bluuebunny/crossref_metadata_2025 huggingface dataset:
Slow outward tail currents in molluscan bursting pacemaker neurons: two components differing in temperature sensitivity
Authors: Thompson S , Smith SJ , Johnson JW
Published: March 2018

Bursting pacemaker neurons of Tritonia and Aplysia were studied in voltage-clamp experiments to determine the ionic requirements of the slow outward tail current. This current is important for terminating bursts and hyperpolarizing the neuron during the interval between bursts and therefore contributes to the timing of pacemaker activity. A method for rapid extracellular perfusion of the neuron soma with solutions of different ionic composition was used to study the ionic dependence of the tail current. Measurements of the current-voltage relationship were made at different times during the decay of the tail current to determine the reversal potential in different ionic solutions. These experiments showed that 2 or more slow outward currents contribute to the tail current. Temperature had a large effect on these currents. At cold temperatures (10–15 degrees C), the tail current was predominantly a K current. At warm temperatures (20–23 degrees C), both a K current and a K-insensitive outward current were seen. Bursting pacemaker activity occurred throughout this temperature range. Both the K current and the K-insensitive current required Ca influx for activation. These findings help to reconcile conflicting reports in the literature concerning the ionic dependence of the slow outward tail current and suggest a mechanism for temperature compensation of bursting activity in these pacemaker cells.
Electrophysiological interactions of isomers of cyclazocine with the phencyclidine antagonist metaphit in rat cerebellar Purkinje neurons
Authors: Wang Y , Palmer MR , Freedman R , Rice KC , Lessor RA , Jacobson AE , Hoffer BJ
Published: March 2018

Metaphit, 1-(1-(3-isothiocyanatophenyl) cyclohexyl) piperidine, an analog of phencyclidine (PCP) has been shown previously to selectively block PCP receptors and to irreversibly antagonize the depressant effect of PCP in cerebellum. In this study, we examined the electrophysiological interactions of metaphit and naloxone with stereoisomers of cyclazocine, an agent known to have analgesic and psychotomimetic activity in behavioral studies, effects that have been ascribed to opiate and PCP receptor activity. A dose-dependent and reversible slowing of Purkinje neuron discharge was seen with local application of (+)- or (-)-cyclazocine. We found that the blockade of (- )-cyclazocine effects required both high doses of naloxone and the presence of metaphit, whereas the responses to (+)-cyclazocine were blocked by metaphit alone on most cerebellar Purkinje neurons. These findings suggest that the depressant reaction of (+)-cyclazocine in cerebellar Purkinje neurons is primarily mediated through PCP receptors. (-)-Cyclazocine responses, on the other hand, appear to be due to activity at both PCP and kappa opioid receptors.
Replacement of synaptic terminals in lamina II and Clarke's nucleus after unilateral lumbosacral dorsal rhizotomy in adult cats
Authors: Murray M , Goldberger ME
Published: March 2018

Evidence from previous light-microscopic studies suggested that lumbosacral dorsal rhizotomy in cats elicits sprouting of converging undamaged systems into partially deafferented Clarke's nucleus and lamina II. We therefore applied quantitative electron-microscopic methods to determine whether this sprouting is associated with replacement of synaptic terminals (reactive reinnervation). We used stereological and morphometric methods to estimate terminal number per cross section in right and left lamina II and Clarke's nucleus in adult cats after acute and chronic unilateral (right-sided) lumbosacral deafferentation. Planimetric measurements of area indicated no significant shrinkage of either region as a result of the deafferentation; an increase in area occupied by glial cytoplasm (gliosis) equaled the decrease occupied by axonal components. The gliosis appears to persist indefinitely, although the degenerative debris stainable with conventional light-microscopic methods does not persist. Analysis of the synaptic population of lamina II reveals that the large central or “scalloped” terminals comprise a substantial fraction (greater than 40%) of the total area occupied by terminals in control material and that this population is largely lost upon deafferentation, leaving a large population of small terminals with spherical vesicles. Nevertheless, estimates of total terminal number indicate no difference between control and deafferented lamina II, suggesting a rapid and virtually complete replacement of lost dorsal root terminals by small terminals containing spherical vesicles. Terminal number in Clarke's nucleus also remains constant despite the loss of the dorsal root input. We conclude that there is also a virtually complete and rapid replacement of lost terminals in Clarke's nucleus by terminals containing spherical vesicles. These data provide an example of a case in which axonal sprouting demonstrated with light- microscopic methods is associated with electron-microscopic evidence of reactive reinnervation.
Noradrenergic neurons from the locus ceruleus in dissociated cell culture: culture methods, morphology, and electrophysiology
Authors: Masuko S , Nakajima Y , Nakajima S , Yamaguchi K
Published: March 2018

We have developed a dissociated primary cell culture of noradrenergic neurons from the locus ceruleus of postnatal (1- to 5-d-old) mice or rats. Slices of the brain stem were made on a Vibratome. Then the region of locus ceruleus, which was identified by observing the slices under a dissecting microscope, was dissected out from the slices. The removed fragments of brain slices were dissociated and cultured up to 3 weeks on a non-neuronal feeder layer, which consisted predominantly of astroglial cells, or on a fibronectin-treated collagen substratum. After 2 weeks of culture, about 70% of total neuronlike cells revealed positive catecholamine histofluorescence, indicating that they were probably noradrenergic neurons. About 98% of large- and medium-sized cultured neurons (soma diameter greater than or equal to 20 microns) was histofluorescence positive. The fluorescence-positive cells had long processes rich in varicosities, and the shape of their soma was either multipolar or fusiform. Electron microscopy using permanganate fixation revealed that the varicosities along their processes had small granular vesicles, which may contain norepinephrine. Physiological properties of these noradrenergic neurons were investigated with intracellular microelectrodes or with the whole-cell version of the patch clamp. We observed that many cells were producing spontaneous firing. Many of these spontaneously firing cells had no obvious contact with neighboring cells. The neurons were depolarized when glutamate was applied by pressure ejection. They also responded to GABA and glycine with either hyperpolarization or depolarization, and these responses were antagonized by picrotoxin and strychnine. Application of substance P generally produced depolarization with an increase in input resistance. The neurons responded with hyperpolarization to somatostatin, beta-endorphin, and enkephalin. This culture system will become a useful tool for elucidating the cellular and molecular properties of the central noradrenergic neurons.
Somatostatin mRNA: regional variation in hybridization densities in individual neurons
Authors: Uhl GR , Sasek CA
Published: March 2018

Somatostatin mRNA is detected by in situ hybridization of 35S-labeled single-stranded cDNA probes to coronal sections of the rat brain that include the periventricular nucleus of the hypothalamus. Features supporting hybridization specificity include its anatomic distribution, the results of studies using multiple cDNA probes, RNAase experiments, competition studies, and correlations with patterns of somatostatin peptide immunostaining in adjacent sections. The hybridization densities vary strikingly from region to region, with highest densities in the periventricular nucleus and more modest levels in areas such as the cerebral cortex and the striatum. On the basis of the results of in situ and immunohistochemical approaches, we suggest that this variation is due to regional differences in the density of hybridization per positive cell, as well as to regional variation in the densities of somatostatinergic perikarya.
The emergence of tuning in newly generated tuberous electroreceptors
Authors: Zakon HH
Published: March 2018

Tuning curves of afferent electroreceptive fibers in the anterior lateral line nerve of the weakly electric fish, Sternopygus macrurus, indicate that the tuberous electroreceptors of each individual are well- tuned to its own electric organ discharge (EOD) frequency. In order to study how receptor tuning may develop, new receptor organs were induced to form in regenerating cheek skin, and their tuning properties were compared with those of intact receptors from the same fish. At 3 weeks after the onset of regeneration, new receptors of a given fish were broadly tuned with best frequencies (BFs) lower than that fish's EOD frequency and the BFs of its own intact tuberous receptors. Three weeks later, regenerated receptors of the same fish were indistinguishable from intact receptors in BF, although tuning curves were occasionally slightly broader than normal. To determine if the presence of an ongoing electric field is necessary for the genesis of proper tuning, receptors were allowed to regenerate in fish deprived of their EODs. At 6 weeks, tuning curves of these receptors also had BFs that were tuned similarly to intact receptors and to each individual's characteristic EOD frequency (determined by recordings of the pacemaker nucleus in the medulla). Thus, as regenerating receptors mature, they gradually become more sharply tuned and tuned to progressively higher frequencies until reaching the correct BF, which matches the EOD frequency; however, tuning to the appropriate EOD frequency occurs without reference to the ongoing electric field.
Expression of the weaver gene in dopamine-containing neural systems is dose-dependent and affects both striatal and nonstriatal regions
Authors: Roffler-Tarlov S , Graybiel AM
Published: March 2018

In an earlier report we presented evidence pointing to a differential effect of the mutant gene weaver on the dopamine-containing fiber systems innervating the striatum. In mice homozygous for the weaver mutation, there is a severe loss of dopamine in the caudoputamen, the main target of the nigrostriatal system. By contrast, dopamine is entirely conserved in the nucleus accumbens, a target of the mesolimbic system, and is moderately affected in the olfactory tubercle. The present study shows that these defects in dopamine are gene dose- dependent, that they are established by the end of the first month of life, and that the losses are permanent and not progressive. As in homozygous weavers, the greatest defects in striatal dopamine in heterozygous weavers occur in the dorsolateral caudoputamen and the lateral olfactory tubercle. The abnormalities in the striatal dopamine content of weaver mice are not accompanied by abnormalities in the turnover of dopamine, judging from measurements of the dopamine metabolite dihydroxyphenylacetic acid. Norepinephrine content is also normal in each striatal region. No deficits in striatal dopamine occur in mice homozygous for the mutant genes staggerer and Purkinje cell degeneration, which, like the weaver mutation, result in ataxia and cerebellar pathology. A survey of nonstriatal regions in the weaver mice showed that the effects of the weaver gene on the dopamine- containing innervation of the forebrain are not confined to striatal targets but also extend to the septum and the hypothalamus. By contrast, dopamine in the frontal cortex, the amygdala, the olfactory bulb, and the retina is entirely spared. The pattern and extent of loss of dopamine in the weaver forebrain is thus region- and system- specific. In confirmation of our initial findings, a ca. 30% depletion of dopamine occurs in the weaver midbrain, the region containing the cell bodies of origin of the mesostriatal dopamine systems. A comparison of histofluorescent sections through weaver and control midbrains revealed a reduction of catecholamine-containing neurons in the pars compacta of the weaver animals. These results point to a subpopulation of dopamine-containing neurons as primary targets of the weaver gene or as being closely associated with such primary targets. As a gene-dose effect has also been shown for the cerebellar granule cell loss in the weaver, the mutant gene must have at least 2 cellular targets. We suggest that the cerebellar and mesostriatal pathologies may be linked by a common molecular mechanism.
A system of indoleamine-accumulating neurons in the rabbit retina
Authors: Sandell JH , Masland RH
Published: March 2018

The indoleamine-accumulating neurons of the rabbit retina were labeled by intraocular injection of 5,7-dihydroxytryptamine (5,7-DHT). The retinas were fixed with 2.5% paraformaldehyde and 0.2% glutaraldehyde and inspected by fluorescence microscopy. Five kinds of cell accumulated the indoleamine. They were labeled to essentially the same brightness and remained so despite variations in the concentration at which 5,7-DHT had been applied or the duration of its application. Experiments in which 5,7-DHT was applied to retinas incubated in vitro gave identical results. To see the whole shape of the cells, we visually guided micropipettes to the fluorescent cell bodies and injected the cells with Lucifer yellow CH. To study the cells as a population, we used a new method in which the fluorescence of 5,7-DHT is photochemically converted to an insoluble diaminobenzidine product. The dendrites of all of the indoleamine-accumulating cells were then simultaneously visible. Used together, these techniques revealed an interrelated system of indoleamine-accumulating neurons. All of the cells contribute processes to a dendritic plexus that lies at the inner margin of the inner plexiform layer. The plexus is roughly 4 micron thick. It is pierced by the stalks of the Muller cells and is occasionally interrupted by ganglion cell bodies, where they extend above the average margin of the ganglion cell layer. Otherwise it fills much of the space at the junction of the plexiform and ganglion cell layers. The type 1 and type 2 cells are amacrine cells with cell bodies at the inner margin of the inner nuclear layer. They have 5–8 radially branching primary dendrites which extend horizontally across the inner plexiform layer before descending to join the dendritic plexus. They differ from each other in cell body shape, dendritic morphology, and the course of their dendrites within the inner plexiform layer. Each has a “displaced” counterpart, with a morphology similar to the type 1 or type 2 cell but with a cell body located in the ganglion cell layer. The displaced cells are separate functional elements because, in contrast to the type 1 and type 2 cells, they have no dendrites (and hence can have no synaptic connections) in the outer part of the inner plexiform layer. The fifth kind of cell (type 3) appears not to have been described before. Its cell body is located at the outer margin of the inner nuclear layer.(ABSTRACT TRUNCATED AT 400 WORDS)
Cell surface changes accompanying the neural differentiation of an embryonal carcinoma cell line
Authors: Levine JM , Flynn P
Published: March 2018

The murine embryonal carcinoma cell line P19S18O1A1 develops into neuronlike cells after treatment with retinoic acid (Edwards and McBurney, 1983). We have analyzed the expression of cell surface carbohydrate antigens and intracellular cytoskeletal antigens in differentiating O1A1 cells in order to identify the cell types present in the cultures and to characterize the differentiation process. Undifferentiated O1A1 cells express the SSEA-1 antigen, GD3 ganglioside, and the D1.1 ganglioside antigen, carbohydrate markers that are found on early embryonic cells and neuroepithelial germinal cells in vivo. The cells also bind tetanus toxin, cholera toxin, and monoclonal antibody A2B5, probes that bind to gangliosides found on the surfaces of neurons and immature astrocytes in vivo and in vitro. They contain vimentin-type intermediate filament antigens but have no detectable neurofilament or glial filament protein antigens. After aggregation of the cells in medium containing retinoic acid followed by growth in a serum-free chemically defined medium, over 80% of the cells differentiate into neurons as determined by immunofluorescent labeling with antibodies against neurofilament protein antigens. The differentiated cells no longer express either the embryonic or neuroepithelial carbohydrate antigens, but they continue to express the cell surface markers characteristic of neurons. These changes in the expression of cell surface antigens are accompanied by changes in ganglioside metabolism, including a shift towards the synthesis of more complex gangliosides. Thus, the retinoic acid-induced changes in O1A1 cells in vitro resemble the in vivo development of neurons. This establishes the O1A1 cell line as a relevant model system for studies of the molecular basis of neuronal differentiation and development.
The primary olfactory projection has two chemically distinct zones
Authors: Schwob JE , Gottlieb DI
Published: March 2018

The sensory neurons of the olfactory epithelium form an anatomically uniform population but are differentially excited by odorants. We have discovered an unexpected biochemical heterogeneity within this population that extends to its axonal projection onto the olfactory bulb. This heterogeneity is recognized by a newly generated monoclonal antibody, designated RB-8, that differentially stains the primary olfactory projection in rats and divides it into 2 nonoverlapping zones. With light-microscopic immunohistochemistry, RB-8 densely labels the fascicles of the olfactory nerve from the ventral and lateral parts of the olfactory epithelium, where there is also some epithelial staining. This area, which we designate RB-8-positive, comprises about two-thirds of the epithelial sheet. RB-8 labeling of the other third of the epithelium, which includes the dorsal recess and medial tips of the dorsal turbinals, is not detectable, and the fascicles from these RB-8- negative areas are only weakly stained. These RB-8-negative areas form a contiguous zone on flattened maps of the epithelial sheet. In the olfactory bulb, RB-8 staining of the glomeruli in the ventrolateral part is correspondingly dense, while that in the dorsomedial glomeruli is undetectable or very light. In the labeled glomeruli, the RB-8 staining is precisely coextensive with anti-olfactory marker protein staining, which serves as a marker for the olfactory axons and terminals. In addition, knife-cut lesions of the olfactory nerve totally eliminate the RB-8 staining in the glomeruli where the destruction of the olfactory terminals is complete. There is also a good correlation between the staining patterns in the bulb and epithelium and what is known from tract-tracing studies of the arrangement of the axonal projection of the epithelium onto the bulb. This evidence strongly suggests that, in the olfactory nerve and glomeruli, RB-8 stains the olfactory axons and their terminals. A survey of the CNS and peripheral tissues demonstrates that staining with RB-8 is nervous system-specific; not all components of the CNS and PNS are stained. The antigen recognized by RB-8 was characterized in immunoblots and by use of a direct radioimmunoassay (RIA) which assessed binding of 125I-RB-8. With this assay, the RB-8 binding sites in whole brain are shown to be membrane-associated, saturable, immunologically specific for RB-8, and trypsin-sensitive. In SDS-PAGE immunoblots of membrane proteins, the antigen in rat forebrain and in the olfactory nerve is a protein of 125 kDa Mr, which comigrates in mixtures of membranes from the 2 sources.(ABSTRACT TRUNCATED AT 400 WORDS)
Selectivity for interaural time difference in the owl's midbrain
Authors: Takahashi T , Konishi M
Published: March 2018

The barn owl uses the interaural difference in the timing of sounds to determine the azimuth of the source. When the sound has a wide frequency band, localization is precise. When localizing tones, however, the barn owl errs in a manner that suggests that it is confused by phantom targets. We report a neural equivalent of these phenomena as they are observed in the space-specific neuron of the owl's inferior colliculus. When stimulated with a tone, the space- specific neuron discharges maximally at interaural time differences (ITDs) that differ by one period of the stimulus tone. Changing the stimulus frequency changes the period of the ITD-response functions, but 1 ITD evokes, in most neurons, a maximal response, regardless of frequency. This ITD is the characteristic delay (CD). When stimulated with noise, there is a maximal response only to ITDs at or near the CD. Thus, the space-specific neuron can unambiguously signal the CD, provided that the signal contains more than 1 frequency. The preferential response to a single ITD, which is observed with noise stimuli, was also observed when the summed waveform of the best frequency and another tone, F2, was presented. The response of the space-specific neuron to these 2-tone stimuli could not be accounted for by the summing or averaging of the ITD-response functions obtained with the best frequency or F2 alone, suggesting that nonlinear neural processes are involved.
Migratory patterns of sympathetic ganglioblasts and other neural crest derivatives in chick embryos
Authors: Yip JW
Published: March 2018

While it is well established that neural crest cells migrate to give rise to a variety of cell types and tissues, the migratory patterns among individual neural crest derivatives have not been fully worked out. Such knowledge is important not only in relation to the mechanisms of neural crest migration and differentiation, but also in understanding the ontogeny of the PNS. Of special interest in the latter regard is the origin of sympathetic ganglion cells and their innervation, since studies on ganglion cell innervation have suggested that the matching between pre- and postganglionic neurons might be based on their early positions along the neuraxis (see preceding paper). In the present study, I have used the quail-chick transplantation technique to examine the migratory patterns of the precursors of sympathetic ganglion cells, dorsal root ganglion cells, and melanocytes that originate from 1 or 2 segments of the trunk neural crest. My results indicate that precursors of sympathetic ganglion cells, on average, migrate 2 segments rostrally and 3 segments caudally. Thus, individual sympathetic ganglia comprise neurons that originate from up to 6 segments of the neuraxis. Precursors of melanocytes also migrate along the rostrocaudal axis, but to an even greater extent than sympathetic ganglioblasts. In contrast, precursors of dorsal root ganglion cells do not migrate at all along the rostrocaudal axis (although they migrate along a ventromedial pathway to give rise to the dorsal root ganglia of the same segmental level). The number of neural crest segments contributing to each sympathetic ganglion is approximately equal to the number of spinal cord segments contributing to its innervation.(ABSTRACT TRUNCATED AT 250 WORDS)
Differential sensitivity of tetrodotoxin of nociceptive neurons in 4 species of leeches
Authors: Johansen J , Kleinhaus AL
Published: March 2018

We have recently shown that the nociceptive neurons (N cells) in the leech can be segregated into a medial type (Nmed) and a lateral type (Nlat) according to their physiological and pharmacological properties. We now report that the Na-dependent action potential (AP) of the 2 cell types in Macrobdella have differential sensitivities to tetrodotoxin and that the APs of the N-cell homologs from different leech species also vary considerably in their response to TTX. The normal AP of the N cells in the 4 leech species was exclusively Na-dependent; its overshoot varied logarithmically with [Na]0, was unaffected by the presence of Mn and absence of Ca, and could not be elicited in Na-free Ringer's solutions. The decrease in the maximal rate of depolarization (Vmax) of the Na-dependent AP of the Nmed cell in Macrobdella produced by TTX followed a reverse Langmuir curve for bimolecular reaction with an ED50 of 9 microM. Almost complete blockage was obtained at 50 microM. In contrast, TTX inhibition of Vmax in the Nlat cell reached saturation at a level of only 55% reduction in Vmax even at doses in excess of 200 microM TTX. The simplest interpretation of these data is that there are 2 types of Na channels in the Nlat cell. This is further supported by the fact that the dose-response data could be well fitted by a Langmuir curve, assuming that the Nlat cell possesses 2 populations of Na conductances--one insensitive to TTX and one (like the Nmed cell) with an ED50 to TTX of 9 microM, the 2 populations being present in a 0.45:0.55 ratio, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
Guidance of neuronal growth cones in the grasshopper embryo. I. Recognition of a specific axonal pathway by the pCC neuron
Authors: Bastiani MJ , du Lac S , Goodman CS
Published: March 2018

The selective affinities that growth cones display for specific axonal surfaces give rise to stereotyped patterns of selective fasciculation. Previous studies on cell recognition by neuronal growth cones in the grasshopper embryo led to the proposal and initial experimental testing of the labeled-pathways hypothesis. Here we report on a further experimental analysis of this hypothesis, using the first 3 longitudinal axon fascicles, which initially contain only the axons of 7 identified neurons. We describe and experimentally test the selective affinity of the pCC growth cone for the MP1 and dMP2 axons in the MP1/dMP2 fascicle. The pCC growth cone appears to demonstrate an absolute, rather than hierarchical, preference for the MP1/dMP2 fascicle, as compared with other longitudinal axon fascicles, which supports the notion that the surfaces of the MP1 and dMP2 axons have some special distinguishing label that guides the pCC growth cone onto and along them.
Synaptic organization of claustral and geniculate afferents to the visual cortex of the cat
Authors: LeVay S
Published: March 2018

Claustral and geniculate afferents to area 17 were labeled by anterograde axonal transport of peroxidase-conjugated wheatgerm agglutinin, and examined in the electron microscope. A peroxidase reaction protocol that led to labeling in the form of minute holes in the EM sections was used. Both types of afferents formed type 1 (presumed excitatory) synapses exclusively. In agreement with previous reports the great majority of geniculate afferents to layers 4 and 6 contacted dendritic spines. The claustral afferents to layers 1 and 6 also predominantly contacted spines. In layer 4, however, claustral afferents contacted spines and dendritic shafts about equally. The results suggest a substantial claustral input to smooth-dendrite cells in layer 4, which are thought to be inhibitory in function. This may be the circuit by which the claustrum helps to generate end-stopped cortical receptive fields (Sherk and LeVay, 1983).
Distribution of stage-specific neurite-associated proteins in the developing murine nervous system recognized by a monoclonal antibody
Authors: Yamamoto M , Boyer AM , Crandall JE , Edwards M , Tanaka H
Published: March 2018

A monoclonal antibody, 4D7, obtained with embryonic rat brain as an immunogen, recognizes an epitope on 3 protein species of 150–160, 100– 110, and 80 kDa, present in mouse and rat brain during the fetal period. Vital immunostaining of dissociated cultures of fetal forebrain indicates that the antigen is localized largely on the external plasma membrane of a subpopulation of neurites. Immunocytochemistry reveals that the distribution of the antigen in vivo is restricted to the nervous system. Immunoreactivity is concentrated primarily in the pathways of a limited set of CNS and PNS axon systems during early stages of their development, as delineated by staining with the neurofilament antibody, C2. Depending on the particular axon system, immunoreactivity with 4D7 persists only for one to several days of prenatal or perinatal development. In the spinal cord, stage-specific- neurite-associated proteins (SNAP) expression occurs first along motor axon pathways on embryonic day (E) 10 and then within the nerve trunks of dorsal root ganglia and the commissural fiber system on E11. Immunoreactivity is detectable among most cranial nerves starting in the interval from E11 to E13. Within the brain, the onset of SNAP expression within several discrete axon tracts occurs in the interval E14–16, including the lateral olfactory tract, anterior commissure, corpus callosum, fasciculus retroflexus, and fornix. Immunoreactivity within the embryonic intermediate zones of some structures matches the location of certain other axon systems. Sites of 4D7 staining which do not correspond to the location of axon populations include the internal portion of the external granular layer of the postnatal cerebellum and the cortex of the reeler mutant mouse. The predominant localization of the 4D7 antigen among axon systems and its precisely regulated spatio- temporal pattern of expression are consistent with the possibility that the SNAP antigens play a significant role in the early stages of growth of axonal tracts in vivo.
Corticofugal influences of S1 cortex on ventrobasal thalamic neurons in the awake rat
Authors: Yuan B , Morrow TJ , Casey KL
Published: March 2018

Corticofugal influences on the responses of 39 ventrobasal (VB) thalamic neurons to repetitive stimuli were studied in awake rats by focally suppressing the evoked and spontaneous electrocortical activity of the primary (S1) somatosensory cortex with magnesium or lidocaine plus magnesium. Suppression of the S1 cortex reduced the number of spikes discharged by 19 (66%) of 29 VB units in response to each of 25 electrical stimuli delivered to the medial lemniscus; 9 units were unaffected and 1 showed an increased response to 1–10 Hz stimuli. The responses of 6 (38%) of 16 VB units to electrical somatic stimuli were also reduced following S1 cortical suppression; 9 units were unaffected and 1 showed an increased response to 20–40 Hz stimuli. A comparison of the reduced responsiveness of 5 units studied during medial lemniscal and somatic stimulation did not reveal any additional response attenuation attributable to subthalamic corticofugal influences. We conclude that, in the awake rat, somatosensory transmission to VB thalamic neurons is primarily facilitated by S1 corticothalamic neurons.
Suppression of the membrane defect by divalent cations in the Drosophila mutant shibire
Authors: Costello WJ , Salkoff LB
Published: March 2018

The single-gene mutant shibire (shits) is temperature-sensitive. It causes reversible paralysis at heat pulses greater than 29 degrees C by blocking synaptic transmission. The synapses of these heat-pulsed flies are depleted in vesicles but contain numerous cisternae. We report that such alterations in synaptic physiology and morphology of heat-pulsed flies can be suppressed by internal perfusion of salines with high concentrations (10–18 mM) of the divalent cations Ca2+ or Mg2+. Synaptic morphology in these perfused flies remains normal even when exposed to nonpermissive temperatures (greater than 29 degrees C); in addition, synaptic transmission maintains a high resistance to failure both in frequency and stimulus duration. We also observed many cisternae in close association with extrajunctional as well as with postsynaptic regions of the sarcolemma in heat-pulsed shits flies not perfused with the increased concentrations of divalent cations. Flies perfused with increased amounts of divalent cations lacked such cisternae in the sarcolemma. The evidence suggests that the divalent cations can mitigate an overall membrane defect expressed by the shits gene, perhaps by influencing lipid-phase transition behavior.
Coexistence of galanin-like immunoreactivity with catecholamines, 5- hydroxytryptamine, GABA and neuropeptides in the rat CNS
Authors: Melander T , Hokfelt T , Rokaeus A , Cuello AC , Oertel WH , Verhofstad A , Goldstein M
Published: March 2018

The coexistence of galanin (GAL)-like immunoreactivity (LI) with markers for catecholamines, 5-hydroxytryptamine (5-HT), GABA, or some neuropeptides was mapped in the rat CNS by using adjacent sections, as well as by elution-restaining and double-labeling immunocytochemistry. Many instances of coexistence were observed, but there were also numerous GAL-positive cell body populations displaying distributions similar to those of these markers but without apparent coexistence. In the hypothalamic arcuate nucleus GAL-LI was found in a large proportion of tyrosine hydroxylase (TH)-positive cell bodies (A12 cells), both in the dorsomedial and ventrolateral subdivisions, with a higher number in the latter. GAL-LI coexisted in glutamic acid decarboxylase (GAD)- positive somata in the posterior aspects of the arcuate nucleus and at all rostrocaudal levels in fibers in the external layer of the median eminence. In the anterior hypothalamus, a large population of the cells of the parvocellular and magnocellular paraventricular nuclei contained both GAL-LI and vasopressin-LI. Moreover, somata containing both GAD- and GAL-LI were seen lateral to the mammillary recess in the tuberal and caudal magnocellular nuclei. Some of the neurons of the caudal group were shown to project to the occipital cortex using combined retrograde tracing and immunofluorescence. With regard to mesencephalic and medullary catecholamine neurons, GAL-LI coexisted in a large proportion of the noradrenergic locus coeruleus somata (A6 cell group) and in the A4 group dorsolateral to the fourth ventricle, as well as in the caudal parts of the A2 group in the dorsal vagal complex. However, in more rostral parts of the latter, especially in the medial subdivision of the solitary tract nucleus, a very large population of GAL-IR small cell bodies was seen intermingling with catecholamine neurons, but they did not contain TH-LI. Furthermore, GAL-IR cell bodies coextensive with, but not coexisting in, TH-IR somata were seen in the C1 (epinephrine) horea in the ventrolateral medulla at the level of area postrema and in the most rostral aspects of the C1 group. Finally, 5-HT-positive cell bodies of the mesencephalic and medullary raphe nuclei and a subpopulation of coarse 5-HT nerve fibers in the hippocampus co-contained GAL-LI. The present results demonstrate that a GAL-like peptide is present in many systems containing other neuroactive compounds, including dopamine, norepinephrine, 5-HT, GABA, and vasopressin.(ABSTRACT TRUNCATED AT 400 WORDS)
m-Azido-phencyclidine covalently labels the rat brain PCP receptor, a putative K channel
Authors: Sorensen RG , Blaustein MP
Published: March 2018

Phencyclidine (PCP) is a schizophrenomimetic drug of abuse. PCP binds with high affinity (apparent dissociation constant, KD less than 10(-6) M) to rat brain membranes and blocks, selectively, a voltage-gated, noninactivating K channel found in rat brain synaptosomes (presynaptic nerve terminals). Thus, it has been proposed that the high-affinity PCP receptor in brain is this K channel. Consistent with this hypothesis, we now show that several K channel blockers displace 3H-PCP from the rat brain receptor. Additionally, we have used a photolabile analog of PCP, m-azido-PCP (Az-PCP), to identify the brain PCP receptor/putative K channel. In the dark, Az-PCP bound reversibly to 2 classes of sites on rat brain synaptic membranes [KD = 0.14 +/- 0.01 microM (n = 5) for high-affinity binding, and KD = 255 +/- 55 microM for low-affinity binding]. Competitive binding studies between Az-3H-PCP and nonlabeled PCP analogs, and between Az-PCP and several tritiated PCP analogs, indicated that the high-affinity Az-PCP binding site is the high- affinity PCP receptor. Several amino-pyridines (APs) and tetraalkylamines (TAAs), which are known to block K channels in excitable cells, were also found to displace 3H-PCP from its high- affinity binding site on rat brain synaptic membranes. The rank order of potency for displacement of 3H-PCP from this site for the APs was 4- AP approximately equal to 3,4-diAP greater than 2-AP much greater than 3-AP; for the TAAs it was TBA greater than TEA much greater than TMA (the tetra-butyl, ethyl, and methyl amines, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)
Perturbed pattern of catecholamine-containing neurons in mutant Drosophila deficient in the enzyme dopa decarboxylase
Authors: Budnik V , Martin-Morris L , White K
Published: March 2018

We have initiated a study of catecholamine-containing neurons in Drosophila melanogaster because of the potential, with this organism, to perturb catecholamine metabolism using genetic tools. The major objectives of this study were (1) to define the pattern of catecholamine-containing neurons and (2) to determine the effect of the absence of dopa decarboxylase (DDC) enzyme activity on the catecholamine-containing neurons. We chose to analyze the catecholamine- containing neurons in the ventral ganglion of the larval CNS. To define the catecholamine-containing neurons, CNSs were dissected and reacted with glyoxylic acid. The catecholamine histofluorescence (CF) neuronal pattern (normal-CF neurons) in the wild-type ventral ganglion is stereotypic. In the mutant ventral ganglia, in the absence of DDC enzyme activity, most normal-CF neurons still exhibit CF, probably indicating the presence of accumulated L-dopa. Interestingly, in the mutant CNSs, additional novel neuronal subsets also exhibit CF. Analysis of CNSs from early developmental stages revealed that the novel-CF neurons become fluorogenic earlier than the normal-CF neurons in the mutant CNS. To determine whether neuronal subsets, in addition to the normal-CF, neurons are able to sequester catecholamines, CNSs from wild-type larvae were incubated in exogenous catecholamine (L-dopa or dopamine). Incubations in L-dopa or dopamine revealed normally nonfluorogenic neurons that are able to take up the amine and become fluorogenic. Among the neurons able to sequester L-dopa or dopamine are subsets that are similar to the novel-CF neurons in the mutant CNS. This similarity is best characterized by a major novel-CF neuronal cluster in the subesophageal-thoracic region. These results suggest that in the absence of DDC activity, subsets of normally nonfluorogenic neurons capable of sequestering L-dopa or dopamine accumulate the fluorogenic catecholamine. Hypotheses that might explain the mode of accumulation of the catecholamine within the novel-CF neurons are considered.
Selective growth of rat Schwann cells in neuron- and serum-free primary culture
Authors: Needham LK , Tennekoon GI , McKhann GM
Published: March 2018

A serum-free medium, designated S4 [consisting of Eagle's Minimum Essential Medium supplemented with the following components (mg/liter): endothelial mitogen, 100; vitamin C, 1; insulin, 5; cholera toxin, 0.01; putrescine, 8; sodium selenite, 0.0043; MgCl2 X 6H2O, 40; and albumin, fatty acids, amino acids, vitamins, and trace elements], has been established to select for the growth of Schwann cells from primary cultures of dissociated neonatal rat sciatic nerve in the absence of neurons and without exposure of the cells to serum or antimitotic agents. By immunological criteria, confluent primary cultures, grown on a substrate of extracellular matrix derived from cultured bovine aorta endothelial cells, consist of approximately 90% Schwann cells after maintenance in this serum-free medium. Secondary cultures of at least 99% purity may be established after treatment of primary cultures maintained in S4 with antiserum against a fibroblast surface antigen, Thy 1.1, in conjunction with complement. Such secondary Schwann cells continue to proliferate in S4, with no decrease in purity, through at least 4 passages.
In vitro neuronal differentiation of Drosophila embryo cells
Authors: Salvaterra PM , Bournias-Vardiabasis N , Nair T , Hou G , Lieu C
Published: March 2018

Early gastrula-stage Drosophila embryo cells will differentiate in vitro to form several cell types, including neurons. We report here the morphological appearance of cultured embryo cells, the pattern of DNA synthesis, and the expression of neurotransmitter-metabolizing macromolecules. The cells initially exhibit no overt morphological differentiation, and all cells incorporate 3H-thymidine following a 1 hr pulse-labeling period. As cells undergo morphological differentiation, fewer total cells as well as qualitatively different cell types incorporate label. By the time cells are 8 or 9 hr old, no myocytes or myotubes are labeled. In contrast, some neurons are labeled with a thymidine pulse as late as 18 hr. We have also stained cultured cells of various developmental ages with the insect neuron-specific antibody: anti-HRP. Some positive cells can be detected as early as 5 hr, when no overt morphological differentiation is apparent. As the cells differentiate, the staining is limited to the small, round neuronal type and its processes. These findings suggest that this neuron-specific cell marker is expressed very early in cultured gastrula-stage cells and may be used to identify neuronal precursor cells. We have studied the patterns of expression of several macromolecules involved in acetylcholine metabolism using these cultures. The appearance of choline acetyltransferase (ChAT), the biosynthetic enzyme for ACh production, is first detected in 5-hr-old cells. There is an initial phase of low-level expression, followed by a rapid rise in activity shortly after the differentiating neuron clusters make contact with one another. ChAT activity reaches a plateau in 36–48-hr-old cells. Acetylcholinesterase activity can be detected several hours before ChAT and also shows a period of low-level expression followed by a rapidly increasing phase, reaching a plateau at around 36–48 hr. 125I-alpha-bungarotoxin binding appears in cells about 4 hr old and rapidly approaches maximum levels by about 36 hr. The in vitro expression pattern for ChAT and AChE is similar to that seen in vivo. AChE activity has been localized histochemically to the neurons and their processes in vitro. The normal in vitro expression pattern for ChAT and AChE can be altered by adding various cholinergic drugs to the culture medium during cell differentiation. Medium conditioned by older cultures can also result in lower levels of ChAT and AChE expression.(ABSTRACT TRUNCATED AT 250 WORDS)
An ultrastructural study of GABA-immunoreactive neurons and terminals in the septum of the rat
Authors: Onteniente B , Geffard M , Campistron G , Calas A
Published: March 2018

The fine structure and types of contact made by GABAergic elements in the septal nuclei were studied at the electronmicroscopic level by means of peroxidase immunocytochemistry, using anti-GABA antibodies. Observations were made on normal and colchicine-injected rats. GABA- immunoreactivity was distributed within somata, dendrites, axonal varicosities and terminals, and myelinated axons. The peroxidase reaction product was diffuse in the cytoplasm; cytoplasmic organelles were generally devoid of immunoreactivity, while showing a strong reaction on the outer surface of their membrane. GABA-immunoreactive (GABA-I) neurons were small (10 microns on average) to medium (20 microns) in size, with round or multipolar cell bodies. Additionally, labeled large (30 microns) cells were observed within the myelinated fibers of the medial septal nucleus after intraseptal administration of colchicine. No difference in the ultrastructural features and distribution of the immunoreactivity of the 2 kinds of cell was noticed, except for a higher number of synaptic contacts on large neurons of the medial septum. GABA-I cells of the medial and lateral nuclei received synapses on their soma and dendrites, made by both immunonegative and GABA-I terminals. Nonimmunoreactive boutons contacting GABA-I cell bodies were of 2 types: those containing small, clear synaptic vesicles and those that additionally contained large dense vesicles. Synaptic vesicles of GABA-I boutons were rarely labeled internally, but showed varying electron densities. Synapses made by GABA-I boutons on GABA-I or unlabeled somata and dendrites were always of symmetrical type. Synapses made by non-GABA-I boutons on GABA-I cells were either symmetrical or asymmetrical.
Functional properties of parietal visual neurons: radial organization of directionalities within the visual field
Authors: Steinmetz MA , Motter BC , Duffy CJ , Mountcastle VB
Published: March 2018

Parietal visual neurons (PVNs) were studied in waking monkeys as they executed a simple fixation-detection task. Test visual stimuli of varied direction, speed, and extent were presented during the fixation period; these stimuli did not control behavior. Most PVNs subtend large, bilateral receptive fields and are exquisitely sensitive to stimulus motion and direction but insensitive to stimulus speed. The directional preferences of PVNs along meridians are opponently organized, with the preferred directions pointing either inward toward or outward away from the fixation point. Evidence presented in the preceding paper (Motter et al., 1987) indicates that opponent directionality along a single meridian is produced by a feed-forward inhibition of 20 degrees-30 degrees spatial extent. The observations fit a double-Gaussian model of superimposed but unequal excitatory and inhibitory receptive fields: When the former is larger, inward directionality results; when smaller, outward directionality results. We examine here the distribution of the meridional directional preferences in the visual field. Tests showed that opponent organization is not produced by differences in local directional properties in different parts of the receptive field. The distribution of response intensities from one meridian to another is adequately described by a sine wave function. These data indicate a best radial direction for each neuron with a broad distribution of response intensities over successive meridians. Thus, any single PVN, with rare exceptions, cannot signal radial stimulus direction precisely. We then determined how accurately the population response predicted radial stimulus direction by the application of a linear vector summation model. The resulting population vector varied from stimulus direction by an average of 9 degrees. Whether or not the perception of the direction of motion depends upon a population vector remains uncertain. PVNs are especially sensitive to object movement in the visual surround, particularly in the periphery of the visual field. This, combined with their large receptive fields and their wide but flat sensitivity to stimulus speed, makes them especially sensitive to optic flow. This is discussed in relation to the role of the parietal visual system in the visual guidance of projected movements of the arm and hand, in the guidance of locomotion, and in evoking the illusion of vection.