bluuebunny/crossref_metadata_2025 huggingface dataset:
Horizontal cells in the retina of the rabbit
Authors: Dacheux RF , Raviola E
Published: March 2018

The light responses, morphology, and connections of horizontal cells (HCs) were studied in the retina of the rabbit using intracellular recordings and the injection of visible markers. Two types of HCs were identified, axonless and axon-bearing HCs. Axonless HCs and the somatic end of axon-bearing HCs respond to white light of varying intensity with graded hyperpolarizations; both display a transient superimposed on the sustained hyperpolarization at stimulus initiation and a small rod aftereffect at the cessation of high intensity stimuli. Anatomically, both are connected to cones, but their responses also suggest rod influence. Both summate stimuli from a retinal area which is much larger than their respective fields. However, only axonless HCs transfer a fluorescent, low molecular weight dye to adjoining, homologous cells. The axon terminal of axon-bearing HCs has response properties different from those of the cell body: the transient at stimulus initiation is absent; furthermore, at high levels of illumination, the rod aftereffect becomes equal in amplitude to the primary hyperpolarization. Anatomically, it is connected to rods, but its responses also suggest cone influence. Its receptive field approximates in diameter its anatomical spread and it does not transfer fluorescent dye to its neighbors.
The ultrastructural localization of serotonin immunoreactivity within the nucleus of the solitary tract of the cat
Authors: Maley B , Elde R
Published: March 2018

Using a modification of the peroxidase-antiperoxidase technique, serotonin immunoreactivity was localized at the ultrastructural level in the nucleus of the solitary tract of the cat. Structures containing serotonin immunoreactivity included unmyelinated axons, varicosities (0.5 to 2 micrometers in diameter), and synaptic terminals. The serotonin-containing synaptic terminals were found less frequently than axons or varicosities. Within unmyelinated axons and varicosities, the immunoreactivity was associated mainly with large granular vesicles (80 to 150 nm). While large granular vesicles were found in all immunoreactive structures, greater numbers were observed in axons and nonsynaptic varicosities. Serial sections of several nonsynaptic serotonin-immunoreactive varicosities indicated the lack of synaptic specializations associated with these structures. In a typical section, only one or two granular vesicles were in synaptic terminals which contained numerous small clear vesicles. Serotonin-immunoreactive terminals formed asymmetrical contacts with dendrites and spines. No synaptic contacts involving immunoreactive terminals were found on cell bodies or other axonal structures. Serotonin-containing neuronal perikarya within the nucleus of the solitary tract were never observed. The abundance of nonsynaptic varicosities containing large granular vesicles suggests a possible neurohumoral role for serotonin within the feline nucleus of the solitary tract. This is discussed in relation to previous reports concerning the paucity of genuine synaptic contacts involving serotonin in other regions of the central nervous system. The presence of serotonin-immunoreactive terminals in the nucleus of the solitary tract also suggests its function as a putative neurotransmitter.
Metabolic changes in deafferented central neurons of an insect, Acheta domesticus. I. Effects upon amino acid uptake and incorporation
Authors: Meyer MR , Edwards JS
Published: March 2018

Chronic cercal deafferentation of the terminal ganglion in developing crickets (Acheta domesticus), which is known to suppress normal development of giant interneuron dendritic arborizations is shown here to reduce [3H]leucine uptake and incorporation into ganglion proteins. Short term deafferentation of adult crickets, in contrast, does not depress amino acid uptake and incorporation significantly. Following unilateral long term deafferentation of the terminal ganglion, a comparison was made of the [3H]leucine incorporation into primary dendritic processes and somata of deafferented and normally innervated medial giant interneurons (MGIs) within the same ganglion by means of quantitative autoradiography. Grain densities within dendrites of deafferented MGIs were significantly lower than in paired control MGIs' grain densities within somata of deafferented MGIs also were reduced, although the effects of deafferentation were less pronounced in somata than in target dendrites. These results imply a specific influence of afferent innervation on protein metabolism during growth and development of target postsynaptic elements.
Functional synapse elimination in the developing avian cochlear nucleus with simultaneous reduction in cochlear nerve axon branching
Authors: Jackson H , Parks TN
Published: March 2018

We studied the chick auditory system to determine whether there is a normal developmental reduction in the number of cochlear nerve axons innervating individual cochlear nucleus (nucleus magnocellularis, NM) neurons. We also examined the preterminal branching patterns of cochlear nerve axons during development. The number of cochlear nerve axons innervating individual NM neurons was estimated by counting the increments in the postsynaptic response as the intensity of cochlear nerve electrical stimulation was varied gradually; this number fell from a mean of 4.0 on embryonic day 13 (E13) to a mean of 2.2 on E17 and E18 and the 4th day after hatching. This highly reliable decline in functional convergence was accompanied by a decrease in the number of preterminal branches of cochlear nerve fibers innervating the NM. On E13 and E14, most axons stained by iontophoretic injections of horseradish peroxidase showed two distinct preterminal branches in the NM. By E17 and E18 and thereafter, cochlear nerve axons were unbranched and terminated with a characteristic single large calycine ending in the NM. There are about twice as many cochlear nerve axons as neurons in the NM and the number of fibers in the nerve appears to decline only slightly between E13 and E17. The 50% decline in the number of cochlear nerve axons making functional synapses on individual NM neurons therefore is associated principally with the concurrent elimination of cochlear nerve axon branching in the NM.
Structural changes during transmitter release at synapses in the frog sympathetic ganglion
Authors: Dickinson-Nelson A , Reese TS
Published: March 2018

Interactions between synaptic vesicles and the presynaptic membrane which accompany transmitter release were examined at excitatory, cholinergic synapses in bullfrog sympathetic ganglia. Ganglia were fixed at rest or during electrical stimulation of the preganglionic axons and then were either thin-sectioned or freeze-fractured. Release of transmitter for brief periods is accompanied by selective depletion of four-fifths of the synaptic vesicles aligned at active zones, an overall loss of half of the synaptic vesicles in the terminals, and synaptic vesicle openings at active zones. These findings are consistent with the hypotheses that synaptic vesicles which are ready to be released are aligned at active zones and that these vesicles fuse with and add their membrane to the presynaptic membrane as they release transmitter. Larger vesicles with dense cores also contact and open onto the presynaptic membrane at the active zone, appearing to release their contents by exocytosis. The arrangement of intramembrane particles at fractured postsynaptic specializations resembles that at other excitatory, cholinergic synapses.
Modification of retrograde degeneration in transected spinal axons of the lamprey by applied DC current
Authors: Roederer E , Goldberg NH , Cohen MJ
Published: March 2018

In the spinal cord of the lamprey, regeneration of giant reticulospinal axons occurs following transection. We show that partial degeneration of the proximal axonal segment, or “die-back,” also occurs following spinal transection and it precedes regenerative outgrowth. The die-back during the first 5 days post-transection is reduced significantly by application of a 10-microA DC current across the site of transection, with the cathode distal to the lesion. Reversing the polarity of the applied current (cathode proximal to the lesion) increases the extent of axonal die-back relative to the sham-treated controls. Following spinal transection, saline-filled wick electrodes were implanted in the body musculature on either side of the lesion. Electrically treated animals received current across the lesion for 5 days, while the sham- treated controls received no current. After 5 days, several giant axons in each preparation were injected intracellularly in the spinal cord with the dye Lucifer Yellow. The extent of axonal die-back in the proximal cord stump was determined in the filled fibers by measuring the distance of the axon end from the site of lesion. The mean distances of axonal die-back were as follows: controls, 1750 microns +/- 45 SEM; cathode-distal, 740 microns +/- 33 SEM; cathode-proximal, 2820 microns +/- 60 SEM. These differences between the treatment groups proved to be significant using the Wilcoxon rank sum test. We propose that die-back is caused by the entry of cations driven into the cut surface of the cord by the endogenous injury current. The applied DC current interacts with the endogenous current of injury to either decrease or increase the flow of cations into the cord, depending on the direction of applied current flow across the lesion. This in turn causes a corresponding reduction or enhancement of the axonal die-back.
Somatostatin regulates dopamine release in rat striatal slices and cat caudate nuclei
Authors: Chesselet MF , Reisine TD
Published: March 2018

The effects of somatostatin on the release of tritiated dopamine (DA) formed continuously from tritiated tyrosine were studied in vitro in superfused striatal slices and in vivo in both caudate nuclei and both substantiae nigrae of halothane-anesthetized cats using a push-pull cannula technique. Somatostatin (3 X 10(-10) to 3 X 10(-7) M) increased the spontaneous tritiated dopamine release from rat striatal slices. This effect was dose dependent and was completely prevented by tetrodotoxin (5 X 10(-7) M). When applied for 30 min in one cat caudate nucleus, somatostatin (10(-7) M) immediately increased the local release of tritiated DA, while a gradual inhibition of the tritiated amine's efflux was observed in the contralateral caudate nucleus. No changes in tritiated dopamine were seen in either substantia nigra during or after the peptide's application in the caudate nucleus. These results suggest that somatostatin in the striatum may play a role in the local and the distal control of dopamine release from the terminals of dopaminergic nigrostriatal neurons.
Nerve growth factor-inducible large external (NILE) glycoprotein: studies of a central and peripheral neuronal marker
Authors: Salton SR , Richter-Landsberg C , Greene LA , Shelanski ML
Published: March 2018

The PC12 clone of pheochromocytoma cells undergoes neuronal differentiation in the presence of nerve growth factor (NGF). Concomitant with this is a significant induction in the incorporation of radiolabeled fucose or glucosamine into a 230,000-dalton cell surface glycoprotein named the NGF-Inducible Large External, or NILE, glycoprotein (GP) (McGuire, J. C., L. A. Greene, and A. V. Furano (1978) Cell 15: 357–365). In the current studies NILE GP was purified from PC12 cells using wheat germ agglutinin-agarose affinity chromatography and SDS-polyacrylamide gel electrophoresis (PAGE). Polyclonal antisera were raised against purified NILE GP and were found to selectively immunoprecipitate a single 230,000-dalton protein from detergent extracts of PC12 cells metabolically labeled with either [3H]fucose, [3H]glucosamine, or [35S]methionine. These antisera stained the surfaces of PC12 cells by indirect immunofluorescence and were cytotoxic to PC12 cells in the presence of complement. Limited treatment of PC12 cells with either trypsin or pronase produced a fucosylated 90,000-dalton immunoreactive fragment of NILE GP which remained in the membrane. Using quantitative immunoelectrophoresis, the action of NGF on NILE GP was represent an increase in the amount of protein, rather than a selective increase in carbohydrate incorporation. Immunofluorescent staining of primary cell cultures and tissue whole mounts revealed that immunologically cross-reactive NILE GP appears to be expressed on the cell surfaces (somas and neurites) of most if not all peripheral and central neurons examined. Immunoprecipitation of radiolabeled cultures showed that the cross- reactive material had an apparent molecular weight by SDS-PAGE of 225,000 to 230,000 in the peripheral nervous system and 200,000 to 210,000 in the central nervous system. NILE-cross-reactive material was also found to a small extent on Schwann cell surfaces, but not at all on a variety of other cell types. These results suggest that immunoreactive NILE GP is distributed widely and selectively on neural cell surfaces.
Principles of motor organization of the monkey cervical spinal cord
Authors: Jenny AB , Inukai J
Published: March 2018

The organization of spinal cord motor columns innervating 18 selected macaque forelimb muscles was studied with the technique of retrograde transport of horseradish peroxidase. The reliability of the method was evaluated in the cat hindlimb. Motor columns innervating forearm muscles with similar actions on the hand appear to overlap in the anterior horn. Extensor motoneurons are generally positioned ventral and/or lateral to flexor motoneurons. Motoneurons controlling hand movement are located primarily in segments C8 and T1.
The laminar organization of the lateral geniculate body and the striate cortex in the squirrel monkey (Saimiri sciureus)
Authors: Fitzpatrick D , Itoh K , Diamond IT
Published: March 2018

The organization of the projection from the lateral geniculate body to the striate cortex in the squirrel monkey has been re-examined using the anterograde and retrograde transport of horseradish peroxidase (HRP) and wheat germ agglutinin conjugated to HRP. The results confirm earlier findings that the projections of the magnocellular and parvocellular layers of the lateral geniculate body terminate in separate sublaminae of layer IV of striate cortex; a more superficial projection of the parvocellular layers to a narrow strip at the base of layer III (IVA in Brodmann's terminology) has also been confirmed. In addition to these well characterized pathways, our results show that the projections of the lateral geniculate body terminate in more superficial levels of layer III and sparsely in layer I of striate cortex. The projections to the upper portion of layer III terminate in distinct patches which coincide precisely with patches of cytochrome oxidase activity previously identified in this zone. The projections to the patches originate primarily from small, pale-staining cells of the “intercalated layers” which surround the magnocellular layers of the lateral geniculate body. A comparison of the organization of the geniculo-cortical projections in the squirrel monkey with that of the cat, Galago, and Tupaia suggests that, despite marked species differences in the laminar organization of the lateral geniculate body and striate cortex, there are striking similarities in the pathway which terminates in the most superficial layers of striate cortex.
The electromotor system of the stargazer: a model for integrative actions at electrotonic synapses
Authors: Bennett MV , Pappas GD
Published: March 2018

The electric organs of Astroscopus are modified from extraocular muscles and innervated by the enlarged oculomotor nuclei. The electromotor neuron somata are contacted by fine processes with which they form gap junctions. Presynaptic vesicles and active zones are also present, although physiological data give no indication of chemically mediated transmission. Antidromic stimulation produces long lasting graded depolarizations in the electromotor neurons. The latency is sufficiently short to indicate that the cells are electrotonically coupled, which was confirmed by direct measurement. Antidromic invasion may normally fail and is easily blocked by hyperpolarization revealing initial segment and axon spikes. Spinal stimulation evokes postsynaptic potentials (PSPs) and orthodromic impulses; the PSPs are not smoothly graded in amplitude. A medullary nucleus innervates the electromotor nucleus; the medullary cells also show short latency graded antidromic depolarizations and presumably are electrotonically coupled. Their coupling accounts for the variability in PSPs evoked by spinal stimulation. Apparent time constants differ greatly for direct stimulation of a single cell, decay of afterhyperpolarization, electrotonic spread from one cell to a neighbor, and decay of PSPs and graded antidromic depolarizations. The differences can be accounted for in terms of a highly interconnected network of electrotonically coupled cells, which was simulated computationally. Because of the long membrane time constant graded antidromic depolarizations summate. Because antidromic invasion is facilitated by depolarization, the antidromic depolarizations can show pronounced facilitation. The observed “plasticity” within this electrotonically coupled system provides a model for integrative actions at other sites of coupling.
Coupling between neurons of the developing rat neocortex
Authors: Connors BW , Benardo LS , Prince DA
Published: March 2018

We have estimated the prevalence of coupling between neurons of the rat neocortex during postnatal development. Single intracellular injections of the fluorescent dye Lucifer Yellow CH resulted in dye coupling among 70% of neurons from 1 to 4 days of age. Dye coupling dropped to 30 to 40% by 10 to 18 days and occurred in 20% of injected adult neurons. The number of neurons per dye-coupled aggregate also decreased. Whereas three to seven coupled neurons were common in cortex of 1 to 4 days, aggregates of more than two neurons were exceptionally rare in adults. The frequency of dye coupling did not vary systematically with cortical depth at any age. When chemical synaptic activity was blocked, most 4- day neurons exhibited short latency antidromically evoked depolarizations which were relatively insensitive to repetitive activation and membrane polarization. These depolarizations may represent electrotonically conducted spikes from coupled neurons. No such potentials were found in adult neurons. The results suggest that neuronal coupling is extensive in immature rat neocortex, but that coupling declines at a time just before the numbers of chemical synapses increase most rapidly.
A calcium/calmodulin-dependent protein kinase from mammalian brain that phosphorylates Synapsin I: partial purification and characterization
Authors: Kennedy MB , McGuinness T , Greengard P
Published: March 2018

A calcium/calmodulin-dependent protein kinase, which phosphorylates a synaptic vesicle-associated protein designated Synapsin I, has been shown to be present in both soluble and particulate fractions of rat brain homogenates. In the present study, the particulate activity was solubilized by washing with a low ionic strength solution, and the enzymes from the two fractions were partially purified by ion exchange chromatography and calmodulin-Sepharose affinity chromatography. By each of several criteria, the partially purified enzymes from the two sources were indistinguishable. These criteria included specificity for various substrate proteins, concentration dependence of activation by calcium and calmodulin, pH dependence, and apparent affinities for the substrates Synapsin I and ATP. The mild conditions that released the particulate enzyme indicated that it was not tightly bound to the membrane and suggested that it may exist in a dynamic equilibrium between soluble and particulate-bound states. The partially purified enzyme preparations from both the soluble and particulate fractions contained three proteins that were phosphorylated in the presence of calcium and calmodulin, a 50-kilodalton (Kd) protein and two proteins in the 60-Kd region. When compared by phosphopeptide mapping and two- dimensional gel electrophoresis, the proteins were indistinguishable from three proteins of corresponding molecular weights that were shown by Schulman and Greengard (Schulman, H., and P. Greengard (1978) Nature 271: 478–479) to be prominent substrates for calcium/calmodulin- dependent protein kinase in a crude particulate preparation from rat brain. The 50-Kd substrate was the major Coomassie blue staining protein in both partially purified enzyme preparations. The peak of this protein coincided with that of enzyme activity during DEAE- cellulose and calmodulin-Sepharose chromatography. These results suggest that the 50-Kd phosphoprotein may be an autophosphorylatable subunit of the Synapsin I Kinase or may exist in a complex with it.
Axonal regeneration in lamprey spinal cord
Authors: Yin HS , Selzer ME
Published: March 2018

Spinal cords of sea lamprey larvae were transected at one of two levels: (a) rostral, at the last gill, or (b) caudal, at the cloaca. Following various recovery times, regeneration of the posteriorly projecting giant reticulospinal axons (RAs) was demonstrated by intra- axonal injection of horseradish peroxidase (HRP). Regeneration of axons of anteriorly projecting dorsal cells (DCs) and giant interneurons (GIs) was demonstrated by intrasomatic HRP injection into cells located just below the transection scar. After 40 days of recovery, 55% of proximally transected RAs (rostral cut) regenerated at least as far as the center of the scar, whereas only 15% of distally transected RAs (caudal cut) did so. Maximum distance of regeneration was 5.3 mm beyond the scar for proximally transected RAs but only 38 u for distally transected RAs. Proximally transected RAs also branched more profusely than distally transected ones. These data (when combined with others in the literature) suggest that the regenerative capacity of RAs may decrease with distance of axotomy from the cell body. Distance of regeneration and degree of branching of proximally transected RAs peaked between 40 and 100 days. Thereafter, there appeared to be a tendency toward neurite retraction. Of axotomized GIs, 76% regenerated anteriorly at least as far as the center of a caudal transection scar (GIs are located only in the caudal part of the cord). The maximum distance of regeneration was 1.3 mm beyond the scar. Of DC axons, 56% regenerated anteriorly at least as far as the transection site. The maximum distance was 1.1 mm beyond the scar. DCs located just below a caudal transection regenerated at least as well as those located below a rostral transection. Axonal regeneration was also demonstrated for a few lateral cells, edge cells, and crossed caudally projecting interneurons.
Presynaptic calcium diffusion and the time courses of transmitter release and synaptic facilitation at the squid giant synapse
Authors: Zucker RS , Stockbridge N
Published: March 2018

At the squid giant synapse, a presynaptic action potential is accompanied by an influx of calcium ions that continue to be detectable with arsenazo III microspectrophotometry for several seconds. Nevertheless, transmitter release occurs phasically, lasting only about 2 msec. If a second action potential follows within about 100 msec after the first, it releases more transmitter. In this paper, we present a mathematical model of intracellular calcium diffusion with binding to fixed cytoplasmic sites, active extrusion at the surface, and influx during an action potential, to predict the distribution of intracellular calcium following an action potential. With a square law relation between submembrane calcium and transmitter release, the model predicts the phasic release of transmitter and the magnitude and time course of synaptic facilitation following an action potential, as well as the relatively long persistence of free intracellular calcium.
Regulation of alpha-bungarotoxin receptor accumulation in chick retina cultures: effects of membrane depolarization, cyclic nucleotide derivatives, and Ca2+
Authors: Betz H
Published: March 2018

The regulation of a putative neuronal nicotinic acetylcholine receptor, the alpha-bungarotoxin (alpha-Btx)-binding protein, was investigated in primary cultures of chick embryo retina. Depolarization of the cells by veratrum alkaloids, or by a high K+ concentration of the culture medium, reduced the accumulation of cell surface alpha-Btx receptors in these cultures. In contrast, the addition of a blocker of voltage- dependent Na+ channels, tetrodotoxin, increased alpha-Btx receptor accumulation in differentiated retina cultures. Derivatives of cyclic adenosine 3′:5′-monophosphate also increased the number of alpha-Btx receptor sites, whereas dibutyryl cyclic guanosine 3′:5′-monophosphate, but not 8-bromo-cyclic guanosine 3′:5′-monophosphate, had the opposite effect. The blocker of voltage-dependent Ca2+ channels, D600, and media containing a reduced Ca2+ concentration also increased alpha-Btx receptor levels. All of these different culture conditions altered the rate of receptor loss after blocking protein synthesis by cycloheximide. These results show that the synthesis of the neuronal alpha-Btx receptor is regulated by membrane depolarization, cyclic nucleotides, and Ca2+ in a fashion analogous to the regulation of muscle acetylcholine receptor.
Frequency-specific projections of individual neurons in chick brainstem auditory nuclei
Authors: Young SR , Rubel EW
Published: March 2018

Nucleus magnocellularis and nucleus laminaris in the avian brainstem contain second- and third-order auditory neurons thought to be homologous to the mammalian anteroventral cochlear nucleus and medial superior olivary nucleus, respectively. Nucleus laminaris in the chicken is a tonotopically organized sheet of bipolar neurons; each of these neurons receives spatially segregated bilateral innervation from the two magnocellular nuclei. In the present study, this projection was studied at the single cell level by analyzing the pattern of terminal arborizations of individual horseradish peroxidase-filled axons. Reconstruction of the terminal arborizations of nucleus magnocellularis axons revealed that each axon forms an elongated band of endings, the long axis of which is parallel to the physiologically defined isofrequency bands. Within a band, the individual terminal collaterals form distinct patches separated by areas without endings. We suggest that the elongated terminal fields provide the basis of the tonotopic organization observed in nucleus laminaris and that the trajectories of the ipsilateral and contralateral axons may provide differential conduction delays that are important for binaural integration of acoustic information.
Release of dopamine from coaggregate cultures of mesencephalic tegmentum and corpus striatum
Authors: Shalaby IA , Kotake C , Hoffmann PC , Heller A
Published: March 2018

Dissociated dopamine (DA) neurons from 14-day fetal mice were dissected from the rostral mesencephalic tegmentum (RMT) and were allowed to reaggregate in vitro with cells from the corpus striatum (CS). As previously demonstrated under these conditions, DA neurons develop punctate fluorescent varicosities and the capacity to synthesize, accumulate, and retain DA (Kotake, C., P. C. Hoffmann, and A. Heller (1982) J. Neurosci. 2: 1307–1315). After 17 to 22 days in culture, the RMT-CS coaggregates were assessed for their ability to release DA. Coaggregates were incubated in 5.6 x 10(-6) M [3H]DA, washed, and then superfused at 100 microliters/min for 2 hr. Fractions were collected every 2 min. Basal efflux of [3H]DA/2 min was 1% of tissue stores of 3H. K+, 70 mM infused for 8 min induced a peak release of 5.87% of tissue stores of 3H, and 50 mM K+ induced a peak release of 2.13%. The potassium-induced release of [3H]DA was calcium dependent. When d- amphetamine was infused for 12 min, 100 microM solutions induced a peak release of 8.91%, 10 microM induced a peak release of 4.36%, and 1 microM induced a peak release of 1.85% of tissue stores of 3H. Substance P at 100 microM induced a peak release of 2.19% of tissue stores of 3H. Tetrodotoxin (0.5 and 2.5 microM) decreased basal efflux by 40%, blocked substance P-induced release, but did not affect either potassium- or d-amphetamine-induced release of [3H]dopamine.
Secretin: specific binding to rat brain membranes
Authors: Fremeau RT , Jensen RT , Charlton CG , Miller RL , O'Donohue TL , Moody TW
Published: March 2018

The binding of [125I]secretin to rat brain membranes was investigated. Radiolabeled secretin bound with high affinity (KD = 0.2 nM) to a single class of noninteracting sites. Binding was specific, saturable, and reversible. Regional distribution studies indicated that the specific binding was greatest in the cerebellum, intermediate in the cortex, thalamus, striatum, hippocampus, and hypothalamus, and lowest in the midbrain and medulla/pons. Pharmacological studies indicated that only secretin, but not other peptides, inhibits binding of [125I]secretin with high affinity. Also, certain guanine nucleotides inhibited high affinity binding. These data indicate that rat brain membranes possess high affinity binding sites specific for secretin and that with the use of [125I] secretin the kinetics, stoichiometry, specificity, and distribution of secretin receptors can be directly investigated.
Glutamic acid decarboxylase-immunoreactive neurons and horseradish peroxidase-labeled projection neurons in the ventral posterior nucleus of the cat and Galago senegalensis
Authors: Penny GR , Fitzpatrick D , Schmechel DE , Diamond IT
Published: March 2018

Immunocytochemical methods were used to identify neurons in the ventral posterior nucleus of the cat and Galago senegalensis that contain glutamic acid decarboxylase (GAD), the synthetic enzyme for the inhibitory neurotransmitter, GABA. In both species GAD-immunoreactive neurons make up about 30% of the total neurons in the ventral posterior nucleus and form a distinct class of small cells. After cortical injections of horseradish peroxidase (HRP), GAD-immunoreactive cells are not labeled with HRP and may, therefore, be GABAergic local circuit neurons. Comparison of the dendritic morphology of GAD-immunoreactive neurons with that of HRP-filled projection neurons reveals that the morphology of the GAD-containing neurons is distinct and, in particular, that the GAD-immunoreactive neurons display fewer primary dendrites. The relay neurons, in turn, can be divided into classes based on dendritic morphology and cell body size.
Immunocytochemical studies of actin localization in the central nervous system
Authors: Goldman JE
Published: March 2018

The contractile protein, actin, is a major component of the central nervous system (CNS). Its accumulation at presynaptic and postsynaptic sites has been suggested by biochemical and ultrastructural studies. The present work examines directly the distribution of actin within the CNS by immunocytochemical means. Sections of rat CNS were stained with an antiserum against chicken gizzard actin, using a peroxidase- antiperoxidase technique. In Vibratome sections, reaction product appeared widely distributed in the perikaryal cytoplasm of neurons, in axons and dendrites, and in astrocytes and blood vessels. In paraffin sections, however, much of the cytoplasmic reaction was not observed. Instead, the reaction product was concentrated in processes within the neuropil and in discrete densities about the perimeters of neurons. This latter distribution, observed in areas in which large axosomatic synapses are known to occur, was consistent with a concentration at synaptic sites. Immunocytochemistry performed at the ultrastructural level, using a pre-embedding technique, revealed a diffuse cytoplasmic staining, with increased accumulation under the neuronal plasma membrane, particularly at postsynaptic densities.
Laser microbeam surgery: ultrastructural changes associated with neurite transection in culture
Authors: Gross GW , Lucas JH , Higgins ML
Published: March 2018

The exposure of neuronal and glial cell processes to a large number (up to 300) of 12-nsec laser pulses at a wavelength of 337 nm and energy densities below the threshold for nonlinear absorption results in a gradual, gentle process transection in the laser focus. Within 10 to 20 sec after cessation of firing, the process pinches in the target area. During this time, mitochondria become swollen and bleached, the plasma membrane develops an obvious tautness, microtubules disappear, and organelles accumulate to either side of the process constriction. Depending on the irradiation parameters, a local pinching may proceed to a transection in about 30 sec or it may reverse to yield a normal- appearing process in approximately 5 min. Severe process pinching is accompanied by a sudden depolarization that may last for 2 to 5 min and is usually followed by a repolarization to the original resting potential even if the process has transected. Spiral retraction of cut processes and cytoplasmic spillage observed after mechanical transections are not seen with this laser method. Process stretching is minimized or eliminated. Extensive vacuolization often associated with mechanical transections does not develop unless substrate involvement in the form of shock waves is apparent. For the performance of cell surgery in culture, this method appears to offer a reliable approach to morphological alteration of single cells and to the tailoring of two- dimensional neuronal networks. It should also allow more quantitative and better-controlled studies of axonotmesis, degeneration, and regeneration on the single cell level, and it may be used as a probe for the investigation of cytoskeletal dynamics. A mechanism describing the cytoskeletal changes associated with laser-induced cell process transection is proposed.
Analysis of fast excitatory postsynaptic currents in bullfrog parasympathetic ganglion cells
Authors: Connor EA , Parsons RL
Published: March 2018

Excitatory postsynaptic currents (EPSCs), miniature excitatory postsynaptic currents (MEPSCs), and acetylcholine-induced current fluctuations (noise) have been studied in voltage-clamped bullfrog parasympathetic ganglion cells of the atrial septum. EPSCs were also recorded from voltage-clamped sympathetic B cells and, in general, it was found that the basic properties of the EPSCs are similar in both parasympathetic and sympathetic B cells. For parasympathetic cells, the EPSC reached a peak amplitude of several nanoamperes within 3 msec and decayed exponentially. For 31 cells voltage-clamped to -50 mV (22 to 23 degrees C), peak amplitude was -4.3 +/- 1.4 nA (mean +/- SD) and the decay time constant, tau, was 5.6 +/- 1.0 msec. tau was independent of EPSC amplitude at a set voltage but increased with hyperpolarization, the coefficient of voltage dependence being -0.0070 +/- 0.0025 mV-1 in 13 cells (21 to 23 degrees C). The EPSC amplitude-voltage relationship was linear between -30 and -90 mV. The reversal potential, determined by interpolation, was -4.0 +/- 6.7 mV (n = 11). The EPSC tau had a Q10 equal to 2.9. Blocking the acetylcholinesterase with methane sulfonyl fluoride (MSF) pretreatment prolonged EPSC decay but decreased EPSC amplitude. In addition, EPSC decay after MSF treatment deviated from a single exponential function. MEPSCs exhibited decay characteristics very similar to those of EPSCs recorded at the same voltage and temperature. Acetylcholine-induced current fluctuations were well described by a single Lorentzian function with the estimated mean channel open time (tau noise) very similar to the EPSC decay time constant.(ABSTRACT TRUNCATED AT 250 WORDS)
Olfactory bulb increases marker protein in olfactory receptor cells
Authors: Chuah MI , Farbman AI
Published: March 2018

Olfactory mucosa was excised from the heads of embryonic day 15 (E15) rat fetuses, prior to the appearance of olfactory marker protein (OMP), and cultured with or without the presumptive olfactory bulb. Specific immunohistochemical staining for the OMP was detected in both groups of explants 6 days after explantation. At no time do the supporting cells and basal cells show positive OMP staining. Results of radioimmunoassay on the two groups of cultures showed that about twice as much OMP was present in organ cultures grown with presumptive olfactory bulb. Estimates of the area of olfactory mucosa in the explants revealed no significant differences between those cultured with or without the bulb. Cell counts revealed that the increased amount of OMP in explants with the bulb was due to a higher number of OMP-positive receptor neurons. Explants of olfactory mucosa cultured with other tissues (cerebrum, cerebellum, cervical spinal cord, and heart) contained approximately the same amount of OMP as explants of mucosa alone; i.e., these other tissues did not enhance OMP levels. Similarly, there was no enhancement of OMP level when the olfactory mucosa was separated from the bulb by a Millipore filter of 0.45 micron pore size. Thus the results of the study indicate that the presumptive olfactory bulb specifically increases OMP level by a mechanism that requires direct contact between the receptor cells and bulb.
Injury-induced neuronotrophic activity in adult rat brain: correlation with survival of delayed implants in the wound cavity
Authors: Nieto-Sampedro M , Manthrope M , Barbin G , Varon S , Cotman CW
Published: March 2018

Mechanical or chemical injury to adult rat brain elicited the accumulation in the affected area of trophic activity for cultured parasympathetic, sympathetic, and sensory neurons. Neuronotrophic activity was relatively low both in noninjured brain tissue extracts and in extracts prepared from the tissue surrounding an injury immediately after the lesion was made. However, trophic titers increased considerably over time, first in the brain tissue that formed the walls of the wound and then in the Gelfoam filling the wound cavity. In the tissue adjacent to the injury, trophic titers began to rise immediately after the lesion, reached a maximum 10 days later, and decayed thereafter. In the wound cavity, occupied by Gelfoam, neuronotrophic activity began to increase 6 days postlesion, reached a maximum at day 16 after injury, and decreased at later times. The levels of induced trophic activity appeared to be proportional to the size of the wound. Injury to various brain areas including temporal, entorhinal, occipital, parietal, and frontal cortices, hippocampus, corpus striatum, and cerebellum, all induced a similar increase in neuronotrophic factor(s). Damage to the myelinated fibers of the corpus callosum did not. High trophic titers decayed rapidly with distance from the wound except in areas heavily deafferented by the lesion, where activity also reached high levels. Extracts from all of the above- mentioned brain areas contained toxic activity for cultured spinal cord neurons. The level of neuronotoxic activity was similar both before the lesion and 15 days postlesion, with the possible exception of the corpus callosum. Intraventricular injections of kainic acid at doses which destroy areas CA4, CA3, and part of CA1 of the hippocampus also induced a time-dependent rise of neuronotrophic activity in this structure, comparable to that achieved by mechanical damage. Both kainic acid treatment and mechanical injury cause extensive glial proliferation in the injured and/or deafferented area. The apparent concurrence of glial reaction and increase in neuronotrophic activity suggests that glial cells may be a major source of the induced trophic activity. As an in vivo correlate of cell culture data, the survival of striatal transplants into host cortical wounds was examined. Fragments of embryonic corpus striatum did not survive when transplanted into a freshly made cavity in the entorhinal/occipital cortex of adult rats. Survival was enhanced by introducing a delay between the time at which the wound cavity was made and that at which the striatal tissue was implanted in it.(ABSTRACT TRUNCATED AT 400 WORDS)
Reciprocal changes in the firing rate of neostriatal and dorsal raphe neurons following local infusions or systemic injections of D- amphetamine: evidence for neostriatal heterogeneity
Authors: Rebec GV , Curtis SD
Published: March 2018

A local infusion of d-amphetamine (d-AMPH) into the dorsal raphe nucleus (DRN) inhibited neuronal activity in this site and produced a mirror-image excitation in the ventrolateral, but not dorsomedial, neostriatum. This effect, which was mimicked by 5-methoxy-N,N- dimethyltryptamine, a serotonin autoreceptor agonist, was not altered by pretreatment with alpha-methyl-p-tyrosine. Similar regional differences in neostriatal activity were obtained following an electrolytic lesion of the DRN or an intraperitoneal injection of d- AMPH. In fact, whereas 1.0 mg/kg of d-AMPH accelerated ventrolateral activity and inhibited dorsomedial neurons, 7.5 mg/kg produced the opposite effect. At both doses, however, DRN activity was inversely related to firing rate in the ventrolateral, but not dorsomedial, neostriatum. These results indicate that only certain regions of the neostriatum are responsive to changes in DRN activity and that these regions respond differently to systemic injections of d-AMPH than other neostriatal sites.
Reciprocal interactions between the position sense representations of the two forearms
Authors: Lackner JR , Taublieb AB
Published: March 2018

Muscle spindle afferents are known to influence the conscious appreciation of limb position. If a limb muscle is mechanically vibrated while the limb is physically restrained from moving under the action of the resulting tonic vibration reflex, illusory motion of the limb will be experienced in the direction that would be associated with stretch of the vibrated muscle. Two experiments are presented that explore how grasping with the other hand the forearm of an arm whose biceps or triceps muscle is being vibrated affects the apparent position of the two arms. If the vibrated arm is grasped before the onset of vibration, then illusory motion of both arms is experienced during vibration. The magnitude of this apparent motion and displacement is less than that experienced when the vibrated arm is not held. If the vibrated arm is grasped after the onset of vibration and when illusory displacement is being experienced, then its apparent motion is suppressed and the grasping arm is correctly perceived as stationary. The existence of this reciprocal, position sense interaction between the two arms suggests that the conscious awareness of limb position is not dependent solely on afferent and efferent information about individual limbs in isolation, but potentially involves coordinating spatial information about the configuration of the entire body.
Quantitative features of synapse formation in the fly's visual system. I. The presynaptic photoreceptor terminal
Authors: Frohlich A , Meinertzhagen IA
Published: March 2018

Photoreceptors of the adult fly's compound eye each form a population of stereotyped output synapses distributed over the surface of their terminal. The formation of this class of afferent synapses during development has been followed from serial electron microscopy of the same eye region in four pupal and several adult stages, all of female Musca domestica. These synapses, or tetrads, have an invariant postsynaptic composition of four members and so may provide a model for multiple-contact synapses in general. In the adult fly the four postsynaptic elements of each synapse are contributed by two interneurons, L1 and L2, and, usually, two alpha processes of an amacrine cell. These postsynaptic elements assemble at individual developing synapses by selective sequential addition. Assembly starts with L1 or L2, subsequent elements of the final tetrad adding in all conceivable permutations, at least as fast as one per 7 hr. They rarely (only once) incorporate incorrect or supernumerary elements, however. The synaptic population as a whole was also sampled during development to analyze the possible factors determining the normal precision of the size of the adult population. The number of synapses per terminal increases gradually until 74% pupal development. Thereafter it decreases so that the final number of synapses in each receptor's population is the consequence of a net loss. Synapses enlarge with age, chiefly by incorporating new elements, but the loss of synaptic sites is only partially offset by the increase in size of those that remain. Throughout all stages examined in pupal and adult life, total synaptic area is linearly proportional to the surface area of the axon terminal. Thus, from the 74% pupal development stage onward, a population of many small synapses closely spaced, on average, over the terminal's surface transforms into one characteristic of the adult with progressively fewer, larger, more widely spaced synapses.
The development of peptidergic neurons in the foregut of the chick
Authors: Epstein ML , Hudis J , Dahl JL
Published: March 2018

We have used antisera specifically directed against Met-enkephalin (met- ENK), somatostatin (SOM), substance P (SP), and vasoactive intestinal peptide (VIP), to study the development of neurons containing these peptides in the foregut of the chick. All four peptides were detected early in ontogeny, at 4 to 9 days of incubation (d.i.), and were localized primarily to cell bodies in the primitive myenteric plexus. There were differences in the times at which they were first detected and in the sequence of their appearance in the proventriculus, gizzard, and duodenum. The differentiation of these peptidergic neurons in the duodenum was examined in some detail. Cell bodies containing these peptides were first detected in the primitive myenteric plexus at 5 to 7 d.i. and increased in number from 7 to 11 d.i. Processes containing varicosities became prominent between 11 and 13 d.i. VIP was the first of the peptides to appear in the submucosal plexus and was found in more proximal regions of the duodenum at 5 d.i. Shortly thereafter, SOM- and SP-containing cell bodies were seen; met-ENK-containing cell bodies were never detected in the submucosal plexus. At 13 d.i., the circular smooth muscle contained a number of VIP-immunoreactive and a smaller number of SOM-immunoreactive processes. met-ENK- and SP-immunoreactive processes appeared in the circular smooth muscle between 17 and 21 d.i.; VIP- and SP-immunoreactive processes appeared in the mucosal plexus at 17 to 21 d.i. Our results suggest that neuropeptides appear very early in the ontogeny of enteric neurons, at the same time or even before cholinergic and serotonergic neurons express their phenotypes. These findings argue against a sequential developmental order in which peptidergic neurons appear after those containing acetylcholine and serotonin.