bluuebunny/crossref_metadata_2025 huggingface dataset:
Opiate and peptide inhibition of transmitter release in parasympathetic nerve terminals
Authors: Gray DB , Pilar GR , Ford MJ
Published: March 2018

Somatostatin, morphine, and opioids inhibit transmitter release at intact neuromuscular junctions between ciliary ganglion neurons and the choroidal smooth muscle of the chick eye. Somatostatin and morphine, however, have no effect on release from terminals on the striated muscle target of the ciliary ganglion, the iris. In neuronal terminals of both the choroid and the iris, a high-affinity Na+-dependent choline uptake-mediated ACh synthesis is present at hatching. Both tissues exhibit a basal release of 3H-ACh which is potentiated severalfold during a 5 minute incubation in 55 mM K+ Tyrodes. Fifty percent of the basal release and 100% of the stimulated release are Ca2+ dependent and probably mediated through N-like voltage-dependent Ca2+ channels. Co- incubation of the choroid with 10 microM morphine sulfate blocks approximately 90% of the stimulated release. The same effect is seen with 100 nM somatostatin, 10 microM dynorphin, and 100 microM met- enkephalin arginine phenylalanine. Preincubation of the excised choroid with pertussis toxin (200 ng/ml) reverses the inhibitory effects of both morphine and somatostatin. In contrast, 3H-ACh release from terminals in the striated iris is not affected by either morphine or somatostatin at micromolar levels. These results suggest that both opiate and somatostatin receptors are present in the choroid target and that they may act through a final common pathway to modulate ACh release via G proteins. Second messengers such as cyclic AMP or diacylglycerol do not appear to mediate these effects; neither increasing cAMP levels in terminals nor activation of protein kinase C affects evoked release or its inhibition by morphine or other neuromodulators. It is unclear whether endogenous neuromodulation occurs in this system, although somatostatin-like immunoreactivity can be demonstrated in terminals of choroid neurons.
Mechanisms of contour perception in monkey visual cortex. I. Lines of pattern discontinuity
Authors: von der Heydt R , Peterhans E
Published: March 2018

We have studied the mechanism of contour perception by recording from neurons in the visual cortex of alert rhesus monkeys. In order to assess the relationship between neural signals and perception, we compared the responses to edges and lines with the responses to patterns in which human observers perceive a contour where no line or edge is given (anomalous contour), such as the border between gratings of thin lines offset by half a cycle. With only one exception out of 60, orientation-selective neurons in area V1 did not signal the anomalous contour. Many neurons failed to respond to this stimulus at all, others responded according to the orientation of the grating lines. In area V2, 45 of 103 neurons (44%) signaled the orientation of the anomalous contour. Sixteen did so without signaling the orientation of the inducing lines. Some responded better to anomalous contours than to the optimum bars or edges. Preferred orientations and widths of tuning for anomalous contour and bar or edge were found to be highly correlated, but not identical, in each neuron. Similar to perception, the neuronal responses depended on a minimum number of lines inducing the contour, but not so much on line spacing, and tended to be weaker when the lines were oblique rather than orthogonal to the border. With oblique lines, the orientations signaled were biased towards the orientation orthogonal to the lines, as in the Zollner illusion. We conclude that contours may be defined first at the level of V2. While the unresponsiveness of neurons in V1 to this type of anomalous contour is in agreement with linear filter predictions, the responses of V2 neurons need to be explained. We assume that they sum the signals of 2 parallel paths, one that defines edges and lines and another that defines anomalous contours by pooling signals from end-stopped receptive fields oriented mainly orthogonal to the contour.
Mechanisms of contour perception in monkey visual cortex. II. Contours bridging gaps
Authors: Peterhans E , von der Heydt R
Published: March 2018

We have studied the mechanism of contour perception by recording from neurons in the visual cortex of alert rhesus monkeys. We used stimuli in which human observers perceive anomalous contours: A moving pair of notches in 2 bright rectangles mimicked an overlaying dark bar. For control, the notches were closed by thin lines so that the anomalous contours disappeared or half of the figure was blanked, with a similar effect. Orientation-selective neurons were studied. With the receptive fields centered in the gap, 23 of 72 (32%) neurons tested in area V2 responded to the moving “bar” even though the stimulus spared their response fields, and when the notches were closed, their responses were reduced or abolished. Likewise, when half of the figure was removed, the neurons usually failed to respond. Neurons with receptive fields within 4 degrees of the fovea signaled anomalous contours bridging gaps of 1 degree-3.5 degrees. The anomalous-contour responses were compared quantitatively with response field profiles and length-summation curves and found to exceed the predictions by linear-summation and summation- to-threshold models. Summation models also fail to explain the effect of closing lines which add only negligible amounts of light. In V1, only one of 26 neurons tested showed comparable responses, and only with a narrow gap. The others responded only when the stimulus invaded the response field and did not show the effect of closing lines, or failed to respond at all. The contour responses in V2, the nonadditivity, and the effect of closure can be explained by the previously proposed model (Peterhans et al., 1986), assuming that the corners excite end-stopped fields orthogonal to the contour whose signals are pooled in the contour neurons.
Appearance and transient expression of oxytocin receptors in fetal, infant, and peripubertal rat brain studied by autoradiography and electrophysiology
Authors: Tribollet E , Charpak S , Schmidt A , Dubois-Dauphin M , Dreifuss JJ
Published: March 2018

The development of oxytocin (OT) receptors in the rat brain and spinal cord was studied by in vitro light microscopic autoradiography and by electrophysiology. OT receptors were labeled using a monoiodinated OT antagonist in tissue sections from animals aged between embryonic day 12 (E12) and postnatal day 90 (PN90); the response of ongoing spike activity to the addition of OT was assessed in neurons located in the dorsal motor nucleus of the vagus nerve of the neonate. Specific binding was detected first at E14 in a region that later differentiated into the dorsal motor nucleus of the vagus nerve. Many other regions were progressively labeled between E20 and PN5. From PN5 to PN16, the distribution of binding sites remained essentially unchanged but differed markedly from that characteristic of the adult. The change- over from the “infant pattern” to the “adult pattern” occurred in 2 stages: the first change took place between PN16 and PN22, a time corresponding to the preweaning period; the second change occurred after PN35 and thus coincided with the onset of puberty. During the first transition period, binding was reduced or disappeared in several areas intensely labeled at earlier stages, in particular, in the cingulate cortex and the dorsal hippocampus. At the same time, binding sites appeared in the ventral hippocampus. At puberty, high densities of OT binding sites appeared in the ventromedial hypothalamic nucleus and the olfactory tubercle. Electrophysiological activity was recorded from vagal neurons in slices obtained from animals sacrificed at PN1- PN12. OT and a selective OT agonist reversibly increased the firing rate of these neurons in a concentration-dependent manner. The neuronal responsiveness was similar to that reported previously in the adult. These results suggest that OT binding sites detected by autoradiography in the developing rat brain represent, at least in some areas, functional neuronal receptors.
Acetylcholine in the crayfish optic lobe: concentration profile and cellular localization
Authors: Wang-Bennett LT , Pfeiffer C , Arnold J , Glantz RM
Published: March 2018

The crayfish optic lobe contains high levels of acetylcholine (ACh) and choline as measured with a chemiluminescent assay in small fragments of optic lobe tissue. The highest concentrations were found in the medulla externa and medulla interna (second and third optic neuromeres), which have ACh concentrations of 270 pmol/mg tissue. This concentration is about 16 times that measured in the photoreceptors and lamina ganglionaris (the first optic neuromere). Immunocytochemistry (based upon antisera to choline-glutaryl-BSA) revealed low levels of ACh-like reactivity in the lamina ganglionaris associated with the terminal arbors of centrifugal and/or tangential neurons. The most intense ACh- like reactivity was observed in monopolar neurons of the medulla externa and medulla interna. One monopolar neuron/medullary column (or about 2500 neurons/medullary neuropile) exhibited reactivity and an estimated cytoplasmic concentration of 8.1 mM.
The relationship between the number of synaptic vesicles and the amount of transmitter released
Authors: Koenig JH , Kosaka T , Ikeda K
Published: March 2018

The relationship between the number of synaptic vesicles and the amount of transmitter released from identified synapses was investigated in the dorsal longitudinal flight muscle (DLM) of the temperature- sensitive endocytosis mutant of Drosophila melanogaster, shibirets- 1(shi). In the shi fly at 29 degrees C, vesicle recyling is blocked, but transmitter release proceeds normally. Thus, by inducing transmitter release at 29 degrees C, shi synapses gradually become depleted of synaptic vesicles. In this way it was possible to regulate the number of vesicles in a synapse. Intracellular recordings were made from individual fibers of the DLM in shi flies after various periods at 29 degrees C while stimulating at 0.5 Hz. The amplitude of the evoked excitatory junction potential (ejp), gradually decreased with longer exposure and was brought to various levels. The fiber was then rapidly fixed for electron microscopy. The number of vesicles per synapse was compared with the amplitude of the ejp at the time of fixation. It was observed that the smaller the ejp amplitudes became, the fewer vesicles were in the synapses. Also, as the ejp amplitude decreased, an increased number of synapses contained no vesicles. It is concluded that synaptic vesicles are directly involved in the release process.
Differential localization of microtubules in cerebellar cells
Authors: Ginzburg I , Behar L , Littauer UZ , Griffin WS
Published: March 2018

The distribution and subcellular localization of microtubules in rat brain cerebellum was analyzed by immunohistochemistry with antibodies prepared against 3 synthetic peptides corresponding to the C-terminal region of beta-tubulin. The peptides used correspond to amino acid positions 416–425 (peptide 1), 416–431 (peptide 2), and 426–445 (peptide 4). The antibodies thus obtained displayed a remarkable specificity in reacting with different cell types in the rat cerebellum. Antibodies directed against peptide 1 primarily stained Purkinje cells and their dendrites and axons. Peptide 2 antibodies preferentially stained the glomeruli, while antibodies directed against peptide 4 preferentially stained Bergmann glial fibers. These results are discussed in terms of dissimilarities in microtubule organization and masking of epitopes by microtubule-associated proteins (MAPs) in individual cerebellar cells, which may be related to specific functional properties.
Effects of DIDS on the chick retinal pigment epithelium. II. Mechanism of the light peak and other responses originating at the basal membrane
Authors: Gallemore RP , Steinberg RH
Published: March 2018

4,4′-diisothiocyanostilbene-2,2′-disulfonate (DIDS) appears to block a Cl- conductance in the basal membrane of the chick retinal pigment epithelium (RPE) (Gallemore and Steinberg, 1989). The present paper describes the effect of DIDS, as well as effects of anion movement blockade with other pharmacological agents, on 3 responses that originate as depolarizations of the RPE basal membrane: the light peak of the DC electroretinogram, the response to sodium azide, and the response to a small transtissue hyperosmotic gradient. Intracellular and extracellular potentials, and measurements of subretinal [K+]o were obtained from an in vitro preparation of chick retina-RPE-choroid. Choroidal perfusion with each of 4 anion transport and Cl- channel blockers (DIDS, 4-acetamido-4′-isothiocyanostilbene, phenylanthranylic acid, and thiocyanate) suppressed the light peak. DIDS, the most potent agent, abolished the basal membrane light-peak depolarization. Choroidal perfusion with azide depolarized the basal membrane and decreased its apparent resistance, Rba, as in cat (Linsenmeier and Steinberg, 1987). The basal membrane depolarization, recorded extracellularly as an increase in the transtissue potential (TTP), was blocked by DIDS. Azide also increased RPE c-wave amplitude, which could be explained by the decrease in Rba, and it suppressed the light peak. As previously described (Shirao and Steinberg, 1987), a 25 mOsm transtissue hyperosmotic load (retinal side hyperosmotic) increased the TTP, increased the c-wave and decreased the transtissue resistance. DIDS suppressed each of these effects. We conclude that the light peak, the response to azide, and the response to a retinal hyperosmotic load may each originate as an increase in a basal membrane anion conductance that may be permeable to chloride.(ABSTRACT TRUNCATED AT 250 WORDS)
Peripheral target regulation of dendritic geometry in the rat superior cervical ganglion
Authors: Voyvodic JT
Published: March 2018

Dendritic arborizations of neurons in the adult rat superior cervical ganglion were measured in control ganglia and in ganglia innervating peripheral targets that were relatively larger or smaller than normal. The relative size of the target--the submandibular gland in these experiments--was manipulated during development by changing the ratio between the amount of target tissue and the number of innervating ganglion cells. Thus, ligating the submandibular salivary duct reduced the size of the gland, whereas partially denervating the gland produced a relatively larger target by making a smaller number of ganglion cells innervate a gland of normal size. Neurons innervating targets that were smaller than normal had significantly smaller dendritic arborizations and cell bodies than control cells. Conversely, neurons projecting to relatively larger than normal targets had larger dendritic arborizations and cell bodies, and more primary dendritic branches. Such cells were also innervated by a larger than normal number of preganglionic inputs. A similar change in dendritic geometry was observed when relative target size was increased after cutting the cervical sympathetic trunk, showing that target regulation of dendritic geometry is not dependent on ganglion cell activity or the presence of presynaptic innervation. Dendrites in the superior cervical ganglion normally grow in parallel with body size throughout life (Purves et al., 1986a; Voyvodic, 1987a). The present results imply that an important aspect of dendritic growth is an ongoing responsiveness of ganglion cells to feedback signals arising from the peripheral targets they innervate.
Distinct autophosphorylation sites sequentially produce autonomy and inhibition of the multifunctional Ca2+/calmodulin-dependent protein kinase
Authors: Lou LL , Schulman H
Published: March 2018

The multifunctional Ca2+/calmodulin-dependent protein kinase (multifunctional CaM kinase) may be an important mediator for neurotransmitters and hormones that utilize Ca2+ as a “second messenger.” We examined the ability of autophosphorylation to convert the multifunctional CaM kinase to a Ca2+/calmodulin-independent (autonomous) form to determine whether autophosphorylation is a mechanism for short- or long-term enhancement of Ca2+ action. As the kinase incorporates phosphate during continuous stimulation by Ca2+/calmodulin, its ability to phosphorylate exogenous substrates becomes increasingly autonomous. Withdrawal of Ca2+ after a critical level of phosphate incorporation is reached leads to a “burst” or rapid increase in Ca2+-independent autophosphorylation. The “burst” of autophosphorylation is distinct from the initial Ca2+-dependent autophosphorylation, however, since it inhibits substrate phosphorylation. Both Ca2+-dependent and Ca2+-independent substrate phosphorylation are inhibited by this autonomous autophosphorylation. Thus, autophosphorylation has a dual role in modulating the activity of multifunctional CaM kinase. It initially enables the kinase to continue phosphorylating substrates after Ca2+ levels decline, but it eventually suppresses this autonomous activity. Tryptic phosphopeptide mapping demonstrates that appearance of phosphothreonine-containing peptides is common to several conditions used to generate an autonomous enzyme. Sequencing reveals the critical “autonomy” site to be threonine286. The inhibitory mode of autophosphorylation involves 3 additional phosphopeptides containing a serine and a threonine residue.
Afferent basis of visual response properties in area MT of the macaque. I. Effects of striate cortex removal
Authors: Rodman HR , Gross CG , Albright TD
Published: March 2018

The middle temporal area (MT) of the macaque monkey is a region of extrastriate cortex involved in the analysis of visual motion. MT receives strong projections from striate cortex and from area V2, which is dependent on striate for visual responsiveness. Accordingly, the visual properties of MT neurons have been thought to reflect the further processing of its input from striate cortex. We examined the dependence of MT activity on pathways deriving from striate cortex by recording from MT neurons following removal of their striate input. Repeated recordings in area MT were made in 4 hemispheres of anesthetized macaques following either partial or total ablations of striate cortex. Cells in MT were tested for responsiveness, selectivity for direction of motion and direction tuning, and ocular dominance. Receptive fields were also plotted. In an additional animal, we recorded from MT neurons during reversible cooling of the central representation in striate cortex. We found that striate cortex removal or inactivation did not abolish the visual responsiveness of the majority of MT cells. Although the residual responses were generally much weaker than in the intact animal, direction selectivity and binocularity were still present. Moreover, receptive field size and overall topography appeared unaltered.
High-affinity uptake of L-kynurenine by a Na+-independent transporter of neutral amino acids in astrocytes
Authors: Speciale C , Hares K , Schwarcz R , Brookes N
Published: March 2018

L-Kynurenine (KYN), an intermediary product in the kynurenine pathway of tryptophan metabolism, is the common precursor from which are formed both quinolinic acid, a potent endogenous “excitotoxin,” and kynurenic acid, a nonselective antagonist of excitotoxins. The present work examines 3H-KYN transport in primary astrocyte cultures derived from the cerebra of newborn mice. Influx and efflux of 3H-KYN were attributable almost entirely to carrier-mediated transport. The tritium recovered in uptake experiments was identifiable as 3H-KYN, indicating a low rate of KYN metabolism during incubations up to 30 min. KYN uptake decreased in the presence of extracellular Na+, at least in part because KYN efflux was accelerated. Marked trans stimulation of KYN efflux by extracellular KYN provided evidence of the exchanging nature of the carrier. Saturation curves for the initial velocity of KYN uptake conformed to a 1-component saturable system with Km of 32 microM and Vmax of 2.1 nmol mg-1 protein min-1. KYN was notably concentrated by the astrocytes, with an estimated steady-state distribution ratio of 180-fold for 1 microM KYN. Analog inhibition studies showed that the KYN transporter exhibited a clear preference for large neutral amino acids; leucine, tryptophan, and phenylalanine were recognized with relatively higher affinity than KYN. In summary, KYN is concentratively transported into astrocytes by a Na+-independent exchanger with high affinity for branched-chain and aromatic neutral amino acids. The substrate specificity and high affinity of this transport system resemble the properties of neutral amino acid transport across the blood-brain barrier in the rat and human.
Synaptogenesis in the rat suprachiasmatic nucleus demonstrated by electron microscopy and synapsin I immunoreactivity
Authors: Moore RY , Bernstein ME
Published: March 2018

Synaptogenesis was studied in the rat suprachiasmatic nucleus (SCN) using quantitative ultrastructural analysis and synapsin I immunohistochemistry. SCN neurons are formed between embryonic days 13 and 17 (E13-E17), and the SCN is a distinct hypothalamic nucleus by E18. At E19 the nucleus is very immature and there are few synapses. Synaptogenesis proceeds slowly until P4 but increases rapidly between P4 and P10. At P10 the number of synapses per unit area is the same as in the adult SCN and all synaptic types present in the adult are evident. However, the SCN continues to increase in volume to the adult with approximately 30% of total synaptic number added between P10 and a young adult age. The appearance of synapsin I immunoreactivity correlates very precisely with the development of synapses in the SCN as shown by ultrastructural analysis between E19 and P6. The pattern of appearance of synapsin I immunoreactivity demonstrates that synaptogenesis in the SCN is significantly delayed in comparison to adjacent hypothalamic nuclei. Synapsin I immunohistochemistry is a reliable marker of synapse formation in the developing SCN. A correlation of these anatomical data with prior functional studies suggests that SCN neurons are born as individual circadian oscillators that undergo a rapid development in the first 10 days after birth to form a functional neural network subserving circadian rhythm generation and regulation.
Distribution and lateral mobility of GABA/benzodiazepine receptors on nerve cells
Authors: Velazquez JL , Thompson CL , Barnes EM , Angelides KJ
Published: March 2018

The distribution and mobility of GABA/benzodiazepine receptors has been examined on living nerve cells by fluorescence digital imaging and fluorescence photobleach recovery with 2 novel fluorescent derivatives of the benzodiazepines Ro7–1986 and 1012S. These fluorescent derivatives retain their high affinity for the GABA/benzodiazepine receptor complex with Ks of 79 and 85 nM, respectively. Digital fluorescence imaging of GABA/benzodiazepine receptors on rat spinal cord neurons maintained in culture reveal that, even in the absence of observable synaptic contact, GABA/benzodiazepine receptors are localized on the cell body in clusters of very high density. Receptor density on processes is lower than on cell bodies, although occasional areas of intense fluorescence were observed. Fluorescence photobleach recovery measurements show that 70% of the GABA/benzodiazepine receptors on the cell body and greater than 85% on the processes of rat spinal cord neurons are immobile. In comparison, voltage-gated sodium channels are diffusely distributed and very mobile on the cell body (D = 10(-9) cm2/sec). The results suggest that even in the absence of nerve-nerve cell contact, there are specific mechanisms that segregate GABA/benzodiazepine receptors to the neuronal cell body and control their lateral mobility.
Phasic activation of lateral geniculate and perigeniculate thalamic neurons during sleep with ponto-geniculo-occipital waves
Authors: Steriade M , Pare D , Bouhassira D , Deschenes M , Oakson G
Published: March 2018

Ponto-geniculo-occipital (PGO) waves are spiky field potentials generated in cholinergic nuclei of the dorsolateral mesopontine tegmentum just prior to and during rapid-eye-movement (REM) sleep and transferred toward thalamic nuclei. These events are commonly regarded as physiological correlates of oneiric behavior. We have examined the PGO-related discharges of physiologically identified neurons located in the dorsal lateral geniculate (LG) nucleus and perigeniculate (PG) sector of the reticular thalamic complex in chronically implanted, naturally sleeping cats. PGO focal waves and associated unit discharges were simultaneously recorded by the same microelectrode. PGO waves herald the other signs of REM sleep (EEG desynchronization and muscular atonia), appearing 30–90 sec before REM sleep over the EEG-synchronized activity of slow-wave sleep (pre-REM epoch). (1) Most PG neurons discharged bursts of action potentials in relation to PGO waves during both pre-REM and REM sleep. (2) The PGO-related activity of LG neurons was quite different. During the pre-REM stage, PGO waves correlated with a short (7–15 msec), high-frequency (300–500 Hz) spike burst of LG neurons, followed by a long (0.2–0.4 sec) train of single spikes, whereas during REM sleep, the PGO-related activity lacked the initial burst and consisted of a spike train that only slightly exceeded the tonically increased background firing of LG cells. The stereotyped characteristics of the PGO-related spike bursts during the pre-REM epoch suggest that they are the extracellular reflection of a low- threshold spike deinactivated by the tonic membrane hyperpolarization of LG cells associated with the EEG-synchronized sleep state. Such bursts are inactivated during the tonic depolarization of LG cells that occurs in REM sleep. The synchronous spike bursts discharged by LG cells in relation with the PGO waves of the pre-REM epoch probably underlie the much larger amplitude of the PGO waves of the pre-REM epoch as compared with those of the REM-sleep state. Since LG neurons have relatively low spontaneous firing rates during the EEG- synchronized pre-REM epoch, the PGO-related activity of this transitional stage leads to a higher signal-to-noise ratio in the visual thalamocortical channel than during REM sleep. We suggest that the PGO-related activity during the pre-REM epoch is related to vivid imagery during this stage of sleep.
Regulation of subtypes of beta-adrenergic receptors in rat brain following treatment with 6-hydroxydopamine
Authors: Johnson EW , Wolfe BB , Molinoff PB
Published: March 2018

The technique of quantitative autoradiography has been used to localize changes in the densities of subtypes of beta-adrenergic receptors in rat brain following treatment with 6-hydroxydopamine. Previously reported increases in the density of beta 1-adrenergic receptors in the cerebral cortex were confirmed. The anatomical resolution of autoradiography made it possible to detect changes in the density of beta 2-adrenergic receptors in the cortex and in a number of other brain regions. The density of beta 1-adrenergic receptors increased from 30 to 50% depending on the region of the cortex being examined. The increase in the somatomotor cortex was greater than that in the frontal or occipital cortex. The increase in the density of beta 2- adrenergic receptors in the cortex was not as widespread as that of beta 1-adrenergic receptors and occurred primarily in frontal cortex, where the density of receptors increased by 40%. The densities of both beta 1- and beta 2-adrenergic receptors increased in a number of forebrain, thalamic, and midbrain structures. Selective changes in the density of beta 1-adrenergic receptors were observed in the superficial gray layer of the superior colliculus and in the amygdala. The density of beta 2-adrenergic receptors increased in the caudate-putamen, the substantia nigra, and the lateral and central nuclei of the thalamus, whereas the density of beta 1-adrenergic receptors did not change in these regions. The densities of both subtypes of beta-adrenergic receptors increased in the hippocampus, the cerebellum, the lateral posterior nucleus of the thalamus, and the dorsal lateral geniculate.(ABSTRACT TRUNCATED AT 250 WORDS)
Macrophage function during Wallerian degeneration of rat optic nerve: clearance of degenerating myelin and Ia expression
Authors: Stoll G , Trapp BD , Griffin JW
Published: March 2018

This study examined Wallerian degeneration (WD) of rat optic nerve (ON) with 2 goals: to determine which cell types are involved in myelin degradation and clearance and to evaluate the extent to which Ia antigen, a product of the immune response genes, is expressed. We examined immunostained 1-micron and ultrathin cryosections of rat ON at 1, 8, and 16 weeks after nerve transection. Serial 1-micron cryosections were stained with monoclonal antibodies to rat Ia antigen (Ox6) and with antibodies that identify astrocytes (GFAP) and monocytes/macrophages (ED1). In normal ON, ED1-positive cells were not found. A few ED1-positive monocytes/macrophages were present in the transected ON at one week. Macrophages were prominent throughout the ON at 8 weeks; by 16 weeks their number was decreasing. These cells contained myelin debris in various stages of digestion. GFAP-positive astrocytes did not contain myelin debris in their cytoplasm. At all 3 times a subpopulation of the ED1-positive monocytes/macrophages expressed Ia antigen. Ia antigen was not detected in endothelial cells or GFAP-positive astrocytes. In ultrathin cryosections stained by immunogold procedures, Ia immunoreactivity was found exclusively in cells containing multiple vacuoles and myelin debris and lacking intermediate filaments. The same cell type was labeled by ED1 antibodies. Our results indicate that macrophages remove the vast majority of debris during Wallerian degeneration in the CNS; a proportion of these macrophages concomitantly express Ia antigen. These results suggest that the Ia expression by macrophages observed in other CNS disorders does not necessarily reflect specific local immune events, but in some instances can represent a nonspecific response to CNS damage.
Metabolic specialization in fast and slow muscle fibers of the developing rat
Authors: Nemeth PM , Norris BJ , Solanki L , Kelly AM
Published: March 2018

Individual fibers of prospective fast (extensor digitorum longus; EDL) and slow (soleus) muscles of rats have been analyzed to determine the profiles of key energy-generating enzymes at successive stages of postnatal development. Mean activities of lactate dehydrogenase (LDH) and adenylokinase (AK), 2 enzymes associated with contractile function, are significantly different in the 2 fiber populations at birth; furthermore, wide variations in enzyme activities exist among the individual fibers. There is a progressive refinement of enzyme levels in the soleus into a more uniform fiber population, while the fibers in the EDL progressively diverge into 2 distinct phenotypes. Changes in EDL and soleus are punctuated by periods of rapid change, with the period between 10 and 21 d being most eventful. Generally, the maturation profiles of LDH and AK coincide with the transition from neonatal to adult fast myosins and closely reflect the timing of energy demands imposed by contractile activity patterns. In contrast, activities of the oxidative enzymes malate dehydrogenase and beta- hydroxyacyl CoA dehydrogenase are similar in both muscles at birth and steadily increase during the first 3 weeks, suggesting a progressive adaptation to the aerobic extrauterine environment. After 30 d, there are differential changes in the oxidative profiles of enzymes for fatty acid and glucose metabolism. The profiles follow dietary changes associated with weaning, which suggests a phenotypic dependence of neonatal muscle on the particular available energy substrate. All enzymes are low in all fibers of EDL and soleus at birth, indicating their modest metabolic capacity.(ABSTRACT TRUNCATED AT 250 WORDS)
Nasotemporal overlap of crossed and uncrossed retinal ganglion cell projections in the Japanese monkey (Macaca fuscata)
Authors: Fukuda Y , Sawai H , Watanabe M , Wakakuwa K , Morigiwa K
Published: March 2018

The nasotemporal overlap of crossed and uncrossed retinal ganglion cell projections were studied in 11 Japanese monkeys (Macaca fuscata) using HRP and fluorescent dyes (DAPI and RITC) as retrograde tracers and by physiological recordings of antidromic field potentials. A strip of nasotemporal overlap ran orthogonal to the horizontal meridian in all the whole-mount retinas studied. In HRP-labeled retinas of 6 monkeys, the width of the overlap gradually increased from 0.6 degrees in the central retina up to 5 degrees at eccentricity of 5 mm, and to 15 degrees at the extreme periphery. We also noted a clear asymmetric distribution of crossed and uncrossed retinal ganglion cell projections particularly in the perifoveal region; ipsilaterally projecting cells encircled the nasal edge of the fovea, whereas few contralaterally projecting cells were observed in the temporal edge. Soma-size analysis revealed that crossed projections in the temporal portion of the overlap arose mainly from large and small cells (presumably P alpha and P gamma cells, respectively); uncrossed projections in its nasal portion arose from medium-sized cells (presumably P beta cell). Direct evidence of the overlap as well as of the asymmetry was obtained in subsequent fluorescent dye experiments in 3 monkeys. Physiological studies on 2 additional monkeys confirmed the widening of the nasotemporal overlap towards the upper and lower parts of the retina. Moreover, in the nasal portion of the overlap, only slow potentials, which presumably reflect activities of P beta cells, were recorded after stimulation of the ipsilateral LGN as expected from the morphological study. The findings are discussed in relation to clinical observations of macular sparing and splitting, and with regard to the functional differences between P alpha and P beta cell systems on which binocular stereoscopic vision along the midsagittal plane may be based.
Embryonic development of axon pathways in the Drosophila CNS. I. A glial scaffold appears before the first growth cones
Authors: Jacobs JR , Goodman CS
Published: March 2018

Three classes of glial cells are present early in embryogenesis and appear to play a major role in axon pathway formation in the Drosophila CNS. Six longitudinal glial (LG) cells are present over the longitudinal connective on each side of each segment. Six midline glia (MG) cells surround the anterior and posterior commissures of each segment. Finally, the intersegmental nerve root is covered by a glial cell: the segment boundary cell (SBC). All 3 classes of glial cells are present in their final position before axon outgrowth and their pattern prefigures the first axon pathways. The pioneer growth cones that establish the first axon pathways in the longitudinal connective and intersegmental nerve extend along the elongate surface of the LG and SBC glial cells; the pioneer growth cones for the anterior and posterior commissures extend toward and make close contact with the end feet of the MG glial cells. Later, all 3 classes of glial cells enwrap the axon tracts in much the same way as vertebrate oligodendrocytes. The results suggest that these early glial cells provide guidance cues for the first growth cones in the Drosophila CNS. More than simply providing a permissive substrate, the differential extension of specific early growth cones towards either the MG cells or along the LG cells suggests an active role for these glia in growth cone guidance.
Embryonic development of axon pathways in the Drosophila CNS. II. Behavior of pioneer growth cones
Authors: Jacobs JR , Goodman CS
Published: March 2018

We have identified the neurons that pioneer the major CNS axon tracts in the Drosophila embryo and determined their trajectory and fasciculation choices using serial section electron microscopy. Although Drosophila pioneer neurons make choices similar to those of their grasshopper homologs, there are interesting differences that reflect the much smaller nervous system size and the much faster rate of development characteristic of Drosophila. For example, where 2 longitudinal tracts are pioneered independently in grasshopper, only one is formed in Drosophila. This change is due to a change in fasciculation affinity of the pCC growth cone. Additionally, the intersegmental (IS) nerve is pioneered by a different neuron in Drosophila (aCC) than in the grasshopper (U1) because the smaller Drosophila CNS places the IS nerve within filopodial reach of the aCC soma, while in the grasshopper it is not. Drosophila growth cones explore a much more confined neuropil volume than do grasshopper growth cones but can also sample a larger fraction of the CNS as well. For this reason, some cell-cell recognition events critical to pathfinding in the grasshopper embryo may not be as essential in Drosophila. Nevertheless, many specific cellular affinities have been retained through the evolutionary divergence of these 2 species.
Characterization of nicotinic receptors in chick retina using a snake venom neurotoxin that blocks neuronal nicotinic receptor function
Authors: Loring RH , Aizenman E , Lipton SA , Zigmond RE
Published: March 2018

Nicotinic receptor function has been described in the retinas of a variety of vertebrate species. Neuronal bungarotoxin (NBT, also known as bungarotoxin 3.1, toxin F, or kappa-bungarotoxin) blocks nicotinic receptors in several neuronal preparations, while the neuromuscular antagonist alpha-bungarotoxin (BGT) fails to block most of these receptors. NBT (100 nM), but not BGT (10 microM), substantially blocks nicotinic function on ganglion cells in intact chick retina. 125I-NBT binds to 2 sites in homogenates of chick retina; one site that is shared with BGT (Kd = 5–7 nM, Bmax approximately 500 fmol/retina) and one which is not (Kd = 2–3 nM, Bmax approximately 100 fmol/retina). 125I-NBT binding to the NBT-specific site (binding in the presence of 1 microM unlabeled BGT) is localized to 2 bands in the inner plexiform layer, corresponding to regions richly innervated by neurons containing immunoreactivity for choline acetyltransferase. Furthermore, this binding is blocked by competitive nicotinic agonists and antagonists, but nicotine or other nicotinic agonists do not displace 125I-NBT binding with very high affinity relative to the displacement of 3H- nicotine reported by others in brain. Thus, of the 2 NBT binding sites, the site not recognized by BGT most likely represents functional nicotinic receptors in the chick retina, but these receptors have relatively low affinity for nicotinic agonists, similar to nicotinic receptors found in autonomic ganglia.
Columnar specificity of intrinsic horizontal and corticocortical connections in cat visual cortex
Authors: Gilbert CD , Wiesel TN
Published: March 2018

A prominent and stereotypical feature of cortical circuitry in the striate cortex is a plexus of long-range horizontal connections, running for 6–8 mm parallel to the cortical surface, which has a clustered distribution. This is seen for both intrinsic cortical connections within a particular cortical area and the convergent and divergent connections running between area 17 and other cortical areas. To determine if these connections are related to the columnar functional architecture of cortex, we combined labeling of the horizontal connections by retrograde transport of rhodamine-filled latex microspheres (beads) and labeling of the orientation columns by 2- deoxyglucose autoradiography. We first mapped the distribution of orientation columns in a small region of area 17 or 18, then made a small injection of beads into the center of an orientation column of defined specificity, and after allowing for retrograde transport, labeled vertical orientation columns with the 2-deoxyglucose technique. The retrogradely labeled cells were confined to regions of orientation specificity similar to that of the injection site, indicating that the horizontal connections run between columns of similar orientation specificity. This relationship was demonstrated for both the intrinsic horizontal and corticocortical connections. The extent of the horizontal connections, which allows single cells to integrate information over larger parts of the visual field than that covered by their receptive fields, and the functional specificity of the connections, suggests possible roles for these connections in visual processing.
Voltage-dependent calcium channels regulate melatonin output from cultured chick pineal cells
Authors: Harrison NL , Zatz M
Published: March 2018

Chick pineal cells maintained in primary culture display a circadian rhythm of melatonin production and release, and the nocturnal increase in melatonin output is enhanced by elevating extracellular K+. The divalent cations, Co2+, Cd2+, and Mn2+, each reduce nocturnal melatonin output. Nitrendipine and nifedipine also prevent the nocturnal rise in melatonin output, while Bay K 8644 increases it, suggesting a role for voltage-dependent Ca2+ channels in regulating melatonin output. The whole-cell patch-clamp technique was used to record from individual chick pineal cells. Under conditions designed to isolate currents through voltage-dependent Ca2+ channels, biphasic inward currents are elicited by large depolarizing commands (e.g., to 0 mV) from a holding potential of -90 mV; from a holding potential of -40 mV, only a sustained inward current is elicited by steps to 0 mV. Both components of the inward current are blocked by Co2+ or Cd2+. The sustained current is increased in amplitude by Bay K 8644 and blocked by nifedipine, while the transient current is unaffected. Since there is no evidence for vesicular release of melatonin, the “L-type” calcium channels mediating the sustained calcium current appear to be involved in the pathways regulating melatonin synthesis in chick pineal cells.
Ultrastructural basis for interactions between central opioids and catecholamines. II. Nuclei of the solitary tracts
Authors: Pickel VM , Chan J , Milner TA
Published: March 2018

Interactions between central opioids and catecholamines are thought to underlie the ability of adrenergic agonists both to lower blood pressure and alleviate certain symptoms of opiate withdrawal. We examined the cellular substrate for interactions between neurons containing enkephalin-like opioid peptides and catecholamines in cardiovascular portions of the medial nuclei of the solitary tracts (m- NTS) of adult rats. Single sections were dually labeled using a double- bridged peroxidase method for the localization of a monoclonal leucine (Leu5)-enkephalin-antibody and immunoautoradiography for the localization of polyclonal antibodies against the catecholamine- synthesizing enzyme tyrosine hydroxylase (TH). Light microscopy revealed a few perikarya and numerous varicosities containing Leu5- enkephalin-like immunoreactivity (LE-LI). These were distributed among TH-labeled perikarya and processes throughout the rostrocaudal NTS. Electron microscopy of the m-NTS at the level of the area postrema further established the single as well as dual localization of TH and LE-LI in individual perikarya, dendrites, and axon terminals. Silver grains indicative of TH-labeling were usually distributed throughout the cytoplasm, whereas the peroxidase reaction product for LE-LI was localized principally to large (80–150 nm), dense-core vesicles. Immunoautoradiographic labeling for TH was detected in 118 terminals within a series of sections containing 183 terminals with LE-LI. Of these, 26% of the TH-labeled terminals and 32% of the enkephalin- containing terminals formed symmetric synapses with unlabeled dendrites, while only 7% of each type formed symmetric synapses with TH- labeled dendrites. In favorable planes of sections, the unlabeled as well as TH-labeled dendrites received convergent input from both types of terminals. A few of the remaining terminals that contained either TH or LE-LI formed asymmetric junctions with unlabeled distal dendrites; the others were without recognizable synaptic specializations within the plane of section. Approximately 20% of the TH-labeled terminals and 6% of the terminals containing LE-LI were dually labeled for both antibodies. These were invested with astrocytic processes characterized by bundles of intermediate filaments. We conclude that within cardiovascular portions of the m-NTS, opioid peptides and catecholamines contained within the same or separate terminals modulate the activity of target neurons through direct symmetric, probably inhibitory, synaptic junctions and may additionally modulate the activity of neighboring astrocytes through exocytotic release from large dense-core vesicles.(ABSTRACT TRUNCATED AT 400 WORDS)
Neural maps of interaural time and intensity differences in the optic tectum of the barn owl
Authors: Olsen JF , Knudsen EI , Esterly SD
Published: March 2018

This report describes the binaural basis of the auditory space map in the optic tectum of the barn owl (Tyto alba). Single units were recorded extracellularly in ketamine-anesthetized birds. Unit tuning for interaural differences in timing and intensity of wideband noise was measured using digitally synthesized sound presented through earphones. Spatial receptive fields of the same units were measured with a free field sound source. Auditory units in the optic tectum are sharply tuned for both the azimuth and the elevation of a free field sound source. To determine the binaural cues that could be responsible for this spatial tuning, we measured in the ear canals the amplitude and phase spectra produced by a free field noise source and calculated from these measurements the interaural differences in time and intensity associated with each of 178 locations throughout the frontal hemisphere. For all frequencies, interaural time differences (ITDs) varied systematically and most strongly with source azimuth. The pattern of variation of interaural intensity differences (IIDs) depended on frequency. For low frequencies (below 4 kHz) IID varied primarily with source azimuth, whereas for high frequencies (above 5 kHz) IID varied primarily with source elevation. Tectal units were tuned for interaural differences in both time and intensity of dichotic stimuli. Changing either parameter away from the best value for the unit decreased the unit's response. The tuning of units to either parameter was sharp: the width of ITD tuning curves, measured at 50% of the maximum response with IID held constant (50% tuning width), ranged from 18 to 82 microsecs. The 50% tuning widths of IID tuning curves, measured with ITD held constant, ranged from 8 to 37 dB. For most units, tuning for ITD was largely independent of IID, and vice versa. A few units exhibited systematic shifts of the best ITD with changes in IID (or shifts of the best IID with changes in ITD); for these units, a change in the value of one parameter to favor one ear shifted the best value of the other parameter in favor of the same ear, i.e., in the direction opposite to that expected from “time-intensity trading.” Overall sound intensity had little or no effect on ITD tuning, but did increase the best IIDs of units tuned to nonzero IIDs. The tuning of units for ITD and IID changed systematically along different dimensions of the optic tectum to create coextensive, independent neurophysiological maps of ITD and IID.(ABSTRACT TRUNCATED AT 400 WORDS)