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M Constantine-Paton

Publications and source records attributed to M Constantine-Paton.

At least 19 recordsLinked to original sources
PubMed1995

Analysis of synaptic distribution within single retinal axonal arbors after chronic NMDA treatment.

Activation of the NMDA subtype of glutamate receptor has been implicated in structural synaptic plasticity in many developing sensory systems. In the frog retinotectal system, chronic exposure of the optic tectum to NMDA, which decreases the effectiveness of NMDA receptors (Debski et al., 1991), results in the pruning of the branches of retinal terminal arbors (Cline and Constantine-Paton, 1990). However, it is difficult from these studies to relate the involvement of NMDA receptors to changes in synapse distribution. In this study, we have developed an EM sampling procedure to quantitatively compare the amount and the distribution of synaptic contact within single retinal arbors. We report that within each retinal arbor, synaptic contact gradually increases from the main branches to the end branches of the arbor. Chronic NMDA treatment, however, significantly reduces the total amount of synaptic contact within each arbor. This reduced synaptic contact appears to be due to the pruning of the end branches, and the synapses these branches bear. The results are consistent with the hypothesis that NMDA receptor is an integral part of the mechanism that stabilizes coactive synapses, and that maintenance of an axonal branch requires a minimum density of synapses that are correlated with converging neighbors.

Animals↗
PubMed1994

Regulation of N-methyl-D-aspartate (NMDA) receptor function during the rearrangement of developing neuronal connections.

There is evidence from a number of studies that the molecular and biophysical properties of NMDA receptors are altered during normal development. A temporal correlation with changes in NMDA receptor efficacy and periods of synaptic plasticity has been demonstrated in several systems, suggesting that NMDA receptors have a critical function in determining periods of synaptic plasticity. Data from our laboratory demonstrate reduced NMDA sensitivity of the tectal evoked potential following chronic application of NMDA to the tadpole tectum, a treatment that may mimic a naturally occurring mechanism for limiting neuronal plasticity to certain stages of development. Our analysis of the expression pattern of mRNA coding for various glutamate receptor subunits in the rat retinocollicular system establishes that differential regulation of NMDA receptor subunits at the mRNA level could be a molecular basis for changes in biophysical and pharmacological properties of the NMDA receptor complex. However, even though the NMDA receptor is the best studied candidate to function as a 'plasticity switch', there are large gaps in our understanding of the complete set of factors that control the ability of synapses to rearrange during development.

Animals↗
PubMed1994

Effects of NMDA receptor antagonists on the developing brain.

This overview describes two effects of N-methyl-D-aspartate (NMDA) receptor antagonists that are not strictly speaking toxic: There are no signs that cells are dying. Nevertheless, these antagonists, if applied for prolonged periods in young children, could permanently cripple normal brain function. Disturbing the function of the NMDA channel during development can severely disrupt the wiring of defined neural circuits. It also can disrupt the developmental upregulation of the receptor protein itself and possibly many other molecular components of the synapse.

Animals↗
PubMed1993

The development of non-retinal afferent projections to the frog optic tectum and the substance P immunoreactivity of tectal connections.

Accessibility in early development and the presence of a retinotopic map have made the amphibian optic tectum a popular system for exploration of the role of synaptic function in central map refinement. Although a great deal is known about the development of retinal innervation of the tectum, little information exists about the development of non-retinal tectal inputs. Since these other afferent systems may contribute to the synaptic drive of developing tectal cells and thereby be involved in the activity-dependent refinement of the retinotectal map, we sought to determine whether these inputs are present at the early tadpole stages when the first retinal axons refine their synaptic order within the tectal neuropil. Rhodamine-labelled latex beads, retrogradely transported from injection sites in the optic tecta, were used to identify tectal afferent projections. Projection patterns in very young tadpoles were identical to those found in juvenile frogs and heavily labelled regions included areas of the posteroventral tegmental field, the posterior tuberculum, the ventromedial thalamic nucleus, the ventral part of the ventrolateral thalamic nucleus, the suprachiasmatic nucleus and discrete regions within the central and anterior thalamic nuclei. Ipsilateral nucleus isthmi cells were also labelled, indicating the existence of an isthmo-tectal projection in even the youngest animals examined. Additionally, substance P-like immunoreactive tecto-isthmal fibers were traced from the optic tectum to the nucleus isthmi. The presence of these connections suggests that feedback from the nucleus isthmi and/or input from the other brain areas projecting to the tectum may play a role in modulating the cellular mechanisms that underlie the formation of the visual map.

Afferent Pathways↗
PubMed1993

The contributions of NMDA, non-NMDA, and GABA receptors to postsynaptic responses in neurons of the optic tectum.

Activation of the NMDA subtype of glutamate receptor has been implicated in activity-dependent development and plasticity in several systems, including the retinotectal system of amphibians. To gain a better understanding of the response properties of tectal neurons, with particular emphasis on the role of both non-NMDA and NMDA glutamate receptors, we have developed an in vitro slice preparation of the diencephalon and midbrain of frog (Rana pipiens) tadpoles. In these slices, we electrically stimulated the optic tract and recorded both mono- and polysynaptic responses in single tectal neurons using whole-cell voltage clamp or current clamp. By including biocytin in the recording electrode, we were also able to determine the location and morphology of many of these neurons. Using these techniques, we found that the current-voltage (I-V) relations for both mono- and polysynaptic responses of tectal neurons showed voltage dependence only in the presence of extracellular Mg2+. This dependence reflects the hyperpolarization-dependent block of the NMDA channel by Mg2+. Bath application of 6-cyano-7-nitroquinoxaline-2,3-dione, a non-NMDA glutamate receptor antagonist, reduced both mono- and polysynaptic responses of tectal neurons. Bath application of the NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid (DL-APV) strongly reduced polysynaptic responses. When neurons were depolarized by the voltage clamp, relieving the Mg(2+)-dependent block of the NMDA channel, DL-APV application also reduced monosynaptic responses. Application of the GABAA receptor antagonist (-)bicuculline methiodide significantly increased the polysynaptic responses of tectal neurons, reflecting block of inhibition. We further confirmed the presence of these three types of receptors by examining postsynaptic currents evoked by iontophoretic application of the three agonists, NMDA, (R,S)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA), and GABA. These results confirm that the dominant excitatory transmitter in the tectum appears to be glutamate. Furthermore, the retinotectal synapses (i.e., monosynaptic currents) express functional NMDA receptors that are voltage dependent and are not responsible for the bulk of normal excitatory transmission. Polysynaptic responses, however, are mediated by both non-NMDA and NMDA receptors, and inhibition plays a significant role in sculpting these polysynaptic responses.

2-Amino-5-phosphonovalerate↗
PubMed1992

N-methyl-D-aspartate receptor antagonists disrupt the formation of a mammalian neural map.

The topographic ordering of retinal connections in the rat superior colliculus emerges during early postnatal life from an initially diffuse projection. Disruption of N-methyl-D-aspartate (NMDA) receptor activity in the superior colliculus during this period interferes with map remodeling. In rats chronically treated with NMDA receptor antagonists during the first two postnatal weeks, aberrant axons remain and arborize at topographically incorrect sites. These results indicate that, at a stage preceding visually evoked activity, normal NMDA receptor function is important for the development of an ordered neural map in the mammalian brain.

2-Amino-5-phosphonovalerate↗
PubMed1992

Physiological effects of chronic and acute application of N-methyl-D-aspartate and 5-amino-phosphonovaleric acid to the optic tectum of Rana pipiens frogs.

Visually elicited activity contributes to the formation of orderly connections in the optic tectum of frogs. Glutamate receptors of the N-methyl-D-aspartate class participate in this process. Blocking those receptors interferes with activity-dependent refinement of maps in normal frogs and of ocular dominance bands in surgically produced animals with three eyes. Chronic application of N-methyl-D-aspartate sharpens the bands. The possibility that 5-amino-phosphonovaleric acid depresses tectal responsiveness was motivation for studying the effects of 5-amino-phosphonovaleric acid and N-methyl-D-aspartate applied both chronically and acutely. We evaluated tectal responsiveness to visual input by presenting flashes of light to one eye and recording responses in the ipsilateral tectal lobe. This method reveals the output of the tectal cells contralateral to the stimulated eye. These cells project via the nucleus isthmi to the opposite tectal lobe. We also mapped the receptive field dimensions of the crossed isthmotectal axons. Our results show that acute topical application of 500 microM or 1 mM N-methyl-D-aspartate dramatically increases spontaneous activity, while 100 microM N-methyl-D-aspartate causes little change. Chronic treatment with N-methyl-D-aspartate at a low dose (estimated to be in the micromolar range) shown to influence retinotectal mapping, reduces response latencies but produces no statistically significant changes in tectal cell firing rates or receptive field size. Acute application of 5-amino-phosphonovaleric acid produces complex results: 10 microM produces no changes in firing, 100 microM 5-amino-phosphonovaleric acid decreases firing, and doses of 500-100 microM increase the firing.(ABSTRACT TRUNCATED AT 250 WORDS)

2-Amino-5-phosphonovalerate↗
PubMed1991

Chronic application of NMDA decreases the NMDA sensitivity of the evoked tectal potential in the frog.

The activity-dependent mechanism that refines the topography of the retinotectal projection in frogs is mediated by the NMDA receptor. Earlier studies found that chronic treatment of the optic tectum with the NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid (DL-AP5) desegregated eye-specific stripes in three-eyed frogs, while chronic treatment with NMDA sharpened stripe borders (Cline et al., 1987; Cline and Constantine-Paton, 1990). We now report that this same chronic treatment with NMDA decreases the electro-physiologically measured sensitivity of the optic tectum to applied NMDA: acute application of a given concentration of NMDA decreased the evoked tectal potential to a lesser extent in animals chronically treated with NMDA than it did in normal and sham-treated animals. This is observed as a shift to the right in the NMDA dose-response curves for both the positive and negative postsynaptic components of the evoked tectal response. We believe that this decreased NMDA receptor effectiveness further restricts the intermingling of axon branches from the two eyes by limiting synapse stabilization to areas where afferent activity is most correlated. This would account for the anatomical sharpening of stripe borders (i.e., increased afferent segregation). Quantitative autoradiographic analysis of 3H-glutamate binding to NMDA receptors indicated that binding densities within the tectum did not differ between control groups and NMDA chronically treated groups. We suggest that in the experimental animals the response to NMDA may be decreased by a change in the effectiveness of individual NMDA receptors rather than by decreases in receptor number. This experimentally induced change may be analogous to naturally occurring decreases in receptor function that correlate with the end of some periods of visual plasticity in mammals.

2-Amino-5-phosphonovalerate↗
PubMed1990

Activity-dependent tuning and the NMDA receptor.

The refinement of the topographic map of visual space within the optic tectum of the frog is activity-dependent. The use of the three-eyed frog preparation to assay the operation of this fine-tuning mechanism indicates that this process is mediated by the NMDA receptor: Chronic in vivo treatment with APV, an NMDA antagonist, disrupts the segregation of retinal afferents into eye-specific zones while NMDA treatment sharpens this pattern. This latter effect is accompanied by a decreased sensitivity of the system to applied NMDA. Activation of the NMDA receptor may mediate the fine-tuning mechanism by initiating the stabilization of appropriate synapses. The requirements for NMDA receptor activation necessitate the convergence of terminals carrying correlated activity patterns. Such patterns of activity are provided by ganglion cells whose cell bodies lie near one another in the retina, and who should therefore, in an accurate visual map, terminate near one another in the tectum. Synapses from ganglion cells who do not neighbor one another in the retina have uncorrelated firing patterns and therefore do not activate the NMDA receptor. These synapses then would not be stabilized relative to one another. In addition to organizing the retinal projection, NMDA receptor activation may also modulate retinal ganglion cell arbor morphology, since chronic in vivo APV or NMDA treatments decrease arbor density. These results are discussed in terms of the effect of NMDA receptor activation on branch initiation and the rate of branch retraction.

Animals↗
PubMed1990

The role of the NMDA receptor in the development of the frog visual system.

The importance of patterned retinal activity in visual system development has been recognized since Hubel and Wiesels early experiments (1963). The NMDA receptor is one cellular mechanism which can recognize patterned retinal activity and convey an intracellular message of that activity. It will now be of considerable interest to elucidate the cellular events involved in synapse stabilization subsequent to NMDA receptor activation.

Animals↗
PubMed1990

Evoked pre- and post-synaptic activity in the optic tectum of the cannulated tadpole.

We describe the cannulated Rana pipiens, tadpole preparation that allows for stable recording in the tectum of the extracellular potential elicited by optic nerve stimulation. The largest components of the evoked tectal response consist of two previously identified waves and a major third, long-latency wave of long duration. These components were reversibly eliminated by perfusion of high magnesium/no calcium Ringer's solution or Ringer's solution containing cobalt chloride. In contrast, perfusion of high calcium/no magnesium Ringer's increased the amplitude and area of these components. We conclude that these components represent post-synaptic activity. Additionally, small, short-duration waves were identified as arising from the activity of retinal afferents. They consisted of a short-latency (3.1-7.6 ms) and a long-latency (12-23 ms) group. Waves belonging to both of these classes were still visible in both high magnesium/no calcium Ringer's solution or Ringer's solution containing cobalt chloride and were unaffected by high calcium/no magnesium Ringer's. The average conduction velocities of the short- and long-latency groups matched the conduction velocities of, respectively, edge and convexity detectors in the adult. This indicates that retinal afferent input may already be present in adult patterns at the time that tectal circuitry is developing.

Animals↗
PubMed1990

Granule cell induction of 9-O-acetyl gangliosides on cerebellar glia in microcultures.

In previous studies we have shown that the expression of acetylated gangliosides recognized by the JONES monoclonal antibody is correlated with regions of cell migration in the developing rat nervous system. In this study we have investigated the expression of these gangliosides in two different types of cultures prepared from dissociated postnatal rat cerebella. In the first type, cells are plated after dissociation under conditions where most of the glial cells develop a stellate morphology that anchors neurons but does not support their migration. In the second type of culture, cells are plated in a ratio of four neurons to one glial cell and under these conditions the predominant form of astroglia is an elongate form that supports the migration of granule neurons. Granule neurons express JONES antigens in dissociated cell suspensions and in cultures in which cells are plated either after dissociation or in a 4:1 neuron:glia ratio. On the other hand, glial cells grown in the absence of neurons are JONES negative. In addition, the expression of JONES gangliosides by glial cells is different in the two types of culture. In cultures where the astroglial cells display the stellate morphology only a small proportion show JONES staining. Cultures in which the glial cells assume the elongate morphology have a significantly higher number of JONES-positive astroglia.

Acetylation↗
PubMed1990

The differential influence of protein kinase inhibitors on retinal arbor morphology and eye-specific stripes in the frog retinotectal system.

We investigated retinal axon morphology and eye-specific afferent termination zones in the optic tectum of three-eyed tadpoles that were chronically treated with protein kinase inhibitors. The kinase inhibitors sphingosine, H-7, and phorbol ester, which down-regulates protein kinase C with chronic exposure, were applied to the tecta in a slow release plastic, Elvax. In vivo protein phosphorylation assays in drug-treated tadpoles indicated that the treatments decreased 32P incorporation into some protein bands by as much as 60%. Although the drugs did not cause a desegregation of the eye-specific stripes, treated retinal axon arbors covered about half the area covered by untreated arbors or arbors treated with inactive analogs of the drugs. We conclude that eye-specific segregation can be maintained under conditions that markedly alter retinal ganglion cell axon arbor size and that significantly perturb protein phosphorylation. Furthermore, we conclude that the protein kinase(s) that we blocked with these treatments is involved in the growth of axon arbors.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1990

NMDA receptor agonist and antagonists alter retinal ganglion cell arbor structure in the developing frog retinotectal projection.

The development of sensory maps is thought to require an activity-dependent structural rearrangement of afferent terminal arbors within the CNS which recreates the topographic relations of sensory somata present in the periphery. In the frog retinotectal projection, activation of the NMDA receptor plays a role in this structural plasticity. Exposure of the optic tectum of tadpoles to NMDA receptor antagonists results in a rearrangement of retinal ganglion cell arbors so that their organization into a topographic projection and eye-specific stripes is disrupted (Cline et al., 1987; Cline and Constantine-Paton, 1989). Exposure of the optic tectum to the receptor agonist, NMDA, increases the eye-specific segregation of these arbors (Cline et al., 1987). We examined the projection of the supernumerary retina and the morphology of individual retinal afferent arbors of untreated, NMDA-treated, APV-treated, MK801-treated, and MK801/NMDA-treated 3-eyed tadpoles and young postmetamorphic frogs in an effort to understand how NMDA receptor activation is involved in the growth and ordering of retinal arbors. Treatments with MK801 in combination with NMDA resulted in a desegregation of eye-specific stripes, whereas treatments with MK801 or NMDA alone did not. As reported previously, APV treatment resulted in stripe desegregation without increasing the tangential area (measured from 2-dimensional drawings) of the terminal arbors. However, a detailed analysis revealed that the APV-treated tadpole arbors have 35% reduction in branch density (branch tips/area) compared to untreated 3-eyed tadpole arbors. We treated the optic tectum with a range of concentrations of NMDA prepared in the slow-release plastic Elvax. NMDA at 10(-4) M in Elvax was the optimal concentration to produce the sharpening of stripe borders. Exposure of the tectum to NMDA at 10(-6) M in Elvax produced no change in the stripe pattern, while 10(-2) M NMDA in Elvax resulted in beading of the arbors. At the optimal concentration NMDA treatment results in a 75% reduction in the number of axons crossing from a stripe to an interstripe zone. Drawings of individual HRP-labeled, NMDA-treated arbors demonstrate that they have fewer branch points and fewer branch tips. NMDA treatment reduced arbor density by approximately 50%. Arbors drawn from untreated postmetamorphic frogs have twice the branch density of arbors from untreated tadpoles. NMDA treatment in these animals reduced the branch density by 55%, comparable to the reduction seen in tadpole branch density. Our data support a specific hypothesis for NMDA receptor involvement in the activity-dependent structural refinement process within the developing retinotectal projection.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1989

Monoclonal antibody markers for amphibian oligodendrocytes and neurons.

Few immunocytochemical probes have been developed for cold-blooded vertebrates, thus hampering analyses of cellular processes in these species. Those developed from mammalian and avian tissue often fail either to react or to show similar specificities in poikilotherms. Therefore, we have begun raising monoclonal antibodies (mabs) in mice against frog and tadpole brain tissue. The following analyses of two of these mabs suggest that these antibodies represent specific probes for frog axons and oligodendrocytes. Mab Olig recognizes all the myelinated axon tracts of the mature frog brain and spinal cord, as well as the tracts of the developing tadpole CNS once they have become myelinated. Axons cut in cross section show characteristic o-shaped staining around individual axons when processed with this antibody. Particularly easy to visualize in the tadpole are immunoreactive cell bodies and processes, seen in continuity with the myelin sheath. Occasionally, in this developing tissue, cells with highly branched processes characteristic of immature oligodendrocytes are observed. No other cells or processes within the brain or spinal cord react with this antibody. Mab Linc stains numerous filaments in all axonal projections. Occasionally, a thin rim of filamentous staining is observed in cell somata, but many regions rich in neuronal somata or dendrites are unreactive to this antibody. This in vivo staining pattern suggests that the Linc antigen is differentially distributed within neurons and exhibits a high concentration in axons. Linc immunoreactivity is robust in the processes of a subpopulation of dissociated tectal cells in culture. These Linc-positive cells are characterized as neurons on morphological criteria. Also, intense Linc immunoreactivity is present in the bundles of retinal axons that extend from retinal explants. Olig immunoreactivity, however, has not been detected in tectal cultures or retinal explants. Improved staining following Triton X-100 treatment of tissue sections suggests that neither of the mabs recognizes lipid antigens and that both are probably localized within the cell cytoplasm. Only the Linc mab reacts on Western blots of denatured brain protein. Linc consistently recognizes two Triton X-100-insoluble proteins with apparent molecular weights of 56 and 58 kD. The epitopes recognized by the Olig and Linc mabs have been surveyed in terms of their resistance to optic nerve crush and their consequent value in studies requiring such procedures. Possible homologies to known cell-type-specific molecules are discussed.

Animals↗
PubMed1989

Quantitative autoradiographic localization of NMDA, quisqualate and PCP receptors in the frog tectum.

An organizing role for the N-methyl-D-aspartate (NMDA) receptor/channel has been suggested in the development of the retinotectal projection in Rana pipiens. The regional distributions of NMDA, phencyclidine (PCP) and quisqualic acid (QA) receptors were quantified using in vitro autoradiography in the tectum of normal and surgically produced 3-eyed juvenile frogs. NMDA and QA receptor binding was highest in the pretectum. Of the tectal layers, the superficial retinotectal synaptic zone, layer 9, had the highest amount of NMDA and QA receptor binding. Moderate binding was observed in layer 5, with little binding in the cellular layer 6. No specific [3H]N-(1-[2-thienyl]cyclohexyl) piperidine ([3H]TCP) binding was observed in any of the tectal regions.

Animals↗