bluuebunny/crossref_metadata_2025 huggingface dataset:
Altered patterns of dynorphin immunoreactivity suggest mossy fiber reorganization in human hippocampal epilepsy
Authors: Houser CR , Miyashiro JE , Swartz BE , Walsh GO , Rich JR , Delgado-Escueta AV
Published: March 2018

Dynorphin A(1–17), an opioid peptide that is normally present in the hippocampal mossy fiber system, was localized immunocytochemically in the hippocampal formation of control autopsy and temporal lobe epilepsy (TLE) specimens. In control tissue, dynorphin-like immunoreactive (Dyn- IR) structures were confined to the mossy fiber path and were most highly concentrated in the polymorph (hilar) region of the dentate gyrus. Very few Dyn-IR structures were present in the molecular and granule cell layers of the dentate gyrus. In contrast, in all TLE specimens, Dyn-IR elements were present in these layers. The extent of aberrant staining varied among the TLE specimens, and 2 major patterns were observed. The first was a relatively wide band of reaction product in the inner one-third to one-fourth of the molecular layer (8 cases), and the second was a more limited distribution of immunoreactive fibers and presumptive terminals in the granule cell and immediately adjacent supragranular regions (2 cases). The extent of aberrant Dyn-IR structures appeared to be related to the amount of cell loss in the polymorph and CA3 fields and to dispersion of the granule cell somata. Specimens processed with the Timm's sulfide silver method for heavy metals provided independent evidence for the distribution of mossy fibers. In both control and TLE specimens, the patterns of labeling were virtually identical to those of dynorphin localization. These findings suggest that sprouting of mossy fibers or their axon collaterals has occurred in hippocampal epilepsy and that the reorganized fibers contain at least one of the neuropeptides that are normally present in this system. Such fibers could form recurrent excitatory circuits and contribute to synchronous firing and epileptiform activity, as suggested in studies of experimental models of epilepsy.
Glutamate neurotoxicity in vitro: antagonist pharmacology and intracellular calcium concentrations
Authors: Michaels RL , Rothman SM
Published: March 2018

There is now convincing evidence that excessive accumulation of the excitatory amino acid glutamate (GLU) in the extracellular space is toxic to central mammalian neurons. However, the role of different GLU receptors in producing this toxicity has not been adequately ascertained. There is also no adequate information about the correlation of free intracellular calcium concentration with eventual excitotoxic death. We have used cultured rat hippocampal neurons to address these issues. Approximately 75% of our neurons died after a 20- min GLU exposure. The potent kainate/quisqualate receptor antagonist 6- cyano-7-nitroquinoxaline-2,3-dione did not significantly ameliorate the GLU toxicity, while the selective noncompetitive N-methyl-D-aspartate (NMDA) antagonist methyl-10,11-dihydro-5-H-dibenzocyclohepten-5,10- imine (MK-801) blocked the GLU toxicity for periods of at least 2 hr. Interestingly, kainate was very toxic to the hippocampal neurons, but this toxicity was markedly attenuated by MK-801. These results suggest that the major toxicity of GLU is mediated by NMDA receptors and that under some conditions kainate toxicity reflects nonspecific opening of NMDA channels. The intracellular calcium concentrations in these neurons at the end of exposure to GLU and kainate (in the presence and absence of different antagonists) correlated poorly with eventual survival. Antagonists that limited the rise in calcium were still ineffective in preventing death. These results confirm earlier observations that stressed the importance of NMDA receptors in mediating GLU toxicity. However, they indicate that the relationship between toxicity and neuronal calcium concentration may be very complicated. An unexpected finding of these experiments was that MK- 801, unlike competitive antagonists of GLU, elevated intracellular calcium.
Input and output connections of an anteromedial group of spiking local interneurons in the metathoracic ganglion of the locust
Authors: Nagayama T , Burrows M
Published: March 2018

Intracellular recordings were made from an anteromedial group of spiking local interneurons in the metathoracic ganglion of the locust to determine the input connections that shape their mechanoreceptive fields on a hindleg and the output connections that define their effects on hindleg motor neurons. The receptive fields of these interneurons may contain excitatory and inhibitory regions. An excitatory region on the ipsilateral hind leg is formed by direct excitatory connections of the afferents from exteroceptors. Afferent spikes consistently evoke EPSPs in interneurons with a central synaptic latency of 1.2–1.5 msec. The connections appear to be direct and chemically mediated. An inhibitory region of a receptive field is not formed by direct afferent connections. Instead, spiking local interneurons of a midline group, which also receive direct excitatory inputs from the afferents, make direct inhibitory connections with anteromedial interneurons. Spikes in particular midline interneurons consistently evoke IPSPs in anteromedial interneurons with a central synaptic latency of about 1.0 msec. The connections appear to be direct and chemically mediated. By contrast, spikes in anteromedial interneurons could not be linked to any synaptic potentials in midline interneurons, so that direct connections are unidirectional and inhibitory. The large inhibitory regions to the receptive fields of anteromedial interneurons result from convergent inhibitory inputs of several midline interneurons. Each midline interneuron may make inhibitory output connections with several anteromedial interneurons, but there is a high degree of specificity in these divergent connections. Anteromedial interneurons make excitatory output connections with specific hind leg motor neurons. EPSPs in motor neurons follow spikes in the interneurons with short and consistent synaptic latencies of about 1.0 msec. The connections appear to be direct and chemically mediated. Two or more anteromedial interneurons with overlapping receptive fields may converge onto one motor neuron, and, in turn, one interneuron may make divergent connections with several motor neurons of the same pool. This pattern of connections suggests that these interneurons are essential elements in the local circuitry responsible for the expression of local reflexes of a leg. They provide an excitatory drive to the motor neurons in parallel to that from nonspiking local interneurons when particular arrays of exteroceptors are stimulated.
The time course and amplitude of EPSPs evoked at synapses between pairs of CA3/CA1 neurons in the hippocampal slice
Authors: Sayer RJ , Friedlander MJ , Redman SJ
Published: March 2018

Unitary EPSPs were evoked in CA1 pyramidal neurons by activation of single CA3 pyramidal neurons. Seventy-one EPSPs were recorded. The peak amplitudes of these EPSPs ranged from 30 to 665 microV with a mean of 131 microV. Rise times and half-widths were measured, the means +/- SD being 3.9 +/- 1.8 and 19.5 +/- 8.0 msec, respectively. The time courses of these EPSPs were consistent with a brief synaptic current at a localized electrotonic region of the dendritic tree followed by passive spread of current to the soma. EPSPs varied in amplitude from trial to trial. Sufficient records were collected for 12 EPSPs to demonstrate that this variation was greater than could be accounted for by baseline noise. The amplitude variations of one EPSP were reliably resolved from the background noise, and this EPSP fluctuated between 4 discrete amplitudes (including failures) separated by a quantal increment of 278 microV.
Disruption of experience-dependent synaptic modifications in striate cortex by infusion of an NMDA receptor antagonist
Authors: Bear MF , Kleinschmidt A , Gu QA , Singer W
Published: March 2018

To assess the possibility that NMDA receptors play a special role in visual cortical plasticity, the selective antagonist 2-amino-5- phosphonovaleric acid (APV) was continuously infused into the striate cortex of kittens as the visual environment was manipulated during the critical period. The cortex was studied using single-unit recording from sites between 3 and 6 mm from the infusion cannulae. One week of D,L-APV infusion coincident with monocular deprivation or “reverse suture” produced a concentration-dependent increase in the percentage of neurons that (1) lacked normal orientation selectivity and (2) were responsive to stimulation of the deprived eye. These effects outlasted the presence of the drug in the tissue. APV treatment also prevented the acquisition of selectivity and visual responsiveness that normally results from monocular visual experience after dark-rearing. Lasting effects of chronic APV infusion were not observed in adult striate cortex. The effects of APV on kitten striate cortex depended on the presence of the D stereoisomer as infusion of L-APV was without effect. Estimates of extracellular concentration using 3H-APV indicated that significant effects could be obtained with concentrations as low as 20 microM D,L-APV. Recordings from units during infusion indicated that visual responses were reduced by APV. Nonetheless, a normal percentage of visually responsive neurons was found at sites greater than or equal to 3 mm from the infusion cannula. There was no evidence that chronic APV infusion affected the sampling frequency of recorded neurons or disrupted cytoarchitecture at the sites further than 3 mm from the infusion cannula. Taken together, the data indicate that the effects of APV on kitten striate cortex are likely due specifically to the blockade of NMDA receptors. These data are considered in relation to several hypotheses concerning the role of NMDA receptors in the experience-dependent development of striate cortex.
A vasoactive intestinal peptide-like cotransmitter at cholinergic synapses between rat myenteric neurons in cell culture
Authors: Willard AL
Published: March 2018

Intracellular recording and immunochemical techniques were used to study synaptic transmission between individual pairs of rat myenteric plexus neurons in cell culture. This report describes the synaptic connections made by “dual function” presynaptic neurons that evoked slow postsynaptic depolarizations (slow EPSPs) in the same neurons in which they also evoked fast nicotinic cholinergic EPSPs. The slow EPSPs occurred only when presynaptic neurons were stimulated at frequencies of 5 Hz or higher. During the slow EPSPs, slope input resistance increased. The slow EPSPs were not detectably voltage-dependent, and they reversed sign at the estimated K+ equilibrium potential, suggesting that they resulted from a synaptically mediated decrease in resting K+ conductance. Several lines of evidence suggested that dual- function neurons evoke slow EPSPs by releasing a vasoactive intestinal peptide (VIP)-like cotransmitter. (1) Immunocytochemical staining revealed VIP-like immunoreactivity in all physiologically identified dual-function neurons. (2) Responses to exogenously applied VIP mimicked the slow EPSPs. (3) Superfusion of cultures with anti-VIP antisera blocked the slow EPSPs reversibly, as did application of desensitizing doses of VIP. These findings suggest that during periods of increased activity, subsets of cholinergic myenteric neurons release a VIP-like cotransmitter that enhances postsynaptic excitability. The effects of the cotransmitter may help to compensate for decreases in nicotinic EPSPs that occur during increased presynaptic activity.
Excitatory amino acid release and free radical formation may cooperate in the genesis of ischemia-induced neuronal damage
Authors: Pellegrini-Giampietro DE , Cherici G , Alesiani M , Carla V , Moroni F
Published: March 2018

Excessive stimulation of excitatory amino acid (EAA) receptors and abnormal production of oxygen-derived free radicals have repeatedly been implicated in the series of events linking brain hypoxia or ischemia to neuronal death. We report here that in rat hippocampal slices the KCl-stimulated output of labeled D-3H aspartate or of endogenous aspartate and glutamate significantly increased under in vitro simulated hypoxic, hypoglycemic, or ischemic conditions. In particular, when the slices were incubated for 10 min at 32 degrees C under “ischemic” conditions (namely, lack of oxygen and glucose), endogenous aspartate and glutamate in the supernatant increased by 10 and 20 times, respectively. Since radical scavengers (D-mannitol), drugs reducing free radical formation (indomethacin, corticosteroid), or enzymes able to metabolize them (catalase and superoxide dismutase) significantly reduced this output, it was supposed that free radicals caused EAA release. A direct demonstration of this concept was obtained by showing a significant release of EAA after incubation of hippocampal slices with enzymes and substrates known to cause the formation of free radicals, such as xanthine plus xanthine oxidase or arachidonic acid plus prostaglandin synthase. Neither ischemia nor the enzymatic reactions leading to free radical production increased the activity of the cytoplasmic enzyme lactate dehydrogenase in the incubation medium, thus ruling out a nonspecific cellular lysis. It appears therefore that during ischemic states, brain production of reactive molecules (free radicals) causes an increased output of EAA. This may trigger a series of events which could help to explain the delayed loss of neurons after a transient ischemic period.
Afferent basis of visual response properties in area MT of the macaque. II. Effects of superior colliculus removal
Authors: Rodman HR , Gross CG , Albright TD
Published: March 2018

In a previous study (Rodman et al., 1989), we found that many neurons in the middle temporal area (MT) of the macaque monkey remain visually responsive and directionally selective after striate cortex lesions or cooling. In the present study, we examined the effects of superior colliculus (SC) lesions and combined lesions of striate cortex and the SC on the visual properties of MT neurons. Removal of the SC alone had no effect on the proportion of visually responsive cells, strength of direction selectivity and direction tuning, orientation tuning, receptive field size, or binocularity in MT. There was, however, a slight increase in response strength to both stationary and moving slit stimuli. In contrast to the minor effects of SC lesions alone, addition of an SC lesion to striate cortex damage abolished all visual responsiveness in area MT. The results indicate that pathways damaged by the SC lesion are not necessary for most of the properties of MT neurons found in the intact animal, although these pathways are capable of sustaining considerable visual responsiveness and direction selectivity when striate input is removed.
NMDA receptor-mediated synaptic excitation selectively inhibited by ethanol in hippocampal slice from adult rat
Authors: Lovinger DM , White G , Weight FF
Published: March 2018

The effect of ethanol (EtOH) on synaptic transmission mediated by N- methyl-D-aspartate (NMDA) and non-NMDA glutamate receptors was investigated in slices from adult rat hippocampus. Synaptic responses were elicited by stimulation of stratum radiatum and were recorded in CA1 stratum radiatum or stratum pyramidale. Population EPSPs (pEPSPs) mediated by NMDA receptor activation were isolated by application of a solution containing the kainate/quisqualate receptor antagonist 6,7- dinitroquinoxaline-2,3-dione and either low (0.1 mM) Mg2+ or 100 microM bicuculline. Increasing concentrations of EtOH produced increasing inhibition of NMDA receptor-mediated pEPSPs with EtOH concentrations between 1 and 50 mM. At a concentration of 50 mM, EtOH inhibited NMDA receptor-mediated pEPSPS by 43%; the inhibition by 100 mM EtOH was not significantly different from that produced by 50 mM. Methanol and 1- butanol also inhibited the NMDA receptor-mediated pEPSPs; the potency of the alcohols for inhibition of NMDA receptor-mediated pEPSPs was 1- butanol greater than ethanol greater than methanol. pEPSPs mediated by non-NMDA glutamate receptors were isolated by the application of the NMDA receptor antagonist d,1–2-amino-5-phosphonovaleric acid in the presence of 1.5 mM Mg2+. These pEPSPs were not significantly affected by 50 mM EtOH, whereas 100 mM EtOH reduced the amplitude of these pEPSPs by 9%. The observations indicate that synaptic excitation mediated by NMDA receptors in tissue from adult rat is inhibited by intoxicating concentrations of EtOH. The data are consistent with the hypothesis that EtOH-induced inhibition of EPSPs mediated NMDA receptors may contribute to the intoxicating effects of EtOH.
Neuronal properties and trophic activities of immortalized hippocampal cells from embryonic and young adult mice
Authors: Lee HJ , Hammond DN , Large TH , Roback JD , Sim JA , Brown DA , Otten UH , Wainer BH
Published: March 2018

The hippocampal formation elaborates trophic factors such as nerve growth factor (NGF) to support the cholinergic innervation it receives from the septal region. To further study the trophic interactions of this pathway, hippocampal cells from embryonic day 18 and postnatal day 21 mice were immortalized via somatic cell fusion to N18TG2 neuroblastoma cells. The hippocampal cell lines exhibit morphological and cytoskeletal features which are typical of their neuronal parents but which are not expressed by the neuroblastoma parent. When differentiated with retinoic acid, the hippocampal cell lines exhibit electrophysiological features similar to cultured hippocampal neurons. Many of the lines constitutively express high levels of NGF, and at least one cell line exerts a non-NGF trophic effect on the expression of choline acetyltransferase by septal neurons in vitro. These cell lines are potentially useful for investigating the neurochemical and excitable properties of hippocampal neurons and identifying novel trophic activities that promote the development and maintenance of the septohippocampal pathway.
Differentiation of IKA in amphibian spinal neurons
Authors: Ribera AB , Spitzer NC
Published: March 2018

We have examined the development of an inactivating outward current in embryonic amphibian neurons differentiating in culture. On the basis of ionic selectivity, voltage dependence of activation and inactivation and pharmacological sensitivity, it is similar to A currents described in other neurons. In these embryonic neurons the A current appears later than other voltage-dependent currents studied previously. Furthermore, its maturation extends to times later than those required for the differentiation of another potassium current, the voltage- dependent delayed rectifier. These changes in A current are consistent with parallel changes in the action potential and excitability of the developing neurons.
Axonogenesis in the brain of zebrafish embryos
Authors: Chitnis AB , Kuwada JY
Published: March 2018

We analyzed the pattern and development of the earliest tracts and followed pathfinding by the growth cones of an identified cluster of neurons in the brain of zebrafish embryos. Neurons were labeled with an antibody which labels many embryonic neurons, a lipophilic axonal tracer dye, and intracellular dye injections. The embryonic brain is extremely simple, and at 28 hr of development, the forebrain and midbrain consist of 8 main axonal tracts which are arranged as a set of longitudinal tracts connected by commissures. Each tract is established by identified clusters of approximately 2–12 neurons found in discrete regions of the brain. Many identified clusters of neurons project axons in a defined direction appropriate for the cluster and have axons with stereotyped trajectories, suggesting that their growth cones follow cell-specific routes. This was confirmed with intracellular dye injections for neurons of the nucleus of the posterior commissure. The growth cones of these neurons arrive at a site in the anterior tegmentum where 4 tracts meet. At this site, they could, in principle, turn in a number of directions but always extend posteriorly into one of the tracts. The pattern of pathfinding by these growth cones suggests the testable hypothesis that the growth cones of identified clusters of neurons establish the simple set of early tracts by selecting cluster-specific pathways at such intersections in order to reach their targets in the brain.
Interspecific comparison of a Drosophila gene encoding FMRFamide- related neuropeptides
Authors: Taghert PH , Schneider LE
Published: March 2018

In order to identify functionally important regions of a neuropeptide gene in Drosophila melanogaster, we have studied its occurrence in related species and have characterized the structure of a homologous gene in Drosophila virilis. The melanogaster gene encodes a precursor that contains 13 neuropeptides related to the molluscan tetrapeptide FMRFamide (Nambu et al., 1988; Schneider and Taghert, 1988). Using the melanogaster gene as a probe in Southern blot analysis, related sequences were detected in DNA from each of 7 species tested. D. virilis, which is estimated to have diverged from D. melanogaster between 60 and 80 million years ago (Throckmorton, 1975), was chosen for more detailed study. Immunocytochemical staining using an antibody to authentic FMRFamide revealed a similar set of immunoreactive neurons in the CNS of larvae from the 2 Drosophila species. Using a melanogaster gene probe, overlapping clones were isolated from a virilis genomic library; DNA sequence analysis indicated the presence of a homologous gene. Comparisons of the genes and deduced proteins between the 2 species revealed the following points. (1) Both genes are divided into 2 exons: in D. melanogaster the exons are 106 and 1352 bp long; in D. virilis, they are 169 and at least 1232 bp long; in both species, the intron is approximately 2.5 kb long. (2) The sequence of exon I has largely diverged, and in neither species are exon I sequences translated. In this vicinity of the gene, sequence conservation is limited to a 67 bp region that spans the TATA box and the RNA start site. (3) The deduced neuropeptide precursors have very similar sizes (347 vs 339 amino acids) and the presumed signal sequences are perfectly conserved. (4) While the melanogaster precursor contains 13 FMRFamide-related peptides, the virilis precursor contains only 10. (5) The sequences of some but not all of the FMRFamide-like peptides are perfectly conserved. (6) In the rest of the precursor, significant sequence conservation is found only in the N-terminal portion; immediately downstream of the final FMRFamide-like peptide, the protein sequences are highly divergent. (7) 5′ to the RNA start sites (1.2 kb of melanogaster DNA and 1.8 kb of virilis DNA), 17 small (9–52 base pairs) regions are evolutionarily conserved (greater than 80% sequence conservation). We discuss neuropeptide biosynthesis, the functions and evolution of FMRFamide-like neuropeptides in insects, and the cell-specific regulation of neuropeptide gene expression in the contexts of these results.
Morphological plasticity in efferent pathways to the urinary bladder of the rat following urethral obstruction
Authors: Steers WD , Ciambotti J , Erdman S , de Groat WC
Published: March 2018

Partial urethral ligation in female Wistar rats produces changes in the neural control of the lower urinary tract including bladder hyperactivity and facilitation of a spinal micturition reflex pathway. To gain insight into the mechanisms underlying these changes, axonal tracing studies were conducted to examine the postganglionic efferent limb of the micturition reflex pathway which originates in the major pelvic ganglion (MPG). Forty microliters of the tracer Fluoro-Gold (4%) were injected into the right side of the bladder in urethral-obstructed (n = 10) and control (n = 4) rats 6 weeks after urethral ligation or sham surgery. As a control Fast blue (40 microliters, 5%) was injected into the colon to label neurons in the MPG innervating the intestine. Obstructed rats exhibited a 6-fold increase (p less than 0.001) in bladder weight (0.848 gm) compared to controls (0.148 gm). A significant increase (p less than 0.001) in the size of labeled bladder postganglionic neurons in the MPG was noted in obstructed rats (576.4 microns 2, n = 4) as compared to controls (299.6 microns 2). However, labeled, colon postganglionic neurons in the MPG in obstructed (312.9 microns 2) rats were not enlarged compared to controls (359.4 microns 2). Neuronal hypertrophy was not associated with a change in the number of labeled MPG neurons in control and obstructed groups.(ABSTRACT TRUNCATED AT 250 WORDS)
The role of the divergent amino and carboxyl domains on the inactivation properties of potassium channels derived from the Shaker gene of Drosophila
Authors: Iverson LE , Rudy B
Published: March 2018

Several products generated from the Drosophila Shaker gene by alternative splicing predict a group of similar proteins with an identical central and variable amino and carboxyl domains. We have constructed 9 Sh cDNAs combining 3 different 5′ domains with 3 different 3′ domains. RNA transcribed from 6 of these cDNAs induce K+ currents in Xenopus oocytes. All currents share similar properties of voltage dependence, potassium selectivity, and block by 4-AP, TEA, and charybdotoxin. These properties presumably result from a channel core formed by the identical central region of the proteins. The currents differ in macroscopic inactivation kinetics. Five RNAs induced K+ currents which inactivate, each at distinct rates, during short depolarizations. The sixth RNA induces a current that essentially does not inactivate unless depolarized for many seconds. This raises the possibility that Sh may encode nontransient as well as transient K+ currents. Analysis of currents produced by the various combinations suggests that the divergent amino domains influence the stability of a first, nonabsorbing, inactivated state. This results in striking differences in the probability of channel reopening, as observed in single-channel recordings, of those channels with identical carboxyl but different amino domains. Furthermore, based on macroscopic analysis of the currents, we suggest that the primary role of the carboxyl domains is to influence the relative stability between the first and a second inactivated state. The second inactivated state is essentially absorbing, and recovery from this state is very slow. The observed differences in the rates of recovery from inactivation of channels containing different carboxyl domains reflect differences in the rates at which they enter this second inactivated state.
Neuromedin B and gastrin-releasing peptide mRNAs are differentially distributed in the rat nervous system
Authors: Wada E , Way J , Lebacq-Verheyden AM , Battey JF
Published: March 2018

The bombesin-like peptides are a family of structurally related amidated peptide ligands that are known to have a variety of potent pharmacological actions on various cells, including neurons in the rat brain. Two mammalian representatives of the bombesin family of peptides have been identified, gastrin-releasing peptide (GRP) and neuromedin B (NMB). Previously, we cloned the rat preproGRP gene and determined the locations of neurons expressing this gene using in situ hybridization. In this study, we describe the structure and sequence of the rat preproNMB gene, and the first detailed cellular localization of preproNMB mRNA in rat brain using in situ hybridization. Nucleotide sequence analysis of cDNA and genomic clones reveals a 117 amino acid precursor whose overall structure is similar to that described for human preproNMB. Sequence similarity between the rat NMB and GRP genes is observed only over a limited 10 amino acid sequence encoding the carboxy termini of the GRP and NMB peptides, the region shown to be necessary and sufficient for high-affinity receptor binding. In situ hybridization studies performed with cRNA probes specific for NMB or GRP mRNA show that the distribution of cells expressing either mRNA in brain is very distinct. NMB mRNA is found most prominently in the olfactory bulb, dentate gyrus, and dorsal root ganglion. In contrast, the highest levels of GRP mRNA are observed in the forebrain (isocortex and hippocampal formation). This heterogeneity of mRNA distribution for these peptides suggests that these 2 structurally related peptides may have very distinct functions as neuropeptides in the rat nervous system.
Neuronal development and migration in explant cultures of the adult canary forebrain
Authors: Goldman SA
Published: March 2018

The vocal control nucleus (HVc) of the songbird forebrain undergoes neurogenesis in adulthood, as ventricular zone precursor cells divide, and their daughter cells migrate into the subjacent forebrain and differentiate into neurons. A procedure is now described by which the migration and development of these new neurons can be directly observed in vitro, in explant cultures of the adult canary forebrain. Cultured explants have yielded extensive outgrowth of new neurons from 2 neurogenic regions of the adult songbird brain: the HVc and its adjacent dorsomediocaudal neostriatum. Immunocytochemically identified neurons were determined by 3H-thymidine autoradiography to have been newly generated in vivo, in the days preceding explantation. These cells were immunoreactive for a variety of neuron-selective antigens, including MAP-1B, MAP-2, neuron-specific enolase, synaptic vesicle protein-2, N-CAM, and the tetanus toxin and A2B5 ligands. Combined immunostaining and autoradiography of cultures derived from HVcs exposed to 3H-thymidine in vivo revealed a population of newly generated, MAP-2/3H-thymidine-positive neurons. The neurons identified in these adult explant outgrowths comprised new, recently generated postmitotic cells which had arisen in the neostriatal ventricular zone and were in the early stages of their parenchymal migration at the time of tissue explantation. Accordingly, nonneurogenic control regions, including the anterior hyperstriatum, cerebellum, and optic tectum, did not display neuronal outgrowth in culture. Only forebrain regions exhibiting neurogenesis in vivo manifested neuronal outgrowth and maturation in vitro.
Cerebral synthesis and release of kynurenic acid: an endogenous antagonist of excitatory amino acid receptors
Authors: Swartz KJ , During MJ , Freese A , Beal MF
Published: March 2018

Excitatory amino acid (EAA)-mediated synaptic transmission is the most prevalent excitatory system within the mammalian brain. Activation of EAA receptors has been postulated to contribute to neuronal cell death in stroke, epilepsy, hypoglycemia, and Huntington's disease. Kynurenic acid is an endogenous substance that inhibits EAA receptors and may therefore influence important physiologic and pathologic processes. The release of intracerebrally synthesized kynurenic acid into the extracellular fluid (ECF), where it may act at EAA receptors, has not been established in vivo. We studied the synthesis and release of kynurenic acid in the rat striatum using intracerebral microdialysis coupled with high performance liquid chromatography and fluorescence detection. The basal ECF concentration of kynurenic acid in the rat corpus striatum was 17.1 +/- 1.1 nM. Peripheral administration of the immediate biosynthetic precursor of kynurenic acid, L-kynurenine, resulted in marked dose-dependent increases in striatal ECF concentrations of kynurenic acid, peaking at 2-2.5 hr. The highest dose of L-kynurenine (100 mg/kg), administered peripherally, resulted in a 108-fold increase in plasma kynurenic acid levels and a 37-fold increase in cerebral ECF levels. Peripheral administration of kynurenic acid, at a dose that caused plasma levels to increase 430-fold, resulted in only 4-fold increases in striatal ECF concentrations. The precursor responsiveness of striatal ECF kynurenic acid to peripherally infused L-kynurenine was blocked by the central application (via the dialysis probe) of aminooxyacetic acid, an inhibitor of the immediate synthetic enzyme for kynurenic acid, kynurenine aminotransferase. Administration of L-tryptophan was less effective than L-kynurenine in increasing ECF kynurenic acid concentrations and did so at a considerably later time interval (6 hr).(ABSTRACT TRUNCATED AT 250 WORDS)
The differentiation of excitability in embryonic chick limb motoneurons
Authors: McCobb DP , Best PM , Beam KG
Published: March 2018

The well-documented role of neuromuscular activity as a regulator of motoneuron and muscle development raises important questions about the differentiation of excitability in motoneurons. We have recently described changes in expression of voltage-dependent calcium currents that take place during neuromuscular development in the chick embryo (McCobb et al., 1989). We now report similar analyses, using whole-cell patch-recording methods, of the major currents underlying action potential generation in the same motoneurons. Studies were conducted on identified hindlimb motoneurons isolated from the spinal cord at 3 very different stages of chick hindlimb development. Motoneurons could generate overshooting action potentials at the earliest stage studied [embryonic day 4 (E4)]. However, large changes in densities of several voltage-dependent ionic currents occurred thereafter. E6 and E11 motoneurons had progressively larger INa densities and, consequently, greater action potential amplitudes. Densities of 2 potassium currents, Ik and IA, increased on separate schedules. The relatively late and much larger increase in IA resulted in a substantial developmental decline in action potential duration. These changes, which will greatly affect motoneuron output to muscle by affecting Ca2+ entry through voltage-gated channels, occur at the same time that activity-dependent developmental changes occur in the neuromuscular system.
The avian tectobulbar tract: development, explant culture, and effects of antibodies on the pattern of neurite outgrowth
Authors: Kroger S , Schwarz U
Published: March 2018

The tectobulbar tract is the first long-distance projecting fiber pathway to appear during the development of the avian optic tectum (dorsal half of the mesencephalon). Immunologically stained wholemounts of the E3 mesencephalon reveal that the early tectobulbar axons course in a dorsal-to-ventral direction and abruptly turn in a caudal direction shortly before reaching the ventral midline. During subsequent development, more tectobulbar axons are generated that form a parallel array of thick fascicles coursing ventrally within the mesencephalon. At this later stage of development, the tectobulbar tract bifurcates into an ipsilateral and contralateral pathway, both growing in a caudal direction near the mesencephalic ventral midline. Bifurcation and change in direction of growth is accompanied by a complete loss of the fasciculated growth pattern. Each tectobulbar axon is thus divided into a proximal fasciculated and a distal unfasciculated segment. Tectobulbar fascicles occupy the most superficial surface layer of the mesencephalon at early stages and are displaced deeper into the tissue beginning at embryonic day 5. In both of these locations, tectobulbar axons express molecules involved in axon-axon and axon-substrate interactions like the G4 antigen, neural cell adhesion molecule (N-CAM), neurofascin, and T61 antigen as revealed by immunohistochemistry and immunoblotting. Stripes of the mesencephalon explanted onto a basal lamina substratum show vigorous outgrowth of neurites. These processes grow in fascicles at a growth rate of 40 microns/h. Staining of the neurites with specific antibodies, as well as the position of the retrogradely labeled cell bodies, is in agreement with these processes being tectobulbar axons. This in vitro explant system was used to investigate the expression and possible functional involvement of N-CAM, neurofascin, G4 protein, and T61 antigen in the growth of these axons. The presence of antigen- binding fragments of polyclonal anti-G4 antibodies completely blocks fasciculation of the neurites but has no influence on their rate of elongation. Antibodies against N-CAM and neurofascin have no detectable effects. The number and length of the in vitro growing axons are reduced by the monoclonal T61 antibody. This effect is reversible. The elucidation of the exact course in vivo and the accessibility to the axons growing in vitro make the tectobulbar tract an excellent model system for the investigation of the role of these and other proteins in axonal growth and guidance during the development of the CNS.
1B1075: a brain- and pituitary-specific mRNA that encodes a novel chromogranin/secretogranin-like component of intracellular vesicles
Authors: Ottiger HP , Battenberg EF , Tsou AP , Bloom FE , Sutcliffe JG
Published: March 2018

The rat 1B1075 mRNA encodes a 533-residue novel chromogranin/secretogranin-like acidic protein that contains an apparent secretion signal, several pairs of tandem basic residues, and internally repeated sequence elements. 1B1075 transcripts are detected, by blotting and in situ hybridization, at the highest levels in the neocortex, hippocampus, cerebellar cortex, selected pontine and diencephalic nuclei, and presumptive pituitary corticotrophs, at lower levels in specific nuclei in most other brain regions, but in none of several other tissues. Utilizing antisera to several nonoverlapping synthetic peptide fragments of the predicted protein sequence, we detect a brain- and pituitary-specific 57-kDa protein in cellular processes and fiber tracts, generally consistent with axonal transport from the cell bodies identified by in situ hybridization. Ultrastructural studies demonstrate that this protein is a component of intraneuronal vesicles in axons and vesicle-like structures in dendrites. Based on these data, we suggest the name Secretogranin III for the 1B1075 gene product. In related collaborative studies, a mouse deleted for the 1B1075-homologous gene has been produced that should allow assessment of its physiological role.
Relationships between physiological and morphological properties of retinocollicular axons in the hamster
Authors: Mooney RD , Rhoades RW
Published: March 2018

Intracellular recording and HRP injection were used to characterize retinocollicular axons in the hamster. Eighteen retinotectal axons were filled with HRP. The type of axon recovered most often (N = 10) had a receptive field with center-surround organization, gave sustained responses to flashed spots, had linear spatial summation, and responded only to more slowly moving stimuli. These fibers had a mean axonal diameter of 0.58 microns (SD = 0.21) and an average conduction latency of 3.4 msec (SD = 1.2) to optic chiasm (OX) stimulation. They gave rise to circular or vertically elongated arbors with an average cross- sectional area of 33,238 microns2 (SD = 12,763) and were almost completely restricted to the stratum griseum superficiale (SGS). We recovered 6 fibers with very different structural and functional properties. The receptive fields of these axons also had center- surround organization, but gave phasic responses to flashed stimuli, had nonlinear spatial summation, and responded to rapidly moving stimuli. These axons had an average diameter of 0.84 micron (SD = 0.09) and an average latency of 2.1 msec (SD = 0.40) to OX stimulation. They had elongated arbors with an average cross-sectional area of 78,045 microns2 (SD = 14,252) and innervated the SGS, the stratum opticum (SO), and upper stratum griseum intermediale (SGI). We also recovered 2 fibers that had thin (0.3 and 0.6 micron) axons and latencies of 4.2 and 3.6 msec to OX stimulation. They both gave rise to horizontally oriented arbors in the stratum zonale (SZ) and upper SGS.
Voltage dependence of NMDA-activated macroscopic conductances predicted by single-channel kinetics
Authors: Jahr CE , Stevens CF
Published: March 2018

The conductance activated in many mammalian CNS neurons by the glutamate analog NMDA is inhibited at hyperpolarized potentials by extracellular magnesium. Whole-cell recordings from hippocampal neurons in culture were used to determine the voltage dependence of the NMDA conductance in the presence of extracellular magnesium concentrations from 1 microM to 10 mM. The conductance-voltage data are well fitted by a gating function derived from rate constants determined in an earlier study of the kinetic behavior of single channels activated by NMDA. The results are consistent with the assumption that magnesium inhibits current through the NMDA-activated channel by directly blocking the ion pore. In addition, another voltage-dependent blocking or flicker- producing mechanism has to be invoked to account for the behavior of the conductance at both the single-channel and whole-cell level, especially at low concentrations of extracellular magnesium.
Magnocellular and parvocellular contributions to responses in the middle temporal visual area (MT) of the macaque monkey
Authors: Maunsell JH , Nealey TA , DePriest DD
Published: March 2018

Many lines of evidence suggest that the visual signals relayed through the magnocellular and parvocellular subdivisions of the primate dorsal LGN remain largely segregated through several levels of cortical processing. It has been suggested that this segregation persists through to the highest stages of the visual cortex, and that the pronounced differences between the neuronal response properties in the parietal cortex and inferotemporal cortex may be attributed to differential contributions from magnocellular and parvocellular signals. We have examined this hypothesis directly by recording the responses of cortical neurons while selectively blocking responses in the magnocellular or parvocellular layers of the LGN. Responses were recorded from single units or multiunit clusters in the middle temporal visual area (MT), which is part of the pathway leading to parietal cortex and thought to receive primarily magnocellular inputs. Responses in the MT were consistently reduced when the magnocellular subdivision of the LGN was inactivated. The reduction was almost always pronounced and often complete. In contrast, parvocellular block rarely produced striking changes in MT responses and typically had very little effect. Nevertheless, unequivocal parvocellular contributions could be demonstrated for a minority of MT responses. At a few MT sites, responses were recorded while magnocellular and parvocellular blocks were made simultaneously. Responses were essentially eliminated for all these paired blocks. These results provide direct evidence for segregation of magnocellular and parvocellular contributions in the extrastriate visual cortex and support the suggestion that these signals remain largely segregated through the highest levels of cortical processing.
Distribution of photoreceptor subtypes in the retina of diurnal and nocturnal primates
Authors: Wikler KC , Rakic P
Published: March 2018

We have used antibodies specific to either the red/green-or blue- sensitive cones in order to compare their ratio and distributions to that of the rods in the retinae of 3 primate species that differ in their capacity for color vision. We have found that the monoclonal antibody CSA-1 (Johnson and Hageman, 1988) and the polyclonal antibody 4942A, specific to the red- and green-cone opsin (Lerea et al., 1989), applied to retinal whole-mounts labeled approximately 90% of all cones in the diurnal Old-World rhesus monkey (Macaca mulatta) and all of the cones in the nocturnal New-World owl monkey (Aotus trivirgatus) and nocturnal prosimian bushbaby (Galago garnetti). The polyclonal antibody 108B, specific to the blue-cone opsin (Lerea et al., 1989), labeled about 10% of the cones across the entire surface of the rhesus monkey retina, but failed to label any cones in the retina of the 2 nocturnal species. Only the retina of the rhesus monkey possessed an all-cone foveola in which the density of cone inner segments was 17-fold greater than that in the fovea of the owl monkey or bushbaby retina. Surprisingly, the density of cones per unit area outside of the fovea was comparable in all 3 species. Rod density in the dorsal retina was elevated in all animals examined, but was 2-3 times greater in the nocturnal species than in the rhesus monkey retina. Application of the photoreceptor-class-specific antibodies may provide further insights into the evolution and development of wavelength sensitivity in the retina, as well as enhance our understanding of normal and abnormal color vision in humans.
Retrosplenial cortex: possible role in habituation of the orienting response
Authors: Kwon SE , Nadeau SE , Heilman KM
Published: March 2018

A patient with a unilateral lesion that included the retrosplenial area had a propensity to attend to contralateral stimuli. To determine whether the retrosplenial-area lesion was inducing this defect, unilateral retrosplenial lesions were produced in rats by surgical aspiration. Animals were assessed preoperatively and postoperatively for orientation and habituation to bilateral simultaneous stimulation in 3 sensory modalities: visual, tactile, and auditory. At each session, the orientation test was terminated upon completion of 5 trials per modality, and the habituation test ended after 4 consecutive response failures or upon completion of 15 trials per modality. Postoperatively, contralateral orientation was not significantly different from ipsilateral orientation, suggesting that there was no neglect. However, there was a significant delay in habituation to contralateral stimulation in postoperative weeks 2 and 3-5. Our results suggest that rats with retrosplenial area lesions have normal orientation but fail to habituate to contralateral stimuli.
Activity of centrally administered galanin fragments on stimulation of feeding behavior and on galanin receptor binding in the rat hypothalamus
Authors: Crawley JN , Austin MC , Fiske SM , Martin B , Consolo S , Berthold M , Langel U , Fisone G , Bartfai T
Published: March 2018

Synthetic fragments of galanin 1–29 were administered intraventricularly or into the paraventricular nucleus of the hypothalamus for analysis of the critical amino acid sequence necessary to stimulate feeding behavior in rats. Galanin 1-29 and galanin fragment 1-16 significantly increased feeding at doses of 6 nmol microinjected into the lateral ventricles and 1 nmol microinjected into the hypothalamus. There was no significant effect of D-TRP2 galanin 1– 16 microinjected into the hypothalamus, and no significant effect of galanin fragments 1–9, 10–20, 12–29, 17–29, or 21–29 microinjected intraventricularly, on food consumption. Synthetic fragments of galanin 1–29 were assayed for displacement of 125I-galanin 1–29 binding to rat hypothalamic membranes. The efficacies of the galanin fragments in the feeding paradigm were consistent with the relative affinities of these fragments for the hypothalamic galanin receptor in equilibrium binding experiments. The first 16 N-terminal amino acids appear to contain galanin agonist activity on increasing food consumption and to bind to the galanin receptor in the rat hypothalamus.