bluuebunny/crossref_metadata_2025 huggingface dataset:
Damage to the perirhinal cortex exacerbates memory impairment following lesions to the hippocampal formation
Authors: Zola-Morgan S , Squire LR , Clower RP , Rempel NL
Published: March 2018

Recent work has been directed at identifying the critical components of the medial temporal lobe that, when damaged, produce severe memory impairment. The H+A+ lesion includes the hippocampal formation, the amygdala, and the adjacent entorhinal, parahippocampal, and perirhinal cortices. A more restricted medial temporal lobe lesion that includes the hippocampal formation and parahippocampal cortex (the H+ lesion) produces less severe memory impairment. Previous work demonstrated that extending the H+ lesion forward to include the amygdala did not exacerbate the impairment. Here, we tested the hypothesis that extending the H+ lesion forward to include the perirhinal cortex (the H++ lesion), but sparing the amygdala, should produce a more severe memory impairment and one that would approximate the level of memory impairment associated with the H+A+ lesion. Monkeys with the H++ lesion were severely impaired on two of three amnesia-sensitive tasks (delayed nonmatching to sample and delayed retention of object discrimination). On the third amnesia-sensitive task (concurrent discrimination learning), two of the monkeys in the H++ group obtained poorer scores than all seven normal monkeys, although the overall group comparison was not significant. The memory impairment following H++ damage was more severe overall than the impairment associated with the H+ lesion and approached the level of impairment associated with the H+A+ lesions. Quantitative measurement of damage in each anatomical component of the lesion indicated that the perirhinal cortex was the only brain region that was more extensively damaged in the H++ group than in the H+ group. These findings emphasize the importance of the perirhinal cortex in the anatomy of the medial temporal lobe memory system.
Time course of extracellular dopamine and behavioral sensitization to cocaine. II. Dopamine perikarya
Authors: Kalivas PW , Duffy P
Published: March 2018

Cocaine was administered daily (15 mg/kg, i.p. x 1 d followed by 30 mg/kg, i.p. x 5 d) to produce behavioral sensitization. Using microdialysis in the ventral tegmental area and medial substantia nigra, the effect of repeated cocaine was examined on the extracellular levels of dopamine. One day after discontinuing repeated cocaine injections, an acute challenge with cocaine (15 mg/kg, i.p.) produced a significant elevation in extracellular dopamine compared to rats pretreated with daily saline (x6 d). The augmentation in extracellular dopamine persisted longer than the sensitized behavioral response. In contrast, 14 d after discontinuing daily cocaine, the increase in extracellular dopamine produced by an acute cocaine challenge was not augmented, although behavioral sensitization was present. In separate animals, the basal concentration of dopamine in the ventral tegmental area/medial substantia nigra was measured by determining the concentration of dopamine at which no net flux occurred across the dialysis membrane in vivo. One day after discontinuing daily treatments, the basal level of extracellular dopamine in the cocaine pretreated rats was significantly elevated over the level in saline- pretreated animals (1.3 nM vs. 0.8 nM). By 14 d after the last daily injection, the basal levels of dopamine were equivalent in cocaine- and saline-pretreated animals. It is concluded that daily cocaine injections produce a transient alteration in the regulation of somatodendritic dopamine release. While such changes are not responsible for the long-term behavioral sensitization produced by repeated cocaine administration, they may be involved in the initiation of behavioral sensitization.
Prenatal development of excitability in cat retinal ganglion cells: action potentials and sodium currents
Authors: Skaliora I , Scobey RP , Chalupa LM
Published: March 2018

The development of precise retinofugal projections is dependent on activity-mediated events, but as yet nothing is known about the ontogeny of excitable membrane properties in retinal ganglion cells (RGCs). In order to begin to understand how functional maturity is attained in these neurons, whole-cell patch-clamp recordings were obtained from acutely dissociated RGCs of fetal and postnatal timed- pregnant cats. Current-clamp recordings revealed a pronounced developmental increase in the proportion of RGCs capable of generating action potentials. At embryonic day 30 (E30), 5 weeks before birth and during a time when RGCs are still being generated, electrical stimulation elicited spikes in only a third of the cells. None of these neurons were capable of multiple discharges in response to maintained depolarization. The proportion of spiking neurons increased during ontogeny, such that by E55 all RGCs could be induced to generate action potentials, with the majority manifesting repetitive spiking patterns. Application of tetrodotoxin abolished spike activity of all fetal RGCs, indicating that sodium-mediated action potentials are present very early in development. At the same time, voltage-clamp recordings revealed significant ontogenetic modifications in several key properties of the sodium currents (INa). These were (1) a twofold increase in Na current densities; (2) a shift in the voltage dependence of both activation and steady state inactivation: with maturity, sodium currents activate at more negative potentials, while steady state inactivation of INa occurs at less negative potentials; and (3) a decrease in decay time constants of the Na current, at membrane potentials negative to -15 mV. These developmental changes were largely restricted to the period of axon ingrowth (E30-E38), suggesting that maturation of INa is not the limiting factor for the onset of activity- dependent restructuring of retinofugal projections.
Gonadotropin-releasing hormone-containing neurons change size with reproductive state in female Haplochromis burtoni
Authors: White SA , Fernald RD
Published: March 2018

In the preoptico-hypothalamic area (POA) of teleost fish, neurons containing gonadotropin-releasing hormone (GnRH) regulate reproduction through direct projections to pituitary gonadotropes. Here we show that these GnRH-containing cells change size depending on the reproductive and maturational state in female Haplochromis burtoni. We selected animals prior to, during, and after the reproductive portion of their life history, in both brooding and spawning states. Immunocytochemical staining of GnRH-containing neurons in the POA revealed that these cells are up to twice as large in females that have never spawned or are in the act of spawning than they are in females that are carrying broods. Older, postreproductive females have the largest cell sizes. Previous work on male H. burtoni has shown that soma sizes of the homologous neurons change according to social status, with dominant fish having larger cells than subordinates. Since reproductively active females have no apparent social hierarchy and are all exposed to approximately the same external stimuli, the primary factor(s) controlling GnRH-immunoreactive (irGnRH) neuron size appears to be internal reproductive state. Thus, while irGnRH neurons are pleiomorphic in both males and females, cell size change is differently regulated in each.
Axonal regeneration contributes to repair of injured brainstem-spinal neurons in embryonic chick
Authors: Hasan SJ , Keirstead HS , Muir GD , Steeves JD
Published: March 2018

Recent results have demonstrated complete anatomical and functional repair of descending brainstem-spinal projections in chicken embryos that underwent thoracic spinal cord transection prior to embryonic day 13 (E13) of the 21 d developmental period. To determine to what extent axonal regeneration was contributing to this repair process, we conducted experiments using a double retrograde tract-tracing protocol. On E8-E13, the upper lumbar spinal cord was injected with the first fluorescent tracing dye to label those brainstem-spinal neurons projecting to the lumbar cord at that time. One to two days later (on E10-E15), the upper to mid-thoracic spinal cord was completely transected. After an additional 7-8 d, a different second fluorescent tracing dye was injected into the lumbar cord at least 5 mm caudal to the site of transection. Finally, 2 d later on E19 to postnatal day 4, the CNS was fixed and sectioned. Brainstem and spinal cord tissue sections were then viewed with epifluorescence microscopy. In comparison to nontrasected control animals, our findings indicated that there were relatively normal numbers of double-labeled brainstem-spinal neurons after a transection prior to E13, whereas the number of double- labeled and second-labeled brainstem-spinal neurons decreases after an E13-E15 transection. In addition, at each subsequent stage of development from E10 to E12, there was a greater number of double- labeled brainstem-spinal neurons (indicating regeneration of previously severed axons) than cell bodies labeled with the second fluorescent tracer alone (indicating subsequent development of late brainstem- spinal projections). Assessment of voluntary open-field locomotion (hatchling chicks) and/or brainstem-evoked locomotion (embryonic or hatchling) indicated that functional recovery of animals transected prior to E13 was indistinguishable from that observed in control chicks (sham operated or unoperated). Taken together, these data suggest that regeneration of previously axotomized fibers contributes to the observed anatomical and functional recovery after an embryonic spinal cord transection.
Functional studies of Alzheimer's disease tau protein
Authors: Lu Q , Wood JG
Published: March 2018

In vitro assays were used to monitor and compare the kinetic behavior of bovine tubulin polymerization enhanced by tau proteins isolated from Alzheimer's disease (AD) and nondemented (ND) age-matched control brains. Tau from AD cases induced slower polymerization and a steady state turbidity value approximately 50% of that stimulated by tau from control cases. Tau from the most severe AD case was least effective at promoting polymerization. Dark-field light microscopy of the control samples revealed abundant microtubule formation and many microtubule bundles. Microtubule assembly was observed in AD samples as well, but bundling was not obvious. These results were confirmed by negative- stain electron microscopy. Morphological analysis showed that AD tau- induced microtubules were longer than control microtubules. Furthermore, our initial results suggest that the reduction of AD tau activity is correlated with neurofibrillary pathology in AD brains. Earlier reports indicated that AD tau is modified by phosphorylation (Grundke-Iqbal et al., 1986; Wood et al., 1986; Iqbal et al., 1989; Brion et al., 1991a,b; Lee et al., 1991). Our results support the hypothesis that tau modification compromises its function by altering its ability to nucleate and bundle microtubules.
Modulation of N-type calcium channels in bullfrog sympathetic neurons by luteinizing hormone-releasing hormone: kinetics and voltage dependence
Authors: Boland LM , Bean BP
Published: March 2018

Inhibition of Ca channel current by luteinizing hormone-releasing hormone (LHRH) was studied in freshly dissociated bullfrog sympathetic ganglion neurons using whole-cell recording. LHRH inhibited up to 80% of the high-threshold Ca channel current with a half-maximally effective concentration of about 20 nM. LHRH inhibited omega-conotoxin- sensitive but not nimodipine-sensitive current and also did not inhibit Bay K 8644-enhanced currents, suggesting that LHRH inhibits N-type but not L-type channels. Inhibition was faster at higher concentrations of LHRH, reaching a limiting time constant of 2 sec at 0.3-3 mM LHRH. The rate of recovery from block (tau approximately 19 sec) was independent of LHRH concentration. Inhibition of N-type current by LHRH was highly sensitive to the gating state of the channel. Though strongly effective if applied when channels were mostly in the resting state, LHRH had little effect if applied rapidly during a long depolarization that opened the channels. Inhibition could be relieved if channels were activated by short, large test depolarizations or by long, smaller depolarizations. The state-dependent properties of LHRH block could be simulated by a model that assumes that inhibition by LHRH results from activated G-proteins binding to N-type channels and that (1) G-protein binding stabilizes closed gating states and (2) activation of G-protein- bound channels destabilizes the binding of the G-protein to the channel.
Dystrophin in a membrane skeletal network: localization and comparison to other proteins
Authors: Dmytrenko GM , Pumplin DW , Bloch RJ
Published: March 2018

We studied the location, relative abundance, and stability of dystrophin in clusters of ACh receptors (AChRs) isolated from primary cultures of neonatal rat myotubes. Although variable amounts of dystrophin were found at receptor clusters, dystrophin was always associated with organized, receptor-rich domains (AChR domains). Dystrophin was occasionally seen in focal contact domains, but never in clathrin-coated domains. Dystrophin was also present in a diffuse, punctate distribution in regions of myotube membrane that did not contain AChR clusters. Immunogold labeling at the ultrastructural level localized dystrophin in a spectrin-rich filamentous network closely applied to the cytoplasmic surface of the cell membrane at AChR domains. Dystrophin was not associated with overlying actin filaments. Semiquantitative immunofluorescence studies indicated that dystrophin was present in relatively small amounts in these preparations, with only one molecule of dystrophin for every approximately 5 AChR, 43 kDa and 58 kDa molecules, and for every approximately 20-35 beta-spectrin molecules. Clusters were disrupted, but the total amount of dystrophin was not significantly reduced, when myotubes were incubated with sodium azide or in Ca(2+)-free medium, and when isolated AChR clusters were extracted at low ionic strength, at high pH, or in 6 M urea. These treatments extract other peripheral membrane proteins from AChR clusters. Labeling for dystrophin was completely eliminated when clusters were incubated with chymotrypsin, however. Thus, dystrophin forms part of a membrane skeleton at AChR clusters, but it is more difficult to remove than other proteins in the network. This suggests that dystrophin attaches to cluster membrane in a unique way.
Overexpression of Ca2+/calmodulin-dependent protein kinase II in Neuro2a and NG108-15 neuroblastoma cell lines promotes neurite outgrowth and growth cone motility
Authors: Goshima Y , Ohsako S , Yamauchi T
Published: March 2018

To elucidate the functional role of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) in neuronal cells, we studied the phenotypic effects of overexpression of the CaM kinase II wild-type alpha subunit and a mutant enzyme alpha isoform (Ala-286), in which formation of the Ca(2+)-independent form by autophosphorylation is markedly suppressed by replacement of Thr-286 with Ala, using Neuro2a (Nb2a) and NG108-15 neuroblastoma cell lines. The cDNAs inserted into the EcoRI site of pEF321 expression vector were introduced into Nb2a and NG108-15 cells with pEF321-neo (neo). Stable clones were obtained by G418 selection. The specific activities of CaM kinase II in alpha and Ala-286 transfectants were two to four times higher than those in non- transfectants and in cells transfected with neo alone. Indirect immunofluorescence using a monoclonal antibody specific to the CaM kinase II alpha isoform revealed that CaM kinase II was mainly localized in the perikaryal and dendritic cytoplasm of the alpha and Ala-286 transfectants. Immediately after plating, Nb2a and NG108-15 cells transfected with neo, alpha and Ala-286 cDNAs appeared round. Several hours after plating, alpha transfectants showed cell flattening and initiation of neurite outgrowth, and thereafter extended numerous long and branching neurites. Numerous filopodia protruded from flat growth cones, some of which were accompanied by extensive veil formation. Non- and neo transfectants remained round. In Ala-286 transfectants, however, the phenotypic changes were remarkably less than in alpha transfectants.(ABSTRACT TRUNCATED AT 250 WORDS)
Examination of TEA-induced synaptic enhancement in area CA1 of the hippocampus: the role of voltage-dependent Ca2+ channels in the induction of LTP
Authors: Huang YY , Malenka RC
Published: March 2018

The role of voltage-dependent Ca2+ channels (VDCCs) in the induction of long-term potentiation (LTP) in the CA1 region of rat hippocampus was determined by examining the relationship between LTP and the long- lasting synaptic enhancement induced by extracellular application of tetraethylammonium (TEA). Consistent with previous findings (Aniksztejn and Ben-Ari, 1991), the TEA-induced synaptic enhancement did not require NMDA receptor activation. It was blocked by the L-type VDCC antagonist nifedipine or by intracellular injection of the Ca2+ chelator 1,2-bis(2-amino-phenoxy)ethane N,N,N′,N′-tetra-acetic acid (BAPTA) and could be mimicked by direct activation of VDCCs with repetitive depolarizing current pulses. In contrast to its effect on TEA-induced synaptic enhancement, nifedipine had no effect on the magnitude or duration of NMDA receptor-dependent LTP. Saturation of NMDA receptor-dependent LTP reduced the magnitude of the TEA-induced synaptic enhancement. Similarly, increasing synaptic strength by initial application of TEA reduced the magnitude of the subsequent tetanus-induced LTP. Like LTP, the TEA-induced synaptic enhancement did not significantly affect paired-pulse facilitation. These results suggest that dihydropyridine-sensitive VDCCs do not normally contribute to the induction of NMDA receptor-dependent LTP even though their repetitive activation can generate an increase in synaptic strength. The mutual occlusion of LTP and TEA-induced synaptic enhancement suggests that they share a common expression mechanism and perhaps are generated by activation of common Ca(2+)-dependent intracellular processes.(ABSTRACT TRUNCATED AT 250 WORDS)
The epsilon subunit confers fast channel gating on multiple classes of acetylcholine receptors
Authors: Camacho P , Liu Y , Mandel G , Brehm P
Published: March 2018

During vertebrate skeletal muscle development, multiple forms of long- open-time (slow-type) ACh receptor channels are replaced by at least two different types of short-open-time (fast-type) ACh receptors. Expression of ACh receptors in Xenopus oocytes indicates that the substitution of an epsilon subunit for a gamma subunit may account for both types of fast-gated channel types in adult muscle. Unlike the various forms of the embryonic receptor, in which functional diversity is achieved through alterations in subunit composition, the two major fast-gated forms expressed in oocytes have identical subunit composition. These findings provide a structural basis for both types of short-open-time ACh receptor types found in adult muscle.
Confocal microscopy reveals coordinated calcium fluctuations and oscillations in synaptic boutons
Authors: Melamed N , Helm PJ , Rahamimoff R
Published: March 2018

Calcium ions are one of the main factors regulating quantal transmitter release and thus synaptic transmission in the nervous system. Using confocal microscopy, fluorescent imaging with the calcium indicator Rhod-2, and time series analysis, we show that the levels of calcium ions inside single synaptic boutons of the lizard neuromuscular junction are not constant at rest, but undergo coordinated fluctuations in the space domain, which cover a large fraction of the synaptic bouton. Furthermore, oscillations in intracellular calcium were frequently observed in the time domain. Control experiments showed no coordinated fluctuations or oscillations at locations outside the synaptic boutons. Edge detection analysis showed that the coordinated fluctuations and oscillations were not due to movement artifacts. No coordinated fluctuations and oscillations were seen when similar measurements and analyses were performed on artificial fluorescent beads. A variance analysis was performed on artificial fluorescent beads and on synaptic boutons. The variance of the fluorescent signal at the synaptic boutons was larger than the variance in artificial beads with the same mean fluorescence. This extra variance was greatly reduced when the extracellular calcium concentration was decreased from 2.0 mM to 0.4 mM. We conclude that the coordinated fluctuations and oscillations in the calcium-induced fluorescence at the synaptic boutons are genuine biological phenomena and may be of significance in the regulation of transmitter release.
Inactivation of NMDA channels in cultured hippocampal neurons by intracellular calcium
Authors: Legendre P , Rosenmund C , Westbrook GL
Published: March 2018

Calcium-dependent inactivation of NMDA channels was examined on cultured rat hippocampal neurons using whole-cell voltage-clamp and cell-attached single-channel recording. An ATP regeneration solution was included in the patch pipette to retard current “rundown.” In normal [Ca2+]o (1-2 mM) and 10 microM glycine, macroscopic currents evoked by 15 sec applications of NMDA (10 microM) inactivated slowly following an initial peak. At -50 mV in cells buffered to [Ca2+]i < 10(- 8) M with 10 mM EGTA, the inactivation time constant (tau inact) was approximately 5 sec. Inactivation did not occur at membrane potentials of +40 mV and was absent at [Ca2+]o < or = 0.2 mM, suggesting that inactivation resulted from transmembrane calcium influx. The percentage inactivation and tau inact were dependent on [Ca2+]o. The tau inact was also longer with BAPTA in the whole-cell pipette compared to EGTA, suggesting that tau inact reflects primarily the rate of accumulation of intracellular calcium. Inactivation was incomplete, reaching a steady state level of 40–50% of the peak current. At steady state, block of open NMDA channels with MK-801 ((+)-5-methyl-10,11-dihydro-5H- dibenzo[a,d]cyclohepten-5,10-imine) completely blocked subsequent responses to NMDA, suggesting that “inactivated” channels can reopen at steady state. Inactivation was fully reversible in the presence of ATP but was not blocked by inhibiting phosphatases or proteases. In cell- attached patches, transient increases in [Ca2+]i following cell depolarization also resulted in inactivation of NMDA channels without altering the single-channel conductance. This suggests that Ca(2+)- dependent inactivation occurs in intact cells and can be triggered by calcium entry through nearby voltage-gated calcium channels, although calcium entry through NMDA channels was more effective. We suggest that [Ca2+]i transients induce NMDA channel inactivation by binding to either the channel or a nearby regulatory protein to alter channel gating. This mechanism may play a role in downregulation of postsynaptic calcium entry during sustained synaptic activity.
Spatiotemporal responses of astrocytes, ramified microglia, and brain macrophages to central neuronal infection with pseudorabies virus
Authors: Rinaman L , Card JP , Enquist LW
Published: March 2018

We examined the responses of astrocytes, ramified microglia, and brain macrophages to CNS neuronal infection with virulent or attenuated strains of a swine alpha herpesvirus (pseudorabies virus, PRV). After PRV inoculation of the rat stomach or pancreas, the temporal course of viral replication and induced pathology of infected neurons were assessed in the dorsal motor nucleus of the vagus (DMV) and amygdala using an antiserum generated against PRV. Specific monoclonal antibodies against glial fibrillary acidic protein (GFAP), OX42, and ED1 and morphological criteria were used to classify non-neuronal cells. Both PRV strains infected DMV and motor neurons and then passed transneuronally to infect brainstem neurons that innervate the DMV. However, the onset of neuronal infection produced by the attenuated strain occurred approximately 20 hr later than infection with the virulent strain. Animals infected with the attenuated strain also survived longer, permitting transneuronal passage of virus into forebrain areas of the visceral neuraxis. Neuronal infection with both PRV strains produced consistent alterations in astrocytes, ramified microglia, and brain macrophages that correlated spatially and temporally with progressive stages of viral replication and neuronal pathology. Early stages of infection were characterized by increases in immunoreactivity for astrocytic GFAP and microglial OX42 that preceded overt signs of neuronal pathology. At later stages, GFAP immunoreactivity decreased dramatically in focal areas of neuronal infection while OX42 immunoreactivity continued to increase. Subsequently, ED1-immunoreactive brain macrophages infiltrated these infected areas. Double immunocytochemical labeling demonstrated that some astrocytes and brain macrophages were immunopositive for viral antigens but ramified microglia were not. The responses of glia and brain macrophages are consistent with a proposed role in restricting extracellular spread of virus by isolating or phagocytosing infected cells. These phenomena may contribute to the specific transneuronal transport exhibited by PRV.
Hypothalamic lesions that induce female precocious puberty activate glial expression of the epidermal growth factor receptor gene: differential regulation of alternatively spliced transcripts
Authors: Junier MP , Hill DF , Costa ME , Felder S , Ojeda SR
Published: March 2018

Injury of the nervous system triggers a complex series of repair mechanisms that include production of neurotrophic and mitogenic factors by cells neighboring the injured area. While trauma of most parts of the brain results in loss of function, lesions of certain regions of the female hypothalamus enhance the secretory activity of a group of specialized neurons that produce luteinizing hormone-releasing hormone (LHRH), the neuropeptide that controls sexual development. The increased output of LHRH causes sexual precocity by prematurely activating the neuroendocrine reproductive axis. Recent studies have implicated transforming growth factor alpha (TGF alpha) produced by reactive astrocytes in the process by which lesions hasten sexual maturation, and have suggested that the stimulatory actions of TGF alpha on LHRH neurons require the intermediacy of epidermal growth factor receptors (EGFRs). In the present study, we examined the changes in EGFR gene expression following lesions of the preoptic-anterior hypothalamic area (POA-AHA) of immature female rats, identified the cell types where EGFR synthesis increases, and assessed the biochemical activity of the newly formed EGFR protein. RNase protection assays demonstrated that the lesion significantly increased the levels of a predominant mRNA transcript encoding the full-length, membrane-spanning EGFR, but did not affect those of a much less abundant, alternatively spliced mRNA that encodes a truncated, presumably secreted form of EGFR. Following lesions, antibody-induced EGFR kinase activity increased twofold. Antibodies directed against a peptide sequence contained within the carboxy terminus of EGFR showed intense EGFR immunoreactivity in cells surrounding the lesion site; double immunohistochemistry identified these cells as astrocytes since EGFR immunoreactivity was colocalized with that of glial fibrillary acidic protein, an astrocytic marker. That these changes result from an increase in EGFR gene expression was indicated by the elevated levels of EGFR mRNA detected by in situ hybridization in cells of the same area. Although POA-AHA lesions did not result in appearance of EGFR in LHRH neurons themselves, EGFR-positive cells and processes were seen in close proximity to LHRH neurons and their nerve terminals, particularly in the area surrounding the lesion. Since TGF alpha gene expression is also increased in reactive astrocytes of POA-AHA lesions and blockade of EGFR prevented the advancing effect of the lesion on puberty (Junier et al., 1991b), the present results support the concept that, in lesioned animals, TGF alpha stimulates LHRH secretion indirectly via a paracrine mechanism that involves its interaction with EGFRs located on astroglial cells.
Independent guidance of retinal axons in the developing visual system of Drosophila
Authors: Kunes S , Wilson C , Steller H
Published: March 2018

The development of the adult visual system of Drosophila requires the establishment of precise retinotopic connections between retinal photoreceptor cell axons and their synaptic partners in the optic lobe of the brain. To assess the role of axon-axon interactions in retinal axon guidance, we used genetic methods to disrupt the normal spatiotemporal order of retinal axon ingrowth. We examined retinal axon projections to the developing first optic ganglion, the lamina, in two mutants in which reduced numbers of ommatidia develop in the eye imaginal disk. We find that in the developing lamina of these mutants, sine oculis and Ellipse, retinal axons project to proper dorsoventral positions despite the absence of the usual array of neighboring retinal axons. In a second approach, we examined animals that were somatic mosaics for the mutation, glass. In glass- animals, retinal axons project aberrantly and the larval optic nerve is absent. We find that in the developing lamina of glass mosaic animals, wild-type retinal axons project to proper dorsoventral positions despite the misrouted projections of neighboring glass- retinal axons. In addition, wild-type retinal axons project normally in the absence of the larval optic nerve, indicating that the latter is not an essential pioneer for retinal axon navigation. Our observations support the proposal that axon fascicles can make at least some pathfinding decisions independently of other retinal axon fascicles. We suggest that positional guidance cues that might label axon pathways and target destinations contribute to retinotopic pattern formation in the Drosophila visual system.
Inhibition of Ca2+ currents by a mu-opioid in a defined subset of rat sensory neurons
Authors: Schroeder JE , McCleskey EW
Published: March 2018

Activation of the endogenous opioid system can suppress pain without affecting other sensations, but the cellular mechanism of this selectivity is unclear. The analgesia might be due to inhibitory synapses arranged only on neurons whose activity leads to pain sensations. Alternatively, opioids might be released broadly, with neurons involved in pain sensation being especially sensitive. Therefore, we asked whether different subsets of rat dorsal root ganglion (DRG) sensory neurons vary in their sensitivity to opioids. Dissociated neurons were subdivided according to the spinal laminae to which they likely had projected, and whether they had innervated muscle. Using the patch-clamp method, we measured the inhibition of Ca2+ current by DAGO (Tyr-D-Ala-Gly-MePhe-Gly-ol), a peptide that selectively activates the mu (morphine) receptor. We also investigated the presence of different types of Ca2+ channels. In DRG neurons chosen at random, Ca2+ currents were inhibited by DAGO to widely varying degrees, with an average inhibition of 38%. Ca2+ currents in neurons in a subset that projects to laminae I and II had a lower average inhibition, and unlike the randomly selected cells, the responses were predictable and tightly distributed about the mean. This indicates that the variability of opioid sensitivity among DRG neurons reflects the presence of different subsets of cells. Since neurons projecting to laminae I and II, the projection site of nociceptive neurons, did not show high opioid sensitivity, there is no evidence that nociceptive neurons have stronger responses to opioids. But a firm conclusion is impossible because projection site does not strictly define sensory modality.
Excitation of rat locus coeruleus neurons by adenosine 5'-triphosphate: ionic mechanism and receptor characterization
Authors: Shen KZ , North RA
Published: March 2018

ATP and several congeners were applied to locus coeruleus neurons in slices cut from rat pons. Whole-cell recording of membrane current showed that ATP caused an inward current at -60 mV. Effective concentrations (applied by superfusion) were 3-300 microM, and the peak current was about 150 pA at -60 mV. 2-Methylthioadenosine 5′- triphosphate was slightly more potent than ATP, adenosine 5′- diphosphate was about equipotent with ATP, alpha,beta-methylene adenosine 5′-triphosphate was slightly less potent than ATP, and beta,gamma'-methylene adenosine 5′-triphosphate and adenosine 5′- monophosphate had little or no effect. Adenosine (100 microM) caused small outward currents (40 pA). By changing the ionic composition of the pipette and extracellular solution, it was shown that the inward current resulted from both an increase in conductance to sodium ions and a reduction in conductance to potassium ions. It is concluded that rat locus coeruleus neurons express P2 purinoceptors, activation of which depolarizes the cells predominantly by increasing a conductance that allows sodium ions to enter the cell.
Corticosteroids regulate brain hippocampal 5-HT1A receptor mRNA expression
Authors: Chalmers DT , Kwak SP , Mansour A , Akil H , Watson SJ
Published: March 2018

Using in situ hybridization techniques, the expression of 5-HT1A receptor mRNA was measured within the hippocampal formation after bilateral adrenalectomy (ADX). After 24 hr ADX, 5-HT1A receptor mRNA expression was significantly increased in all hippocampal subfields in ADX animals relative to sham-operated controls (SHAM). The magnitude of the increase was most pronounced within CA2 (127%) and CA3/4 (94%)- subfields of dorsal hippocampus, intermediate in the dentate gyrus (73%), and least within CA1 (60%). Administration of exogenous corticosterone (CORT) at the time of ADX maintained the level of 5-HT1A receptor mRNA expression within the range of SHAM animals. In vitro receptor autoradiographic analysis of 5-HT1A receptors in adjacent sections from the same animals indicated a simultaneous increase in 5- HT1A binding throughout the hippocampus in response to ADX. 5-HT1A binding increased to a similar extent (approximately 30%) in CA subfields and dentate gyrus but remained within SHAM levels in CORT- replaced animals. 5-HT1A receptor mRNA levels were also increased in hippocampal subregions of 1 week ADX animals relative to SHAM animals. Within both CA1 and CA2 subfields, the increments were approximately double those observed after 1 d ADX. 5-HT1A receptor binding was increased in every hippocampal subfield to a similar extent as that observed after 1 d ADX. Increases in both 5-HT1A receptor mRNA expression and 5-HT1A receptor binding were preventable by administration of exogenous CORT at the time of ADX. Hippocampal 5-HT1C receptor mRNA and D1 receptor mRNA expression were not significantly altered by either acute or chronic ADX treatment. These data indicate that adrenal steroids may selectively regulate hippocampal 5-HT1A receptors at the level of 5-HT1A receptor mRNA expression.
Protein phosphatase 1-deficient mutant Drosophila is affected in habituation and associative learning
Authors: Asztalos Z , von Wegerer J , Wustmann G , Dombradi V , Gausz J , Spatz HC , Friedrich P
Published: March 2018

The learning and memory of Drosophila melanogaster strains carrying the Su-var(3)6(01) mutation, which is known to affect the structural gene of a protein phosphatase 1 isoenzyme, PP1(87B), were studied in various behavioral paradigms. Three lines of Drosophila comprising the Su- var(3)6(01) mutation in different genetic backgrounds were shown to have diminished protein phosphatase 1 activity and behavioral anomalies. Associative olfactory learning and visual conditioning were impaired. Olfactory acuity for the odorants used and response to electric shock were largely unchanged in the mutant lines. The motility and flight activity of the mutants were reduced. Habituation of the landing response, a nonassociative learning process, was more pronounced in heterozygotes of the mutants than in the wild-type control strains. Taken together with earlier data, the results indicate that protein phosphatase PP1(87B), while affecting several cellular processes, is also part of the biochemical machinery of various forms of neuromodulation in Drosophila.
HSV-1 vector-mediated gene transfer of the human nerve growth factor receptor p75hNGFR defines high-affinity NGF binding
Authors: Battleman DS , Geller AI , Chao MV
Published: March 2018

A series of recombinant herpes simplex virus (HSV-1) vectors have been constructed that encode either the full-length cDNA of the human p75 NGF receptor (p75hNGFR) or truncated forms of the receptor. Infection of cultured fibroblast cells with viral stocks results in abundant expression of all three cDNAs, as detected by affinity cross-linking, immunoblot analysis, and equilibrium binding. Furthermore, viral infection of primary neuronal cultures gives easily detectable p75 expression by immunofluorescence and affinity cross-linking. When p75 was introduced by viral infection into fibroblast cells expressing the trk proto-oncogene, a new binding site was created, consistent with high-affinity NGF binding. This site is not created by the coexpression of truncated forms of p75 that lack either the extracellular ligand binding domain or the cytoplasmic domain of the receptor, suggesting that both of these regions of the receptor are required for the formation of the high-affinity NGF binding site. Hence, these HSV-1 vectors give rise to appropriate NGF receptor binding after viral infection. The application of these HSV-1 constructs to primary neuronal culture and in vivo models of p75NGFR function is discussed.
Topographic organization of corticospinal projections from the frontal lobe: motor areas on the lateral surface of the hemisphere
Authors: He SQ , Dum RP , Strick PL
Published: March 2018

We examined the topographic organization of corticospinal neurons in the primary motor cortex and in the two premotor areas on the lateral surface of the hemisphere [i.e., the dorsal premotor area (PMd) and the ventral premotor area (PMv)]. In two macaques, we labeled corticospinal neurons that project beyond T7 or S2 by placing crystals of HRP into the dorsolateral funiculus at these segmental levels. In another seven macaques, we labeled corticospinal neurons that project to specific segmental levels of the spinal cord by injecting the fluorescent tracers fast blue and diamidino yellow into the gray matter of the cervical and lumbosacral segments. In one set of experiments (n = 2), we defined the representations of the arm and leg in each cortical motor area by injecting one of the two fluorescent tracers into lower cervical segments (C7-T1) and the other fluorescent tracer into lower lumbosacral segments (L6-S1) of the same animal. In another set of experiments (n = 5), we defined the representations of distal and proximal parts of the forelimb in each cortical motor area by injecting one of the two fluorescent tracers into lower cervical segments (C7-T1) and the other tracer into upper cervical segments (C2-C4) of the same animal. In the primary motor cortex and the PMd, cortical regions that project to lower cervical segments were largely separate from those that project to lower lumbosacral segments. In the PMv, few neurons were labeled after tracer injections into lower cervical segments or lower lumbosacral segments. However, corticospinal neurons were labeled in the PMv after tracer injections into upper cervical segments and after HRP placement in the dorsolateral funiculus at T7. The region of the PMv that projects to upper cervical segments was separate from that which projects below T7. Cortical regions that project to upper and lower cervical segments of the spinal cord overlapped considerably in the primary motor cortex and in the PMd. Despite this overlap, we found that the regions of the primary motor cortex and PMd that project most densely to upper cervical segments were largely separate from those that project most densely to lower cervical segments. Furthermore, we found two separate regions within area 4 that send corticospinal projections primarily to the lower cervical segments. One of these regions was located within the classical “hand” area of the primary motor cortex. The other was located at the medial edge of arm representation in the primary motor cortex.(ABSTRACT TRUNCATED AT 400 WORDS)
Modulation of dopamine efflux in the striatum following cholinergic stimulation of the substantia nigra in intact and pedunculopontine tegmental nucleus-lesioned rats
Authors: Blaha CD , Winn P
Published: March 2018

The effects of microinjections of cholinergic agents into the substantia nigra pars compacta on dopamine (DA) efflux in the anterior dorsomedial striatum of urethane-anesthetized rats were investigated using in vivo chronoamperometry and intracerebral microdialysis techniques. A dose-dependent augmentation of DA efflux as evidenced by increases in the chronoamperometric signals was observed in the striatum following nigral microinjections of the cholinergic agonists nicotine or carbachol. Enhancing extracellular concentrations of ACh in the substantia nigra by intranigral infusions of the cholinesterase inhibitor neostigmine also resulted in an increase in the chronoamperometric signal corresponding to DA overflow in the striatum. These stimulatory effects of neostigmine on DA efflux in the striatum were confirmed using in vivo microdialysis. Compared to sham-operated control animals, quinolinic acid lesions of the pedunculopontine tegmental nucleus (PPTg) resulted in an attenuation of the stimulatory effects of intranigral neostigmine on DA efflux in the striatum. In contrast, these treatments resulted in an enhancement of striatal DA efflux in response to nigral infusions of the direct ACh receptor agonist nicotine. Combined, these data suggest that PPTg cholinergic neurons are indirectly involved in regulating the activity of the striatum by modulating the activity of DA neurons in the substantia nigra of the rat.
Increased plasma ACTH responses to stress in nonhandled compared with handled rats require basal levels of corticosterone and are associated with increased levels of ACTH secretagogues in the median eminence
Authors: Viau V , Sharma S , Plotsky PM , Meaney MJ
Published: March 2018

Postnatal handling of rat pups is known to alter permanently hypothalamic-pituitary-adrenal (HPA) responses to a wide variety of stressors. As adults, handled (H) and nonhandled (NH) animals also differ in sensitivity to the delayed negative-feedback effects of glucocorticoids on stress-induced HPA activity. However, it is not understood how handling alters neuroendocrine responses to stressful stimuli, and how differences in feedback sensitivity might account for changes in HPA activity both during and following stress. In the present studies, plasma ACTH responses to both restraint and ether stress were significantly greater in NH compared with H animals. Administration of 100 micrograms/kg corticosterone (B) immediately prior to restraint stress significantly decreased subsequent plasma ACTH responses to restraint to the same extent in H and NH animals, suggesting that H and NH animals do not differ in glucocorticoid fast feedback. H and NH animals adrenalectomized (ADX) 5 d prior to testing did not differ in plasma ACTH responses to restraint stress, demonstrating that the differences between the groups are dependent upon the presence of circulating B. However, the handling effect was apparent in ADX animals provided with a low level of B replacement (approximately 5-6 micrograms/dl). B replacement significantly decreased plasma ACTH levels under both basal conditions and in response to restraint stress in H/ADX rats. In contrast, although B replacement also decreased basal ACTH levels in ADX/NH rats, there were no differences in plasma ACTH responses to restraint between NH/ADX and NH/ADX+B animals.(ABSTRACT TRUNCATED AT 250 WORDS)
Anatomical and functional changes in the organization of the cuneate nucleus of adult rats after fetal forelimb amputation
Authors: Rhoades RW , Wall JT , Chiaia NL , Bennett-Clarke CA , Killackey HP
Published: March 2018

A previous study has shown that fetal forelimb removal in the rat results in an increase in the size of the hindlimb representation in primary somatosensory cortex and suggested that this anomalous cortical organization may have resulted from alterations in the primary afferent innervation of the dorsal column nuclei (Killackey and Dawson, 1989). The present study used both anatomical and electrophysiological techniques to examine the effects of fetal forelimb amputation on the dorsal column nuclei. Rats sustained forelimb removals on embryonic day 16 and were used in terminal experiments when they reached adulthood (> 60 d of age). Analysis of cytochrome oxidase-stained sections demonstrated that the cuneate nucleus ipsilateral to the lesion decreased in volume by an average of 36.7% (N = 7, p < 0.001, paired t test), but there was no corresponding increase in the volume of the gracile fasciculus and nucleus. Bilateral application of HRP to the sciatic nerves demonstrated that axons that innervate only the gracile nucleus on the intact side of the brainstem were present in the cuneate nucleus on the deafferented side. Injection of HRP into the skin overlying the point of the amputation (the stump) indicated that axons innervating this region filled most of the dorsal one-half of the shrunken cuneate nucleus and overlapped with the sciatic nerve afferents innervating the cuneate on this side. Mapping the receptive fields of multiple unit clusters demonstrated that most recording sites in the shrunken cuneate nucleus were activated by inputs from the stump and adjacent skin. In addition, 9.1% (N = 30) of such unit clusters (N = 328) could also be excited by stimulation of the hindlimb. These were observed in only three of the nine experiments. Unit clusters with split receptive fields including the skin overlying the stump and the hindlimb were located throughout the rostrocaudal extent of the cuneate nucleus. These results indicate that fetal forelimb amputation results in anatomical expansion of the central projections of hindlimb afferents into the cuneate nucleus. This anatomical organization appears weakly expressed in the receptive fields of cuneate neurons.