Showing papers in "The Journal of Neuroscience in 1983"
TL;DR: The cortical and subcortical connections of the middle temporal visual area of the macaque monkey were investigated using combined injections of [3H]proline and horseradish peroxidase within MT to determine a hierarchical arrangement of visual areas.
Abstract: The cortical and subcortical connections of the middle temporal visual area (MT) of the macaque monkey were investigated using combined injections of [3H]proline and horseradish peroxidase within MT. Cortical connections were assigned to specific visual areas on the basis of their relationship to the pattern of interhemispheric connections, revealed by staining for degeneration following callosal transection. MT was shown to be reciprocally connected with many topographically organized cortical visual areas, including V1, V2, V3, and V4. These pathways link regions representing corresponding portions of the visual field in the different areas. In addition, MT has reciprocal connections with two previously unidentified cortical areas, which we have designated the medial superior temporal area (MST) and the ventral intraparietal area (VIP). The laminar distribution of terminals and cell bodies in cortical areas connected with MT follows a consistent pattern. In areas V1, V2, and V3, the projections to MT arise largely or exclusively from cells in supragranular layers, and the reciprocal connections from MT terminate mainly in supragranular and infragranular layers. In contrast, the projections to MST and VIP terminate mainly in layer IV, and the reciprocal pathways originate from cells in both superficial and deep layers. On the basis of this pattern, each connection can be designated as forward or feedback in nature, and a hierarchical arrangement of visual areas can be determined. In this hierarchy, MT is at a higher level than V1, V2, and V3, and at a lower level than MST and VIP. Subcortical projections were seen from MT to the claustrum, the putamen, the caudate nucleus, the inferior and lateral subdivisions of the pulvinar complex, the ventral lateral geniculate nucleus, the reticular nucleus of the thalamus, the superior colliculus, and the pontine nuclei.
1,410 citations
TL;DR: It is found that individual neurons in the cat primary visual cortex can communicate over suprisingly long distances horizontally, in directions parallel to the cortical surface, by combining the techniques of intracellular recording and injection of horseradish peroxidase with three- dimensional computer graphic reconstructions.
Abstract: The intrinsic connections of the cortex have long been known to run vertically, across the cortical layers. In the present study we have found that individual neurons in the cat primary visual cortex can communicate over suprisingly long distances horizontally (up to 4 mm), in directions parallel to the cortical surface. For all of the cells having widespread projections, the collaterals within their axonal fields were distributed in repeating clusters, with an average periodicity of 1 mm. This pattern of extensive clustered projections has been revealed by combining the techniques of intracellular recording and injection of horseradish peroxidase with three- dimensional computer graphic reconstructions. The clustering pattern was most apparent when the cells were rotated to present a view parallel to the cortical surface. The pattern was observed in more than half of the pyramidal and spiny stellate cells in the cortex and was seen in all cortical layers. In our sample, cells made distant connections within their own layer and/or within another layer. The axon of one cell had clusters covering the same area in two layers, and the clusters in the deeper layer were located under those in the upper layer, suggesting a relationship between the clustering phenomenon and columnar cortical architecture. Some pyramidal cells did not project into the white matter, forming intrinsic connections exclusively. Finally, the axonal fields of all our injected cells were asymmetric, extending for greater distances along one cortical axis than along the orthogonal axis. The axons appeared to cover areas of cortex representing a larger part of the visual field than that covered by the excitatory portion of the cell9s own receptive field. These connections may be used to generate larger receptive fields or to produce the inhibitory flanks in other cells9 receptive fields.
955 citations
TL;DR: Two types of glial fibrillary acidic protein-positive (GFAP+) astrocytes were found in cultures of developing rat optic nerve and these ligands cannot be used on their own as neuron- specific markers in central nervous system cultures.
Abstract: Two types of glial fibrillary acidic protein-positive (GFAP+) astrocytes were found in cultures of developing rat optic nerve. Type 1 astrocytes had a fibroblast-like morphology, did not bind tetanus toxin or the monoclonal antibody A2B5 (both of which bind to specific polysialogangliosides), and were stimulated to divide by an extract of bovine pituitary and by epidermal growth factor (EGF). Type 2 astrocytes had a neuron-like morphology, bound tetanus toxin and A2B5 antibody, and were not stimulated to divide by bovine pituitary extract or by EGF. Although both types of astrocytes were present in cultures of white matter, only type 1 astrocytes were found in cultures of gray matter. Astrocytes did not convert from one type to the other in culture: while many type 1 astrocytes adopted a neuron-like morphology when exposed to dibutyryl cyclic adenosine 3':5'-monophosphate, or pituitary or brain extracts, especially in serum-free medium, such morphologically altered cells did not bind tetanus toxin or A2B5 antibody. Although small numbers of tetanus toxin-binding, A2B5+, GFAP+ cells were present in suspensions of freshly dissected, neonatal optic nerves, most of the type 2 astrocytes in cultures of such optic nerves developed from tetanus toxin-binding, A2B5+, GFAP- cells, which were induced to express GFAP by the culture conditions. Since type 2 astrocytes have a neuron-like morphology and bind tetanus toxin and A2B5 antibody, these ligands cannot be used on their own as neuron-specific markers in central nervous system cultures.
755 citations
TL;DR: It is suggested that the time-dependent development of A9 DA neuron inactivation induced by repeated neuroleptic treatment may provide a mechanism for understanding the delayed onset of extrapyramidal side effects often observed with antipsychotic drugs.
Abstract: Extracellular single unit recording techniques were used to study the effects of both acute and repeated oral neuroleptic administration on the in vivo activity of rat A9 and A10 dopaminergic (DA) neurons. All antipsychotic drugs examined acutely (haloperidol, l-sulpiride, chlorpromazine, and clozapine) increased the number of spontaneously firing DA neurons in both A9 and A10 compared to controls. Repeated (21 day) treatment with haloperidol, l-sulpiride, and chlorpromazine (antipsychotic drugs which can cause extrapyramidal side effects) markedly reduced the number of active DA cells below control levels in both regions. The "silent" DA neurons were in an apparent state of tonic depolarization inactivation since they could be induced to discharge by the microiontophoretic application of the inhibitory neurotransmitter gamma-aminobutyric acid, but not the excitatory amino acid glutamate. The depolarization inactivation observed may be specific for antipsychotic drugs since a non-neuroleptic phenothiazine (promethazine), the inactive isomer of sulpiride (d-sulpiride), and a tricyclic antidepressant (desmethylimipramine) neither increased DA activity when given acutely nor induced depolarization inactivation when administered repeatedly. In contrast to the other drugs tested, repeated treatment with clozapine (an effective antipsychotic drug which does not produce extrapyramidal side effects) resulted in the depolarization inactivation of A10 neurons but not A9 cells. These data suggest that neuroleptics which can induce extrapyramidal side effects produce depolarization inactivation of both A9 and A10 neurons whereas antipsychotic drugs which lack this property inactivate only A10 neurons. It is suggested that the time-dependent development of A9 DA neuron inactivation induced by repeated neuroleptic treatment may provide a mechanism for understanding the delayed onset of extrapyramidal side effects often observed with these drugs.
654 citations
TL;DR: Evidence supports the interpretation that the effect is produced by a central influence of the systems controlling directed visual attention and the angle of gaze upon those linking the retinae to the parietal lobe.
Abstract: The responses of parietal visual neurons are markedly increased during attentive fixation, as compared to those evoked in relaxed wakefulness, an effect specific for directed attention and unrelated to putative differences in the general level of arousal. Those responses are also strongly influenced by the angle of gaze, an effect observed only during directed visual attention. The change in response is smoothly graded along a meridian for about one-half the neuron population; the average spatial gradient from maximum to minimum is 78% response for a 20 degrees shift in eye position. No lateral preference was observed. For the remaining half, responses were either maximal or minimal for fixations dead ahead, and changes occurred with deviations in any direction. Angle of gaze effects were observed for neurons with foveal as well as eccentrically located receptive fields, all of which were organized in retinotopic not spatial coordinates. Control experiments showed that the effect was not produced by changes in visual background with changes in the angle of gaze, nor to changes in fixation distance, nor to variations in the intensity of stimuli viewed from different angles. The effect depends upon the position of the eye in the orbit, but is unlikely due to a direct central action of changes in nonretinal orbital afferent activity at different angles of gaze, for the effect was rarely observed with changes in the angle of gaze during relaxed wakefulness without directed visual attention. The evidence supports the interpretation that the effect is produced by a central influence of the systems controlling directed visual attention and the angle of gaze upon those linking the retinae to the parietal lobe.
607 citations
TL;DR: The dependences of the response kinetics on displacement and on temperature suggest that the thermal interconversion between open and closed transduction channels is limited by an enthalpy of activation of about 12 kcal/mol.
Abstract: The receptor current of hair cells from the bullfrog's sacculus was measured by voltage clamp recording across the isolated sensory epithelium. Several hundred hair cells were stimulated en masse by moving the overlying otolithic membrane with a piezoelectrically activated probe. As measured by optical recording of otolithic membrane motion, the step displacement stimuli reached their final amplitudes of up to 1 micrometer within 100 microseconds. The relationship between displacement and steady-state receptor current is an asymmetric, sigmoidal curve about 0.5 micrometer in extent. The time constant of the approach to steady state depends upon the magnitude of the hair bundle displacement and ranges from 100 to 500 microseconds at 4 degrees C; the time course is faster with larger displacements or at higher temperatures. Both the displacement-response curve and the kinetics of the response are changed by alterations in the Ca2+ concentration at the apical surface of the cells. The characteristics of the response are not consistent with simple models for the transduction process that involve enzymatic regulation of channel proteins or diffusible second messengers. Mechanical stimulation is instead posited to act directly by altering the free energy difference between the open and closed forms of the transduction channel, thereby inducing a redistribution between these states. The dependences of the response kinetics on displacement and on temperature suggest that the thermal interconversion between open and closed transduction channels is limited by an enthalpy of activation of about 12 kcal/mol.
530 citations
TL;DR: The auditory response properties of units in one of the telencephalic nuclei controlling song have recently been shown to respond to acoustic stimuli are investigated using a technique that permitted great flexibility in manipulating complex stimuli such as song.
Abstract: Songbirds such as the white-crowned sparrow memorize the song of conspecific adults during a critical period early in life and later in life develop song by utilizing auditory feedback. Neurons in one of the telencephalic nuclei controlling song have recently been shown to respond to acoustic stimuli. I investigated the auditory response properties of units in this nucleus using a technique that permitted great flexibility in manipulating complex stimuli such as song. A few of the units exhibited considerable selectivity for the individual9s own song. In wild-caught birds, song-specific units exhibited intradialect selectivity. In those birds that sang abnormal songs due to laboratory manipulation of song exposure during the critical period for song learning, units were selective for the abnormal songs. By systematic modification of a song, and by construction of complex synthetic sounds mimicking song, the acoustic parameters responsible for the response selectivity were identified. Song-specific units responded to sequences of two song parts but not to the parts in isolation. Modification of the frequencies of either part of the sequence, or increasing the interval between the parts, varied the strength of the response. Thus, temporal as well as spectral parameters were important for the response. When sequences of synthetic sounds mimicking song were effective in evoking an excitatory response, the response was sensitive to the aforementioned manipulations. Wih these techniques it was possible to elucidate the acoustic parameters required to excite song-specific units. All songs of the repertoire eliciting a strong excitatory response contained the appropriate parameters, which were missing from all weakly effective, ineffective, or inhibitory songs.
509 citations
TL;DR: It is proposed that off- cells must pause if the tail flick is to occur, and that this pausing allows the transmission of nociceptive input through spinal reflex loops.
Abstract: Neurons of the rostral ventromedial medulla (RVM) have been implicated in the modulation of nociceptive transmission. In order to further analyze their role in pain behavior, we studied their activity while eliciting the tail flick reflex with noxious heat. Recording sites were regions in the RVM from which microstimulation (less than or equal to 10 microA, 400 mu sec, 50 Hz continuous pulse trains) inhibited the tail flick reflex. Extracellular unit activity and tail temperature were recorded, stored, and plotted with reference to either the time of tail flick or the time when the stimulating temperature reached 45 degrees C. Neuronal discharges were found to be either increased (on-cells), decreased (off-cells), or unchanged around the time of the tail flick. The decreases in discharge were more closely correlated with the tail flick behavior than with the temperature of the stimulus. These off-cells were located at sites of lowest threshold for tail flick inhibition and tended to be ventral to on-cells. We propose that off-cells must pause if the tail flick is to occur, and that this pausing allows the transmission of nociceptive input through spinal reflex loops.
491 citations
TL;DR: The wide ranging, patchy distributions of single afferents in 4Calpha suggest that individual 4C alpha axons supply more than one ocular dominance stripe, and these properties are consistent with derivations from somata in the parvocellular and magnocellular divisions of the LGN.
Abstract: We used horseradish peroxidase (HRP) to orthogradely label afferent axons in macaque striate cortex. Of the 38 axons that we recovered, nine were recorded intracellularly before being filled with HRP. Light microscope and computer reconstructions of filled processes reveal highly stereotyped patterns of arborization and suggest that there are at least five discrete populations of lateral geniculate nucleus (LGN) afferent axon: (1) those to layer 4C beta, which have extremely circumscribed, dense terminal fields (small branches of which occasionally intrude into 4C alpha) but which have not been shown to project to other laminae; (2) afferents to layer 4A, which in some cases send fine ascending collaterals into layer 2–3 and which do not, apparently, send collaterals to other laminae; (3) afferents to layer 1, which are fine, extend over large distances horizontally, and send collaterals to layer 6A; (4) afferents to the lower two-thirds of layer 4C alpha, which have few or no collaterals in layer 6; and (5) afferents to the upper half of layer 4C alpha, which have arborizing collaterals in layer 6B. Of the nine axons that were recorded intracellularly, those with projections to layer 4C beta (two axons) and to layer 1 (one axon) had color-selective properties, whereas those (six axons) which arborized in 4C alpha all had transient, broad band and highly contrast-sensitive receptive fields. These properties are consistent with derivations from somata in the parvocellular and magnocellular divisions of the LGN, respectively. Afferents to 4C alpha were found to cover approximately 6 times as much surface area as afferents to 4C beta. The preterminal trunks of all axons were found to follow tortuous paths through the neuropil--paths that may derive from axon segregation during development. The wide ranging, patchy distributions of single afferents in 4C alpha suggest that individual 4C alpha axons supply more than one ocular dominance stripe. In one case where the terminal arborization of a 4C alpha axon was mapped against the transneuronally determined pattern of ocular dominance, three separate patches of terminal boutons were indeed found to coincide with the bands of one eye.
424 citations
TL;DR: C corticotropin-releasing factor, like many other neurohormones and peptides, may participate in neuroendocrine regulation as well as play a role as a neurotransmitter-like substance in numerous extrahypothalamic circuits.
Abstract: The discovery of a 41-amino acid peptide with potent corticotropin-releasing factor properties has prompted a search for neurons that contain this substance and potentially utilize it in intercellular communication. The present study utilized immunohistochemical methods and an antiserum directed against a synthetic replica of ovine corticotropin-releasing factor. The rat hypothalamus was found to contain striking immunoreactive groups of neuronal perikarya within the paraventricular, periventricular, and anterior hypothalamic nuclei, some of which are likely to project to the external layer of the median eminence and thereby comprise a hypophysiotropic system. Certain other hypothalamic nuclei, as well as many other regions of the central nervous system, were found to contain corticotropin-releasing factor-immunoreactive neurons. Among the most prominent of these were neurons in the bed nucleus of stria terminalis, the central nucleus of the amygdala, the region of the dorsal raphe, locus ceruleus, the external cuneate nucleus, and the medullary reticular formation. Thus, corticotropin-releasing factor, like many other neurohormones and peptides, may participate in neuroendocrine regulation as well as play a role as a neurotransmitter-like substance in numerous extrahypothalamic circuits.
416 citations
TL;DR: The anatomical organization and electrophysiological characteristics of a projection from the nucleus accumbens to anteroventral parts of the globus pallidus and to a subpallidal region that includes the substantia innominata, the lateral preoptic area, and anterior part of the lateral hypothalamic area were investigated in the rat.
Abstract: The anatomical organization and electrophysiological characteristics of a projection from the nucleus accumbens to anteroventral parts of the globus pallidus and to a subpallidal region that includes the substantia innominata (SI), the lateral preoptic area (LPO), and anterior parts of the lateral hypothalamic area (LHA) were investigated in the rat. Autoradiographic experiments, with injections of 3H-proline into different sites in the nucleus accumbens and adjacent caudoputamen, indicate that the descending fibers are organized topographically along both mediolateral and dorsoventral gradients, although labeled fibers from adjacent regions of the nucleus accumbens overlap considerably in the ventral globus pallidus and subpallidal region. Injections confined to the caudoputaman only labeled fibers in the globus pallidus. Retrograde transport experiments with the marker true blue confirmed that only the nucleus accumbens projects to the subpallidal region and that the caudoputamen projects upon the glubus pallidus in a topographically organized manner. In electrophysiological recording experiments single pulse stimulation (0.1 to 0.7 mA; 0.15 msec duration) of the nucleus accumbens changed the discharge rate of single neurons in the ventral globus pallidus and in the SI, LPO, and LHA. Typically, the responses were inhibition of neuronal discharge with latencies of 6 to 18 msec. Single pulse stimulation of the dorsolateral caudoputamen altered the discharge rate of single neurons in dorsal regions of the globus pallidus, with inhibition being the most frequently observed response. The results of these anatomical and electrophysiological experiments are complementary and indicate that fibers from the nucleus accumbens innervate the anteroventral region of the globus pallidus as well as the subpallidal region, while most fibers of the caudoputamen innervate the globus pallidus but not the subpallidal region. It appears, therefore, that these two components of the striatum have different output connections. The possible functional significance of these findings is discussed in relation to the projections of the subpallidal region, which may include an output to the mesencephalic locomotor region, and in relation to the nucleus accumbens afferents from the amygdala and hippocampal formation.
TL;DR: The apparent concurrence of glial reaction and increase in neuronotrophic activity suggests that glial cells may be a major source of the induced trophic activity.
Abstract: Mechanical or chemical injury to adult rat brain elicited the accumulation in the affected area of trophic activity for cultured parasympathetic, sympathetic, and sensory neurons. Neuronotrophic activity was relatively low both in noninjured brain tissue extracts and in extracts prepared from the tissue surrounding an injury immediately after the lesion was made. However, trophic titers increased considerably over time, first in the brain tissue that formed the walls of the wound and then in the Gelfoam filling the wound cavity. In the tissue adjacent to the injury, trophic titers began to rise immediately after the lesion, reached a maximum 10 days later, and decayed thereafter. In the wound cavity, occupied by Gelfoam, neuronotrophic activity began to increase 6 days postlesion, reached a maximum at day 16 after injury, and decreased at later times. The levels of induced trophic activity appeared to be proportional to the size of the wound. Injury to various brain areas including temporal, entorhinal, occipital, parietal, and frontal cortices, hippocampus, corpus striatum, and cerebellum, all induced a similar increase in neuronotrophic factor(s). Damage to the myelinated fibers of the corpus callosum did not. High trophic titers decayed rapidly with distance from the wound except in areas heavily deafferented by the lesion, where activity also reached high levels. Extracts from all of the above- mentioned brain areas contained toxic activity for cultured spinal cord neurons. The level of neuronotoxic activity was similar both before the lesion and 15 days postlesion, with the possible exception of the corpus callosum. Intraventricular injections of kainic acid at doses which destroy areas CA4, CA3, and part of CA1 of the hippocampus also induced a time-dependent rise of neuronotrophic activity in this structure, comparable to that achieved by mechanical damage. Both kainic acid treatment and mechanical injury cause extensive glial proliferation in the injured and/or deafferented area. The apparent concurrence of glial reaction and increase in neuronotrophic activity suggests that glial cells may be a major source of the induced trophic activity. As an in vivo correlate of cell culture data, the survival of striatal transplants into host cortical wounds was examined. Fragments of embryonic corpus striatum did not survive when transplanted into a freshly made cavity in the entorhinal/occipital cortex of adult rats. Survival was enhanced by introducing a delay between the time at which the wound cavity was made and that at which the striatal tissue was implanted in it.(ABSTRACT TRUNCATED AT 400 WORDS)
TL;DR: The results of these studies suggest that cells in GPe, GPi, and STN may be involved in the control of movement parameters.
Abstract: We describe the relations between the direction, amplitude, and velocity of step-tracking arm movements and the frequency of single cell discharge in the external (GPe) and internal (GPi) segments of the globus pallidus and the subthalamic nucleus (STN) of the behaving monkey. Statistically significant relations to the direction, amplitude, and peak velocity of the movement were found in all structures studied predominantly during the movement but also during the reaction time. For movements in a particular direction, the discharge rate was frequently a linear function of the movement amplitude and/or peak velocity. The slopes of this relation differed for different cells and comprised both positive and negative values. STN differed from both GPe and GPi in that (a) a larger proportion of neurons in STN showed significant relations to the direction of movement and (b) the onset times of changes in neural activity related to movement occurred earlier in STN than in GPe or GPi. The results of these studies suggest that cells in GPe, GPi, and STN may be involved in the control of movement parameters. Loss of the basal ganglia output related to the amplitude or velocity of movement might account for the impairments of step movements observed in Parkinsonian patients. On the other hand, deranged or excessive output related to amplitude or velocity control might result in the excesses of movement observed in other disorders, such as chorea and hemiballismus. These studies also provide direct evidence that the STN exerts a specific influence on basal ganglia output related to the control of movement parameters.
TL;DR: The depletions of spinal cord substance P and somatostatin produced by neonatal capsaicin treatment were similar to those produced by dorsal rhizotomy, and Capsaicin does not appear to be specific for primary afferents containing either substance P or som atostatin.
Abstract: The numbers of myelinated and unmyelinated fibers were counted in dorsal roots of adult rats treated neonatally with capsaicin in doses ranging from 5 to 100 mg/kg. Substance P and somatostatin levels in the spinal cord, dorsal roots, and sensory ganglia also were determined in control and treated animals. Capsaicin administration lead to the loss of both small myelinated and unmyelinated fibers from dorsal roots. However, whereas a near total loss, up to 94%, of unmyelinated fibers was achieved after high doses of capsaicin, the reduction of myelinated fibers, even of the smallest caliber, did not exceed 40%. The degree of fiber loss showed a clear dose dependency, with little detectable damage to myelinated fibers at doses of less than 50 mg/kg and with an ED50 for damage to unmyelinated fibers of 5 to 10 mg/kg. In all of the structures examined, particularly the dorsal roots, a roughly parallel decrease of substance P and somatostatin was found with capsaicin dose. The depletions of spinal cord substance P (55%) and somatostatin (20%) produced by neonatal capsaicin treatment were similar to those produced by dorsal rhizotomy. Capsaicin does not appear to be specific for primary afferents containing either substance P or somatostatin.
TL;DR: Neurons in the rat central nervous system were examined for their content of both the acetylcholine-synthesizing enzyme, choline acetyltransferase (ChAT), and the transmitter-degrading enzyme, acetylCholinesterase (AChE).
Abstract: Neurons in the rat central nervous system (CNS) were examined for their content of both the acetylcholine-synthesizing enzyme, choline acetyltransferase (ChAT; acetyl-CoA, choline O-acetyl-transferase, EC 2.3.16), and the transmitter-degrading enzyme, acetylcholinesterase (AChE). ChAT was localized immunohistochemically and AChE was localized histochemically in normal, colchicine-treated, or diisopropylfluorophosphate (DFP)-treated rats, either in neighboring sections by standard procedures or simultaneously in the same sections by immunofluorescence for ChAT, followed by photography, followed by histochemistry for AChE, followed by brightfield rephotography of the same neurons. This combination of fluorescence and brightfield procedures allows both to be carried out at maximum intensity without fear of interference and enables the unambiguous visualization of all neurons that might be expected with either procedure. Based on previous examination of singly stained neighboring sections, five CNS areas were examined in detail in DFP-treated rats using the simultaneous procedure for localizing both ChAT and AChE in the same neurons in the same sections. Several hundred neurons were thus examined in each area. These areas were the caudate putamen, nucleus basalis of Meynert, medial septum, nucleus of the diagonal band of Broca (vertical and horizontal limbs), and zona incerta. In the caudate putamen and nucleus basalis magnocellularis of Meynert, all ChAT-positive neurons contained AChE and vice versa. In the medial septum and nucleus of the diagonal band of Broca, all ChAT-positive neurons contained AChE, but these neurons containing both ChAT and AChE were intermingled with neurons positive only for AChE.(ABSTRACT TRUNCATED AT 250 WORDS)
TL;DR: In the cat, as in other mammals, development of the retinogeniculate pathway is broadly characterized by an initial period of overlap followed by a period of segregation that gives rise to the adult pattern of afferent input.
Abstract: The prenatal development of connections between the retina and the lateral geniculate nucleus (LGN) was studied by means of the anterograde axonal transport of 3H-amino acids or horseradish peroxidase injected intraocularly in fetal cats older than embryonic day 27 (E27) and in newborn cats. (Gestation is 65 days.) A retinothalamic pathway exists as early as E28, when label can be seen in both ipsilateral and contralateral optic tracts. Afferents from the contralateral eye are the first to invade the anlage of the LGN by E32 with those from the ipsilateral eye following about 3 days later. Initially, the pattern of labeling within the nucleus is uniform, suggesting that the two sets of afferents must share a good deal of territory at early ages. By E47, however, gaps appear in the labeling pattern contralaterally, indicating that afferents from the two eyes are beginning to segregate from each other. Segregation continues so that by E54 it is possible to identify unambiguously regions of the LGN destined to comprise ipsilateral and contralateral eye layers. By birth, afferent input appears adult-like in organization, with the two sets of afferents almost completely segregated from each other into their appropriate layers. Cellular lamination of the nucleus has just commenced, however, thereby lagging the onset of afferent segregation by about 2 weeks. Prenatal development could be followed much more easily in the horizontal than in the coronal plane of section due to the finding here that the LGN is displaced approximately 90 degrees in the horizontal plane between E40 and E60. Measurements of the area occupied by the ipsilateral and contralateral afferents within the LGN indicated that even prior to segregation, the two sets of afferents are not completely intermixed within the LGN. On the contrary, those from the contralateral eye retain almost exclusive control of some territory throughout development. This detail contrasts with development in primates, in which intermixing of afferents from the two eyes is thought to be complete early on (Rakic, P. (1976) Nature 261: 467-471). Nevertheless, in the cat, as in other mammals, development of the retinogeniculate pathway is broadly characterized by an initial period of overlap followed by a period of segregation that gives rise to the adult pattern of afferent input.
TL;DR: The demonstration that following reinnervation, catecholamine synthetic capacity is restored suggests that the juxtaglomerular dopamine neurons remain in the bulb and that afferent receptor input is required for expression of TH enzyme.
Abstract: Peripheral afferent input regulates the expression of dopaminergic properties in a population of local circuit intrinsic neurons of the rodent olfactory bulb Lesions of the olfactory receptor neurons produced in the mouse by intranasal irrigation with either ZnSO4 or Triton X-100 and in the rat by surgical deafferentation or axotomy are associated with a decrease in the levels of dopamine (DA), the DA metabolite 3,4-dihydroxyphenylacetic acid (DOPAC), the activity of the enzyme tyrosine hydroxylase (TH), bulb weight and an increase in norepinephrine (NE) levels in the olfactory bulb The anatomical correlates of the biochemical sequelae of deafferentation of olfactory bulb DA neurons were studied using immunohistochemical techniques to localize TH Within 3 to 4 weeks all lesions produced a dramatic and uniform reduction in TH staining of the juxtaglomerular DA neurons and their processes which was paralleled by a reduction in DA and DOPAC levels and bulb weight Seven weeks following reversible chemical lesions produced by Triton X-100, DA and DOPAC levels and tissue weight as well as TH staining in the juxtaglomerular neurons returned to control levels These observations suggested that DA neurons remained present even when not demonstrable with TH antibodies Additional evidence for the continued presence of the DA neurons was the ability of the olfactory bulbs from both lesioned mouse and rat to synthesize DA from exogenously administered L-3,4-dihydroxyphenylalanine (l-DOPA) These data suggested that the decrease in DA levels and TH staining in the olfactory bulb following lesions of the olfactory receptor neurons were produced by transneuronal mechanisms since there was no direct injury of the bulb Furthermore, the demonstration that following reinnervation, catecholamine synthetic capacity is restored suggests that the juxtaglomerular dopamine neurons remain in the bulb and that afferent receptor input is required for expression of TH enzyme
TL;DR: It is reported that prenatal treatment of females with testosterone propionate (TP) significantly increases the number of SNB neurons found in adulthood, and that androgenized females did not display the postural correlates of male copulatory behavior more often than did control females.
Abstract: The spinal nucleus of the bulbocavernosus (SNB) and its target muscles are reduced or absent in normal female rats (Breedlove, S. M., and A. P. Arnold (1980) Science 210: 564–566). We now report that prenatal treatment of females with testosterone propionate (TP) significantly increases the number of SNB neurons found in adulthood. Dihydrotesterone propionate (DHTP) treatment just after but not before birth also masculinizes the number of SNB neurons in females. SNB soma size is significantly masculinized, i.e., enlarged, by administration of androgen prenatally or as late as 7 to 11 days after birth, even though this late postnatal treatment has no effect on the number of SNB cells. Following TP treatment in adulthood, the androgenized females did not display the postural correlates of male copulatory behavior more often than did control females. From these results we infer the following. (1) Androgens act both before and after birth to influence the sexually dimorphic development of the SNB system. (2) There are different sensitive periods for the masculinization of SNB neuronal number and neuronal size, indicating that these two dimorphic characteristics of the SNB are masculinized by somewhat independent mechanisms. (3) TP and DHTP may act via separate mechanisms to alter the number of SNB neurons. (4) Aromatized metabolites of testosterone are not necessary for masculinization of the SNB system. (5) Virilization of the SNB system does not ensure the masculinization of the traditionally defined measures of male copulatory behavior in rodents.
TL;DR: Aplysia hemolymph enhances cell survival, the initiation of neurite outgrowth from multiple sites on the cell body surface, the linear growth of the processes, and the amount of branching by those processes.
Abstract: Neurons from the abdominal ganglion of the mollusc Aplysia californica regenerate neurite processes in dissociated cell culture. Both the nature of neurite outgrowth and the morphology of the cells are influenced by the presence of adult Aplysia hemolymph in the growth medium and the presence of a portion of a cell's original axonal process. Aplysia hemolymph enhances cell survival, the initiation of neurite outgrowth from multiple sites on the cell body surface, the linear growth of the processes, and the amount of branching by those processes. Hemolymph also decreases the diameter of the outgrowing neurite fascicles and the diameter of the individual neurites within the fascicles. The presence of a cell's original axon reduces the time required for the initiation of neurite outgrowth and restricts the formation of multipolar processes. In addition, the presence of an initial axonal segment is essential for neurite regeneration from large adult neurons.
TL;DR: The hypothesis that perinatal androgens normally direct the sexually dimorphic development of the SNB and its target muscles is supported.
Abstract: Prenatal treatment of male rats with the anti-androgen, flutamide (FL), demasculinizes the sexually dimorphic spinal nucleus of the bulbocavernosus (SNB) by reducing the number of SNB neurons, the size of the somas and nuclei of SNB neurons, and the size of their target muscles in adulthood. However, FL does not affect mounting or the traditional, postural measure of intromission, indicating that the SNB system does not play a major role in the mediation of these particular behaviors. Postnatal testosterone propionate (TP) treatment of male rats castrated on the day of birth results in more male copulatory behaviors in adulthood and masculinizes all measures of the SNB system. The postnatal masculinization by TP is more pronounced in males treated prenatally with FL, for morphological but not behavioral measures. The combined treatment of prenatal FL and day 1 castration without TP therapy results in a male with a completely demasculinized SNB system. Specifically, such males have SNB neurons that are as scarce and as small as those of females and, like females, they lack the target muscles of the SNB. These results support the hypothesis that perinatal androgens normally direct the sexually dimorphic development of the SNB and its target muscles.
TL;DR: One of the antibodies was used to localize immunohistochemically known cholinergic structures in the rat brain, including motor neurons, basal forebrain neurons, and neostriatal neurons.
Abstract: Immunohistochemical localization of choline acetyltransferase (ChAT) in cholinergic neurons has been difficult to achieve because of problems encountered in producing specific antisera. Here we describe the production and characterization of several distinct monoclonal antibodies to ChAT. Each of the monoclonal antibodies exhibits one of three general patterns of cross-species reactions; one pattern shows reactivity limited mainly to bovine ChAT, a second pattern shows reactivity only to ChAT from higher mammals including humans, and the third pattern shows reactivity to ChAT from all mammals tested. The antibodies bound specifically to two closely related bovine proteins of 68,000 and 70,000 daltons using the Western blotting technique. One of the antibodies was used to localize immunohistochemically known cholinergic structures in the rat brain, including motor neurons, basal forebrain neurons, and neostriatal neurons.
TL;DR: It is suggested that the growth of nerve fibers on fibronectin substrates results from direct interaction with a specific portion of the fibronECTin molecule and that this interaction can be inhibited by heparin and possibly other glycosaminoglycans.
Abstract: In an initial report (Carbonetto, S. T., M. M. Gruver, and D. C. Turner (1982) Science 216: 897–899) we described the use of 2- hydroxyethylmethacrylate (HEMA) in preparing defined culture substrates for studying nerve fiber growth. In those studies fibronectin and collagen were conspicuous, among a variety of HEMA-embedded proteins, in supporting fiber growth of embryonic neurons from chick dorsal root ganglia. Here we further document and extend our preliminary studies, especially with regard to the interaction of growing nerve fibers with fibronectin. HEMA substrates were prepared with proteolytic fragments of fibronectin, each of which had one or more of the functional sites of the intact molecule. Only those fragments of fibronectin that retained the region of the molecule known to mediate myoblast attachment were active in supporting nerve fiber growth. When added in excess to the culture medium, the smallest of the active fragments inhibited fiber growth on substrates that contained intact fibronectin. In culture medium depleted of serum fibronectin, HEMA gels containing collagens purified from connective tissues (types I and III) or from basement membranes (type IV) were about as effective as HEMA substrates containing fibronectin in supporting fiber growth. Nerve fiber growth on collagen substrates proceeded in the absence of fibronectin in the culture medium. Several glycosaminoglycans (heparin, chondroitin sulfate, hyaluronic acid) were ineffective as substrates for fiber growth. Treatment of HEMA/ fibronectin gels with heparin or incorporation of heparin along with fibronectin in the gel profoundly diminished the efficacy of fibronectin in supporting nerve fiber growth. Our studies suggest that the growth of nerve fibers on fibronectin substrates results from direct interaction with a specific portion of the fibronectin molecule and that this interaction can be inhibited by heparin and possibly other glycosaminoglycans.
TL;DR: In this article, anteroventral cochlear nerve recordings were made from cells in brain slices of the antero-vocal cortex of the auditory nerve and two types of electrical characteristics were found: linear currentvoltage relationships in the sub-threshold voltage range and large, regularly firing action potentials in the suprathreshold range.
Abstract: Intracellular recordings were made from cells in brain slices of the anteroventral cochlear nucleus. Responses to electrical stimulation of the stump of the auditory nerve were: (1) all-or-none, following the stimulus with no delay, and insensitive to the removal of extracellular Ca2+, probably representing the firing of directly stimulated auditory nerve fibers, or (2) graded, excitatory postsynaptic potentials, with synaptic delays of about 0.7 msec, which were often followed by graded, inhibitory synaptic potentials with synaptic delays of 1.2 msec or longer. The excitatory and inhibitory synaptic potentials were abolished by the removal of extracellular Ca2+. The result that delays of inhibitory synaptic responses were at least 2 times as long as those of excitatory ones indicates that probably an additional synapse was interposed. Responses to intracellularly injected current pulses show that cells in the anteroventral cochlear nucleus have one of two types of electrical characteristics. Type I properties are characterized by linear current-voltage relationships in the subthreshold voltage range and large, regularly firing action potentials in the suprathreshold range. Type II properties are characterized by nonlinear current- voltage relationships; suprathreshold depolarization elicits only one or two small action potentials. Type II characteristics are particularly well suited for maintaining the information contained in the timing and patterns of firing of the auditory nerve fibers.
TL;DR: A map of the system of serotonin-immunoreactive cell bodies, fibers, and nerve endings is constructed, and a dense plexus of nerve endings showing serotonin-like immunoreactivity surrounds each of the thoracic second roots in the vicinity of groups of peripheral neurosecretory neurons.
Abstract: Serotonin exerts a wide range of physiological actions on many different lobster tissues. To begin the examination of the role of serotonin in lobsters at a cellular level, we have used immunohistochemical methods to search for presumptive serotonergic neurons, their central and peripheral projections, and their terminal fields of arborization. Whole mount preparations of the ventral nerve cord and various peripheral nerve structures have been used for these studies. With these tissues, more than 100 cell bodies have been found that show serotonin-like immunoreactivity. Although a few of the cell bodies are located peripherally (near the pericardial organs, a well known crustacean neurohemal organ), the vast majority are located in central ganglia. Every ganglion in the ventral nerve cord contains at least one immunoreactive cell body. The projections of many of the neurons have been traced, and we have constructed a map of the system of serotonin-immunoreactive cell bodies, fibers, and nerve endings. In addition, a dense plexus of nerve endings showing serotonin-like immunoreactivity surrounds each of the thoracic second roots in the vicinity of groups of peripheral neurosecretory neurons. These peripheral nerve plexuses originate from central neurons of the ventral nerve cord. In some cases we have been able to trace processes from particular central cell bodies directly to the peripheral nerve root plexuses; in other cases we have traced ganglionic neuropil regions to these peripheral endings.
TL;DR: The sources of ascending input to the medial geniculate body of the cat were studied using the retrograde transport of horseradish peroxidase to provide evidence for multiple parallel auditory pathways through the thalamus.
Abstract: The sources of ascending input to the medial geniculate body (MGB) of the cat were studied using the retrograde transport of horseradish peroxidase (HRP). HRP injections were made iontophoretically through micropipettes which were also used to record physiological properties at the injection sites. This technique produced small injections which appeared to be restricted to single subnuclei. The tectothalamic projection of the auditory system was found to consist of at least four distinct and separate pathways. The ventral division of the MGB receives a topographical projection from the central nucleus of the inferior colliculus (ICC) which preserves tonotopicity and provides short latency, sharply frequency-tuned responses. The medial part of the ICC projects to the deep dorsal nucleus, which contains only units tuned to high frequencies. The major inputs to the caudodorsal nucleus (DC) stem from nucleus sagulum and the pericentral nucleus of the inferior colliculus (ICP). Units in DC and the ventrolateral nucleus, which also receive input from ICP, have very broad tuning properties and late, habituating responses. Injections of HRP into the medial division (MGM) produced labeled cells scattered throughout the external nucleus of the inferior colliculus and the ventral part of ICC. This widespread input is reflected in the wide range of auditory responses found in MGM. Auditory responses in the suprageniculate nucleus were poorly defined and many units did not respond to tonal stimuli; following HRP injections no filled cells were found in the inferior colliculus, but labeled cells were found in the deeper layers of the superior colliculus and in the interstitial nucleus of the brachium of the inferior colliculus. Together with recent findings on the auditory thalamocortical projection, these results provide evidence for multiple parallel auditory pathways through the thalamus.
TL;DR: It is suggested that glycine may be a neurotransmitter mediating binaural inhibition in certain SOC neurons and that the projection to the lateral superior olivary nucleus from the medial nucleus of the trapezoid body may be glycinergic.
Abstract: The present study seeks to identify neurotransmitters mediating binaural inhibition in lateral superior olivary nucleus neurons. Neurons in this auditory structure receive inputs from both ears and are thought to code for localization of sound in space. Iontophoretic application of glycine during monaural stimulation was found to mimic the inhibition observed with binaural stimulation. Binaural inhibition was blocked by application of the glycine receptor antagonist, strychnine, as were the effects of iontophoretic application of glycine. The post-strychnine recovery time course for return of synaptically mediated binaural inhibition and recovery of the effects of iontophoretic glycine application were identical. Although the superior olivary complex (SOC) neurons displaying binaural inhibition could in some cases be inhibited by GABA, the binaural inhibition rarely was blocked by iontophoretic application of the GABA receptor antagonist, bicuculline. These findings suggest that glycine may be a neurotransmitter mediating binaural inhibition in certain SOC neurons and that the projection to the lateral superior olivary nucleus from the medial nucleus of the trapezoid body may be glycinergic.
TL;DR: The results are discussed in terms of physiological functions associated with CCK, presynaptic receptors, and axonal flow of CCK receptors in the guinea pig and rat central nervous system.
Abstract: Cholecystokinin (CCK) receptor binding sites have been localized by autoradiography in the guinea pig and rat central nervous system. [125I]CCK-triacontatriapeptide labeled the sites in brain slices with an observed association constant equal to 0.041 min-1 and a dissociation constant equal to 0.008 min-1. CCK-triacontatriapeptide (CCK-33) and the C-terminal octapeptide of CCK-33 (CCK-8) potently inhibited [125I]CCK-33 binding with Ki's of 2 nM, whereas desulfated CCK-8 (CCK8-ds) and the C-terminal tetrapeptide of CCK-33 (CCK-4) were much weaker. Receptors were concentrated in the olfactory bulb, in the superficial laminae of the primary olfactory cortex, in the deep laminae of the cerebral cortex, and in the pretectal area. Substantial numbers of sites were also found in the basal ganglia, in the amygdala, and in the hippocampal formation. [125I]CCK-33 binding sites appear to be located on fibers of the optic tract and probably on olfactory tract fibers as well. These results are discussed in terms of physiological functions associated with CCK, presynaptic receptors, and axonal flow of CCK receptors.
TL;DR: The results suggest that neuronal coupling is extensive in immature rat neocortex, but that coupling declines at a time just before the numbers of chemical synapses increase most rapidly.
Abstract: We have estimated the prevalence of coupling between neurons of the rat neocortex during postnatal development. Single intracellular injections of the fluorescent dye Lucifer Yellow CH resulted in dye coupling among 70% of neurons from 1 to 4 days of age. Dye coupling dropped to 30 to 40% by 10 to 18 days and occurred in 20% of injected adult neurons. The number of neurons per dye-coupled aggregate also decreased. Whereas three to seven coupled neurons were common in cortex of 1 to 4 days, aggregates of more than two neurons were exceptionally rare in adults. The frequency of dye coupling did not vary systematically with cortical depth at any age. When chemical synaptic activity was blocked, most 4- day neurons exhibited short latency antidromically evoked depolarizations which were relatively insensitive to repetitive activation and membrane polarization. These depolarizations may represent electrotonically conducted spikes from coupled neurons. No such potentials were found in adult neurons. The results suggest that neuronal coupling is extensive in immature rat neocortex, but that coupling declines at a time just before the numbers of chemical synapses increase most rapidly.
TL;DR: The results suggest that the growth cones of the G and C neurons are determined to recognize and extend upon labeled axons, leading us to propose the “labeled pathways” hypothesis.
Abstract: In the previous paper (Raper, JA, M Bastiani, and CS Goodman (1983) J Neurosci 3:20-30) we showed that the growth cones of two sibling neurons, the G and C cells, follow the same route in the developing grasshopper neuropil until they reach a stereotypic choice point Here their growth cones diverge from each other as G turns and extends anteriorly and C turns and extends posteriorly In this paper we show that the G and C growth cones fasciculate and extend in opposite directions upon a specific bundle of four axons This occurs even though many other axons are within filopodial reach of the G and C growth cones We identified the four neurons (the A1, A2, P1, and P2 cells) whose axons form the bundle that G and C extend upon by filling individual axons with Lucifer Yellow and viewing the filled cells in living embryos and by filling individual neurons with HRP and reconstructing the axon bundle from electron micrographs The G neuron extends anteriorly in the bundle containing these four axons; the C neuron extends posteriorly in the same bundle only after several other axons have joined in These results suggest that the growth cones of the G and C neurons are determined to recognize and extend upon labeled axons, leading us to propose the "labeled pathways" hypothesis
TL;DR: The results suggest that the 50-Kd phosphoprotein may be an autophosphorylatable subunit of the Synapsin I Kinase or may exist in a complex with it.
Abstract: A calcium/calmodulin-dependent protein kinase, which phosphorylates a synaptic vesicle-associated protein designated Synapsin I, has been shown to be present in both soluble and particulate fractions of rat brain homogenates. In the present study, the particulate activity was solubilized by washing with a low ionic strength solution, and the enzymes from the two fractions were partially purified by ion exchange chromatography and calmodulin-Sepharose affinity chromatography. By each of several criteria, the partially purified enzymes from the two sources were indistinguishable. These criteria included specificity for various substrate proteins, concentration dependence of activation by calcium and calmodulin, pH dependence, and apparent affinities for the substrates Synapsin I and ATP. The mild conditions that released the particulate enzyme indicated that it was not tightly bound to the membrane and suggested that it may exist in a dynamic equilibrium between soluble and particulate-bound states. The partially purified enzyme preparations from both the soluble and particulate fractions contained three proteins that were phosphorylated in the presence of calcium and calmodulin, a 50-kilodalton (Kd) protein and two proteins in the 60-Kd region. When compared by phosphopeptide mapping and two-dimensional gel electrophoresis, the proteins were indistinguishable from three proteins of corresponding molecular weights that were shown by Schulman and Greengard (Schulman, H., and P. Greengard (1978) Nature 271: 478-479) to be prominent substrates for calcium/calmodulin-dependent protein kinase in a crude particulate preparation from rat brain. The 50-Kd substrate was the major Coomassie blue staining protein in both partially purified enzyme preparations. The peak of this protein coincided with that of enzyme activity during DEAE-cellulose and calmodulin-Sepharose chromatography. These results suggest that the 50-Kd phosphoprotein may be an autophosphorylatable subunit of the Synapsin I Kinase or may exist in a complex with it.