bluuebunny/crossref_metadata_2025 huggingface dataset:
Ultrastructural localization of D2 receptor-like immunoreactivity in midbrain dopamine neurons and their striatal targets
Authors: Sesack SR , Aoki C , Pickel VM
Published: April 2018

Potential cellular substrates for functions ascribed to the dopamine D2 receptor were examined in rat brain using immunoperoxidase for localization of a D2 receptor peptide and immunogold staining for the catecholamine biosynthetic enzyme tyrosine hydroxylase (TH). Specificity of the rat polyclonal antiserum, raised against a 15 amino acid fragment from the third intracellular loop of the D2 receptor, was shown by immunoblot analysis and by selective labeling of cultured Chinese hamster ovary cells permanently transfected with the cDNA for the D2 receptor. Although the light microscopic distribution of immunolabeling for the D2 peptide was diffuse, it was selectively localized to regions containing dopamine cells (substantia nigra and ventral tegmental area) or their forebrain projections (dorsal and ventral striatum, nucleus accumbens, and olfactory tubercles). Electron microscopic examination of the medial substantia nigra and ventral tegmental area revealed readily detectable peroxidase immunoreactivity for the D2 peptide, primarily associated with the smooth endoplasmic reticulum and plasmalemmal surfaces of dendrites. Many D2 peptide- immunoreactive dendrites also contained immunogold labeling for TH, although some dendrites were singly labeled for either marker. In the medial and dorsolateral striatum, immunoperoxidase product for the D2 peptide was localized most extensively in dendrites, with the greatest intensity of immunolabeling seen in spines. A number of striatal dendrites exhibiting D2 peptide labeling were contacted by axon terminals immunoreactive for TH. Additionally, D2 peptide immunoreactivity was distributed to some synaptic vesicles and portions of the plasmalemmal surface in unmyelinated axons and in axon terminals. Most D2 peptide-immunoreactive terminals either lacked detectable membrane specializations, or formed thin, symmetric synapses in single sections. A few D2 peptide-labeled terminals formed asymmetric junctions on dendritic spines. In dually labeled sections, most D2 peptide-immunoreactive terminals lacked detectable immunolabeling for TH. However, in fortunate planes of section, peroxidase product for D2 peptide immunoreactivity was occasionally seen in pre-terminal portions of axons whose terminal varicosities contained immunogold labeling for TH. These ultrastructural results are consistent with the localization of a dopamine D2 receptor-like protein that is strategically positioned to subserve (1) autoreceptor functions at the level of dendrites in the midbrain and presynaptic axon terminals in the striatum, as well as (2) postsynaptic actions on striatal spiny dendrites and other nondopamine terminals.
Extracellular cGMP in the hippocampus of freely moving rats as an index of nitric oxide (NO) synthase activity
Authors: Vallebuona F , Raiteri M
Published: April 2018

The nitric oxide (NO) synthase/cGMP pathway has been studied in vivo in the adult rat hippocampus by monitoring the levels of extracellular cGMP during microdialysis in conscious unrestrained animals. The basal cGMP efflux was concentration-dependently reduced upon local infusion of the NO synthase inhibitor NG-nitro-L-arginine (NARG; 10 microM to 1 mM). The NO donors hydroxylamine and S-nitroso-N-penicillamine, perfused through the dialysis probe at 1 mM, increased by about 200% the extracellular levels of cGMP. The glutamate receptor agonist NMDA (125–500 microM) produced concentration-dependent cGMP responses that were abolished by the selective receptor antagonist D-2-amino-5- phosphonovaleric acid or by NARG. Local perfusion of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX; 1 mM) produced a steady eightfold increase of extracellular cGMP levels. The effect of IBMX was highly sensitive to NARG. The inhibition by NARG of the IBMX-induced cGMP response was reversed when the NO synthase substrate L-arginine was administered. It is concluded that cGMP collected during in vivo microdialysis reflects NO synthase activity in the rat hippocampus. The technique may be utilized to investigate the pathophysiology and the pharmacology of the NO/cGMP pathway in the hippocampus of living animals.
Identification of hair cell progenitors and intermitotic migration of their nuclei in the normal and regenerating avian inner ear
Authors: Tsue TT , Watling DL , Weisleder P , Coltrera MD , Rubel EW
Published: April 2018

Postembryonic production of sensory hair cells occurs in both normal and aminoglycoside-damaged avian inner ears. The cellular source and mechanism that results in new differentiated hair cells were investigated in the avian vestibular epithelia using three distinct cell-cycle-specific labeling methods to identify proliferating sensory epithelial cells. First, immunocytochemical detection of the proliferating cell nuclear antigen, an auxiliary protein of DNA polymerase, allowed labeling of cells in late G1, S, and early G2 phases of the cell cycle. Second, a pulse-fix tritiated thymidine autoradiographic protocol was used to identify cells in S phase of the cell cycle. Finally, Hoechst 33342, a fluorescent DNA stain, was used to identify epithelial cells in mitosis. The distribution of cells active in the cell cycle within the normal and ototoxin-damaged vestibular epithelium suggests that supporting cells within the sensory epithelia are the cellular precursors to the regenerated hair cells. Differences between the proliferation marker densities in control and damaged end organs indicate that the upregulation of mitotic activity observed after streptomycin treatment is due primarily to an increase in the number of dividing progenitor cells. The differences between the extent of ototoxic damage and the level of reparative proliferative response suggest a generalized stimulus, such as a soluble chemical factor, plays a role in initiating regeneration. Finally, after DNA replication is initiated, progenitor cell nuclei migrate from their original location close to the basement membrane to the lumenal surface, where cell division occurs. This pattern of intermitotic nuclear migration is analogous to that observed in the developing inner ear and neural epithelium.
Olfactory glomeruli in the zebrafish form an invariant pattern and are identifiable across animals
Authors: Baier H , Korsching S
Published: April 2018

Glomeruli are anatomical and possibly functional modules in the vertebrate olfactory bulb. We investigated the spatial arrangement of glomeruli in the olfactory bulbs of adult zebrafish (Brachydanio rerio). A solution of the lipophilic tracer Dil was injected into the nasal cavities. Axons of sensory neurons projecting from the olfactory epithelium into the bulb were traced anterogradely, thus labeling the whole population of glomeruli. The glomerular distribution was analyzed in detail by confocal laser-scanning microscopy. We find that a typical olfactory bulb contains a small number of about 80 glomeruli that have a stereotyped configuration in all animals investigated. All glomeruli exhibit bilateral symmetry. Twenty-two single glomeruli could be identified from animal to animal by their characteristic position and morphology. The remaining glomeruli either are embedded in glomerular plexus and therefore cannot be delineated reliably, or belong to a densely clustered subpopulation of on average 49 glomeruli in the dorsal olfactory bulb. No sexually dimorphic glomeruli were identified. To test whether glomerular constancy is specific for the zebrafish, we performed similar tracing experiments in the goldfish and found several indications for a similar invariance of glomeruli in this species. The remarkable stereotypy of this pattern is reminiscent of the insect olfactory system and has been demonstrated here for the first time in a vertebrate. It will now be possible to examine whether these identifiable glomeruli are functionally specialized in terms of odor processing. If so, zebrafish may emerge as a tractable model system for studies on olfactory coding.
Fate of myelin lipids during degeneration and regeneration of peripheral nerve: an autoradiographic study
Authors: Goodrum JF , Earnhardt T , Goines N , Bouldin TW
Published: April 2018

Four weeks after labeling myelin lipids with an intraneural injection of 3H-acetate, sciatic nerves were crushed, and the distribution of radiolabeled myelin lipids was followed by autoradiography from 1 d to 10 weeks later. Just prior to crush, silver grains were localized to the myelin sheath. Three days after crush, axons were degenerating and myelin sheaths were breaking down; silver grains appeared over lipid droplets within Schwann cells, fibroblasts, and macrophages. One week after crush the basal-lamina-delimited Schwann-cell tubes (Bungner bands) contained myelin debris, and some tubes already contained regenerating axons. Schwann cells were often displaced to the periphery of the tubes by phagocytes containing heavily labeled myelin debris; extratubal macrophages within the endoneurium contained labeled lipid droplets but no myelin debris. Two weeks after nerve crush silver grains were associated with newly formed myelin around regenerating axons. Many extratubal endoneurial macrophages now contained labeled myelin debris and lipid droplets. By 3 weeks myelination of regenerating axons was advanced, and the myelin sheaths were well labeled. Extratubal macrophages had become the major labeled structure within the nerve because they contained large amounts of labeled myelin debris and lipid droplets. From 4 to 10 weeks after nerve crush the new myelin sheaths continued to thicken and to be well labeled. Debris- laden extratubal macrophages remained the major site of labeled material within the endoneurium. Our results confirm that there is reutilization of myelin cholesterol by Schwann cells to form new myelin, and indicate that some lipid catabolism takes place in Schwann cells and endoneurial fibroblasts prior to infiltration of the nerve by macrophages. However, most of the myelin debris is phagocytized by macrophages within 1–2 weeks following nerve injury. These debris-laden macrophages persist within the nerve for many weeks, indicating that much of the salvaged cholesterol is not reutilized for myelin regeneration.
Long-term growth and remodeling of regenerated retino-collicular connections in adult hamsters
Authors: Carter DA , Bray GM , Aguayo AJ
Published: April 2018

The capacity of regenerating axons for long-term growth and synaptic plasticity was investigated in the visual system of adult hamsters. Four to six and 8–10 months after the eye and the superior colliculus (SC) were linked by a peripheral nerve (PN) graft, the retinal ganglion cell (RGC) axons that had regrown into the SC were examined ultrastructurally. Together with the data from hamsters with similar PN grafts for 2 months (Carter et al., 1989), this study spans most of the life of these animals. The overall findings indicate that (1) the RGC axons extended twice as far into the SC and the number of RGC terminals increased 30-fold between 2 and 4–6 months. These parameters did not change thereafter. The highest density of regenerated RGC terminals observed in the SC was 11.5% of controls. (2) The new RGC terminals acquired most of their normal ultrastructural characteristics by 2 months. (3) The mean size of the terminals was larger than in controls but decreased gradually, and there was a small increase in the size of the regenerated synapses. (4) At all times, the RGC terminals remained confined to the layers of the SC that normally receive retinal inputs, and their synapses were formed in normal proportions with the dendritic shafts and spines of SC neurons. Thus, there is a protracted long-term growth and remodeling of the RGC axons that have regenerated into the SC of these adult mammals.(ABSTRACT TRUNCATED AT 250 WORDS)
Alpha calcium/calmodulin-dependent protein kinase II selectively expressed in a subpopulation of excitatory neurons in monkey sensory- motor cortex: comparison with GAD-67 expression
Authors: Jones EG , Huntley GW , Benson DL
Published: April 2018

In situ hybridization histochemistry and immunocytochemistry, including double immunofluorescence, were used to study the populations of neurons expressing the alpha subunit of type II calcium/calmodulin- dependent protein kinase (CAM II kinase-alpha) or glutamic acid decarboxylase (GAD) in the somatic sensory and motor areas of the macaque monkey cerebral cortex. Sections were subjected to in situ hybridization using radioactive, complementary RNA probes specific for monkey CAM II kinase-alpha or 67 kDa GAD mRNAs. Others were stained immunocytochemically for CAM II kinase-alpha and/or GABA. CAM II kinase- alpha and GAD-67 are expressed in different populations of cells, with no colocalization. CAM II kinase-alpha is expressed in pyramidal cells of layers II-VI, especially layers II and III, as well as in certain small nonpyramidal cells of layer IV in areas 3a, 3b, 1, and 2 and of middle regions of area 4. Both cell types produce excitatory amino acid transmitters. Therefore, as in subcortical regions, CAM II kinase-alpha will be found on the presynaptic side of excitatory synapses but on the postsynaptic side only when these synapses occur on excitatory neurons in the sensory-motor cortex. Quantitative examination showed that CAM II kinase-alpha immunoreactive cells form, on average, approximately 50% of the total neuronal population in each area, while GABA immunoreactive or GAD cRNA hybridized cells form approximately 25–30%. Thus, CAM II kinase-alpha expressing cells cannot account for the total population of non-GABAergic cortical cells, and a certain proportion of the pyramidal cells probably do not express it. In other cortical areas, gene expression for the two molecules is regulated by afferent activity. Therefore, the present results form a necessary basis for studies aimed at determining the role of activity-dependent changes in the balance of excitation and inhibition as a mechanism underlying plasticity of representational maps in the primate sensory-motor cortex.
Cholinergic basal forebrain transplants restore diminished metabolic activity in the somatosensory cortex of rats with acetylcholine depletion [published erratum appears in J Neurosci 1994 Mar;14(3): following table of contents]
Authors: Jacobs SE , Fine A , Juliano SL
Published: April 2018

It has been known for several years that stimulus-evoked metabolic activity is reduced in the somatosensory cortex of animals with basal forebrain lesions that deplete the neocortex of acetylcholine (ACh). During 2-deoxyglucose (2-DG) experiments, animals with unilateral basal forebrain lesions demonstrate a decreased response to somatic stimulation, while background metabolic activity in the surrounding cortical regions remains normal. In an attempt to ameliorate these deficits, we examined the ability of embryonic cholinergic basal forebrain transplants inserted into neocortex to innervate surrounding cortical regions and restore functional 2-DG activity in adult host rats previously depleted of ACh by basal forebrain lesions. To accomplish this goal, a series of experiments were conducted in which we (1) depleted the cerebral cortex of ACh by injecting an excitotoxin into the rat basal forebrain, (2) transplanted embryonic basal forebrain or embryonic neocortical (control) tissue into the ACh- depleted cortex and, (3) 6–12 months later, used the 2-DG metabolic mapping technique to examine effects of the transplants on metabolic activity evoked by whisker stimulation in rat somatosensory (barrel) cortex. Histochemical analysis revealed that acetylcholinesterase (AChE) staining within 2 mm of the basal forebrain transplants was similar in density to the contralateral normal hemisphere. AChE staining farther than 2 mm from the basal forebrain transplants and throughout hemispheres containing neocortical (control) transplants was greatly reduced, with few AChE-positive fibers present, a finding typical of cerebral cortex in basal forebrain-lesioned animals. Stimulus-evoked 2-DG uptake in barrels adjacent to the basal forebrain transplants, and therefore within AChE-rich territory, was similar to that in corresponding barrels identically activated in the contralateral hemisphere. 2-DG activity was reduced, however, in stimulated barrels outside the region of dense AChE-positive staining, as well as in all activated barrels in hemispheres containing control transplants of embryonic neocortex. These results indicate that transplantation of cell suspensions containing embryonic cholinergic basal forebrain, but not neocortex, can ameliorate basal forebrain lesion-induced deficits in functional activity, and that the restoration of activity is influenced by proximity to the transplant.
Primary afferent plasticity following partial denervation of the trigeminal brainstem nuclear complex in the postnatal rat
Authors: Renehan WE , Crissman RS , Jacquin MF
Published: April 2018

Although interaxonal competition is believed to be an essential component of the normal development of numerous mammalian neuronal populations, there is considerable debate regarding the role of competition in the development and maintenance of the somatic sensory system. The results of recent investigations suggest that trigeminal primary afferents may compete for target territory in the brainstem, but it is unclear whether these interactions continue after birth. The present study explored this important issue by examining the response of individual trigeminal primary afferent neurons to partial denervation of the trigeminal brainstem nuclear complex at early postnatal ages. We utilized intracellular recording and HRP injection techniques to label primary afferent central terminal arbors in rats that sustained electrocautery of mystacial vibrissae in rows A, C, and E on the day of birth. A total of 42 low-threshold trigeminal primary afferent neurons were labeled in subnucleus interpolaris. Twenty-eight of these afferents supplied undamaged B or D row vibrissae while 14 supplied lesioned vibrissae. Qualitative and quantitative analyses revealed that the arbors associated with undamaged afferents were enlarged (mean arbor area of 13512 +/- 790.67 microns 2 vs normal area of 6130 +/- 214 microns 2) and were oriented toward the adjacent (partially denervated) territory. There was no significant change in the size of the lesioned afferent arbor area. The perimeter of the lesioned afferent arbors was increased, however, suggesting that the arbor shape had changed. This was confirmed with a form factor calculation that indicated that the circularity of the arbors associated with lesioned vibrissae was significantly reduced. Thus, while the arbors of undamaged afferents were enlarged and oriented in the direction of the partially denervated territory, the lesioned afferent arbors were not enlarged but assumed a flattened/elongate morphology within their appropriate row. The lesion-induced increase in the size of the undamaged afferent arbors was not associated with an increase in the number of bouton-like fiber swellings. The density of boutons was only 25% the value seen in normal animals. Thus, while the area supplied by the undamaged afferent arbors increased, there was no evidence that the absolute amount of terminal arbor was similarly increased (as would be the case if sprouting had occurred; see Renehan et al., 1989). We would therefore conclude that the undamaged afferents had undergone arbor expansion, but not sprouting. These data are consistent with prior suggestions that trigeminal primary afferents utilize some form of competitive interaction(s) to establish their final form and disposition. This competition would appear to continue into early postnatal periods.(ABSTRACT TRUNCATED AT 400 WORDS)
Cogeneration of neurons with a unique molecular phenotype in layers V and VI of widespread lateral neocortical areas in the rat
Authors: Arimatsu Y , Nihonmatsu I , Hirata K , Takiguchi-Hayashi K
Published: April 2018

Monoclonal antibody PC3.1 detects a unique subpopulation of neurons located mainly in layer VI and, to a lesser extent, in layer V within the lateral neocortical areas in the rat. In an attempt to characterize these neurons, we determined the time of their generation in selected neocortical areas by a double-labeling experiment combining quantitative long-survival 3H-thymidine autoradiography and immunohistochemistry for the PC3.1 antigen. We found that the vast majority of PC3.1-positive neurons in both layers V and VI were generated concurrently at embryonic day 15 in all areas examined, demonstrating a strict correlation between the molecular identity of neurons and the time of their generation, irrespective of their final positions along the radial and tangential axes. In contrast, PC3.1- negative neurons, which should represent more diverse phenotypic identities, were generated during a more extended period of cortical development and tended to exhibit radial (inside-to-outside) and tangential (ventral-to-dorsal and rostral-to-caudal) neurogenetic gradients. Our findings indicate that laminar and tangential locations of cortical neurons are not established solely by a combination of mechanisms for the inside-out movement of newly generated neurons in each cortical area and for the broad tangential neurogenetic gradients. The results of this study suggest a distinct way of cortical development in which neurons with a common molecular phenotype are generated concurrently and migrate toward their eventual positions, which are not necessarily located in a single lamina. In addition, our results suggest some kind of tangential heterogeneity in the mechanism involved in neocortical histogenesis, supporting the concept of early regional specification within the neocortex.
Effects of selective neonatal temporal lobe lesions on visual recognition memory in rhesus monkeys
Authors: Bachevalier J , Mishkin M
Published: April 2018

Ten-month-old infant monkeys that had received neonatal ablations of either inferior temporal cortex (area TE) or the medial temporal region were compared with age-matched normal infant monkeys in visual delayed nonmatching-to-sample with trial-unique objects. Both types of early damage caused impairment in visual recognition, but the degree of deficit after early area TE lesions differed sharply from that after early medial temporal removals. Thus, whereas early medial temporal damage yielded a marked decline in visual recognition when the delays and lists were gradually increased, early area TE damage yielded normal recognition up to a delay of 60 sec and only mild impairment at longer delays and lists. The data indicate that, unlike adult monkeys, which suffer severe and nearly equivalent losses in visual object recognition after both types of ablation, the infant monkeys' recognition ability is largely spared after early damage to area TE but not after early damage to the medial temporal lobe. Together with recent clinical reports of profound memory loss in children with early dysfunction of the medial temporal region, the present findings demonstrate that medial temporal lobe structures operate early to sustain visual recognition memory, and recovery from early damage is limited at best. Early damage to higher-order visual cortex, however, can be largely compensated, presumably by one or more of the visual cortical areas that were left intact.
Neural correlates of attentive selection for color or luminance in extrastriate area V4
Authors: Motter BC
Published: April 2018

Rhesus monkeys were trained on a conditional orientation discrimination task in order to assess whether attentive selection for a color or luminance stimulus feature would affect visual processing in extrastriate area V4. The task required monkeys to select a bar stimulus based on its color or luminance and then to discriminate the angular tilt of the selected stimulus. The majority of neurons (74%) were selectively activated when the color or luminance of the stimulus in the receptive field matched the color or luminance of the cue. The activity was attenuated when there was not a match between the stimulus and the cue. The differential activation was based on the presence or absence of the stimulus feature and was independent of spatial location. Across the population of V4 neurons, optimal stimuli that matched the selected color or luminance elicited about twice the activity as stimuli that did not match the selected feature. The feature-selective changes in activity were observed to develop beginning about 200 msec after the stimulus onset and were maintained over the remainder of the behavioral trial. In this task the activity of V4 neurons reflected a selection based on the cued feature and not simply the physical color or luminance of the receptive field stimulus. Under these conditions, the topographic representation of the neural activity in area V4 highlights the potential targets in the visual scene at the expense of background objects. These observations offer a physiological counterpart to psychophysical studies suggesting that stimuli can be preferentially selected in parallel across the visual field on the basis of a unique color or luminance feature
Increased levels of hemoglobin-derived and other peptides in Alzheimer's disease cerebellum
Authors: Slemmon JR , Hughes CM , Campbell GA , Flood DG
Published: April 2018

Several studies point to the importance of peptides and proteolysis in Alzheimer's disease (AD). Because of its ability to study small proteins and peptides, reverse-phase HPLC was employed to study these species in AD. Cerebellum was chosen for these initial studies because it does not show significant neuronal loss but does show some pathology in AD. Examination of over 600 peptide peaks per case revealed 15 that were elevated in AD. Nine were fragments of hemoglobin, and the remainder included two species of calmodulin, two of myelin basic protein, and one each of 67 kDa neurofilament protein and PEP-19. The cleavage sites on hemoglobin were after hydrophobic residues and immunolocalization was seen preferentially around blood vessel walls and granule cells. The elevation of the non-serum-derived peptides was characteristic of general metabolic changes that occurred in AD cerebellum, and the presence of elevated hemoglobin polypeptides indicated either possible disruption of the blood-brain barrier or selective evasion of it by peptidaceous products. Further studies are required to establish whether hemoglobin fragments have a role in neurodegenerative processes such as AD.
Lamina-specific expression and activity-dependent regulation of seven GABAA receptor subunit mRNAs in monkey visual cortex
Authors: Huntsman MM , Isackson PJ , Jones EG
Published: April 2018

Seven monkey-specific GABAA receptor subunit cDNAs were isolated and cloned; radioactive cDNA and cRNA probes derived from them were used for Northern blot analysis and in situ hybridization histochemistry of the primary visual cortex (area 17), with comparative observations on other cortical areas. cDNAs corresponding to alpha 1, alpha 2, alpha 4, alpha 5, beta 1, beta 2, and gamma 2 GABAA receptor subunits were isolated and had sequences unique to the monkey but recognized mRNAs of distinct molecular weights consistent with those reported in other species. mRNAs for the alpha 1, beta 2, and gamma 2 subunits were expressed at much higher levels in area 17 than in motor, somatosensory, or temporal association cortex, possibly reflecting the greater density of GABA cells and synapses in area 17. In areas 17 and 18, each of the seven subunit mRNAs showed individually distinct patterns of laminar distribution. alpha 1, beta 2, and gamma 2 subunit mRNAs, which are thought to form the basis of receptors with the full range of classical GABAA receptor properties in the adult, were particularly enriched in layers II-III, IVC, and VI of area 17, following patterns of receptor distribution previously demonstrated by radioligand binding and immunocytochemistry. alpha 2, alpha 4, alpha 5, and beta 1 transcripts had quite different localization patterns that did not match the antoradiographic or immunocytochemical receptor localization patterns. alpha 2 and alpha 5 subunit mRNAs, which are thought to be the subunits mainly expressed in development, were enriched in layer VI and the underlying white matter, possibly reflecting the involvement of receptors formed from alpha 2 and alpha 5 polypeptides in trophic interactions in the cortical subplate zone during development of the cerebral cortex. Following 8–21 d periods of monocular deprivation induced by intravitreal injection of TTX, levels of alpha 1, beta 2, and gamma 2 subunit mRNAs were substantially reduced in deprived ocular dominance columns of layer IVC in area 17. The effect was greatest for the alpha 1 subunit; for both alpha 1 and gamma 2 subunit mRNAs, it extended into deprived rows of cytochrome oxidase-identified periodicities in other layers. Apart from the alpha 5 subunit mRNA, which showed reduced levels in layer VI, the other subunit mRNAs were unaffected by monocular deprivation. These results demonstrate the heterogeneity of GABAA receptor subunit expression in a complex, multilaminar cortical area. They suggest that receptors with different functional properties may be assembled from different combinations of subunit polypeptides in different layers and show that subunit expression is differentially regulated under activity-dependent conditions.
AMPA-induced excitotoxic lesions of the basal forebrain: a significant role for the cortical cholinergic system in attentional function
Authors: Muir JL , Everitt BJ , Robbins TW
Published: April 2018

The aim of the present study was to clarify the role of the basal forebrain (BF)-cortical cholinergic system in visual attentional function by investigating the effect of alpha-amino-3-hydroxy-5-methyl- 4-isoxazole propionic acid (AMPA)-induced lesions of the basal forebrain on performance of a five-choice serial reaction time task. AMPA lesions in the present study produced a profound effect on performance of the task, as measured by choice accuracy and correct response latency. This deficit was significantly greater than that observed in earlier studies following ibotenate- or quisqualate-induced lesions of the BF. However, detailed histological and biochemical analysis revealed three rather different BF lesions depending upon the batch of AMPA supplied. In one group of animals (BF/1) the deficits in task performance were substantially greater and longer lasting compared to another group of lesioned animals (BF/2), which showed behavioral recovery several months following the lesion. The former sustained severe pallidal damage in addition to marked reductions in cortical ChAT activity. Support for the attentional nature of these deficits was obtained by the ability to improve task performance in BF/1 lesioned animals by increasing the duration of the visual stimulus and thus reducing the attentional load placed on these animals. In contrast, performance deficits could be reinstated in those animals showing behavioral recovery (BF/2) by reducing the duration of the visual stimulus and thus increasing attentional load. In the second experiment more discrete lesions of the magnocellular cholinergic neurons were made, resulting in extensive reduction of cortical ChAT activity with considerably less neuronal loss from the dorsal pallidum compared to the BF/1 lesion group. Once again, deficits on the task were substantially greater than observed previously following either quisqualate- or ibotenate-induced BF lesions. Furthermore, the cholinergic specificity of these deficits was supported by the attenuation of behavioral impairments following administration of the anti-cholinesterase physostigmine. Taken together with our earlier work, which has failed to demonstrate mnemonic deficits following lesions to the magnocellular neurons of the nucleus basalis of Meynert, these results suggest that the most consistent deficit produced following lesions of the BF-cortical cholinergic system is attentional dysfunction Analogous deficits in visual attention are also seen in patients with Alzheimer's disease, which can also be improved by anti- cholinesterase treatment.
Activated mutants of the alpha subunit of G(o) promote an increased number of neurites per cell
Authors: Strittmatter SM , Fishman MC , Zhu XP
Published: April 2018

The high concentration of the GTP-binding protein G(o) in the neuronal growth cone suggests that G(o) activation state may after neurite outgrowth. We find that activation of pertussis toxin-sensitive G- proteins by mastoparan increases neurite outgrowth from neuroblastoma cells. To examine G(o) activation specifically, point mutations homologous to activating, oncogenic mutations in alpha i2 and alpha s were introduced into the alpha subunit of G(o). The stability of the alpha o mutants to tryptic digestion confirms that they are activated. When expressed in PC12 or N1E-115 cells, activated alpha o doubles total neurites length per cell, primarily by increasing the number of neurites per cell. The growth cones of cells expressing activated alpha o are narrower than control growth cones. Expression of wildtype alpha o or the activated alpha subunits of other G-proteins did not affect total neurite length per cell. Thus, factors that lead to activation of G(o) can modulate neurite number per cell.
Patch-clamp recordings reveal powerful GABAergic inhibition in dentate hilar neurons
Authors: Soltesz I , Mody I
Published: April 2018

Whole-cell and cell-attached patch-clamp recordings were used to investigate the nature of GABAA receptor-mediated inhibition in the adult rat dentate gyrus in standard 400-microns-thick horizontal slices. In the presence of the glutamate receptor antagonists D-2-amino- 5-phosphonovaleric acid and 6-cyano-7-nitroquinoxaline-2,3-dione, whole- cell voltage-clamp experiments with chloride-filled electrodes ([Cl-]in = [Cl-]out) revealed a high degree of spontaneous activity (10–60 Hz) in all hilar neurons (HNs) recorded with access resistances lower than 20 M omega. The events were inward at negative holding potentials, reversed at around the Cl- equilibrium potential, and were completely abolished by the specific antagonists of the GABAA receptor channel picrotoxin and SR-95531 in a reversible manner, indicating that they were spontaneous inhibitory postsynaptic currents (sIPSCs) mediated by GABAA receptors. The majority of the sIPSCs were TTX-insensitive miniature currents resulting from the action potential-independent release of GABA. The 10–90% rise times and the monoexponential decay time constants of the sIPSCs were significantly longer in HNs than those found in neighboring granule cells (GCs). Furthermore, the decay time constant of the hilar sIPSCs was not voltage dependent, contrary to the voltage dependency of the decay time constant of the sIPSCs recorded from GCs. As HNs have longer electrotonic length than GCs do, dendritic filtering may contribute to the kinetic differences. Non- stationary fluctuation analysis showed that whereas the number of channels open at the peak of individual sIPSCs was similar, the single- channel conductances significantly differed between the two cell groups. The 21% smaller single-channel conductance and the existence of electrotonically close GABAergic synapses on HNs indicate that dendritic filtering alone cannot explain the differences between HNs and GCs. The distinct subunit composition of the GABAA receptor channels in HNs and GCs may also be responsible for the altered kinetics of IPSCs in HNs. However, the subunit specific benzodiazepine agonist zolpidem (3 microns) prolonged the monoexponential decay time constants in both HNs and GCs. Thus, differences between the GABAA receptors of the two cell types are not due to a simple all-or-none presence/absence of the alpha 5 subunit.(ABSTRACT TRUNCATED AT 400 WORDS)
Antiinflammatory influences of alpha-MSH molecules: central neurogenic and peripheral actions
Authors: Macaluso A , McCoy D , Ceriani G , Watanabe T , Biltz J , Catania A , Lipton JM
Published: April 2018

alpha-Melanocyte-stimulating hormone (alpha-MSH1–13) and its COOH- terminal tripeptide alpha-MSH11–13 (Lys Pro Val) inhibit inflammation when administered systemically. Recent evidence indicates that alpha- MSH1–13 can likewise inhibit inflammation in the skin solely via an action within the brain. Because of the potential importance of this discovery to understanding the control of inflammation and because alpha-MSH molecules might be useful for treatment of inflammation, experiments were performed to learn more about the mechanisms of action of these peptides. In tests on inflammation induced in the mouse ear by intradermal injections of recombinant human interleukin-1 beta, alpha- MSH1–1–13 administered intracerebroventricularly effectively reduced inflammation. This effect of centrally administered alpha-MSH1–13 was inhibited by systemic injection of the nonspecific beta-adrenergic receptor blocker propranolol and by administration of a specific beta 2- adrenergic receptor antagonist; the effect was not altered by blockade of cholinergic, alpha-adrenergic, or beta 1-adrenergic receptors. In mice with inflammation induced in a hind paw and with the spinal cord transected, the antiinflammatory effect of centrally administered alpha- MSH1–13 was prevented, indicating that intact descending neuronal pathways are required for the antiinflammatory influence of the central peptide. Systemic injection of alpha-MSH1–13 in animals with spinal cord transection had a smaller and later antiinflammatory effect, which suggests that the molecule also has an action, albeit lesser, in the periphery. However, alpha-MSH11–13 injected intraperitoneally had marked antiinflammatory activity in animals with spinal cord transection.(ABSTRACT TRUNCATED AT 250 WORDS)
Contrasting subcellular localization of the Kv1.2 K+ channel subunit in different neurons of rat brain
Authors: Sheng M , Tsaur ML , Jan YN , Jan LY
Published: April 2018

In the nervous system, a wide diversity of K+ channels are formed by the oligomeric assembly of subunits encoded by a large number of K+ channel genes. The physiological functions of a specific K+ channel subunit in vivo will be dictated in part by its subcellular location within neurons. We have used a combined in situ hybridization and immunocytochemical approach to determine the subcellular distribution of Kv1.2, a member of the Shaker subfamily of K+ channel genes. In contrast to other characterized K+ channel subunits, Kv1.2 protein shows a complex differential subcellular distribution in neurons of rat brain. In some of these neurons (e.g., hippocampal and cortical pyramidal cells, and Purkinje cells), Kv1.2 is concentrated in dendrites, while in others (e.g., cerebellar basket cells), Kv 1.2 is predominantly, if not exclusively, localized to nerve terminals. Furthermore, Kv1.2 immunoreactivity was also detected in certain axon tracts. We hypothesize that the differential sorting of Kv1.2 could result from association of Kv1.2 with varying heterologous K+ channel subunits in different cell types, with the implication that Kv1.2 may participate in distinct heteromultimeric K+ channels in different subcellular domains. The findings suggest that Kv1.2-containing K+ channels may play diverse functional roles in several neuronal compartments, regulating presynaptic or postsynaptic membrane excitability, depending on the neuronal cell type.
A slow excitatory postsynaptic potential mediated by 5-HT2 receptors in nucleus prepositus hypoglossi
Authors: Bobker DH
Published: April 2018

Intracellular recordings were made from neurons of the nucleus prepositus hypoglossi in slices of guinea pig medulla. 5-HT (serotonin) caused a hyperpolarization followed by a late depolarization. The hyperpolarization was mediated by 5-HT1A receptor activation and could be selectively blocked by pindobind-5HT1A (PBD). 5-HT then caused a depolarization only. A selective 5-HT2 agonist, (+)-1-(2,5-dimethoxy-4- iodophenyl)-2-aminopropane (DOI), also caused a depolarization. Ketanserin and spiperone, 5-HT2 antagonists, blocked the depolarization due to both 5-HT and DOI. Focal electrical stimulation caused an IPSP mediated by 5-HT acting upon 5-HT1A receptors and a slow EPSP (s-EPSP). PBD blocked the IPSP, leaving an isolated s-EPSP. Both spiperone and ketanserin antagonized the s-EPSP, while DOI occluded it. The s-EPSP was from 2 to 10 mV in amplitude and 35–50 sec in duration, and showed voltage dependence consistent with a decrease in potassium conductance. Both the IPSP and the s-EPSP were presynaptically inhibited by the 5- HT1D agonist sumatriptan. These data indicate that the s-EPSP is mediated by 5-HT acting upon 5-HT2 receptors. This represents strong support for the role of 5-HT as an excitatory neurotransmitter in the CNS. Further, it demonstrates that synaptic release of 5-HT can mediate opposing effects on the membrane potential of a single cell.
Distributed input to the tail-siphon withdrawal circuit in Aplysia from neurons in the J cluster of the cerebral ganglion
Authors: Raymond JL , Byrne JH
Published: April 2018

Plasticity in the circuits for the withdrawal reflexes has been correlated with several simple forms of nonassociative and associative learning in Aplysia, and biochemical, biophysical, and molecular mechanisms of plasticity in these circuits have been described. In order to examine network features of this plasticity, we identified and characterized a component of the modulatory circuitry for the tail- siphon withdrawal circuit. Activation of mechanoafferent neurons in the J cluster of the cerebral ganglion produced strong and distributed input to the tail-siphon withdrawal circuit. Stimulation of the J cells led to excitatory and inhibitory effects in the sensory neurons in the pleural ganglion, the tail motor neurons in the pedal ganglion, and several classes of interneurons in the pleural ganglion, including the multifunctional neuron LPI17. Activation of the J cells produced both fast and slow post-synaptic potentials in neurons of the tail-siphon withdrawal circuit. Of particular interest was the ability of the J cells to produce slow EPSPs in the pleural sensory neurons. These slow EPSPs were associated with an increase in the excitability of the sensory neurons, but no effect of the J cells on spike duration in the sensory neurons was observed. The J cells appear to mediate both sensory and modulatory inputs to the circuit for tail withdrawal.
Local modulation of basal forebrain: effects on working and reference memory
Authors: Givens B , Olton DS
Published: April 2018

The functional roles of the medial septal area (MSA) and nucleus basalis magnocellularis (NBM) in memory were investigated to determine (1) their relative contribution to working and reference memory, (2) their operation in spatial and nonspatial memory, (3) the temporal dynamics of the neural activity within these nuclei as they relate to mnemonic processes, (4) the neurochemical regulation of their activity, and (5) the importance of ACh for their function. Working memory was tested in a continuous conditional discrimination (CCD), and reference memory was tested in the CCD and a sensory discrimination (SD). Bipolar recording electrodes in the dentate hilus monitored hippocampal EEG (theta rhythm). Immediately prior to behavioral testing, trained rats were infused with tetracaine, scopolamine, or muscimol into the MSA or NBM, and the subsequent behavioral and physiological changes were measured and correlated. MSA infusions of all three drugs reduced the power of hippocampal theta and impaired choice accuracy in the CCD; the magnitude of both effects was greater for larger doses and steadily decreased over time after the infusion, producing a strong positive correlation between the power of theta and choice accuracy in the CCD. These infusions had no effect on measures of reference memory in the CCD or in the SD. The results demonstrate that rhythmic activity along the septohippocampal pathway reflects processing of nonspatial working, but not reference memory. NBM infusions did not affect hippocampal theta but did reduce choice accuracy in the SD and completely disrupted performance in the CCD. The NBM appears to have a critical role in both working and reference memory.
c-fos expression in vomeronasal pathways of mated or pheromone- stimulated male golden hamsters: contributions from vomeronasal sensory input and expression related to mating performance
Authors: Fernandez-Fewell GD , Meredith M
Published: April 2018

The vomeronasal system projects to the accessory olfactory bulb (AOB), to the medial (Me) and posterior medial cortical nuclei (PMCN) of the amygdala, to the bed nucleus of the stria terminalis (BNST), and to other central structures shown to be important in mating behavior, including the medial preoptic area (MPOA). In these experiments c-fos expression was used as a marker of neural activity to identify the contribution of vomeronasal sensory input during mating behavior in male golden hamsters, either intact or with vomeronasal organs removed (VNX). Inexperienced hamsters were either stimulated with a receptive female and allowed to mate, exposed to female hamster vaginal fluid (HVF), which contains stimuli known to act through the VN system, or placed in a clean cage alone. Densely stained Fos-positive nuclei were evident in mated animals in the central VN pathway [AOB, Me, posterior medial BNST (pmBNST)] and a VN target area (MPOA). HVF-exposed animals showed Fos expression in the AOB, Me, and BNST but not MPOA. Unstimulated animals showed almost no activation. Most VNX animals exposed to females did not mate, but performed intense chemoinvestigation. They had few Fos-positive nuclei in any of these areas except the caudal pmBNST. A few VNX animals that did mate had patterns of Fos activation that were similar but less intense than those of intact mating animals, suggesting a selective activation of VN central pathways during mating regardless of VN sensory input. The main olfactory system showed low levels of Fos expression in all animals (stimulated and unstimulated). Fos expression in the MPOA and rostral pmBNST was seen only in mated animals, suggesting that these regions are concerned with mating performance or its consequences, rather than the chemosensory input that triggers it. Fos expression in the caudal encapsulated pmBNST was evident in all groups of animals that performed chemosensory investigation, regardless of VN status or mating, suggesting that this region either directs or responds to chemosensory investigation.