bluuebunny/crossref_metadata_2025 huggingface dataset:
Serotonin synthesis and distribution in Drosophila dopa decarboxylase genetic mosaics
Authors: Valles AM , White K
Published: March 2018

In Drosophila, the enzyme dopa decarboxylase (DDC) catalyzes the last step in the biosynthesis of serotonin (5HT) and dopamine. In the present study, the distribution of DDC and one of its biosynthetic products (5HT) was determined in CNSs that were genetic mosaics, composed of neurons that have DDC activity and neurons that lack DDC activity. Phenotypic mosaicism, that is, immunoreactive (IR) and non-IR neurons within the same nervous system, was observed for both DDC and 5HT immunoreactivity. DDC-IR neurons were always 5HT-IR, but some 5HT- IR neurons devoid of DDC immunoreactivity were also observed. 5HT-IR neurons devoid of DDC immunoreactivity were always in close apposition to other DDC-IR neurons. We suggest that in vivo uptake mechanisms are responsible for 5HT accumulation in neurons devoid of DDC immunoreactivity.
Chronic levodopa treatment alters basal and dopamine agonist-stimulated cerebral glucose utilization
Authors: Engber TM , Susel Z , Kuo S , Chase TN
Published: March 2018

The effect of chronic levodopa administration on the functional activity of the basal ganglia and its output regions was evaluated by means of the 2-deoxyglucose (2-DG) autoradiographic technique in rats with a unilateral 6-hydroxydopamine lesion of the nigrostriatal pathway. The rates of local cerebral glucose utilization were studied under basal conditions as well as in response to challenge with a selective D1 or D2 dopamine-receptor agonist. Levodopa (100 mg/kg/d, i.p.) was administered for 19 d either continuously via infusion with an osmotic pump or intermittently by twice-daily injections. Following a 3-d washout, glucose utilization was found to be decreased by both levodopa regimens in the nucleus accumbens; intermittent levodopa also decreased glucose utilization in the entopeduncular nucleus, subthalamic nucleus, ventrolateral thalamus, ventromedial thalamus, ventroposterolateral thalamus, and lateral habenula. In control (lesioned and treated chronically with saline) rats, the D1 agonist SKF 38393 (5 mg/kg, i.v.) increased 2-DG uptake in the substantia nigra pars reticulata and entopeduncular nucleus ipsilateral to the lesion by 84% and 56%, respectively. Both continuous and intermittent levodopa blunted the SKF 38393-induced elevation in glucose metabolism in the substantia nigra pars reticulata, while intermittent levodopa also attenuated the increase in the entopeduncular nucleus. The D2 agonist quinpirole (0.4 mg/kg, i.v.) did not increase glucose utilization in any brain region in control animals; following intermittent levodopa treatment, however, quinpirole increased 2-DG uptake by 64% in the subthalamic nucleus and by 39% in the deep layers of the superior colliculus on the ipsilateral side.(ABSTRACT TRUNCATED AT 250 WORDS)
Differential effects of monocular deprivation on glutamic acid decarboxylase and type II calcium-calmodulin-dependent protein kinase gene expression in the adult monkey visual cortex [published erratum appears in J Neurosci 1991 May;11(5):following Table of Contents]
Authors: Benson DL , Isackson PJ , Gall CM , Jones EG
Published: March 2018

Increases in immunocytochemically detectable type II calcium-calmodulin- dependent protein kinase (CaM II kinase) and decreases in immunocytochemically detectable glutamic acid decarboxylase (GAD) are known to occur in the visual cortex of adult monkeys following brief periods of monocular visual deprivation. In the present study, GAD and CaM II kinase gene expression was investigated under these conditions. The polymerase chain reaction (PCR) was used to generate species- specific cDNA clones that were used to make antisense RNA probes. A second form of CaM II kinase alpha, CaM II kinase alpha-33, which contains an additional phosphorylation consensus sequence, was identified. In situ hybridization in normal visual cortex revealed a complex sublaminar organization of GAD-expressing cells within layers IVC and VI and a distribution of CaM II kinase alpha-expressing cells that was greatest in layers II, III, IVB, and VI. In situ hybridization in the cortex from animals that had been monocularly deprived revealed enhanced CaM II kinase mRNA levels in deprived-eye columns of layer IVC and, associated with the deprived eye, cytochrome oxidase-stained periodicities in other layers. In layer IV, the enhancement of labeling in deprived-eye stripes was, on average, 16% greater than in normal-eye stripes. By contrast, GAD, mRNA levels appeared unchanged in all layers, suggesting a posttranscriptional regulatory mechanism.
Patterns of unit activity in the rostral thalamus of cats related to short-latency discrimination between different auditory stimuli
Authors: Woody CD , Gruen E , Melamed O , Chizhevsky V
Published: March 2018

Short-latency auditory-responsive units were found in the rostral thalamus of cats during performance of conditioned eyeblink responses (CRs) elicited discriminatively by a forward-paired, 70-dB-click conditioned stimulus (CS) as opposed to a backward-paired, 70-dB-hiss discriminative stimulus (DS). Discharges in response to the CS or DS were found in 57% of 138 units tested. Forty-one percent of units responding to the CS did so at latencies of less than 40 msec. After conditioning a discriminative CR to click CS, an increase in the ratio of CS-evoked activity to baseline activity was found relative to that before conditioning. This increase was attributable, in part, to a decrease in baseline activity and, in part, to an increase in the magnitude of response to the CS. These responses preceded early components of the electromyographically measured motor responses with latencies sufficient to contribute to initiation of the movement. After acquisition of the CR, the proportion of CS responsive units also increased. We conclude that this area of the thalamus, a region thought to support thalamocortical reverberatory activity, also functions to transmit short-latency auditory signals. Our evidence further suggests that this region may participate in the elicitation of conditioned responses by specific auditory stimuli and in discrimination between auditory stimuli of different significance.
Chemically distinct compartments of the thalamic VPM nucleus in monkeys relay principal and spinal trigeminal pathways to different layers of the somatosensory cortex
Authors: Rausell E , Jones EG
Published: March 2018

The ventral posteromedial nucleus (VPM) of the monkey thalamus was investigated with combined immunocytochemical, histochemical, and connection-tracing techniques. Injections of anterogradely transported tracers were placed selectively in the caudal nucleus of the spinal trigeminal nuclear complex, and retrogradely transported horseradish peroxidase (HRP) or fluorescent dyes were placed on the surface or into the depths of defined parts of the trigeminal representation in the first somatic sensory area (SI) of the cerebral cortex. The results are correlated with those of the preceding paper (Rausell and Jones, 1991), which demonstrated the presence of 2 domains in the nucleus on the basis of different patterns of cytochrome oxidase (CO) staining and calcium-binding protein immunoreactivity. The cells of the CO-defined rod and matrix domains receive inputs from different components of the trigeminal afferent system and project to different layers of SI. The large- and medium-sized relay cells of the CO-rich rods, which are immunoreactive for parvalbumin, all project to middle layers of SI. The small relay cells of the weakly-stained CO-matrix, surrounding and intervening between the rods, are immunoreactive for 28-kDa calbindin and project to superficial layers (I and II) of SI. Anterograde tracing studies reveal that the rod domain in VPM is innervated by fibers arising in the contra- and ipsilateral principal trigeminal nucleus, while the matrix domain (and calbindin-positive domains in adjacent nuclei) are innervated by fibers arising in the caudal nucleus of the spinal trigeminal complex. These results demonstrate the modularity and parallel streaming of the functional components of the trigeminal part of the somatic sensory system and suggest that lemniscal and nonlemniscal elements of the system gain access by separate routes to different layers of the SI cortex.
Autoradiographic localization of putative nicotinic receptors in the rat brain using 125I-neuronal bungarotoxin
Authors: Schulz DW , Loring RH , Aizenman E , Zigmond RE
Published: March 2018

Neuronal bungarotoxin (NBT), a snake venom neurotoxin, selectively blocks nicotinic receptors in many peripheral and central neuronal preparations. alpha-Bungarotoxin (alpha BT), on the other hand, a second toxin isolated from the venom of the same snake, is an ineffective nicotinic antagonist in most vertebrate neuronal preparations studied thus far. To examine central nicotinic receptors recognized by NBT, we have characterized the binding of 125I-labeled NBT (125I-NBT) to rat brain membranes and have mapped the distribution of 125I-NBT binding in brain sections using quantitative light microscopic autoradiography. The binding of 125I-NBT was found to be saturable, of high affinity, and heterogeneously distributed in the brain. Pharmacological studies suggested that more than one population of sites is labeled by 125I-NBT. For example, one component of 125I-NBT binding was also recognized by alpha BT, while a second component, not recognized by alpha BT, was recognized by the nicotinic agonist nicotine. The highest densities of these alpha BT-insensitive, nicotine- sensitive sites were found in the fasciculus retroflexus, the lateral geniculate nucleus, the medial terminal nucleus of the accessory optic tract, and the olivary pretectal nucleus. alpha BT-sensitive NBT binding sites were found in highest density in the lateral geniculate nucleus, the subthalamic nucleus, the dorsal tegmental nucleus, and the medial mammillary nucleus (lateral part). The number of brain regions with a high density of 125I-NBT binding sites, blocked either by alpha BT or by nicotine, is low when compared with results obtained using other approaches to studying the central distribution of nicotinic receptors, such as labeling with 3H-nicotine or labeling with cDNA probes to mRNAs coding for putative receptor subunits. It is proposed that 125I-NBT labels a subpopulation of nicotinic receptors in the rat brain, and that, given its ability to block nicotinic receptors in a variety of neuronal preparations, NBT will be a useful probe for studying the functional properties of these sites.
Hypertension induced by hypothalamic transplantation from genetically hypertensive to normotensive rats
Authors: Eilam R , Malach R , Bergmann F , Segal M
Published: March 2018

The role of the hypothalamus (HTH) in the pathogenesis of genetic hypertension was studied in spontaneously hypertensive rats (SHR). It is currently believed that, in this strain, the genetic defect manifests itself mainly in the HTH. We examined this hypothesis by grafting HTH neurons from embryos of SHR or control Wistar Kyoto (WKY) rats into the HTH of adult normotensive WKY rats. Changes in host systolic blood pressure (SBP) were monitored, and alterations in vasoactive intestinal polypeptide (VIP) gene expression of the host brain were studied. In rats grafted with HTH tissue from SHR embryos (G- SHR), the blood pressure rose by 31% as compared with that in the grafted control group. The blood pressure climbed gradually over a period of 6 weeks to its highest level, which was maintained for at least 3 months following grafting. Along with the elevated blood pressure, the heart weight increased by 80% compared to controls. Behavioral changes were also evident in the G-SHR rats, and these were similar to those of the native SHR strain. In situ hybridization histochemistry showed a 40% elevation in VIP transcripts in the suprachiasmatic nucleus of the host G-SHR brain compared to controls. These studies demonstrate that transplantation of embryonic SHR HTH tissue into brains of adult normotensive rats results in the development of hypertensive characteristics in the host. It thus appears that the HTH is a prime candidate for the source of changes leading to spontaneous hypertension in mammals.
Synaptic responses evoked by mechanical stimulation of the mucosa in morphologically characterized myenteric neurons of the guinea-pig ileum
Authors: Bornstein JC , Furness JB , Smith TK , Trussell DC
Published: March 2018

Recordings were made from myenteric neurons of the guinea-pig ileum during reflexes evoked by mechanical stimulation of the mucosa. Impaled neurons were injected with dye (Lucifer yellow or biocytin), and their shapes were determined. All neurons were 5–12 mm from the stimulus, a brush stroke that deformed the mucosal villi. Neurons were classified as S-neurons or AH-neurons (Hirst et al., 1974). About 40% of S-neurons oral to a stimulus responded with bursts of fast EPSPs (average frequency, 15–40 Hz); these neurons were in ascending reflex pathways. About 60% of S-neurons anal to a stimulus responded with similar bursts of fast EPSPs or slow depolarizations; these neurons were in descending pathways. Only 2 of 48 AH-neurons responded, both in descending pathways. Most S-neurons in either ascending or descending pathways received inputs from at least 2 or 3 other neurons. Action potentials evoked during a response averaged 3–10 Hz in frequency, with occasional bursts at up to 100 Hz. The speed of conduction along the reflex pathways was about 0.5 m/sec. All S-neurons were uniaxonal, but they differed in size, dendritic morphology, and projections. The axons of S- neurons injected with biocytin were followed up to 7 mm within the myenteric plexus. Three S-neurons projected to the tertiary plexus and were probably longitudinal muscle motor neurons; 2 of these were in descending pathways. Five S-neurons projected along the intestine and had varicose collaterals in some ganglia. These neurons were probably interneurons; 3 were descending and 2 ascending, and all responded in the appropriate reflex pathway. Many S-neurons had short axons that entered the circular muscle and were probably circular muscle motor neurons. Others projected several millimeters along the intestine before entering the circular muscle or fading beyond detection. From this study, we have been able to deduce the circuits mediating ascending and descending mucosa-to-muscle reflexes. It is concluded that AH-neurons are primary sensory neurons and S-neurons are interneurons and muscle motor neurons in the circuits.
A novel calmodulin antagonist, CGS 9343B, modulates calcium-dependent changes in neurite outgrowth and growth cone movements
Authors: Polak KA , Edelman AM , Wasley JW , Cohan CS
Published: March 2018

The neurotransmitter 5-HT alters growth cone motility and neurite elongation in neuron B19, isolated from the buccal ganglion of Helisoma trivolvis (Haydon et al., 1984). The effects of 5-HT are mediated by increases in intracellular calcium levels within the growth cones (Cohan et al., 1987). 5-HT causes a receptor-mediated depolarization of the membrane, which results in the opening of voltage-sensitive calcium channels. The resulting calcium influx decreases both the elongation rate and the total outgrowth of neurites. However, the mechanism(s) mediating these calcium-dependent changes is unclear. As many of the intracellular effects of calcium in eukaryotic cells are mediated by the calcium-binding protein calmodulin, we tested the involvement of such an interaction in the regulation of neurite outgrowth. In these experiments, a new, potent calmodulin antagonist with increased selectivity, CGS 9343B (CGS; Norman et al., 1987), was used to inhibit calmodulin activity during the application of 5-HT to neuron B19. The addition of 100 microM 5-HT to the culture medium resulted in a significant decrease in the rate of neurite elongation and total neurite outgrowth. Administration of CGS to the culture medium at a concentration (1.8 microM) equivalent to its IC50 for calmodulin inhibition completely blocked the inhibitory effects of 100 microM 5- HT, on both neurite elongation and total neurite outgrowth. CGS alone caused a slight decrease in elongation rate but had no significant effect on total outgrowth. CGS did not block 5-HT-induced electrical activity, indicating that it was not acting as a 5-HT receptor antagonist.(ABSTRACT TRUNCATED AT 250 WORDS)
Stimulation of somatostatin expression in developing ciliary ganglion neurons by cells of the choroid layer
Authors: Coulombe JN , Nishi R
Published: March 2018

An important component of neuronal development is the matching of neurotransmitter expression with the appropriate target cell. We have examined how peptide transmitter expression is controlled in a simple model system, the avian ciliary ganglion (CG). This parasympathetic ganglion contains 2 distinct types of neurons: choroid neurons, which project to vasculature in the eye's choroid layer and use somatostatin as a co-transmitter with ACh, and ciliary neurons, which innervate the ciliary body and iris and use ACh but no known peptide co-transmitter. We have found that the earliest developmental stage in which neurons with somatostatinlike immunoreactivity (SOM-IR) are consistently found in vivo is stage 30 (embryonic day 6.5), a time shortly after the extension of neurites to targets in the eye's choroid layer. In cell culture, CG neurons expressed SOM-IR in co-culture with choroid cells, but not when cultured with striated muscle myotubes or with ganglion non-neuronal cells. No significant differences in neuronal survival or in ChAT activity were observed under these different co-culture conditions, which suggests that somatostatin expression is independently regulated. The stimulation of somatostatin expression was also specific in that other neuropeptides commonly found in autonomic neurons [neuropeptide Y (NPY), substance P (SP), vasoactive intestinal polypeptide (VIP)] were not induced in the presence of choroid cells. The ability to stimulate SOM-IR was not contact dependent because a macromolecule of greater than or equal to 10 kDa in choroid-conditioned medium (ChCM) was found to stimulate somatostatin expression in a dosage-dependent fashion. The somatostatin-stimulating activity induced SOM-IR in more than 90% of CG neurons, as well as in retrogradely labeled ciliary neurons, which would not normally express SOM-IR. Thus, the expression of somatostatin in cultured CG neurons is regulated by a macromolecule produced by cells in the choroid layer, a target normally innervated in vivo by CG neurons expressing somatostatin.
Calcium ion distribution in nascent pioneer axons and coupled preaxonogenesis neurons in situ
Authors: Bentley D , Guthrie PB , Kater SB
Published: March 2018

The “calcium hypothesis” of regulation of growth cone motility and neurite elongation has derived from analysis of a variety of neurons growing in vitro. It proposes that calcium ion concentration within growth cones is an important regulator of motility and growth. We now extend this analysis by investigating calcium concentrations within growth cones and nascent neurites of identified embryonic neurons growing on their normal substrate in situ. The pair of Ti1 pioneer neurons are the first to extend axons in limb buds of grasshopper embryos. Their growth cones migrate along a stereotyped pathway, where they encounter a series of guidance cues, including preaxonogenesis afferent neurons (guidepost cells). By injecting a pioneer neuron with fura-2 dye, we measured calcium concentration in the injected neuron, and in cells to which it became dye-coupled, at successive stages of pioneer outgrowth through the limb. Pioneer neurons undergoing axonogenesis had calcium concentrations in the above-100-nM range characteristic of a variety of neuron types studied in vitro. In pioneer neurons not yet in contact with guidepost cells, a shallow calcium concentration gradient, highest at the growth cone, was often observed. Guidepost cells that had not begun axonogenesis had lower calcium concentrations, in the 65 nM range. Therefore, an increase in cytosolic calcium concentration may be associated with the onset of axonogenesis. Nascent pioneer neurons were fura-2 dye-coupled to each other and established coupling with contacted guidepost cells. Calcium concentration measurements along pioneer neurites suggest that calcium ions also are transferred from pioneer neurons to these coupled guidepost cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Seasonal changes in avian song nuclei without seasonal changes in song repertoire
Authors: Brenowitz EA , Nalls B , Wingfield JC , Kroodsma DE
Published: March 2018

Seasonal variation in the size of song nuclei in the brains of male songbirds may be related to the ability to learn to sing new songs as adults. This hypothesis was tested with the rufous-sided towhee (Pipilo erythrophthalmus), a species in which song repertoires are stable after 1 yr of life. Towhees were hand raised in the laboratory and tutored with normal towhee songs. After song repertoires were recorded at 1 yr of age, photoperiods were manipulated so that 10 male towhees experienced short days and 10 males experienced long days. Circulating hormone concentrations and anatomical attributes of song nuclei were then measured. Photoperiod-related differences in the song nuclei of these towhees were as large as those seen in “open-ended learners” (i.e., species that continue to learn new songs as adults). Seasonal changes of the adult song system may thus occur without disrupting existing song repertoires and without the development of new songs. The synaptic plasticity provided by such seasonal variation, however, may enable song learning by adult birds.
Non-steady-state measurement of in vivo radioligand binding with positron emission tomography: specificity analysis and comparison with in vitro binding
Authors: Perlmutter JS , Moerlein SM , Hwang DR , Todd RD
Published: March 2018

We previously have developed a non-steady-state method for in vivo measurement of radioligand-receptor binding in brain using positron emission tomography (PET) and 18F-spiperone (18F-SP). This method has proven to be highly sensitive to the detection of decreases in the apparent number of available specific binding sites. The purposes of this investigation are to demonstrate the specificity of this PET assay and compare findings to in vitro binding assays. Three to six studies were performed in each of five male baboons. Each animal was pretreated with either ketanserin [serotonergic (S2)], eticlopride [dopaminergic (D2)], or unlabeled SP to compete with 18F-SP for specific binding sites. Sequential PET scans and arterial-blood samples were collected for 3 hr after intravenous injection of 18F-SP. Data were analyzed with a three-compartment model that considered the accumulation of radiolabeled metabolites in arterial blood. Five baboons were killed, and radioligand-receptor binding in vitro was measured by homogenate techniques. There was no detectable in vitro or in vivo specific binding of SP in cerebellum. The specific binding of SP in striatal tissue in vitro was approximately 74% to D2 sites and 26% to S2 sites, whereas ketanserin displaced all specific binding in frontal cortex. In close agreement, specific binding measured in vivo with PET revealed that 68% of apparent striatal binding could be blocked by pretreatment with eticlopride, and 34% by ketanserin. The small apparent difference between receptor binding in vitro and in vivo may result from the relatively poor resolution of PET.(ABSTRACT TRUNCATED AT 250 WORDS)
Muscarinic receptor binding increases in anterior thalamus and cingulate cortex during discriminative avoidance learning
Authors: Vogt BA , Gabriel M , Vogt LJ , Poremba A , Jensen EL , Kubota Y , Kang E
Published: March 2018

Training-induced neuronal activity develops in the mammalian limbic system during discriminative avoidance conditioning. This study explores behaviorally relevant changes in muscarinic ACh receptor binding in 52 rabbits that were trained to one of five stages of conditioned response acquisition. Sixteen naive and 10 animals yoked to criterion performance served as control cases. Upon reaching a particular stage of training, the brains were removed and autoradiographically assayed for 3H-oxotremorine-M binding with 50 nM pirenzepine (OXO-M/PZ) or for 3H-pirenzepine binding in nine limbic thalamic nuclei and cingulate cortex. Specific OXO-M/PZ binding increased in the parvocellular division of the anterodorsal nucleus early in training when the animals were first exposed to pairing of the conditional and unconditional stimuli. Elevated binding in this nucleus was maintained throughout subsequent training. In the parvocellular division of the anteroventral nucleus (AVp), OXO-M/PZ binding progressively increased throughout training, reached a peak at the criterion stage of performance, and returned to control values during extinction sessions. Peak OXO-M/PZ binding in AVp was significantly elevated over that for cases yoked to criterion performance. In the magnocellular division of the anteroventral nucleus (AVm), OXO-M/PZ binding was elevated only during criterion performance of the task, and it was unaltered in any other limbic thalamic nuclei. Specific OXO-M/PZ binding was also elevated in most layers in rostral area 29c when subjects first performed a significant behavioral discrimination. Training-induced alterations in OXO-M/PZ binding in AVp and layer Ia of area 29c were similar and highly correlated.
Vasopressin generates a persistent voltage-dependent sodium current in a mammalian motoneuron
Authors: Raggenbass M , Goumaz M , Sermasi E , Tribollet E , Dreifuss JJ
Published: March 2018

During the period of life that precedes weaning, the facial nucleus of the newborn rat is rich in 3H-vasopressin binding sites, and exogenous arginine vasopressin (AVP) can excite facial motoneurons by interacting with V1 (vasopressor-type) receptors. We have investigated the mode of action of this peptide by carrying out single-electrode voltage-clamp recordings in coronal brainstem slices from the neonate. Facial motoneurons were identified by antidromic invasion following electrical stimulation of the genu of the facial nerve. When the membrane potential was held at or near its resting level, vasopressin generated an inward current whose magnitude was concentration related; the lowest peptide concentration still effective in eliciting this effect was 10 nM. The vasopressin-induced current, IAVP, was resistant to tetrodotoxin (TTX) and was insensitive to a reduction in extracellular calcium concentration. It was sustained, was inward at all potentials tested (-120 to -25 mV), and increased in magnitude during depolarization. IAVP was not generated by the blockade of a potassium current, because it did not reverse at hyperpolarized potentials, was not affected by a two-fold increase in the transmembrane potassium gradient, and was not modified by the potassium channel blockers tetraethylammonium bromide (TEA), 4-aminopyridin (4-AP), barium, cesium, quinine, glibenclamide, and apamin. Also, IAVP was not affected by changes in the transmembrane chloride gradient. In contrast, it could be reduced by partially substituting extracellular sodium with equimolar N-methyl-D-glucamine or Tris. Our results suggest that vasopressin increases the excitability of facial motoneurons by generating a persistent sodium-dependent membrane current that is voltage gated and TTX resistant.
Chronic quinolinic acid lesions in rats closely resemble Huntington's disease
Authors: Beal MF , Ferrante RJ , Swartz KJ , Kowall NW
Published: March 2018

We previously found a relative sparing of somatostatin and neuropeptide Y neurons 1 week after producing striatal lesions with NMDA receptor agonists. These results are similar to postmortem findings in Huntington's disease (HD), though in this illness there are two- to threefold increases in striatal somatostatin and neuropeptide Y concentrations, which may be due to striatal atrophy. In the present study, we examined the effects of striatal excitotoxin lesions at 6 months and 1 yr, because these lesions exhibit striatal shrinkage and atrophy similar to that occurring in HD striatum. At 6 months and 1 yr, lesions with the NMDA receptor agonist quinolinic acid (QA) resulted in significant increases (up to twofold) in concentrations of somatostatin and neuropeptide Y immunoreactivity, while concentrations of GABA, substance P immunoreactivity, and ChAT activity were significantly reduced. In contrast, somatostatin and neuropeptide Y concentrations did not increase 6 months after kainic acid (KA) or alpha-amino-3- hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) lesions. At both 6 months and 1 yr, QA lesions showed striking sparing of NADPH-diaphorase neurons as compared with both AMPA and KA lesions, neither of which showed preferential sparing of these neurons. Long-term QA lesions also resulted in significant increases in concentrations of both 5-HT and 5- hydroxyindoleacetic acid (HIAA), similar to findings in HD. Chronic QA lesions therefore closely resemble the neurochemical features of HD, because they result in increases in somatostatin and neuropeptide Y and in 5-HT and HIAA. These findings strengthen the possibility that an NMDA receptor-mediated excitotoxic process could play a role in the pathogenesis of HD.
Control of retinal information coding by GABAB receptors
Authors: Pan ZH , Slaughter MM
Published: March 2018

The directional selectivity of amacrine and ganglion cells was studied using conventional intracellular recording techniques and drug application in the superfused retina-eyecup preparation of the tiger salamander. Baclofen, a GABAB receptor agonist, enhanced normal directional responses in some directionally selective third-order neurons. In about 30% of the cells that were not normally directional, baclofen induced direction-selective responses. This effect was particularly marked when 2-amino-4-phosphonobutyrate (APB) was used to isolate the OFF pathway. Comparisons of the effects of APB and baclofen on induced directional cells indicate that directional information in the ON and OFF channels is often handled separately and frequently is not aligned. This tends to obscure the observation of directionality as seen from the soma. Application of picrotoxin blocked both normal directional selectivity and baclofen-induced directional selectivity in some cells. Superfusion of picrotoxin and strychnine together blocked directionality in almost all cells. In both normal and induced directionality, the null direction response varied from cell to cell between a small depolarization, no voltage response, or a hyperpolarization. Injection of positive current often revealed “silent” inhibition. Some induced direction-selective cells did not show any evidence of inhibition in the null direction. The similarities in the response to baclofen, the influence of GABA and glycine antagonists, and the characteristics of the null-direction responses suggest that both normal and induced directionality originate from the same sources or mechanisms. Baclofen also induced orientation selectivity, but this was rarely observed.
Characterization of dopamine release in the substantia nigra by in vivo microdialysis in freely moving rats
Authors: Robertson GS , Damsma G , Fibiger HC
Published: March 2018

Dopamine (DA) is released not only from the terminals of the nigrostriatal projection, but also from the dendrites of these neurons, which arborize in the substantia nigra pars reticulata (SNR). Although striatal DA release has been extensively studied by in vivo microdialysis, dendritic DA release in the SNR has not been characterized by this technique. Extracellular DA was monitored simultaneously in the ipsilateral striatum and SNR. The nigral probe was implanted at a 50 degree angle, permitting 2.5 mm of SNR to be dialyzed. Delivery of the tracer Fluoro-Gold into the striatal probe retrogradely labeled tyrosine hydroxylase-positive cell bodies and dendrites in the vicinity of the nigral probe. Hence, it could be demonstrated that dopaminergic neurons near the nigral probe projected to the vicinity of the striatal probe. Addition of 50 mM KCl to the SNR perfusion solution produced a 3.5-fold increase in DA and a 50% reduction in dihydroxyphenylacetic acid (DOPAC) in the SNR; in contrast, this manipulation in the SNR caused DA release in the striatum to be decreased by 20%, while striatal DOPAC was increased by 50%. Local administration of nomifensine (10 microM) in the SNR produced a sevenfold increase in SNR DA but had no effect on striatal DA. Systemic injection of d-amphetamine (2 mg/kg, s.c.) elevated DA in the SNR and striatum five- to sevenfold, while DOPAC was decreased in both structures by at least 40%. To determine the effect of tetrodotoxin (TTX), basal concentrations of DA in the SNR were first elevated threefold by including nomifensine (1 microM) in the nigral perfusion solution.
Cortical organization of language
Authors: Ojemann GA
Published: March 2018

Recent data obtained by various methods of clinical investigations suggest an organization of language in the human brain involving compartmentalization into separate systems subserving different language functions. Each system includes multiple essential areas localized in the frontal and temporoparietal cortex of the dominant hemisphere, as well as widely dispersed neurons. All components of a system are activated in parallel, possibly by ascending thalamocortical circuits. The features peculiar to cerebral language organization include not only the lateralization of essential areas to one hemisphere, but also a substantial variance in the individual patterns of localization within that hemisphere, a variance that in part relates to individual differences in verbal skills.
Neurotransmitter modulation of calcium channels in rat sympathetic neurons
Authors: Plummer MR , Rittenhouse A , Kanevsky M , Hess P
Published: March 2018

Adrenergic, cholinergic, and a variety of peptide neurotransmitters are known to modulate Ca currents in peripheral neurons. Using a protocol that allows for simultaneous assessment of effects on dihydropyridine (DHP)-sensitive and DHP-insensitive current components, we compared the actions of norepinephrine (NE), bethanechol (BeCh), and neuropeptide Y (NPY) on Ca currents in neonatal rat superior cervical ganglion neurons. Here, we show that these transmitters selectively depress the activity of DHP-insensitive Ca channels. Intracellular application of GTP-gamma-S, an activator of GTP-binding proteins, also exclusively affected the DHP-insensitive current, whereas 1,2-oleoylacetylglycerol (OAG), a protein kinase C (PKC) activator, depressed both the DHP- sensitive and DHP-insensitive currents. Pertussis toxin interrupted the coupling between NE and its effector, whereas three different inhibitors of PKC did not. Thus, we confirmed that the selective actions of the transmitters on Ca current appear to be mediated via GTP- binding proteins, but we found no evidence for direct involvement of PKC and conclude that the observed actions of OAG are distinct from those mediated by the neurotransmitters studied.
A unique neuronal organization in the cat pretectum revealed by antibodies to the calcium-binding protein calbindin-D 28K
Authors: Nabors LB , Mize RR
Published: March 2018

The pretectum is an important center for visual reflexes. However, the location, boundaries, and connections of individual nuclei of the pretectum are incompletely understood. In cat, the traditionally defined nuclear boundaries have been placed in doubt by recent evidence showing that the retinal input to the pretectum forms four continuous projection zones that do not match the cytoarchitectural boundaries of individual pretectal nuclei defined by previous studies. We now show that antibodies to the calcium-binding protein calbindin-D 28K (CaBP) label clusters of neurons within the pretectum that match the zones of retinal termination. Four obvious cell clusters within the pretectum were labeled by CaBP antisera. Computer three-dimensional reconstruction of these cell clusters revealed that they form four distinct but continuous zones that run the rostrocaudal length of the pretectum in a medial-to-lateral direction. By combining anterograde HRP labeling of retinal terminals with CaBP immunocytochemistry, these CaBP-labeled cell clusters were found virtually to overlap the retinal projection zones. The CaBP-labeled neurons included both multipolar and fusiform morphologies, and most were medium- to large-sized cells. HRP retrograde transport studies showed that many CaBP-labeled neurons in the clusters projected to the LGN, while none projected to the inferior olive (IO). GABA-immunoreactive neurons were also found within the CaBP cell clusters, but these neurons were smaller than most CaBP-labeled neurons, and none were retrogradely labeled following HRP injections into the LGN or IO. Two-color antibody double-labeling experiments did not reveal any GABA neurons within the clusters that colocalized CaBP. In summary, calbindin is a precise marker of neuron clusters that overlap the retinal projection zones in the cat pretectum. Many of these CaBP neurons project to the LGN, and none contain GABA.
Long-term increases in the evoked population spike in the CA1 region of rat hippocampus induced by beta-adrenergic receptor activation
Authors: Heginbotham LR , Dunwiddie TV
Published: March 2018

The effects of the selective beta-adrenergic receptor agonist isoproterenol (ISO) were characterized in the CA1 region of the rat hippocampal slice preparation. As has been previously described, 500 nM ISO increased the amplitude of the evoked population spike response without having any effect upon field EPSP (fEPSP) responses. However, the increase in the population spike response was quite persistent and was not reversed by greater than 30 min of washout in the majority of the slices tested; we have termed this prolonged increase beta- adrenergic potentiation (BAP). As with the acute effect of ISO, BAP is confined to an increase in the population spike response and not the fEPSP. In input-output curves, this was clearly observed as a persistent leftward shift in the EPSP-population spike relationship. Similar long-term increases in the population spike could also be elicited by superfusion of the slices for 10 min with 20–25 microM norepinephrine (NE). Although both the acute and the long-term effects of ISO were blocked by pretreatment with timolol, a beta-adrenergic antagonist, the long-term effects were not reversed by superfusion with timolol following ISO treatment, demonstrating that the prolonged effects were not due to slow washout of ISO from the tissue. BAP was not blocked by pretreatment with 50 microM 2-amino-5-phosphonovaleric acid, an NMDA receptor antagonist that blocks hippocampal long-term potentiation and the long-lasting changes in synaptic responses induced in the dentate gyrus by NE and ISO.
Specialized vascularization of the primate visual cortex
Authors: Zheng D , LaMantia AS , Purves D
Published: March 2018

We have analyzed blood vessel distribution in the primary and secondary visual cortices of the squirrel monkey in relation to cortical modules, laminae, and cytoarchitectonic areas. Measurements of microvessel length in tangential sections through the primary visual cortex showed that blobs are more richly vascularized than intervening cortical regions. Thus, the mean total length of microvessel profiles per unit was 42% greater within these cortical modules than within adjacent (interblob) areas. Total microvessel length per unit area in another class of module, the stripes in the secondary visual cortex, was 27% greater than in interstripe regions. Microvessel distribution also varied systematically from layer to layer in the primary visual cortex, being greatest in lamina IVc. Finally, the overall microvessel length per unit area in sections of the primary visual cortex was 26% greater than that in the secondary visual cortex. These observations indicate that the modular, laminar, and regional organization of the primate visual cortex is reflected in the underlying distribution of cortical microvessels. These vascular patterns should be discernable in living animals with vascular contrast agents and appropriate imaging techniques.
Dopamine D2 mechanisms in canine narcolepsy
Authors: Nishino S , Arrigoni J , Valtier D , Miller JD , Guilleminault C , Dement WC , Mignot E
Published: March 2018

Narcolepsy is a sleep disorder characterized by abnormal manifestations of rapid-eye-movement (REM) sleep and excessive daytime sleepiness. Using a canine model of the disease, we found that central D2 antagonists suppressed cataplexy, a form of REM-sleep atonia occurring in narcolepsy, whereas this symptom was aggravated by D2 agonists. The effect on cataplexy was stereospecific for the S(-) enantiomer of sulpiride (a D2 antagonist) and the R(+) enantiomer of 3-PPP (a D2 agonist). There was also a significant correlation between the in vivo pharmacological potency and in vitro drug affinity for D2 receptors (but not for D1 and alpha 2 receptors) among the seven central D2 antagonists tested. Selective D1 compounds were also tested; however, the results were inconsistent because both antagonists and agonists generally suppressed cataplexy. Our current results demonstrate that central D2-type receptors are critically involved in the control of cataplexy and REM sleep. Furthermore, the finding that small doses of D2 antagonists suppressed cataplexy and induced behavioral excitation, while small doses of D2 agonists aggravated cataplexy and induced sedation, suggests that this effect is mediated presynaptically. However, considering the fact that selective dopamine reuptake inhibitors did not modify cataplexy and that our previous pharmacological results demonstrated a preferential involvement of the noradrenergic system in the control of cataplexy, we believe that the effect of D2 compounds on cataplexy is mediated secondarily via the noradrenergic systems.
Extracellular serotonin levels change with behavioral state but not with pyrogen-induced hyperthermia
Authors: Wilkinson LO , Auerbach SB , Jacobs BL
Published: March 2018

Extracellular 5-HT in the anterior hypothalamus/preoptic area (AH/POA) and caudate nucleus of the freely moving cat was measured using in vivo brain microdialysis. Administration of 8-OH-DPAT, a 5-HT1A receptor agonist that decreases 5-HT neuronal activity, decreased extracellular 5-HT in both brain areas. Extracellular 5-HT levels were also examined in relationship to the sleep-wake cycle, because previous data from our laboratory have indicated that behavioral state is the primary determinant of 5-HT neuronal discharge. As with 5-HT neuronal discharge, extracellular 5-HT was increased during active behavioral states and decreased during somnolent periods. These first two sets of findings confirm the ability of the microdialysis technique to measure physiological fluctuations in extracellular 5-HT levels and support the hypothesis that neuronal discharge is a major determinant of extracellular 5-HT levels. Levels of the 5-HT metabolite 5- hydroxyindole acetic acid (5-HIAA) in the AH/POA were also responsive to changes in behavioral state and administration of 8-OH-DPAT, though fluctuations in extracellular 5-HIAA were less robust and temporally delayed. Finally, extracellular 5-HT and 5-HIAA were examined in the AH/POA during fever induced by systemic injection of the synthetic pyrogen muramyl dipeptide. Previous data from our laboratory have indicated that 5-HT neuronal activity is unaffected by this manipulation, though 5-HT has been implicated specifically in thermoregulation. Pyrogen-induced hypothermia produced no specific change in 5-HT efflux, because any changes noted could be accounted for by behavioral state changes. These data are consistent with the hypothesis that the brain serotonergic system is closely linked to the sleep-wake-arousal cycle. However, extracellular 5-HT may be involved in thermoregulatory processes as part of a global role in modulating neuronal activity in coordination with the behavioral state of the animal.
Quantitative pharmacological analysis of 2-125I-iodomelatonin binding sites in discrete areas of the chicken brain
Authors: Siuciak JA , Krause DN , Dubocovich ML
Published: March 2018

We have localized and characterized 2–125I-iodomelatonin binding sites in the chicken brain using in vitro quantitative autoradiography. Binding sites were widely distributed throughout the chicken brain, predominantly in regions associated with the visual system. The specific binding of 2–125I-iodomelatonin to discrete chicken brain areas was found to be saturable, reversible, and of high affinity. The specific binding of 2–125I-iodomelatonin (75 pm) was quantitated for 40 identifiable brain regions. Eight brain regions were chosen for binding characterization and pharmacological analysis: optic tectum, Edinger- Westphal nucleus, oculomotor nucleus, nucleus rotundus, ventral supraoptic decussation, ventrolateral geniculate nucleus, neostriatum, and ectostriatum. These regions showed no rostral-caudal gradient in 2- 125I-iodomelatonin specific binding, and saturation analysis revealed a single class of high-affinity sites with KD values in the range of 33- 48 pM and receptor site density (Bmax) ranging from 31 to 58 fmol/mg protein. Competition experiments carried out with various indoles revealed a similar order of pharmacological affinities in these areas: melatonin greater than 6-chloromelatonin greater than methoxyluzindole greater than N-acetylserotonin greater than luzindole much greater than 5-HT greater than 5-methoxytryptamine. The affinity constants determined by quantitative autoradiography for these compounds to compete for 2–125I-iodomelatonin binding in the optic tectum correlated well with the affinities in chicken brain membranes at 25 degrees C (r = 0.966; slope = 0.845; n = 7) and 0 degree C (r = 0.946; slope = 0.379; n = 7), chicken retinal membranes (r = 0.973; slope = 0.759; n = 7), and the potency or affinity of these compounds to affect the calcium-dependent release of 3H-dopamine from the rabbit retina (r = 0.902; slope = 0.506; n = 6). We conclude that the high-affinity sites labeled by 2–125I-iodomelatonin in various chicken brain areas have identical binding and pharmacological characteristics to the ML-1 melatonin binding site previously described in chicken brain and retinal membranes and to the ML-1 melatonin receptor modulating dopamine release from the retina. In the chicken brain, the ML-1 receptor site may mediate functional responses regulated by melatonin.
Transport of estrogen-induced oxytocin receptors in the ventromedial hypothalamus
Authors: Coirini H , Schumacher M , Flanagan LM , McEwen BS
Published: March 2018

The modulation of oxytocin (OT) receptors (OTRs) by estrogen was investigated in the ventromedial hypothalamus by in vitro receptor autoradiography. Treatment of ovariectomized and adrenalectomized rats with various doses of estradiol benzoate (EB) increased OTR binding not only in the ventromedial nuclei of the hypothalamus (VMN), but also in the area lateral to the nuclei (IVMN). After a single injection of EB, OTRs first were induced within the ventrolateral parts of the VMN, and only hours later they appeared in the IVMN. This is consistent with the interpretation that OTRs are first induced within the estrogen- sensitive neurons of the ventrolateral VMN and then are transported laterally out of the nuclei. Two additional experiments confirmed this interpretation. First, local infusion of a low dose (10 micrograms) of the neuronal transport inhibitor vinblastine blocked the appearance of OTRs in the IVMN but did not prevent the induction of OTRs by EB within the nuclei. Second, a knife cut placed lateral to the VMN prevented the spread of OTRs out of the nuclei. However, even after treatment with a high dose of EB (2 x 10 micrograms), progesterone (P) was required for a maximal extension of the area covered by OTRs. Thus, the OTR is an estrogen-induced neurotransmitter receptor that is transported to its site of action, the lateral ventromedial hypothalamus, where it is modulated by P and where estrogen-induced OT immunoreactivity is found.