bluuebunny/crossref_metadata_2025 huggingface dataset:
The human locus coeruleus: computer reconstruction of cellular distribution
Authors: German DC , Walker BS , Manaye K , Smith WK , Woodward DJ , North AJ
Published: March 2018

Quantitative neuroanatomical techniques were developed to map the distribution of norepinephrine-containing locus coeruleus (LC) neurons in the adult human brain. These neurons reside in the dorsolateral pontine tegmentum and are identifiable by their neuromelanin pigment content. Five brains, ranging in age from 60 to 104 years, were examined. Outlines of coronal or sagittal sections containing the LC were entered into a computer along with the location of each cell, certain neuroanatomical landmarks, and cell size. Sections were aligned with specific neuroanatomical landmarks so that the computer-generated distribution of cells was representative of the in situ distribution of cells. Analysis of (1) the number of cells in sections throughout the rostrocaudal extent of the nucleus, (2) cell size, (3) 3-dimensional reconstructions of the distribution of cells within the brain stem, and (4) 2-dimensional cell-frequency maps, make it possible to quantitatively characterize the distribution of cells within this large nucleus. The total estimated number of LC cells on both sides of the brain ranged from 45,562 to 18,940 (youngest to oldest), and mean soma area ranged from 835 to 718 micron 2 (youngest to oldest). The nucleus is “tube-like” in shape, has a rostrocaudal extent of approximately 16 mm, and is bilaterally symmetrical. Two-dimensional cell-frequency maps were developed to illustrate the regional distribution of cell frequencies at any rostrocaudal/mediolateral point on the horizontal plane; the total unilateral area of the LC ranged from 32.8 to 17.2 mm2 (youngest to oldest). The techniques developed to characterize the 2- and 3-dimensional distributions of LC neurons can be used in future studies to quantitatively examine the effects of aging and disease on this and other brain nuclei.
Calcium-dependent inactivation of a potassium current in the Aplysia neuron R15
Authors: Kramer RH , Levitan IB
Published: March 2018

The endogenously bursting pacemaker neuron R15 of Aplysia exhibits an inwardly rectifying K+ current (IR) that was shown previously to be enhanced by various neurotransmitters via the intracellular second messenger, cyclic AMP (Drummond et al., 1980; Benson and Levitan, 1983; Levitan et al., 1987). Here we present evidence that Ca2+ influx, either caused by spontaneous bursting activity or elicited by depolarizing voltage-clamp pulses, causes a large, long-lasting inactivation of IR. The ionic current inactivated by bursts is identified as IR by several criteria: it activates steeply at membrane potentials more negative than the K+ equilibrium potential, has very fast kinetics, is reduced by lowering external K+ from 10 to 2 mM, and is blocked by adding 1 mM Ba2+, 10 mM Cs+, or 5 mM Rb+ to the bathing medium. The peak inactivation of IR is delayed following a single burst of spikes in R15, such that IR decreases maximally by about 20% after 60–90 sec, and then recovers gradually over more than 10 min. The inactivation caused by many bursts of spikes can reduce IR to less than 50% of its initial amplitude. The delay in onset and slow time course of recovery from inactivation of IR suggest that a complex biochemical mechanism underlies the effect of Ca2+ on IR. The effect of depolarization on IR is due specifically to the influx and intracellular accumulation of Ca2+. Depolarizing voltage-clamp pulses are maximally effective at reducing IR when they elicit a large influx of Ca2+, while pulses approaching the Ca2+ equilibrium potential have little effect.(ABSTRACT TRUNCATED AT 250 WORDS)
Experimental diabetic neuropathy: similar changes of slow axonal transport and axonal size in different animal models
Authors: Medori R , Jenich H , Autilio-Gambetti L , Gambetti P
Published: March 2018

Analysis of slow axonal transport in sciatic and primary visual systems of BB rats with spontaneous diabetes of 2.5–3.5 months duration revealed a delay in transport of the neurofilament (NF) subunits, tubulin, actin, and the 60, 52, and 30 kDa polypeptides in both systems. The polypeptides examined were not affected uniformly. Rather, the transport of the 60, 52, and 30 kDa polypeptides and the rapidly moving component of tubulin, all constituents of the slow component b (SCb) of axonal transport, appeared to be more severely delayed than the transport of polypeptide constituents of the slow component a (SCa), such as NF and the slow-moving tubulin. Transport was not impaired in diabetic BB rats maintained normoglycemic with optimal doses of insulin. A 52 kDa polypeptide constituent of SCb was identified as neuron-specific enolase, and the 30 and 60 kDa polypeptides are likely to be aldolase and pyruvate kinase; all 3 are glycolytic enzymes. Morphometric analysis revealed that the cross- sectional area of sciatic axons was increased proximally at the level of the motor roots and decreased distally at the level of the tibial nerve. The changes in slow transport and caliber observed in central and peripheral axonal systems of diabetic BB rats are virtually identical to those previously described in rats with streptozotocin- induced diabetes, another model of insulin-dependent diabetes. In both models, the alterations of axonal caliber are likely to be secondary to the impairment of axonal transport.(ABSTRACT TRUNCATED AT 250 WORDS)
Metabolic activity in rat tectal grafts is influenced by host sensory innervation
Authors: Dyson SE , Harvey AR , Stone AF , Bell GA
Published: March 2018

It has been shown previously that fetal tectal tissue grafted to the midbrain of newborn host rats grows, differentiates, and receives input from the host brain. In the present study, 4 neuroanatomical techniques have been combined to examine how metabolic activity in tectal transplants is influenced by an identified host sensory pathway. Tectal tissue from E15 pigmented rat embryos was transplanted to the midbrain region of anesthetized newborn rats of the same strain. Six to 22 weeks later, the functional relationship between tectal transplants and the visual system of the host animal was examined by mapping metabolic activity in the grafts and relating this activity to the presence or absence of host retinal innervation. Metabolic activity in tectal grafts was assessed using the radioactive 2-deoxy-glucose (2-DG) method and cytochrome oxidase (CO) histochemistry. Graft regions receiving input from host retinal axons were demonstrated by anterograde labeling after bilateral intraocular injections of HRP or WGA-HRP; all areas in grafts that were homologous to the superficial layers of normal superior colliculus (SC) were identified using AChE histochemistry. The levels of metabolic activity demonstrated with 2-DG and CO varied between animals and within individual grafts. Grafts that did not connect with the host showed only low metabolic activity. In grafts that received host input, localized areas of high metabolic activity were seen with both 2-DG and CO. Highest levels of activity were consistently found in area containing both intense AChE activity and a high density of host retinal innervation.
Localization of cells synthesizing transforming growth factor-alpha mRNA in the mouse brain
Authors: Wilcox JN , Derynck R
Published: March 2018

The distribution of transforming growth factor (TGF)-alpha and TGF-beta 1 mRNA containing cells in adult mouse brain was examined using in situ hybridization histochemistry. There were no detectable TGF-beta 1 mRNA- containing cells found in the brain. TGF-alpha mRNA was localized to cell bodies of the caudate nucleus, dentate gyrus, anterior olfactory nuclei, and a laminar distribution of mitral cells in the olfactory bulb. TGF-alpha-synthesizing cells were localized in brain regions that have been shown to synthesize nerve growth factor, epidermal growth factor (EGF) and enkephalins. The function of local synthesis of TGF- alpha in the brain is as yet unknown.
Origin of the atriopeptin-like immunoreactive innervation of the paraventricular nucleus of the hypothalamus
Authors: Standaert DG , Saper CB
Published: March 2018

The paraventricular nucleus of the hypothalamus (PVH) contains a prominent collection of varicose atriopeptin-like immunoreactive (APir) fibers. We have used immunohistochemistry and fluorescent retrograde tracers to investigate the origin of these fibers. All parts of the PVH contain APir fibers. The densest collections are found in the periventricular area and in the parvocellular components of the nucleus. Somewhat smaller numbers of fibers are found within the borders of the magnocellular part of the PVH. Following the injection of fluorescent retrograde tracers into the PVH and subsequent immunohistochemical staining, numerous retrogradely labeled APir neurons were observed in the anteroventral periventricular nucleus (AVPV), adjacent to the anteroventral tip of the third ventricle. Smaller groups of retrogradely labeled APir neurons were observed in the ventromedial part of the bed nucleus of the stria terminalis, the pedunculopontine and laterodorsal tegmental nuclei, and the medial part of the nucleus of the solitary tract. The APir projection from the AVPV to the PVH is of particular interest in view of the abundant evidence implicating both regions in the regulation of the fluid and electrolyte balance and blood pressure. AP may serve as a central neuromodulator as well as a circulating hormone in cardiovascular regulation.
Periodicity of GABA-containing cells in primate prefrontal cortex
Authors: Schwartz ML , Zheng DS , Goldman-Rakic PS
Published: March 2018

The tangential distribution of GABA-containing cells was examined in the principal sulcus of the frontal lobe in 12 macaque monkeys. Following immunostaining with GABA antisera all immunoreactive cells were charted and their distribution analyzed with both statistical and spectral density methods. In addition, a gapless series of sections was used to generate a 2-dimensional reconstruction of cell disposition in the tangential plane parallel to the pia. Our findings indicate that the GABA cells are not distributed uniformly across the cortex, as is commonly believed, but that their density is characterized by 2 independent sinusoidal fluctuations: a high-frequency component with a period ranging from 150 to 250 micron superimposed upon a lower- frequency component with a period of 1000–1275 micron. The half-cycle of the low-frequency component (roughly 625 micron) is very similar to the dimensions of afferent and efferent columns in the principal sulcus, while the half-cycle of the higher-frequency component (approximately 125 micron) is closer in size to that of the functionally defined columns of neurons found in regions of sensory cortex that share common physiological properties. To our knowledge, these findings are the first indication that inhibitory local circuit neurons are not uniformly or randomly distributed, but exhibit periodicities that may be related to the columnar, functional and architectural organization of the cortex.
Class-specific cell death shapes the distribution and pattern of central projection of cat retinal ganglion cells
Authors: Leventhal AG , Schall JD , Ault SJ , Provis JM , Vitek DJ
Published: March 2018

The development of the nasotemporal division in cat retina was studied. We find that in the normally pigmented neonatal cat significant numbers of ganglion cells of all types in temporal retina project to the contralateral dorsal lateral geniculate nucleus (LGNd); far fewer cells in temporal retina project contralaterally to the LGNd in the normal adult. Thus, most of these cells must be eliminated during development. Experimental interruption of one optic tract in the neonate results in the retrograde degeneration of the ipsilaterally projecting ganglion cells in the temporal retina ipsilateral to the lesion. Consequent to the loss of the ipsilaterally projecting cells in this hemiretina, many of the ganglion cells projecting to the intact contralateral LGNd, which are normally eliminated, survive. Also, unlike in the normal cat, in which very few of the small ganglion cells in temporal retina project contralaterally to the thalamus, in optic tract sectioned (OTX) cats, significant numbers of the smallest ganglion cells in the temporal retina ipsilateral to the lesion project contralaterally to the intact thalamus. In order to make a quantitative comparison of the distributions of ipsilaterally and contralaterally projecting cells in the temporal retinae of normal cats, OTX cats, and neonatal kittens, it was necessary to determine the position of the vertical meridian in all animals. We defined the vertical meridian as the median edge (Stone, 1966). The median edge was determined from the distribution of the most nasally located, ipsilaterally projecting cells in temporal retina. The results indicate that the angle of the vertical meridian (median edge) with respect to the area centralis and optic disc is specified before birth and does not differ in normal cats, OTX cats, or neonatal kittens. Since the location of the vertical meridian does not change with age in postnatal life and is not affected by optic tract section, corresponding regions of retina in the different groups could be compared. A quantitative analysis of ganglion cell density in the temporal retina contralateral to the section, ipsilateral to the intact hemisphere, indicated that there was a reduction in the population of ipsilaterally projecting ganglion cells that was complementary to the abnormally large number of contralaterally projecting cells surviving in the temporal retina ipsilateral to the lesion.(ABSTRACT TRUNCATED AT 400 WORDS)
Extrinsic determinants of retinal ganglion cell structure in the cat
Authors: Leventhal AG , Schall JD , Ault SJ
Published: March 2018

The degree to which a retinal ganglion cell's environment can affect its morphological development was studied by manipulating the distribution of ganglion cells in the developing cat retina. In the newborn kitten there is an exuberant ganglion cell projection from temporal retina to the contralateral lateral geniculate nucleus (LGNd) (Leventhal et al., 1988) and from nasal retina to the ipsilateral LGNd. Neonatal, unilateral optic tract section results in the survival of many of these ganglion cells (Leventhal et al., 1988). The morphology of ganglion cells which survive in regions of massively reduced ganglion cell density was studied. As reported previously (Linden and Perry, 1982; Perry and Linden, 1982; Ault et al., 1985; Eysel et al., 1985), we found that the dendritic fields of all types of ganglion cells on the border of an area depleted of ganglion cells extended into the depleted area. The cell bodies and dendritic fields of alpha and beta cells within depopulated areas, as well as on the borders of the depopulated areas, were larger than normal. The dendritic fields of these cells also exhibited abnormal branching patterns. For alpha and beta cell types the relative increase in size tended to be greatest where the relative change in density was the greatest. In fact, isolated beta cells within the cell-poor area centralis region resembled normal central alpha cells in the cell-rich region of the area centralis in the same retina. Interestingly, in the same regions of reduced density where alpha and beta cells were dramatically larger than normal, the cell body and dendritic field sizes of other cell types (epsilon, g1 and g2 were unchanged. These results indicate that neuronal interactions during development contribute to the morphological differentiation of retinal ganglion cells and that different mechanisms mediate the morphological development of different classes of cells in cat retina.
Locomotion produced in mesencephalic cats by injections of putative transmitter substances and antagonists into the medial reticular formation and the pontomedullary locomotor strip
Authors: Noga BR , Kettler J , Jordan LM
Published: March 2018

The purpose of this study was to determine the distribution of cells in the medial reticular formation (MRF) and the pontomedullary locomotor strip (PLS), which can induce locomotion when activated. Controlled microinjections of neuroactive substances (Goodchild et al., 1982) into the MRF or PLS were made in order to activate cell bodies in those areas. The ability of trigeminal receptive field stimulation to induce locomotion before and after drug infusion into the PLS was also assessed since the PLS and the spinal nucleus of the trigeminal nerve are similar in their anatomical distribution. Experiments were performed on precollicular-postmamillary decerebrate cats walking on a treadmill. Injections of glutamic acid (GA; 500 nmol) into the MRF produced locomotion that was antagonized by infusion of glutamic acid diethyl ester into the same spot. Decreases in the current threshold for locomotion produced by electrical stimulation of the MRF were observed when the MRF was infused with either GA (40–80 nmol), DL- homocysteic acid (DL-HCA; 200 nmol), or picrotoxin (PIC; 15 nmol). Injections of GA (100 nmol), DL-HCA (700 nmol), PIC (10–50 nmol), and substance P (2 nmol) into the PLS also produced locomotion. Locomotion produced by injections of PIC into the PLS was blocked by infusion of equal amounts of muscimol or GABA. Effective PLS injection sites were all confined to the trigeminal spinal nucleus or immediately ventral and medial to this in the adjacent lateral reticular formation. Trigeminal nerve peripheral field stimulation evoked locomotion after microinjection of PIC into the PLS, although this same facial stimulus was not effective prior to drug injection. We conclude that the MRF and PLS regions of the cat brain stem contain cells that produce locomotion when chemically stimulated, and we suggest that the PLS is closely related to or synonymous with the spinal nucleus of the trigeminal nerve. Furthermore, we suggest that stimulation of trigeminal afferents is analogous to stimulation of segmental afferent pathways in the production of locomotion (Sherrington, 1910; Jankowska et al., 1967; Afelt, 1970; Budakova, 1972; Grillner and Zangger, 1979).
Glutamate-like immunoreactivity in identified neuronal populations of insect nervous systems
Authors: Bicker G , Schafer S , Ottersen OP , Storm-Mathisen J
Published: March 2018

Glutamate is considered to be the most likely transmitter candidate at excitatory synapses onto skeletal muscles of insects. We investigated the distribution of glutamate-like immunoreactivity (Glu-LI) in identified motor neurons of glutaraldehyde-fixed metathoracic ganglia of the locust in paraffin serial sections. The presumably glutamatergic fast and slow extensor tibiae motor neurons show Glu-LI, whereas other cells, including the GABAergic common inhibitory motor neurons and the cluster of octopaminergic dorsal unpaired median cells, show rather low levels of staining. Immunoreactivity of the fast extensor tibiae motor neuron is located in soma, neurites, axon, and the terminal arborizations. A double-labeling experiment on sections of the locust metathoracic ganglion showed that antisera against glutamate and GABA discriminate between the presumably glutamatergic and GABAergic motor neurons and that GABA-LI-positive neurons are low in Glu-LI. The results suggest that Glu-LI can be used as a marker for detecting potential glutamatergic neurons in insects under the present conditions. Application of the glutamate antiserum to sections of the honeybee brain revealed Glu-LI in motor neurons but also in certain interneurons. The most prominent populations of Glu-LI-positive cells were the monopolar cells and large ocellar interneurons, which are first-order interneurons of the visual and ocellar system. Several groups of descending interneurons also showed Glu-LI. The distributions of Glu-LI and GABA-LI are complementary in locust and bee ganglia. The high level of Glu-LI in certain interneuronal populations, as well as in identified glutamatergic motor neurons, suggests that insect central nervous systems may contain glutamatergic neuronal pathways.
Zinc alters excitatory amino acid neurotoxicity on cortical neurons
Authors: Koh JY , Choi DW
Published: March 2018

Recent studies have suggested that large amounts of free zinc may be coreleased during excitatory synaptic transmission at glutamatergic synapses, and may act postsynaptically to decrease actions mediated by N-methyl-D-aspartate (NMDA) receptors, while often increasing neuroexcitation mediated by quisqualate receptors. The present study examined the ability of zinc to alter excitatory amino acid (EAA) neurotoxicity. Murine cortical cell cultures were exposed to EAAs for 5 min in defined solutions, and neuronal cell injury was examined the following day both morphologically and by lactate dehydrogenase assay. Inclusion of 30–500 microM zinc in the exposure solution produced a zinc concentration-dependent, noncompetitive attenuation of NMDA- induced neuronal injury, with an ED50 of about 80 microM. In contrast, zinc produced the same concentration-dependent potentiation of quisqualate neurotoxicity; and with 500 microM zinc, a small potentiation of kainate neurotoxicity was suggested. The effect of zinc on the neurotoxicity of the broad-spectrum agonist glutamate was consistent with these effects on specific agonists, as well as with a previous study showing that glutamate neurotoxicity normally depends predominantly on NMDA-receptor activation. Zinc produced a concentration-dependent reduction in glutamate-induced neuronal injury in a fashion similar to that seen with NMDA, but less effectively. In addition, despite this overall protective effect, zinc paradoxically increased the glutamate-induced destruction of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d)-containing neurons, a subpopulation that was shown in the preceding paper (Koh and Choi, 1988) to exhibit resistance to NMDA receptor-mediated neurotoxicity, and vulnerability to non-NMDA receptor-mediated neurotoxicity.(ABSTRACT TRUNCATED AT 250 WORDS)
The overall pattern of ocular dominance bands in cat visual cortex
Authors: Anderson PA , Olavarria J , Van Sluyters RC
Published: March 2018

This study describes the overall arrangement of geniculocortical input representing the system of cortical ocular dominance bands in layer IV of striate cortex in the adult cat. The pattern of ocular dominance bands was revealed by transneuronal transport of the intraocularly injected tracer wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). Our data indicate that this procedure does not damage the retina and that it results in relatively uniform uptake and transport of the tracer. Using previously published techniques (Olavarria and Van Sluyters, 1983, 1985), both cortical hemispheres of each cat were unfolded, flattened and tangentially sectioned. Analysis of the WGA-HRP labeling patterns in these sections revealed a relatively continuous network of irregularly branching bands in layer IV of area 17 in both hemispheres. Because of a systematic difference in the level of interband labeling, ocular dominance bands appear less distinct in the hemisphere contralateral to the injected eye. There is also a tendency for interband labeling to be greater in cortical regions that represent the more peripheral aspects of the binocular portion of the visual field. The width of an individual ocular dominance band in the cat fluctuates, so that it appears to be made up of a series of uniformly sized, roughly circular beads of label. The diameter of these beads averages 667 micron, and preliminary counts indicate that there are 650–675 beads in each striate cortex. Contrary to earlier suggestions, in 4 out of 6 hemispheres analyzed quantitatively there was no tendency for ocular dominance bands to be oriented along a preferred axis in cat striate cortex, including an axis orthogonal to the border between areas 17 and 18. Ocular dominance bands in area 18 appear to be broader than those in area 17, and they seem to have a greater tendency to be oriented orthogonal to the 17/18 border than those in area 17. Compared with the ocular dominance pattern in monkey striate cortex, the ocular dominance pattern in the cat is much less regular. In general, cat ocular dominance bands appear to fluctuate more in width, to change direction more often, and to be less likely to run orthogonal to the 17/18 border. The greater regularity of the primate ocular dominance pattern may be related to differences in the way in which the visual hemifield is mapped onto the striate cortex in these 2 species.
Effect of neonatal infraorbital nerve transection on substance P- and leucine enkephalin-like immunoreactivities in trigeminal subnucleus caudalis of the rat
Authors: Rhoades RW , Chiaia NL , Hess PR , Miller MW
Published: March 2018

The distributions of substance P-like immunoreactivity (SPLI) and leucine-enkephalin-like immunoreactivity (LENKLI) in subnucleus caudalis of normal adult rats were compared with those observed in the adult rats that sustained transection of the infraorbital (IO) nerve either on the day of birth or in adulthood. All immunocytochemical experiments in the neonatally nerve damaged rats were carried out at least 60 d after the nerve transection. In the animals that sustained nerve transections as adults, brains were processed for immunohistochemistry between 7 and 60 d after the lesions. In the rats that sustained IO nerve transections as adults, there was a transient reduction in the density of the SPLI in layers I and II of ipsilateral subnucleus caudalis. It was most apparent about 2 weeks after the nerve transection and returned to near normal values by 60 d after the lesion. In the rats that sustained IO nerve transections on the day of birth, there was no reduction in the density of SPLI in caudalis, and the band of staining on the deafferented side of the brain stem was actually 40% wider than that on the intact side. Neither neonatal nor adult IO nerve transection had appreciable effects upon the distribution of LENKLI in the rat's trigeminal brain-stem complex. In another series of experiments, rats that sustained neonatal transection of the IO nerve had this same nerve recut in adulthood. Twelve days after the second lesion, the brains of these animals were processed for SPLI. There was a marked reduction in the density of the staining for this peptide on the deafferented side. This last result is consistent with the interpretation that the increased distribution of SPLI in the neonatally nerve damaged rats is due, at least partially, to reorganization of primary afferents.
Modulation of cone horizontal cell activity in the teleost fish retina. II. Role of interplexiform cells and dopamine in regulating light responsiveness
Authors: Yang XL , Tornqvist K , Dowling JE
Published: March 2018

Following the destruction of the terminals of the dopaminergic interplexiform cells by intraocular injections of 6-hydroxydopamine (6- OHDA), cone horizontal cells exhibited high light responsiveness in prolonged darkness and their responses to moderate and bright full- field flashes were as large as 60 mV. Furthermore, the light responsiveness of these cells in the 6-OHDA-treated retinas was not enhanced by background illumination. The application of dopamine (50 microM) by superfusion to 6-OHDA-treated retinas resulted in a decrease in light responsiveness and changes in response waveform of the cone horizontal cells. Twenty minutes following dopamine application the responses of the cone horizontal cells closely resembled the response of cells recorded in prolonged dark-adapted retinas. Dopamine caused similar changes in cone horizontal cells recorded in light-exposed retinas, but had no obvious effects on rod horizontal cells. The selective dopamine D1 receptor antagonist, Sch 23390, enhanced cone horizontal cell responsiveness when applied to prolonged dark-adapted retinas, mimicking background illumination. The light responsiveness of cone horizontal cells recorded after application of Sch 23390 was less than that for cells in retinas that had been exposed to background lights, but light responsiveness could not be further enhanced by background illumination. Another dopamine antagonist, (+)-butaclamol, was found to have effects similar to Sch 23390 on cone horizontal cells, but (-)-butaclamol, the inactive enantiomer, did not enhance the light responsiveness of these cells. The results suggest that the dopaminergic interplexiform cells play a crucial role in the regulation of cone horizontal cell responsiveness by prolonged darkness and background illumination. These cells may release dopamine tonically in the dark, which suppresses cone horizontal cell responsiveness. Background illumination may decrease dopamine release and liberate cone horizontal cells from the suppression.
Nonregenerative axonal growth within the mature mammalian brain: ultrastructural identification of sympathohippocampal sprouts
Authors: Crutcher KA , Marfurt CF
Published: March 2018

Damage to septohippocampal neurons in the adult rat results in sprouting of sympathetic axons into the denervated hippocampal formation. However, the distribution of sympathohippocampal fibers has only been assessed with light microscopic techniques, and it is not known if the sprouted fibers leave the blood vessels, along which they migrate into the hippocampal formation, to enter the hippocampal neuropil and, if they do, whether they form synaptic contacts with central neurons. Using the tetramethylbenzidine technique to visualize anterogradely transported wheat germ agglutinin-horseradish peroxidase conjugate, we identified sprouted sympathetic fibers in the hippocampal formation at both the light and electron microscopic level in albino rats receiving medial septal lesions. The majority of labeled fibers were observed within the regions immediately above and below the granule cell layer. Although most of the labeled sprouts were observed in association with intraparenchymal blood vessels, where they were usually apposed to the basal lamina, approximately a third of the labeled profiles were present within the neuropil with no obvious vascular relationships. Most of the profiles were identified as unmyelinated axons or vesicle-filled varicosities. Many of the latter structures contained small dense-cored vesicles, but in our sample none of the labeled profiles were observed to form membrane specializations with adjacent structures, and many were partly surrounded by presumed astrocytic processes. These results document the invasion of the CNS by sprouting axons of peripheral origin indicating that axonal elongation from uninjured neurons can occur within the mature mammalian CNS under certain circumstances. In addition, the presence of significant numbers of sympathetic fibers within the hippocampal neuropil indicates that they may be in a strategic position to influence hippocampal function.
Regional distribution of cGMP-activated ion channels in the plasma membrane of the rod photoreceptor
Authors: Watanabe S , Matthews G
Published: March 2018

In patch-clamp recordings from excised membrane patches, the distribution of cGMP-activated channels in the plasma membrane of the rod photoreceptor was examined. These channels have been shown to be the light-sensitive channels that carry the inward dark current in the rod outer segment, where phototransduction occurs (Matthews, 1987); thus, they are centrally involved in photoreceptor transduction. In the outer segment, cGMP-activated channels were present at high density. In the inner segment, cGMP-activated channels were also present, but their density was much lower than in the outer segment. Calcium-activated potassium channels had the opposite distribution, with a higher density in the inner segment. The results suggest that there is a barrier preventing the unrestricted spread of membrane proteins between the inner and outer segments.
Molecular evidence for early activity-dependent development of hamster motor neurons
Authors: Kalb RG , Hockfield S
Published: March 2018

Monoclonal antibody Cat-301 recognizes a surface antigen on subsets of neurons in many areas of the mammalian CNS, including Y-cells in the cat dorsal lateral geniculate nucleus (LGN) and spinal motor neurons in several species. We have shown previously that the Cat-301 antigen is expressed relatively late in postnatal development, with a time course that matches that of late developmental features of both LGN and spinal cord neurons. The developmental time course of the expression of the Cat-301 antigen in the LGN correlates with the period during which visual deprivation can alter LGN Y-cells. Neonatal visual deprivation (by monocular lid suture or dark rearing) suppresses Cat-301 expression on Y-cells, while deprivation in adult animals has no effect on antigen expression. These results suggested that the onset of Cat-301 immunoreactivity provides a positive molecular marker for the end of a critical period in cat visual system development. To determine if Cat- 301 might be a general marker for experience-dependent development in other areas of the CNS we have examined antigen expression on hamster spinal cord motor neurons. Here, we report that Cat-301 immunoreactivity develops on hamster motor neurons between postnatal days 7 and 14. Altering neuromuscular activity by sciatic nerve crush or thoracic hemicordotomy inhibits Cat-301 expression on motor neurons if performed before the onset of Cat-301 immunoreactivity. Other motor neuron antigens are unaffected by this procedure. In adult animals, nerve crush or cordotomy has no effect on Cat-301 immunoreactivity, demonstrating that Cat-301 expression is not simply dependent on ongoing neural activity. These observations suggest that motor neurons, like LGN neurons, require some pattern of neuronal activity during a critical period in development and, further, that the phenotypic changes in neurons consequent to early activity are reflected by the expression of specific molecules. The identification and characterization of such molecules may yield a description of the molecular mechanisms of experience-dependent development.
Microtubule polarity and distribution in teleost photoreceptors
Authors: Troutt LL , Burnside B
Published: March 2018

We have characterized the polarity orientation of microtubules in teleost retinal photoreceptors. The highly polarized rods and cones contain large numbers of paraxially aligned microtubules and exhibit dramatic cell shape changes. The myoid portion of the inner segments of both rods and cones undergoes contraction and elongation in response to light or circadian signals. Previous studies in our laboratory have demonstrated that in cones but not rods myoid elongation is microtubule- dependent. To determine polarity orientation, we decorated microtubules in photoreceptors of the green sunfish Lepomis cyanellus, with hooks formed from either exogenous or endogenous tubulin subunits. The direction of curvature of the attached hooks in cross section indicates microtubule polarity orientation by allowing one to determine the relative positions within the cell of the plus (fast-growing) and minus (slow-growing) ends of the microtubules. We found that virtually all cytoplasmic microtubules in photoreceptors are oriented with plus ends directed toward the synapse and minus ends toward the basal body at the base of the outer segment. Axonemal microtubules in photoreceptor outer segments are oriented with minus ends toward the basal body as in cilia and flagella. We have suggested previously that cone myoid elongation is mediated by mechanochemical sliding between microtubules. The polarity observations reported here indicate that if microtubules do slide in cones, sliding would necessarily occur between microtubules of parallel orientation as is observed in cilia and flagella.
Oligodendrocytes and CNS myelin are nonpermissive substrates for neurite growth and fibroblast spreading in vitro
Authors: Schwab ME , Caroni P
Published: March 2018

To study the interaction of neurons with CNS glial cells, dissociated sympathetic or sensory ganglion cells or fetal retinal cells were plated onto cultures of dissociated optic nerve glial cells of young rats. Whereas astrocytes favored neuron adhesion and neurite outgrowth, oligodendrocytes differed markedly in their properties as neuronal substrates. Immature (O4+, A2B5+, GalC-) oligodendrocytes were frequently contacted by neurons and neurites. In contrast, differentiated oligodendrocytes (O4+, A2B5-, GalC+) represented a nonpermissive substrate for neuronal adhesion and neurite growth. When neuroblastoma cells or 3T3 fibroblasts were plated into optic nerve glial cultures, the same differences were observed; differentiated oligodendrocytes were nonpermissive for cell adhesion, neurite growth, or fibroblast spreading. These nonpermissive oligodendrocytes were characterized by a radial, highly branched process network, often contained myelin basic protein, and may, therefore, correspond to cells actively involved in the production of myelin-like membranes. Isolated myelin from adult rat spinal cord was adsorbed to polylysine-coated culture dishes and tested as a substrate for peripheral neurons, neuroblastoma cells, or 3T3 cells. Again, cell attachment, neurite outgrowth, and fibroblast spreading was strongly impaired. General physicochemical properties of myelin were not responsible for this effect, since myelin from rat sciatic nerves favored neuron adhesion and neurite growth as well as spreading of 3T3 cells. These results show that differentiated oligodendrocytes express nonpermissive substrate properties, which may be of importance in CNS development or regeneration.
Differential responsiveness of cerebellar Purkinje neurons to GABA and benzodiazepine receptor ligands in an animal model of hepatic encephalopathy
Authors: Basile AS , Gammal SH , Mullen KD , Jones EA , Skolnick P
Published: March 2018

The role of the GABA-benzodiazepine receptor complex in the pathogenesis of hepatic encephalopathy was investigated by recording the electrophysiological responses of single cerebellar Purkinje neurons from rabbits with hepatic encephalopathy due to galactosamine- induced fulminant hepatic failure. Both the GABAmimetic muscimol and the benzodiazepine receptor agonist flunitrazepam were 3–4 times more potent in depressing the spontaneous activity of Purkinje neurons from rabbits with hepatic encephalopathy than from control animals. Furthermore, qualitatively different responses of Purkinje neurons to benzodiazepine receptor antagonists (Ro 15–1788 and Ro 14–7437) were found in controls and rabbits with hepatic encephalopathy. These compounds markedly excited Purkinje neurons from rabbits with hepatic encephalopathy, but had either no effect (Ro 14–7437) or partially suppressed (Ro 15–1788) the spontaneous activity of neurons from control animals. In addition, incubation of Purkinje neurons from rabbits with hepatic encephalopathy with subthreshold concentrations of Ro 14–7437 reduced their sensitivity to muscimol, whereas treatment of control neurons with Ro 14–7437 had no effect on their sensitivity to muscimol. Finally, Purkinje neurons from hepatic encephalopathy and control rabbits displayed no difference in sensitivity to the depressant actions of the alpha-adrenoceptor agonist phenylephrine. These findings demonstrate a differential responsiveness of Purkinje neurons from an animal model of hepatic encephalopathy to ligands that interact with the GABA-benzodiazepine receptor complex. Furthermore, the observations made in this experimental model are consistent with the involvement of the GABA-benzodiazepine receptor complex in mediating hepatic encephalopathy, and provide a potential explanation for the reported efficacy of benzodiazepine receptor antagonists in ameliorating this syndrome.
Electrophysiology and dye-coupling are sexually dimorphic characteristics of individual laryngeal muscle fibers in Xenopus laevis
Authors: Tobias ML , Kelley DB
Published: March 2018

Sex differences at the laryngeal neuromuscular junction of Xenopus laevis were examined by recording intracellularly from muscle fibers in response to nerve stimulation. Male laryngeal muscle contains 2 physiologically distinct fiber types. Type I fibers generate postsynaptic potentials in response to low-magnitude stimulus pulses and action potentials in response to higher-magnitude stimulus pulses. Type II muscle fibers require repetitive stimulation for action potential production, probably because of facilitation. Subthreshold events in type I and II fibers suggest that these neuromuscular synapses have low safety factor junctions. Female laryngeal muscle contains one fiber type (III), which is physiologically distinct from those found in the male. Type III fibers produce an action potential in response to a single-stimulus pulse of suprathreshold voltage delivered to the laryngeal nerve; subthreshold events were not observed. Iontophoretic injection of Lucifer yellow into a single female muscle fiber resulted in as many as 43 labeled fibers. In males, only one fiber was labeled. Dye-coupling was not observed in adult females treated with the androgenic steroid hormone, testosterone. We have previously reported that laryngeal muscle fibers are recruited throughout a stimulus train presented to the laryngeal nerve in males, but are not recruited in females (Tobias and Kelly, 1987). Sex differences in the frequency of electrophysiological fiber types described here may account for sex differences in fiber recruitment. Synchronous activity of dye-coupled fibers may increase the effectiveness of muscle contraction in females.
Morphological basis of short-term habituation in Aplysia
Authors: Bailey CH , Chen M
Published: March 2018

We have explored the morphological basis of the synaptic depression that underlies short-term habituation of the gill-withdrawal reflex in Aplysia by examining the fine structure of the presynaptic terminals of identified sensory neurons--a critical site of plasticity for the biochemical and biophysical changes that underlie this elementary form of learning. The structure of sensory neuron synapses from control (unstimulated) cells was compared with that of sensory neuron synapses from cells in which synaptic transmission had been depressed by repeated activation. We focused our analysis, as we had in an earlier study of long-term memory (Bailey and Chen, 1983), on the morphology of active zones at sensory neuron synapses. We found that both the incidence and size of serially reconstructed active zones were not changed in cells exposed to short-term habituation. This contrasts sharply with the reduction in both the frequency and surface area of sensory neuron active zones that accompanies long-term habituation, and suggests that modulation of active zone number and size may be an anatomical correlate that lies in the long-term domain. A quantitative analysis of the relationship between the active zone and nearby vesicle populations revealed a possible morphological substrate for the homosynaptic depression that underlies short-term habituation. Habituation leads to a depletion of synaptic vesicles immediately adjacent to the active zone. The ratio of this readily releasable pool of vesicles to the total population of vesicles associated with the active zone is 28% for control terminals, but only 11.5% for habituated terminals.(ABSTRACT TRUNCATED AT 250 WORDS)
Monocularly deprived cats: binocular tests of cortical cells reveal functional connections from the deprived eye
Authors: Freeman RD , Ohzawa I
Published: March 2018

Animals that are deprived of vision in one eye during a vulnerable phase of development lose visual function of the eye. Although this phenomenon has been studied extensively, little is known about the mechanism of disconnection of the deprived eye from visual cortex. One fundamental question is whether input remains from that eye. We have examined the hypothesis that there is functional input from a deprived eye to visual cortex that cannot be observed with standard alternate tests of each eye. We have employed a robust visual stimulation procedure in which large sinusoidal gratings are presented to each eye, as well as to both eyes together, at varying relative phases or retinal disparities. Monocular and binocular stimulation was used to test kittens unilaterally deprived for brief, intermediate, or long periods. A fourth group of kittens was studied as normal controls. Standard methods were used to record from single cells in the striate cortex. After initial qualitative exploration of receptive fields, all testing and analysis were quantitative. As expected, monocular tests revealed that, for most cells, the deprived eye was ineffective, i.e., did not activate the unit. This effect was increasingly pronounced as the length of deprivation was increased. However, binocular tests revealed that a large fraction of these cells (30–40%) was clearly influenced by and therefore functionally connected to the deprived eye. This interaction was phase-selective, or suppressive and not selective for phase. There was no indication that the connections that remained were of a specific type, i.e., excitatory or inhibitory. Therefore, excitation and inhibition appear equally resistant to the effects of monocular deprivation. However, with long-term deprivation, we find minimal evidence of functional input from the deprived eye. We conclude that the effects of monocular deprivation occur over a considerably longer time period than was previously thought.
Voltage-gated potassium channels in larval CNS neurons of Drosophila
Authors: Solc CK , Aldrich RW
Published: March 2018

The availability of genetic, molecular, and biophysical techniques makes Drosophila an ideal system for the study of ion channel function. We have used the patch-clamp technique to characterize voltage-gated K+ channels in cultured larval Drosophila CNS neurons. Whole-cell currents from different cells vary in current kinetics and magnitude. Most of the cells contain a transient A-type 4-AP-sensitive current. In addition, many cells also have a more slowly inactivating TEA-sensitive component and/or a sustained component. No clear correlation between cell morphology and whole-cell current kinetics was observed. Single- channel analysis in cell-free patches revealed that 3 types of channels, named A2, KD, and K1 can account for the whole-cell currents. None of these channels requires elevated intracellular calcium concentration for activation. The A2 channels have a conductance of 6–8 pS and underlie the whole-cell A current. They turn on rapidly, inactivate in response to depolarizing voltage steps, and are completely inactivated by prepulses to -50 mV. The KD (delayed) channels have a conductance of 10–16 pS and can account, in part, for the more slowly inactivating component of whole-cell current. They have longer open times and activate and inactivate more slowly than the A2 channels. The K1 channels have a slope conductance, measured between 0 and +40 mV, of 20–40 pS. These channels do not inactivate during 500 msec voltage steps and thus can contribute to the sustained component of current. They exhibit complex gating behavior with increased probability of being open at higher voltages. Although the K1 channels are sufficient to account for the noninactivating component of whole- cell current, we have observed several other channel types that have a similar voltage dependence and average kinetics.