the death of dopaminergic neurons in vitro, which suggests that it may be of possible use in the development of neuroprotective therapies for PD. To determine whether BDNF is neuroprotective for SNpc dopaminergic neurons in the adult brain, we used a rat model of PD in which degeneration of 60–70% of these neurons was induced by an intrastriatal injection of 6- hydroxydopamine (6-OHDA). We report here that intrastriatal grafts of fibroblasts genetically engineered to produce BDNF partially prevent the loss of nerve terminals and completely prevent the loss of cell bodies of the nigrostriatal dopaminergic pathway that is induced by the intrastriatal injection of 6-OHDA. In contrast, the implantation of control fibroblasts that did not produce BDNF failed to protect nerve terminals and cell bodies against 6-OHDA-induced damage. Our observation that grafts of BDNF-producing fibroblasts protect against 6- OHDA-induced degeneration of SNpc dopaminergic neurons in the adult rat brain opens new perspectives for treatments aimed at the prevention of neurodegeneration in PD, using gene therapy and neurotrophic factors such as BDNF.
Regional, cellular, and subcellular variations in the distribution of D1 and D5 dopamine receptors in primate brain
Authors: Bergson C , Mrzljak L , Smiley JF , Pappy M , Levenson R , Goldman-Rakic PS
Published: April 2018

The pathways governing signal transduction in the mesocortical and nigrostriatal dopamine systems of the brain are of central importance in a variety of drug actions and neurological diseases. We have analyzed the regional, cellular, and subcellular distribution of the closely related D1 and D5 subtypes of dopamine receptors in the cerebral cortex and selected subcortical structures of rhesus monkey using subtype specific antibodies. The distribution of D1 and D5 receptors was highly differentiated in subcortical structures. In the neostriatum, both D1 and to a lesser extent D5 antibodies labeled medium spiny neurons, while only D5 antibodies labeled the large aspiny neurons typical of cholinergic interneurons. In the caudate nucleus, D1 labeling was concentrated in the spines and shafts of projection neurons, whereas D5 antibodies predominantly labeled the shafts, and less commonly, the spines of these cells. The D1 receptor was abundantly expressed in the neuropil of the substantia nigra pars reticulata while the D5 antibodies labeled only a few scattered cell bodies in this structure. Conversely, D5 antibodies labeled cholinergic neurons in the basal forebrain more intensely than D1 antibodies. Within the cerebral cortex and hippocampus, D1 and D5 antibody labeling was prominent in pyramidal cells. Double-label experiments revealed that the two receptors were frequently coexpressed in neurons of both structures. Ultrastructurally, D1 receptors were especially prominent in dendritic spines whereas dendritic shafts were more prominently labeled by the D5 receptor. The anatomical segregation of the D1 and D5 receptors at the subcellular level in cerebral cortex and at the cellular level in subcortical areas suggest that these closely related receptors may be preferentially associated with different circuit elements and may play distinct regulatory roles in synaptic transmission.
Increased production of 4 kDa amyloid beta peptide in serum deprived human primary neuron cultures: possible involvement of apoptosis
Authors: LeBlanc A
Published: April 2018

The etiology of the amyloid beta peptide in sporadic Alzheimer's disease (AD) is not known. Amyloid beta peptide (A beta), a proteolytic product of the amyloid precursor protein (APP), is deposited in the senile plaques and cerebrovascular tissues of individuals with either sporadic or familial AD (FAD). Increased A beta production from mutant APPs in FAD fosters the hypothesis that overexpression of A beta plays a primary role in the pathogenesis of AD. The absence of APP mutations in sporadic AD which displays identical pathological features than FAD such as synapse and neuronal loss, senile plaques and neurofibrillary tangles, suggests other causes for overexpression and/or deposition of A beta. To investigate the effect of neuronal death on APP metabolism and A beta secretion, human primary neuron cultures were induced to undergo apoptosis by serum deprivation. Serum deprived neurons display shrunken and rounded morphology, contain condensed chromatine and fragmented DNA, which are characteristic of apoptosis. In serum deprived neurons, metabolism of APP through the nonamyloidogenic secretory pathway is decreased to 20% from 40% in control cultures whereas 4kDa A beta is increased three- to fourfold. The results suggest that human neurons undergoing apoptosis generate excess A beta and indicates a possible mechanism for increased A beta in the absence of APP mutations.
ATP-induced cytoplasmic calcium mobilization in Bergmann glial cells
Authors: Kirischuk S , Moller T , Voitenko N , Kettenmann H , Verkhratsky A
Published: April 2018

ATP receptor mediated Ca2+ signaling was recorded from Bergmann glial cells in cerebellar slices obtained from mice of different ages (postnatal days 6 to 45). To measure the cytoplasmic concentration of Ca2+ ([Ca2+]in), either individual cells were loaded with the Ca(2+)- sensitive probes using the whole cell patch clamp technique or slices were incubated with the dye and the microfluorimetric system was focused on individual cells. Signals were recorded either with single-detector microfluorimetry of the dye fura-2 or by confocal laser scanning microfluorimetry (fluo-3-based recordings). Extracellular application of 100 microns ATP caused a transient elevation of [Ca2+]in, which amplitude was significantly higher in Bergmann glial cell processes as compared with their soma. The rank order of potency for the purinoreceptor agonists was: ADP > or = ATP > UTP> > AMP = adenosine = alpha, beta-methylene-ATP. ATP-triggered Ca2+ transients were reversibly inhibited by the P2 purinoreceptor agonist suramin (100 microM). The involvement of P2 metabotropic receptors is inferred by the observation that ATP mediated cytoplasmic Ca2+ transients were not associated with a measurable change in membrane conductance. The [Ca2+]in increase was due to release from inositol–1,4,5-trisphosphate (InsP3)-sensitive intracellular stores since responses were still observed in Ca(2+)-free extracellular solutions and were irreversibly blocked by the inhibitor of the sarco(endo)plasmic reticulum Ca2+ ATPase, thapsigargin, and by the competitive inhibitor of the InsP3- gated intracellular Ca2+ channels heparin. Intracellular dialysis altered the refilling process of the InsP3-sensitive stores, suggesting that cytoplasmic factors control ATP mediated Ca2+ signalling.
Fibroblast growth factor 2 increases Otx2 expression in precursor cells from mammalian telencephalon
Authors: Robel L , Ding M , James AJ , Lin X , Simeone A , Leckman JF , Vaccarino FM
Published: April 2018

Dissociated primary cultures from rat telencephalon at different developmental stages were used to study the effect of basic fibroblast growth factor (FGF2) on Otx2, Dlx1, and Emx1, three homeobox genes expressed in different regions of the developing mammalian forebrain. At embryonic day (E)13.5. the regional pattern of expression of Otx1, Otx2, Dlx1, Dlx2, Dlx5, and Emx1 is maintained in primary culture, suggesting that cells are already committed to a regional identity at this stage. In these cultures, Otx2 is expressed by precursor cells, whereas Dlx1 and Emx1 are predominantly expressed by postmitotic cells. We found that FGF2 increased Otx2 expression within precursor cells and the total number of Otx2-expressing cells. This effect was gene- specific, dose-dependent, and temporally regulated, with larger effects at earlier stages of development (E11.5). At E13.5, the effect of FGF2 on Otx2 expression was restricted to the basal telencephalon. Our results suggest that a restricted population of neuroblasts respond to FGF2 in a temporally regulated fashion by proliferating and increasing Otx2 expression. This interaction between FGF2 and Otx2 may be important for the regulation of neurogenesis in the forebrain.
Brain-derived neurotrophic factor promotes the survival and sprouting of serotonergic axons in rat brain
Authors: Mamounas LA , Blue ME , Siuciak JA , Altar CA
Published: April 2018

A pathology of brain serotonergic (5-HT) systems has been found in psychiatric disturbances, normal aging and in neurodegenerative disorders including Alzheimer's and Parkinson's disease. Despite the clinical importance of 5-HT, little is known about the endogenous factors that have neurotrophic influences upon 5-HT neurons. The present study examined whether chronic pain parenchymal administration of the neurotrophins brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) or NGF could prevent the severe degenerative loss of serotonergic axons normally caused by the selective 5-HT neurotoxin p-chloroamphetamine (PCA). The neurotrophins (5–12 micrograms/d) or the control substances (cytochrome c or PBS vehicle) were continuously infused into the rat frontoparietal cortex using an osmotic minipump. One week later, rats were subcutaneously administered PCA (10 mg/kg) or vehicle, and the 5-HT innervation was evaluated after two more weeks of neurotrophin infusion. As revealed with 5-HT immunocytochemistry, BDNF infusions into the neocortex of intact (non-PCA-lesioned) rats caused a substantial increase in 5-HT axon density in a 3 mm diameter region surrounding the cannula tip. In PCA-lesioned rats, intracortical infusions of BDNF completely prevented the severe neurotoxin-induced loss of 5-HT axons near the infusion cannula. In contrast, cortical infusions of vehicle or the control protein cytochrome c did not alter the density of serotonergic axons in intact animals, nor did control infusions prevent the loss of 5-HT axons in PCA-treated rats. NT-3 caused only a modest sparing of the 5-HT innervation in PCA-treated rats, and NGF failed to prevent the loss of 5-HT axon density. The immunocytochemical data were supported by neurochemical evaluations which showed that BDNF attenuated the PCA-induced loss of 5-HT and 5- HIAA contents and 3H-5-HT uptake near the infusion cannula. Thus, BDNF can promote the sprouting of mature, uninjured serotonergic axons and dramatically enhance the survival or sprouting of 5-HT axons normally damaged by the serotonergic neurotoxin PCA.
A quantitative analysis of presynaptic calcium dynamics that contribute to short-term enhancement
Authors: Tank DW , Regehr WG , Delaney KR
Published: April 2018

Augmentation and posttetanic potentiation--two forms of short-term synaptic enhancement produced by repetitive presynaptic action potentials--are dependent on the buildup and decay of nerve terminal residual calcium that occurs on the seconds to minutes time scale. With the goal of providing a quantitative understanding of these kinetics, we measured the buildup and decay of calcium ions in nerve terminals at the crayfish neuromuscular junction under a variety of intracellular buffer conditions and stimulation paradigms. The calcium extrusion process in the terminals was characterized by analysis of calcium levels reached during long stimulus trains as a function of action potential frequency. The extrusion was linearly dependent on the free calcium ion concentration. Using this result, we developed a mathematical model and computer simulation of the residual calcium kinetics. The model demonstrates the experimentally observed dependence of decay rate on exogenous calcium buffer concentration, and can be explicitly solved to provide an expression for the limiting exponential time course of calcium decay following trains in terms of calcium buffer and extrusion characteristics. Methods to determine the calcium influx per action potential, characteristics of endogenous buffer, and the rate of calcium extrusion are suggested by our analysis and demonstrated experimentally.
Differential regulation of neuronal nicotinic ACh receptor subunit genes in cultured neonatal rat sympathetic neurons: specific induction of alpha 7 by membrane depolarization through a Ca2+/calmodulin- dependent kinase pathway
Authors: De Koninck P , Cooper E
Published: April 2018

We have examined the regulation of neuronal nicotinic ACh receptor (nAChR) genes and ACh-evoked currents by neonatal rat sympathetic neurons developing in culture. These neurons contain 5 nAChR transcripts: alpha 3, alpha 5, alpha 7, beta 2, and beta 4. When developing in culture, the neurons express 4 of these transcripts, alpha 3, alpha 5, beta 2, and beta 4, at levels similar to those in neurons developing in vivo: alpha 3 mRNA levels increase two- to threefold over the first week, whereas the levels for alpha 5, beta 2, and beta 4 remain essentially constant. In contrast, alpha 7 mRNA levels drop by 60–75% within the first 48 hr and remain low. We show that during the first week, the ACh-evoked current densities on these cultured neurons increase twofold and correlate well with the increase in alpha 3 mRNA levels. Depolarizing the neurons with 40 mM KCl for 1–2 d upregulates the alpha 7 gene; this specific change in alpha 7 mRNA level correlates with an increase in alpha-bungarotoxin (alpha-BTX) binding on the surface of the neurons. Depolarization has little effect on the expression of the other four transcripts, or on the magnitude or kinetics of the ACh-evoked currents. Furthermore, activators or inhibitors of protein kinase A (PKA), protein kinase C (PKC), or tyrosine kinase do not affect nAChR transcript levels in these cultured neurons. The effect of membrane depolarization on alpha 7 expression is a result of Ca2+ influx through L-type Ca2+ channels, and we show that alpha 7 is upregulated through a Ca2+/calmodulin-dependent protein kinase (CaM kinase) pathway. The identification of CaM kinase as a link between activity and neurotransmitter receptor expression may indicate a novel mechanism that underlies some forms of synaptic plasticity.
Role of ventrolateral medulla catecholamine cells in hypothalamic neuroendocrine cell responses to systemic hypoxia
Authors: Smith DW , Buller KM , Day TA
Published: April 2018

Systemic hypoxia stimulates the release of vasopressin (VP) and adrenocorticotropin hormone (ACTH). To examine the involvement of catecholamine cell groups of the ventrolateral medulla (VLM) in the neuroendocrine responses, we have used the c-fos activity mapping technique to compare the effects of hypoxia on VLM catecholamine cells to those on neurosecretory VP and putative corticotropin releasing factor (CRF) containing cells. A limited degree of catecholamine cell activation was evident at predominantly mid-VLM levels at 12% oxygen in the inspired air. Further reduction in inpsirate oxygen levels enhanced recruitment of caudally located VLM catecholamine cells considered to form part of the A1 noradrenergic cell group. Threshold for activation of VP and putative CRF cells occurred at the 10% oxygen level. Unexpectedly, this stimulus also activated neurosecretory oxytocin (OT) cells. With increasing hypoxic severity the number of activated supraoptic VP and OT cells was not significantly different to that observed at the 10% level. However, paraventricular neuroendocrine responses continued to increase with putative CRF containing cells of the medial parvocellular zone having nearly double the level of activity (as measured by the number of cells within this region displaying Fos-like immunoreactivity; FLI) at 6% compared to that apparent to the 10% level of hypoxia. Paraventricular VP cells displaying FLI were also increased at the most severe levels of hypoxia but this effect was much less marked than the medial parvocellular response. Consistent with a role for VLM catecholamine cells in generation of neuroendocrine cell responses to hypoxia, unilateral VLM lesions, restricted to the caudal two thirds of the catecholamine cell column, resulted in significant reductions in the responses of all three cell types. These results, in addition to establishing a role for VLM catecholamine cells in neuroendocrine cell responses to systemic hypoxia, have important general implications for catecholamine cell group involvement in neuroendocrine regulation.
Two voltage-dependent K+ conductances with complementary functions in postsynaptic integration at a central auditory synapse
Authors: Brew HM , Forsythe ID
Published: April 2018

The medial nucleus of the trapezoid body (MNTB) relays auditory information important for sound source localization. MNTB neurons faithfully preserve the temporal patterning of action potentials (APs) occurring in their single giant input synapse, even at high frequencies. The aim of this work was to examine the postsynaptic potassium conductances that shape the transfer of auditory information across this glutamatergic synapse. We used whole cell patch techniques to record from MNTB neurons in thin slices of rat brainstem. Two types of potassium conductance were found which had a strong influence on an MNTB neuron's postsynaptic response. A small low voltage threshold current, Id, limited the response during each EPSP to a single brief AP. Id was specifically blocked by dendrotoxin (DTX), resulting in additional APs during the tail end of the EPSP. Thus DTX degraded the temporal fidelity of synaptic transmission, since one presynaptic AP then led to several postsynaptic APs. A second conductance was a fast delayed rectifier with a high voltage activation threshold, that rapidly repolarised APs and thus facilitated high frequency AP responses. Together, these two conductances allow high frequency auditory information to be passed accurately across the MNTB relay synapse and separately, such conductances may perform analogous functions elsewhere in the nervous system.
Differential effects of glycosaminoglycans on neurite growth on laminin and L1 substrates
Authors: Dou CL , Levine JM
Published: April 2018

Glycosaminoglycans (GAGs), the carbohydrate moieties of proteoglycans, are thought to be positive and negative regulators of axonal growth. The physiological role of GAGs is controversial as some studies have shown that GAGs inhibit cell adhesion and neurite elongation (Exp Neurol 109:111, 1990) whereas other studies have reported a growth stimulatory effect of GAGs (Development 114:17, 1992). These and other studies have examined the effects of GAGs using different types of neurons and different substrate conditions thereby making a direct comparison of the experimental data difficult. To resolve the controversy concerning the ability of exogenous GAGs to modulate neurite growth, we examined the effects of a panel of structurally different GAGs on the growth of postnatal rat cerebellar granule neurons and embryonic rat dorsal root ganglia (DRG) neurons on substrates of either laminin or the L1 glycoprotein. Here we show that chondroitin 4-sulfate (CS4), chondroitin 6-sulfate (CS6), and keratan sulfate (KS) inhibit neurite growth from both cerebellar and DRG neurons on laminin-coated surfaces. On L1 surfaces, however, these GAGs are either extremely weak inhibitors of neurite extension or, in the case of CS4, a modest stimulator of neurite growth. Heparan sulfate (HS) and dermatan sulfate (DS) inhibited the growth of cerebellar neurons but not the growth of DRG neurons on L1-coated surfaces. On laminin surfaces, DS and HS had no effect on neurite growth from both cerebellar and DRG neurons. These results demonstrate a cellular and a substrate specificity to the effects of exogenous GAGs on neurite extension in vitro. They suggest that while CS and KS GAGs may not exert strong negative influences over axonal growth in regions of the developing CNS where the L1 glycoprotein is abundant, these GAGs are capable of inhibiting the growth of axons that extend within an environment rich in laminin.
Nerve growth factor facilitates cholinergic neurotransmission between nucleus basalis and the amygdala in rat: an electrophysiological analysis
Authors: Moises HC , Womble MD , Washburn MS , Williams LR
Published: April 2018

Treatment of rats in vivo with NGF promotes the survival and enhances the neurotransmitter phenotype of basal forebrain cholinergic neurons. We showed recently (Williams et al., 1993) that NGF-induced stimulations of the cholinergic markers ChAT and high-affinity choline uptake are reflected in an enhanced synthesis and release of ACh in terminals fields of basal forebrain cholinergic neurons. The objective of the present study was to determine whether such effects translate into an enhancement in neurotransmission between nucleus basalis neurons and postsynaptic target cells, and therefore are likely to be of physiological significance. Changes in cholinergic neurotransmission after NGF were assessed by comparing the ability of cholinergic pathway activation, produced by electrical stimulation of nucleus basalis or the external capsule, to elicit intracellularly recorded muscarinic responses in basolateral amygdaloid (BLA) neurons in ventral forebrain slice preparations from NGF-treated and control Fischer 344 adult rats. Chronic infusion of NGF for 3 weeks (1.2 micrograms/d, i.c.v.) increased the likelihood of eliciting cholinergic slow depolarizations (slow EPSP) via stimulation of cholinergic pathways in the slice. In addition, the frequency-response curves for generation of the cholinergic slow EPSP by nucleus basalis or external capsule stimulation were shifted approximately twofold to the left and the EF50 values significantly reduced in neurons from NGF-treated slices, compared to those in preparations from vehicle-treated or untreated controls. Treatment with NGF also resulted in a leftward shift in the frequency-response curve for cholinergic pathway-induced blockade of the slow afterhyperpolarization, without change in the maximal inhibitory effect. The NGF-induced enhancement in cholinergic synaptic effectiveness was not accompanied by alterations in the resting membrane properties or intrinsic excitability of BLA pyramidal neurons. Nor did treatment with NGF affect their chemosensitivity or responsiveness to direct postsynaptic applications of the cholinergic carbachol. We conclude from these results that chronic administration of exogenous NGF can facilitate neurotransmission within basal forebrain cholinergic projections in normal adult brain, presumably as a consequence of its ability to stimulate presynaptic mechanisms involved in synthesis and/or release of ACh.
Expression of mu-, delta-, and kappa-opioid receptor-like immunoreactivities in rat dorsal root ganglia after carrageenan-induced inflammation
Authors: Ji RR , Zhang Q , Law PY , Low HH , Elde R , Hokfelt T
Published: April 2018

Recently, antisera that recognize unique epitopes of the cloned mu-, delta-, and kappa-opioid, receptors (MOR, DOR, KOR, respectively) have been developed. In the present study MOR-, DOR-, and KOR-like immunoreactivities (LIs) were examined in rat dorsal root ganglia (DRGs, L4-5) after injection of carrageenan (CAR) into the hindpaw. In normal control rats 20.9%, 13.5%, and 9% of the DRG neurons contained MOR-, DOR-, KOR-LI, respectively. A marked upregulation in MOR-LI was observed in DRG neurons 1 and 3 d after inflammation. In contrast, CAR induced a distinct downregulation in DOR- and KOR-LIs. MOR-, DOR-, and KOR-LIs were preferentially localized in small DRG neurons. MOR-LI was often located in patches in the cytoplasm, and in some cells close to the somatic plasmalemma. However, DOR- and KOR-LIs mainly showed a diffuse staining pattern within cytoplasm. Two or even all three receptors could sometimes be found to coexist in DRG neurons. In the spinal cord, these receptors were mainly confined to the superficial dorsal horn, with a somewhat diffuse staining which was strong for MOR- LI, and weak for KOR-LI. DOR-LI had distinctly punctate, varicose distribution. CAG induced-alterations in opioid receptor staining in spinal cord were much less pronounced than those in the DRGs with a small increase in MOR-LI and a slight decrease in DOR-LI ipsilaterally. There was an accumulation of all three types of receptors in the sciatic nerve both proximal and distal to the ligation site as early as 2 hr, indicating both antero- and retrograde transport of multiple opioid receptors. However, DOR-LI accumulation was stronger than that of MOR- and KOR-LIs. Taken together, these results suggest that all three opioid receptors are involved in the response to inflammation and that they may play different roles in this pathological state. The coexistence of MOR, DOR, and KOR in at least some primary sensory neurons provides a substrate for functional interactions between these receptors.
D1 and D2 dopamine receptor function in the striatum: coactivation of D1- and D2-dopamine receptors on separate populations of neurons results in potentiated immediate early gene response in D1-containing neurons
Authors: Gerfen CR , Keefe KA , Gauda EB
Published: April 2018

D1- and D2-dopamine receptor-mediated regulation of immediate early gene levels in identified populations of neurons in the striatum was examined with quantitative in situ hybridization histochemical techniques. Levels of messenger RNA (mRNA) encoding the immediate early genes zif268 and c-fos were examined in two experiments in rats with unilateral lesions of the nigrostriatal dopamine pathway. In a dose- response study, animals were treated with doses of 0.5, 1.0, and 1.5 mg/kg of the D1 agonist SKF-38393 either alone or in combination with the D2 agonist quinpirole (1 mg/kg). Levels of immediate early gene mRNAs 60 min following drug treatments showed a dose-related increase to the D1 agonist alone and a potentiation to combined D1 and D2 against treatment. In a second experiment, in animals receiving 1 mg/kg SKF-38393 either alone or in combination with 1 mg/kg quinpirole, the level of zif268 mRNA was measured with a double-labeling method in striatal neurons containing enkephalin mRNA, a marker of D2-containing neurons, and in neurons not containing enkephalin, putative D1- containing neurons. In the dopamine-depleted striatum, D1 agonist treatment alone did not affect enkephalin-positive neurons but significantly elevated zif268 mRNA levels in nearly all enkephalin- negative neurons. Combined D1 and D2 agonist treatment further increased zif268 mRNA levels in this population of enkephalin-negative neurons and decreased zif-268 mRNA levels in enkephalin-positive neurons. These data indicate that the synergistic response to combined D1- and D2-receptor stimulation is mediated by interneuronal interactions involving the activation of D1 and D2 receptors on separate populations of striatal neurons.
Direct demonstration of a physiological role for carbon monoxide in olfactory receptor neurons
Authors: Ingi T , Ronnett GV
Published: April 2018

Recent evidence suggests that, like nitric oxide (NO), carbon monoxide (CO), another activator of soluble guanylyl cyclase, may serve as an intercellular messenger in the brain. Heme oxygenase, which converts heme to biliverdin and CO, is abundantly expressed in the brain and is localized to discrete neuronal populations. However, evidence for the actual generation of CO by neurons is lacking. Heme oxygenase-2 immunoreactivity is abundantly present in olfactory receptor neurons where it essentially colocalizes with immunoreactivity to soluble guanylyl cyclase, the target of CO action. To examine the generation of CO by neurons, we measured CO production directly and determined its relationship to cyclic GMP levels in cultured rat olfactory receptor neurons. This system has the advantage of not having measurable NO production, which could confound results since NO is a more potent activator of guanylyl cyclase than CO. Metabolic labeling experiments permitted the direct measurement of 14CO production by neurons in vitro. CO release parallels endogenous cyclic GMP concentrations with its peak at the immature stage of neuronal differentiation in culture. Cyclic GMP production is inhibited by zinc protoporphyrin-9 and zinc deuteroporphyrin IX 2,4-bis glycol, inhibitors of heme oxygenase, indicating that CO is an endogenous regulator of soluble guanylyl cyclase activities in these cells. Transforming growth factor-beta 2, an olfactory neurogenic factor, specifically shows a negative effect on CO release in olfactory receptor neurons. These results indicate that CO may serve as a gaseous neuronal messenger linked to cyclic GMP production and suggests its involvement in developmental processes of the olfactory receptor neuron.
Staurosporine blocks evoked release of FM1-43 but not acetylcholine from frog motor nerve terminals
Authors: Henkel AW , Betz WJ
Published: April 2018

The protein kinase inhibitor staurosporine inhibited, and often abolished, activity-dependent destaining of frog motor nerve terminals that had been preloaded with the fluorescent dye FM1-43. Staurosporine did not, however, block synaptic transmission; staurosporine treated muscles twitched in response to nerve stimulation, and the amplitudes of evoked end plate potentials were reduced only slightly, and in some cases not at all. The blockade of FM1-43 destaining was not reversed by washing, although treatment with black widow spider venom caused complete destaining. Nerve terminal pretreated with staurosporine could subsequently be stained with FM1-43 (and then destained by black widow spider venom). Thus, staurosporine blocked destaining but not staining of nerve terminals. Staurosporine treatment had little effect on the ultra-structure of resting terminals, the main difference we noted being a somewhat closer packing of synaptic vesicles after exposure to staurosporine. However, staurosporine blocked completely the ultrastructural changes produced by prolonged nerve stimulation, such as depletion of synaptic vesicles, appearance of intraterminal cisternae, and the uptake of horseradish peroxidase. The effects of staurosporine were not mimicked by KN-62, H7, calmidozolium, or trifluoroperazine. These and other observations are consistent with, but do not prove the hypothesis that, after exposure to staurosporine, the exocytotic fusion pore behaved like a valve, letting FM1-43 in, but not out, as if staurosporine interfered with the postexocytotic collapse of synaptic vesicles into the surface membrane.