bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in the cortical map of the hand following postnatal median nerve injury in monkeys: modification of somatotopic aggregates
Authors: Wall JT , Huerta MF , Kaas JH
Published: March 2018

Median nerves to the hands of 8–15-d-old marmoset monkeys were transected and precluded from regeneration by ligation. Following periods of 0.4–1.5 years, features of organization in the cortical area 3b hand map were assessed neurophysiologically, and compared to features in normally reared monkeys. Cortical features in monkeys with both histories were similar in certain respects. (1) Receptive field organization was similar in terms of tactile thresholds and receptive field size, continuity, and glabrous-hairy specificity. (2) Somatotopic organization was similar in terms of the continuity of the glabrous representation, and progressions of receptive field shifts across some parts of the hand map. (3) Finally, the overall size of the hand map did not change. In contrast, other cortical features clearly differed following these developmental histories. (1) Neurons at virtually all recording sites in normal hand maps responded to light mechanical stimulation, whereas, following injury, neurons at about 8% of the recording sites responded only to high-intensity stimuli. (2) Somatotopic organization differed in terms of the presence or absence of the representation of skin autonomously innervated by the median nerve, the number and continuity of representations of hairy skin, and the spatial interfacing of representations. (3) Finally, there were differences in the areas and widths of representations of parts of the hand. The overall impression is that there is a correspondence between the cortical features that changed most after injury, and the features that varied most in individual normal monkeys: in both circumstances the most variable features involved properties of spatial patterning across large aggregates of neurons as reflected by the size, shape, continuity, and interfacing of representations. A hypothesis is proposed that suggests that the cortical hand map normally consists of a number of representations that are capable of developing and surviving somewhat autonomously of each other. The features of spatial patterning in the mosaiclike map of these representations are influenced by postnatal availability of inputs from intact hand nerves.
Adaptive adjustment of unit tuning to sound localization cues in response to monaural occlusion in developing owl optic tectum
Authors: Mogdans J , Knudsen EI
Published: March 2018

Bimodal units in the barn owl's optic tectum are tuned to the location of auditory and visual stimuli, and are systematically organized according to their spatial tuning to form mutually aligned maps of auditory and visual space. Map alignment results from the fact that, normally, units are tuned to the values of interaural level difference (ILD) and interaural time difference (ITD) produced by a sound source at the location of their visual receptive fields (VRFs). Monaural occlusion alters the correspondence of ILD and ITD values with locations in space. We investigated the effect that raising owls with a chronic monaural occlusion has on the tuning of tectal units to ILD and ITD. Owls were monaurally occluded beginning at 1 month of age. The effects of monaural occlusion were assessed 2–4 months later by comparing the ILD and ITD tuning of units in monaurally occluded owls with the ILD and ITD tuning of units with equivalent VRFs in normal owls. ILD and ITD tuning was shifted substantially and in the direction of the unoccluded ear (the adaptive direction) in owls raised with a monaural occlusion. In most tecta, the mapped representations of ILD and ITD were shifted systematically. In addition, in some tecta, monaural occlusion induced a change in the topography of the ILD map such that ILD tuning remained essentially constant at values near 0 dB over abnormally large portions of the tectum. Across all recording sites, the average shift in ILD tuning was 9 dB (n = 396) and the average shift in ITD tuning was 40 microseconds (n = 414). In four of five animals, the magnitude of the effect was not equivalent on the two sides of the brain, the adjustments being significantly larger and more systematic on the side ipsilateral to the occlusion. Such differences in the altered ILD and ITD maps on the two sides of the brain in individual animals indicate that, although a component of the adaptive adjustment might be due to regulation of the gain and phase response of the monaural signals early in the auditory pathway, a major component of the adjustment must occur at or beyond the level where the encoding of ILDs and ITDs for left and right space separates.
Vision-independent adjustment of unit tuning to sound localization cues in response to monaural occlusion in developing owl optic tectum
Authors: Knudsen EI , Mogdans J
Published: March 2018

Neurons in the developing optic tectum adjust their tuning to auditory localization cues in response to chronic monaural occlusion so that auditory spatial fields align with visual receptive fields (VRFs). We tested whether this adaptive adjustment of auditory tuning requires visual instruction. Both eyelids were sutured closed at the same time that one ear was occluded in two barn owls that were 1 month old. After 70 and 100 d, respectively, the tuning of units to interaural level difference (ILD) and to interaural time difference (ITD) was measured. These data were compared with equivalent data from 15 normal owls. Unit tuning to ITD was shifted from normal in both of the monaurally occluded owls. In one owl, ILD tuning was also clearly shifted. In the other owl, the map of ILD was flipped upside down and adaptive adjustments in ILD tuning could not be assessed. Instead, adjustments in ILD tuning were observed following removal of the earplug with the eyelids kept closed. Unit tuning was monitored at several sites in the tectum for 1 month after earplug removal using chronically implanted electrodes. Then, ILD tuning was resampled across the entire tectum. Both measures indicated shifts in ILD tuning in response to removal of the earplug in the second blind owl. In both animals, the magnitude of the shifts in ILD tuning and ITD tuning was smaller than has been observed previously in monaurally occluded but sighted owls. The results demonstrate that the brain can make adaptive adjustments in ILD and ITD tuning in response to early monaural occlusion even without the guiding influence of vision.
Interactions between dorsal root axons and their target motor neurons in developing mammalian spinal cord
Authors: Snider WD , Zhang L , Yusoof S , Gorukanti N , Tsering C
Published: March 2018

We have utilized the lipid-soluble tracers Dil and DiA to investigate interactions between group la dorsal root afferent axons and their target motor neurons in developing rat spinal cord. We show here that la axons project toward motor pools in fascicles that exhibit a considerable degree of spatial order. A rough topography is present in that axons that innervate medially located axial motor neurons cross over others in the intermediate zone and follow a separate path along the midline toward their appropriate targets. Surprisingly, we have also found that motor neuron dendritic projections are well established in the transverse plane prior to the arrival of la afferents. Although dendrites from motor pools innervating limb muscles project directly in the path of incoming la afferents, they do not guide afferents to appropriate motor pools. The la afferents pass over the distal dendrites and grow all the way to the border between gray and developing white matter. A significant amount of terminal branching and bouton formation is in the vicinity of motor neuron somata and proximal regions of the dendritic arbors. Few boutons are found near dendrites that project dorsal to the motor pools, and virtually no boutons are found on dendrites in white matter. Our results show that la afferent axons are not guided to appropriate motor pools by random encounters with motor dendrites, and raise the possibility that mechanisms exist that promote an orderly projection of la afferents to particular regions of the ventral horn. The striking lack of innervation of white matter and dorsally directed dendrites by la afferents raises the question of whether descending and intersegmental systems have their initial interactions with these regions of the motor neuron dendritic arbor.
Structure and chromosomal localization of the mammalian agrin gene
Authors: Rupp F , Ozcelik T , Linial M , Peterson K , Francke U , Scheller R
Published: March 2018

Agrin, a component of the synaptic basal lamina, has been shown to induce clustering of ACh receptors on the surface of muscle fibers. Analysis of cDNAs isolated from a rat embryonic spinal cord library demonstrated that agrin contains domains similar to regions of protease inhibitors, laminin and epidermal growth factor. The domain structure of agrin is further revealed here in an analysis of the agrin gene. Two additional internal repeated sequences are defined: one rich in cysteine residues with no homology to other proteins, and another similar to the laminin G domain, which is involved in heparin binding. Alternative RNA splicing at two positions in the gene predicts up to eight possible forms of the agrin protein. The gene (symbol AGRN/Agrn) has been assigned to chromosome 1 region pter-p32 in human and to mouse chromosome 4.
Predicting responses of nonlinear neurons in monkey striate cortex to complex patterns
Authors: Lehky SR , Sejnowski TJ , Desimone R
Published: March 2018

The overwhelming majority of neurons in primate visual cortex are nonlinear. For those cells, the techniques of linear system analysis, used with some success to model retinal ganglion cells and striate simple cells, are of limited applicability. As a start toward understanding the properties of nonlinear visual neurons, we have recorded responses of striate complex cells to hundreds of images, including both simple stimuli (bars and sinusoids) as well as complex stimuli (random textures and 3-D shaded surfaces). The latter set tended to give the strongest response. We created a neural network model for each neuron using an iterative optimization algorithm. The recorded responses to some stimulus patterns (the training set) were used to create the model, while responses to other patterns were reserved for testing the networks. The networks predicted recorded responses to training set patterns with a median correlation of 0.95. They were able to predict responses to test stimuli not in the training set with a correlation of 0.78 overall, and a correlation of 0.65 for complex stimuli considered alone. Thus, they were able to capture much of the input/output transfer function of the neurons, even for complex patterns. Examining connection strengths within each network, different parts of the network appeared to handle information at different spatial scales. To gain further insights, the network models were inverted to construct “optimal” stimuli for each cell, and their receptive fields were mapped with high-resolution spots. The receptive field properties of complex cells could not be reduced to any simpler mathematical formulation than the network models themselves.
Expression of c-fos protein in rat brain elicited by electrical stimulation of the pontine parabrachial nucleus
Authors: Krukoff TL , Morton TL , Harris KH , Jhamandas JH
Published: March 2018

The expression of Fos, the protein product of the primary response gene c-fos, was used metabolically to map the short-term (1 hr) effects of urethane and sodium pentobarbital anesthesia in rat. Subsequently, urethane-anesthetized rats were used to study the integrated response to electrical stimulation (1–1.5 hr) of the pontine parabrachial nucleus (PBN), an important center for relay of autonomic information in the brain. Immunohistochemistry was used to localize Fos-like immunoreactivity (FLI) in the brain. To approximate amounts of FLI in the conscious animal, rats were killed immediately after attaining surgical anesthesia with sodium pentobarbital (50 mg/kg) or urethane (1.2–1.7 gm/kg). No FLI was found in the brains of these rats. In rats killed 1 hr after anesthesia with sodium pentobarbital, FLI was found only in the habenulae. After 1 h of urethane anesthesia, low levels of FLI were found in the following areas: nucleus of the tractus solitarius (NTS); caudal and rostral ventrolateral medulla (VLM); lateral PBN; ventromedial, paraventricular, and supraoptic nuclei (SON) of the hypothalamus; medial preoptic area; central nucleus of the amygdala (ACE); endopiriform cortex; insular cortex; piriform cortex; and islands of Calleja. Electrical stimulation of the PBN (10 sec on, 10 sec off;15–50 microA at 20 Hz for 60–90 min) in rats anesthetized with urethane led to increases in mean arterial pressure (10–30 mm Hg) and to ipsilateral increases of FLI in the lateral PBN, dorsal division of SON, ACE, endopiriform nucleus, insular cortex, piriform cortex, and islands of Calleja. In two animals, ipsilateral increases were found in the ventromedial hypothalamus and medial amygdaloid nucleus.(ABSTRACT TRUNCATED AT 250 WORDS)
Phenotypical characterization of the rat striatal neurons expressing muscarinic receptor genes
Authors: Bernard V , Normand E , Bloch B
Published: March 2018

Neurons expressing the m1, m2, and m4 muscarinic receptor genes in the adult rat striatum were identified and characterized by using several in situ hybridization and immunohistochemical procedures. Combined in situ hybridization for the simultaneous detection of two mRNAs in the same section or in adjacent sections as well as in situ hybridization and immunohistochemistry on adjacent sections permitted us to identify the neurons containing m1, m2, or m4 receptor mRNA. Our observations demonstrate that m1, m2, and m4 receptor genes are expressed in one or several phenotypically distinct neuronal populations. The m1 receptor gene was the most widely expressed (85% of the striatal neurons). Most cholinergic neurons (80% or more) contain m1, m2, and m4 receptor mRNAs. Almost all the substance P neurons contain m1 and m4 receptor mRNA. All enkephalinergic neurons contained m1 receptor mRNA, but only 39% contained m4 receptor mRNA. Most somatostatin and neurotensin neurons expressed the m1 receptor gene, but only a few (15% and 9%, respectively) contained m4 receptor mRNA. The present study offers anatomical evidence that ACh may act directly in complex ways on the main neuronal populations of the striatum through muscarinic receptors. The m1, m2, and m4 receptors may act as autoreceptors to control ACh release and possibly other parameters of ACh neurons. On the other hand, the m1 and m4 receptors may act as heteroreceptors in cholinoceptive efferent neurons (enkephalin and substance P neurons) and other neurons (somatostatin/neuropeptide Y and neurotensin neurons). The presence of m4 receptor mRNA in only parts of the enkephalin, somatostatin, and neurotensin neuronal populations indicates that muscarinic receptor gene expression contributes to the functional and anatomical heterogeneity of the striatum that may relate to higher order of organization, including patch-matrix compartmentalization. The wide expression of m1 and m4 receptor genes in the striatum suggests that ACh may directly influence neurotransmitter release and synthesis in striatal efferent and intrinsic neurons. Our results imply that the specific pattern of expression of the muscarinic receptor genes mediates direct effects of ACh on activities and functions of chemically and topologically defined striatal neuronal populations. Since the expression of muscarinic receptors occurred in the three main neuronal populations of the striatum, namely ACh, enkephalins, and substance P neurons that also express dopamine receptors, it is highly probable that ACh and dopamine may act together at the single-cell level to influence striatal functions.
An electrophysiological correlate of protein kinase C isozyme distribution in cultured cerebellar neurons
Authors: Linden DJ , Smeyne M , Sun SC , Connor JA
Published: March 2018

Protein kinase C (PKC) is a family of at least seven closely related molecules (isozymes) that vary in terms of their requirements for activation and their distribution among cells of the brain. A striking example of this differential distribution is seen in the cerebellum, where Purkinje cells express PKC-I, an isozyme that is strongly activated by both phorbol ester (PE), and low doses of cis-unsaturated fatty acid (c-UFA), while granule cells predominantly express PKC-II, an isozyme that is strongly activated by PE but not c-UFA. Both Purkinje and granule cells have large, easily recorded voltage-gated K currents. These currents are attenuated by PKC activators in several other varieties of neuron. We hypothesized that the effects of these two PKC activators would be predicted by the distribution of the relevant PKC isozyme, and that the delayed outward rectifier current, IK, would be attenuated by both PE and c-UFA in Purkinje cells, but only by PE in granule cells. This hypothesis was confirmed in perforated-patch recordings. The attenuation produced by both activators could be blocked by application of a specific PKC inhibitor, RO-31–8220, and could not be mimicked by inert forms of PE or c-UFA. To our knowledge, this study represents the first report of an electrophysiological correlate of PKC isozyme distribution.
Temporal chimeras produced by hypothalamic transplants
Authors: Vogelbaum MA , Menaker M
Published: March 2018

The expression of locomotor activity by golden hamsters is temporally controlled by circadian oscillators contained within the suprachiasmatic nuclei. A genetic mutation has been found that alters the freerunning period of the locomotor activity rhythm from the wild- type value of approximately 24 to approximately 20 hr in homozygous mutants. It has been shown previously that a transplant of fetal hypothalamic tissue containing the suprachiasmatic nuclei to a host rendered arrhythmic by a complete lesion of the suprachiasmatic nuclei restores rhythmicity with the freerunning period that is normally expressed by the donor genotype. We made partial lesions of the suprachiasmatic nuclei of wild-type hosts, which did not completely abolish their circadian rhythmicity, and then placed hypothalamic implants from homozygous mutant fetal donors into the lesion site. The resulting complex patterns of locomotor activity contain rhythmic components with periods of both host and donor circadian oscillators, and suggest the presence of both stimulatory and inhibitory inputs from the circadian system to the centers controlling locomotor behavior.
A functional anatomical study of unipolar depression
Authors: Drevets WC , Videen TO , Price JL , Preskorn SH , Carmichael ST , Raichle ME
Published: March 2018

The functional neuroanatomy of unipolar major depression was investigated using positron emission tomography to measure differences in regional cerebral blood flow (BF). A relatively homogeneous subject group was obtained using criteria for familial pure depressive disease (FPDD), which are based upon family history as well as upon symptoms and course. Because of the absence of certain knowledge about the pathophysiology of mood disorders and their underlying functional neuroanatomy, we used data obtained from the subtraction of composite images from one-half of depressed and control subjects to identify candidate regions of interest. The major cortical region defined in this manner was statistically tested on a second set of subjects. Using this strategy, we found increased BF in an area that extended from the left ventrolateral prefrontal cortex onto the medial prefrontal cortical surface. Based upon the connectivity between these portions of the prefrontal cortex and the amygdala and evidence that the amygdala is involved in emotional modulation, activity was measured in the left amygdala and found to be significantly increased in the depressed group. A separate group of subjects with FPDD who were currently asymptomatic were also imaged to determine whether these findings represented abnormalities associated with the depressed state, or with a trait difference that might underlie the tendency to become depressed. Only the depressed group had increased activity in the left prefrontal cortex, suggesting that this abnormality represents a state marker of FPDD. Both the depressed and the remitted groups demonstrated increased activity in the left amygdala, though this difference achieved significance only in the depressed group. This suggests that the abnormality involving the left amygdala may represent a trait marker of FPDD, though further assessment in a larger sample size is necessary to establish this. These data along with other evidence suggest that a circuit involving the prefrontal cortex, amygdala, and related parts of the striatum, pallidum, and medial thalamus is involved in the functional neuroanatomy of depression.
Visual projections routed to the auditory pathway in ferrets: receptive fields of visual neurons in primary auditory cortex
Authors: Roe AW , Pallas SL , Kwon YH , Sur M
Published: March 2018

How does cortex that normally processes inputs from one sensory modality respond when provided with input from a different modality? We have addressed such a question with an experimental preparation in which retinal input is routed to the auditory pathway in ferrets. Following neonatal surgical manipulations, a specific population of retinal ganglion cells is induced to innervate the auditory thalamus and provides visual input to cells in auditory cortex (Sur et al., 1988). We have now examined in detail the visual response properties of single cells in primary auditory cortex (A1) of these rewired animals and compared the responses to those in primary visual cortex (V1) of normal animals. Cells in A1 of rewired animals differed from cells in normal V1: they exhibited larger receptive field sizes and poorer visual responsivity, and responded with longer latencies to electrical stimulation of their inputs. However, striking similarities were also found. Like cells in normal V1, A1 cells in rewired animals exhibited orientation and direction selectivity and had simple and complex receptive field organizations. Furthermore, the degree of orientation and directional selectivity as well as the proportions of simple, complex, and nonoriented cells found in A1 and V1 were very similar. These results have significant implications for possible commonalities in intracortical processing circuits between sensory cortices, and for the role of inputs in specifying intracortical circuitry.
The role of NMDA receptor-operated calcium channels in persistent nociception after formalin-induced tissue injury
Authors: Coderre TJ , Melzack R
Published: March 2018

The contribution of intracellular calcium to central sensitization and persistent nociception in response to tissue injury in rats was examined following the subcutaneous injection of formalin into the hindpaw. Formalin injury-induced nociceptive behaviors were enhanced by intrathecal pretreatment with the calcium ionophore A23187 or the calcium channel agonist Bay-K8644. Conversely, formalin nociceptive responses were reduced by intrathecal pretreatment with the calcium chelator Quin 2 or the calcium channel antagonists verapamil and nifedipine. Each of these agents affected the tonic, but not the acute, phase of the formalin response. The enhancement in formalin nociceptive behavior in rats treated with L-aspartate or L-glutamate was reversed by combined pretreatment with the noncompetitive NMDA antagonist MK- 801, but not by nifedipine or the non-NMDA excitatory amino acid antagonist 6-cyano-7-dinitroquinoxaline-2,3-dione. In rats not treated with excitatory amino acids, the analgesic effect of MK-801 was also significantly greater than that produced by nifedipine. Furthermore, combining nifedipine with MK-801 did not produce a significantly greater analgesic effect than MK-801 alone. The results suggest that central sensitization and persistent nociception following formalin- induced tissue injury are dependent on the influx of calcium through predominantly NMDA receptor-operated (and to a lesser extent voltage- gated) calcium channels.
Primary demyelination induced by exposure to tellurium alters Schwann cell gene expression: a model for intracellular targeting of NGF receptor
Authors: Toews AD , Griffiths IR , Kyriakides E , Goodrum JF , Eckermann CE , Morell P , Thomson CE
Published: March 2018

Exposure of developing rats to tellurium results in a highly synchronous segmental demyelination of peripheral nerves with sparing of axons; this demyelination is followed closely by a period of rapid remyelination. Demyelination occurs subsequent to a tellurium-induced block in the synthesis of cholesterol, the major myelin lipid. We utilized the techniques of Northern blotting, in situ hybridization, and immunocytochemistry to examine temporal alterations in Schwann cell gene expression related to demyelination and remyelination. Tellurium- induced demyelination is associated with downregulation of myelin protein expression and a corresponding upregulation of NGF receptor (NGF-R) and glial fibrillary acidic protein (GFAP) expression. Steady- state mRNA levels (expressed on a “per nerve” basis) for P0, the major myelin protein, were decreased by about 50% after 5 d of tellurium exposure, while levels of mRNA for NGF-R and GFAP were markedly increased (about 15-fold). In situ hybridization of teased fibers suggested that the increase in steady-state mRNA levels for NGF-R was primarily associated with demyelinated internodes and not with adjacent unaffected internodes. Although P0 message was almost totally absent from demyelinating internodes, it was also reduced in normal-appearing internodes as well. This suggests that limiting the supply of a required membrane component (cholesterol) may lead to partial downregulation of myelin gene expression in all myelinating Schwann cells. In partially demyelinated internodes, NGF-R and GFAP immunofluorescence appeared largely confined to the demyelinated regions. This suggests specific targeting of these proteins to local areas of the Schwann cell where there is myelin loss. These results demonstrate that demyelination is associated with reversion of the affected Schwann cells to a precursor cell phenotype. Because axons remain intact, our results suggest that these changes in Schwann cell gene expression do not require input from a degenerating axon, but instead may depend on whether concerted synthesis of myelin is occurring.
Anatomy and physiology of multipolar cells in the rat inferior collicular cortex using the in vitro brain slice technique
Authors: Smith PH
Published: March 2018

Coronal brain slices from 21–50-d-old hooded rats were used to characterize intracellular responses of cells in both the external and dorsal cortices of the inferior colliculus (IC). These cells could generate both sodium and calcium spikes. Depending on current amplitude, depolarizing current pulses could elicit either phasic or tonic firing patterns, with spike frequency adaptation. Spiking also occurred at the offset of a hyperpolarizing pulse. These patterns were due primarily to the activation of calcium conductances. Stimulation of the commissural pathway connecting the left and right IC produced a short-latency monosynaptic IPSP followed by an EPSP(s) and a late polysynaptic IPSP(s). Non-NMDA glutamate antagonists eliminated or reduced the amplitude of the EPSP and the late portion of the inhibition, while both IPSPs were blocked by GABAA antagonists. As described previously in guinea pig (Smith, 1986) and rat (Pierson et al., 1989), a large NMDA-mediated depolarizing event (paroxysmal depolarizing shift, or PDS) could be elicited by shocking the commissure of the IC in the presence of picrotoxin or bicuculline, NMDA, 4-aminopyridine, or in 0 Mg2+ Ringer's. The picrotoxin-induced PDS was significantly reduced or abolished in Ringer's containing aminophosphonovalerate. Cells displaying the responses described were labeled with neurobiotin. Those labeled are medium-sized multipolar cells. Their dendrites are usually spiny and can extend superficially up to the cortical surface. Their thin axons give rise to collaterals that branch profusely within the cortex. The main axons project laterally along the circumference of the IC or medially into the commissure separating the collicular hemispheres.
Activation of single neurons in the rat nucleus accumbens during self- stimulation of the ventral tegmental area
Authors: Wolske M , Rompre PP , Wise RA , West MO
Published: March 2018

Single neurons (n = 76) were recorded in the nucleus accumbens septi (NAS) of rats self-stimulating the ipsilateral medial forebrain bundle (MFB) at the level of the ventral tegmental area (VTA). Responses evoked by rewarding trains of stimulus pulses fell into five categories. The first category (40% of the sample) was characterized by a single discharge at invariant latency in response to individual pulses of the train, and hence was termed “tightly time locked” (TTL). Two TTL neurons were collision tested, and both showed collision, suggesting that self-stimulation of the VTA may involve antidromic, and thus direct, activation of a substantial number of NAS axons. The second category (26%) was characterized by discharges that varied in latency from pulse to pulse and hence was termed “loosely time locked” (LTL). Responses of the remainder of the sample showed no coupling to individual pulses but were categorized based on general firing patterns during the train: excited (7%), inhibited (4%), and no change (23%). Irrespective of category, immediately after the self-stimulation session, the likelihood of evoked discharge at monosynaptic latency by single pulse stimulation of the ipsilateral fimbria was reduced (relative to pre-session level), concurrent with elevations in mean firing rate and motor activity. NAS neurons thus exhibit vigorous activation, apparently both antidromically and orthodromically, in response to VTA self-stimulation. The responses of certain LTL and TTL neurons increased as a function of pulse number in the train, suggestive of integrative mechanisms important for brain stimulation reward. Conduction velocities of directly activated (TTL) axons (0.41– 0.65 m/sec) were slower than those previously reported for first-stage, reward-relevant axons. Nonetheless, an implication of direct activation of NAS (and other MFB) axons is that rewarding stimulation triggers action potentials that could invade all axonal branches, including those between the stimulation site and the soma, and send synaptic signals to target neurons. Such signals from NAS neurons could contribute to the increased motor behavior accompanying MFB self- stimulation, and/or could interact with dopamine-mediated signals projected to the NAS from reward circuitry.
Enhanced ACh sensitivity is accompanied by changes in ACh receptor channel properties and segregation of ACh receptor subtypes on sympathetic neurons during innervation in vivo
Authors: Moss BL , Role LW
Published: March 2018

Although presynaptic input can influence the number and distribution of ACh receptors (AChRs) on muscle, the role of cellular interactions in the development of transmitter sensitivity in neurons is less clear. To determine whether presynaptic input modifies neuronal AChR channel function and distribution, we must first ascertain the profile of changes in receptor properties relative to the timing of synapse formation. We have examined the temporal aspects of synaptogenesis in the lumbar sympathetic ganglia of the embryonic chick in anatomical experiments with anterograde 1,1′-dioctadecyl-3,3,3′,3′- tetramethylindocarbocyanine perchlorate labeling of presynaptic inputs and cytochrome oxidase histochemistry. Biophysical studies of sympathetic neurons, within hours of removal from animals at different stages relative to synapse formation, show that both the properties and distribution of AChR channels are modified concurrent with a significant increase in presynaptic input to the neurons. The most striking change in AChR channel distribution is revealed by patching multiple sites on the surface of individual neurons. Following innervation in vivo, many neurons express only one of the four AChR channel subtypes and the AChRs are clustered in discrete, high-activity patches. Furthermore, when neurons at this stage express more than one AChR channel subtype, the different classes are often spatially segregated from one another on the cell surface. This contrasts with patches from neurons removed earlier on, which have lower overall activity, often comprised of multiple channel subtypes. Comparison of the AChR properties of acutely dispersed neurons to those of neurons maintained in vitro indicates that most features of AChR channels are conserved despite their removal from presynaptic and other in vivo influences. These findings are consistent with inductive interactions between pre- and postsynaptic neurons playing an important regulatory role in transmitter receptor expression.
Brain glia release factors with opposing actions upon neuronal survival
Authors: Giulian D , Vaca K , Corpuz M
Published: March 2018

Microglia and astroglia have been thought to govern the survival of neurons after damage to the CNS. To investigate these putative glia- neuron relationships, we examined microglia and astroglia secretion products for effects upon growth of cultured neurons. Activated microglia secrete small neurotoxic factors (< 500 Da), while astroglia constitutively release proteins (> 10 kDa) that promote neuronal growth. Proteins released from astroglia, moreover, attenuate microglial toxicity, suggesting that different glial populations have opposing actions upon neuronal survival. Further study shows that neurotoxins from microglia are heat-stable, protease-resistant molecules with biologic activities blocked by NMDA receptor antagonists. Microglial factors, although toxic for chick ciliary neurons and rat spinal cord neurons, did not reduce numbers of oligodendroglia, astroglia, or Schwann cells in culture. The microglial neurotoxins can be distinguished from cytokines, from free radical intermediates, from the excitatory amino acids glutamate or aspartate, and from the NMDA receptor-mediated toxin quinolinic acid. We propose that secretion products from reactive microglia, but not astroglia, endanger surviving neurons after CNS injury by release of a novel class of neuron-killing molecules.
Activation of metabotropic glutamate receptors increases cAMP accumulation in hippocampus by potentiating responses to endogenous adenosine
Authors: Winder DG , Conn PJ
Published: March 2018

Metabotropic glutamate receptors (mGluRs) are coupled to effector systems through GTP-binding proteins (G-proteins) and appear to mediate slow synaptic responses in the CNS. Although mGluR-mediated increases in phosphoinositide hydrolysis have been well characterized, other mechanisms for signal transduction employed by mGluRs are poorly understood. We recently reported that the selective mGluR agonist 1- aminocyclopentane-1 S,3R-dicarboxylic acid (1S,3R-ACPD) increases cAMP accumulation in rat hippocampal slices. We have now investigated the mechanisms involved in this response. A number of G-protein-linked receptors that are not directly coupled to adenylate cyclase increase cAMP accumulation by potentiating cAMP responses to other agonists. Furthermore, previous studies suggest that glutamate increases cAMP accumulation by a mechanism that is dependent upon the presence of endogenous adenosine. Therefore, we tested the hypothesis that 1S,3R- ACPD-stimulated increases in cAMP accumulation in rat hippocampal slices are dependent upon the presence of endogenous adenosine and are mediated by an mGluR that potentiates cAMP responses to other agonists. We found that adenosine deaminase abolished 1S,3R-ACPD-stimulated cAMP accumulation whereas the adenosine uptake blocker dipyridamole enhanced this response. Additionally, adenosine receptor antagonists blocked mGluR-mediated increases in cAMP accumulation with potencies that were highly correlated with their potencies at A2 adenosine receptors. Furthermore, we performed a series of studies that suggest that 1S,3R- ACPD activates an mGluR subtype that potentiates responses to agonists of other receptors that are coupled to adenylate cyclase and that 1S,3R- ACPD-stimulated increases in cAMP accumulation in hippocampal slices are mediated by potentiation of the cAMP response to low levels of endogenous adenosine that are continuously present extracellularly.
Postural force fields of the human arm and their role in generating multijoint movements
Authors: Shadmehr R , Mussa-Ivaldi FA , Bizzi E
Published: March 2018

When a perturbation displaces the human hand from equilibrium, arm muscles respond by producing restoring forces. When a set of displacements are given at various directions from the same equilibrium position, the resulting restoring forces form a “postural force field.” It is not known whether these postural forces are related to those generated when a reaching movement is executed. However, if a movement is a consequence of a shift of the equilibrium position of the hand toward the target, then, from the postural force field, predictions can be made regarding the nature of the elastic forces acting on the hand during the movement. We have taken the first steps in testing this hypothesis by measuring the postural force field of a subject's arm over relatively large distances, and comparing these forces with the static forces generated at the hand while the subject attempted a reaching movement. Using a robot manipulandum, the hand was displaced at various directions from an equilibrium position. The measured restoring forces were fitted to a nonlinear model to define a postural force field for that equilibrium position. This field was used to predict elastic forces generated when the subject attempted to move the manipulandum from a point on the circumference of a circle to a target at its center--the center corresponded to the equilibrium position at which the postural field was measured. In some of the movement trials, the manipulandum was locked during approximately the first 120 msec of the program for motion and the resulting static “evoked” forces measured. We found that (1) the evoked forces did not point to the target, but were a function of the configuration of the arm and rotated with the shoulder joint, and (2) the magnitude of the evoked forces varied systematically, even though the movements were of the same magnitude. These patterns were remarkably similar to those observed in the postural forces. Our results provide experimental evidence linking maintenance of posture in a multijoint system to that of generating a movement. The evidence is consistent with the hypothesis that the CNS programs a reaching movement by shifting the equilibrium position of the hand toward the target.
Major differences in Ca2+i response to anoxia between neonatal and adult rat CA1 neurons: role of Ca2+o and Na+o
Authors: Friedman JE , Haddad GG
Published: March 2018

Although we and others have previously shown that newborn central mammalian neurons are more tolerant to anoxia than their adult counterparts, we do not know whether neonatal nerve cells accumulate free cytosolic calcium (Ca2+i) less than adults in response to O2 deprivation. In order to determine whether anoxia increases Ca2+i in adult and neonatal neurons, we monitored calcium in CA1 hippocampal neurons using the calcium-sensitive probe fluo-3 and confocal microscopy. These neurons were studied in the dissociated state in order to study their inherent response to anoxia without the influence of modulatory factors such as synaptic input and neurotransmitters. Severe anoxia caused a rapid increase in Ca2+i in adult CA1 hippocampal neurons, followed by swelling and bleb formation. In neonatal neurons, the latency of this calcium rise was about five times longer than in the adult. Removal of extracellular calcium and addition of calcium channel blockers (Co2+) greatly attenuated the increase in Ca2+i in response to anoxia but did not prevent cell swelling and injury. The addition of glutamate antagonists MK-801 and 6-cyano-7-nitroquinoxaline- 2,3-dione did not affect the increase in Ca2+i induced by anoxia. Replacing extracellular sodium with impermeant cations (N-methyl-D- glucamine) prevented anoxia-induced nerve injury. In addition, Ca2+i levels dropped, rather than increased, during the anoxic period in the absence of sodium; Ca2+i returned toward baseline levels upon reoxygenation. Glutamate was also studied, in the presence of O2, to compare with the effects of anoxia. Adult neurons responded with similar latencies to both anoxia and glutamate (10-500 microM). Neonatal neurons, however, exhibited a more rapid increase in Ca2+i in response to glutamate than they did when exposed to anoxia. We conclude (1) that neonatal neurons increase Ca2+i more slowly than adults in response to anoxia and that this response is due to an influx of Ca2+o into the cytosol, (2) that an increase in Ca2+i is not necessary to induce nerve injury when nerve cells are deprived of O2, (3) that the removal of Nao+ prevents cell swelling and blebbing and is associated with a decrease in Ca2+i during anoxia, and finally (4) that glutamate exposure may not be a completely valid model for the direct action of anoxia on neurons.
Novel opioid binding sites associated with nuclei of NG108-15 neurohybrid cells
Authors: Belcheva M , Barg J , Rowinski J , Clark WG , Gloeckner CA , Ho A , Gao XM , Chuang DM , Coscia C
Published: March 2018

Nuclear opioid binding sites have been discovered in NG108-15 neurohybrid cells. Marker enzyme analyses as well as electron and fluorescence microscopy studies attested to the high degree of purity of the nuclear preparations. Immunohistochemical studies on cryostat sections of NG108-15 cells with an antibody to the opioid receptor corroborated a nuclear localization. 3H-[D-Pen2,D-Pen5]enkephalin (3H- DPDPE), 3H-[D-Ala2,D-Leu5]enkephalin (3H-DADLE), and 3H-diprenorphine binding parameters, Kd and Bmax values, and heterologous competition binding and stereospecificity data satisfied criteria for the presence of delta-opioid sites in purified nuclear preparations. Neither mu-([D- Ala2,mephe4,gly-ol5] enkephalin), dihydromorphine, nor kappa-(U69593) specific binding was detectable in purified nuclear preparations. Rates of association and dissociation of 3H-[D-Ser2,L-Leu5]enkephalyl-Thr were comparable to values obtained previously for opioid receptors. Opioid binding was also shown in subnuclear preparations from NG108-15 cell cultures. Agonists, 3H-DADLE and 3H-DPDPE, bind with high affinity to nuclear membranes and with lower affinity to chromatin. In contrast, partial agonist 3H-diprenorphine high-affinity binding sites were predominant in chromatin, while low-affinity binding was found in the nuclear membrane. Accordingly, 5′-guanylylimidodiphosphate sensitivity of 3H-DADLE binding was detected in nuclear membranes but not in chromatin. Both agonist and partial agonist opioid binding to nuclear membrane and chromatin were abolished upon cycloheximide treatment of NG108-15 cells. Taken together, the results suggest that NG108-15 cells contain newly synthesized GTP binding regulatory protein (G-protein)- coupled delta-opioid receptors in nuclear membranes and uncoupled opioid binding sites in chromatin.
Multiple modes of N-type calcium channel activity distinguished by differences in gating kinetics
Authors: Delcour AH , Lipscombe D , Tsien RW
Published: March 2018

In many neurons, N-type Ca2+ channels are a major Ca2+ entry pathway and strongly influence neurotransmitter release. We carried out cell- attached patch recordings (110 mM Ba2+ as charge carrier) to characterize the rapid opening and closing kinetics of N-type Ca2+ channel gating in frog sympathetic neurons. Single channels display at least three distinct patterns of gating, characterized as low-, medium- , and high-rho o modes on the basis of channel open probability (rho o) during depolarizing pulses to -10 mV. Spontaneous transitions from one mode to another are infrequent, with an exponential distribution of dwell times and mean sojourns of approximately 10 sec in each mode. Thus, a channel typically undergoes hundreds or thousands of open- closed transitions in one mode before switching to a different mode. Transitions between modes during a depolarization were occasionally detected, but were rare, as expected for infrequent modal switching. Within each mode, the activation kinetics were well described by a simple scheme (C2-C1-O), as previously reported for other types of Ca2+ channels. Rate constants are strikingly different from one mode to another, giving each mode its own characteristic kinetic signature. The gating behavior at -10 mV ranges from brief openings (approximately 0.3 msec) and long closures (10–20 msec) for low-rho o gating to long openings (3 msec) and brief closures (approximately 1 msec) for high- rho o gating. Intermediate values for mean open durations (approximately 1.5 msec) and mean closed durations (approximately 3 msec) were found for medium-rho o gating. In addition to being kinetically distinct, channel openings in the low-rho o mode often exhibit a unitary current approximately 0.2 pA larger than in the medium- or high-rho o mode. Each mode is characterized by its own voltage dependence: activation occurs at relatively negative potentials and is most steeply voltage dependent in the high-rho o mode, while activation requires very strong depolarizations and is weakly voltage dependent in the low-rho o mode. The proportion of time spent in the individual modes varies greatly from one patch to another, suggesting that modal gating may be subject to cellular control.
Simultaneous pontine and basal forebrain microinjections of carbachol suppress REM sleep
Authors: Baghdoyan HA , Spotts JL , Snyder SG
Published: March 2018

This study was performed to test the hypothesis that cholinoceptive basal forebrain systems can significantly influence cholinoceptive pontine mechanisms known to be important for generating rapid eye movement (REM) sleep. This hypothesis was examined by microinjecting the cholinergic agonist carbachol or saline (vehicle control) into the pons, the basal forebrain, or simultaneously into the pons and basal forebrain, while quantifying the effects on sleep and wakefulness in unanesthetized, chronically instrumented cats. All microinjections were made during wakefulness and were followed by 2 or 4 hr of recording. Polygraphic records were scored for wakefulness, non-REM sleep, REM sleep, and the REM sleep-like state evoked by pontine administration of carbachol (DCarb). Dependent variables quantified following each microinjection included the percentage of recording time spent in each state, the latency to onset of non-REM, REM, and DCarb, the number of episodes per hour of each state, and the duration of the longest episode of each state. A total of 149 microinjections were made into 15 forebrain and 11 pontine sites in eight cats. Basal forebrain administration of carbachol significantly increased wakefulness. Pontine microinjection of carbachol produced a state that polygraphically and behaviorally resembled REM sleep. This REM sleep- like state occurred in amounts significantly greater than natural REM sleep. Pontine carbachol also significantly decreased wakefulness and non-REM sleep. Simultaneous injection of carbachol into the pons and basal forebrain enhanced REM sleep, but the magnitude of this enhancement was significantly less than the increase in REM sleep evoked by carbachol injection into the pons alone. The results show that cholinoceptive regions of the basal forebrain can increase wakefulness and reduce the ability of pontine carbachol to evoke the REM sleep-like state. These findings suggest that basal forebrain administration of carbachol activates an arousal-generating system that can successfully compete with the powerful cholinergic REM sleep- generating system of the pons.