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D L Benson

Publications and source records attributed to D L Benson.

At least 19 recordsLinked to original sources
PubMed2002

Dendritic spine plasticity in hippocampus.

Most excitatory input in the hippocampus and cerebral cortex impinges on dendritic spines. Alterations in dendritic spine density or shape are suspected to be morphological manifestations of changes in physiology or behavior. The links between spine plasticity and physiological responses have probably been best studied in the hippocampus in the context of changes in the circulating levels of steroid hormones or long-term potentiation. Here we review and present data which indicate that both the age of the preparation and the timing of the analysis can dramatically effect the results obtained. Collectively the data suggest that different cellular and morphological strategies may be utilized at different ages and under different circumstances to effect similar physiological responses or behaviors.

Animals↗
PubMed2002

Identification and localization of multiple classic cadherins in developing rat limbic system.

Classic cadherins are multifunctional adhesion proteins that play roles in tissue histogenesis, neural differentiation, neurite outgrowth and synapse formation. Several lines of evidence suggest that classic cadherins may establish regional or laminar recognition cues by virtue of their differential expression and tight, and principally homophilic, cell adhesion. As a first step toward investigating the role this family plays in generating limbic system connectivity, we used RT-PCR to amplify type I and type II classic cadherins present in rat hippocampus during the principal period of synaptogenesis. We identified nine different cadherins, one of which, cadherin-9, is novel in hippocampus. Using in situ hybridization, we compared the cellular and regional distribution of five of the cadherins (N, 6, 8, 9 and 10) during the first two postnatal weeks in hippocampus, subiculum, entorhinal cortex, cingulate cortex, anterior thalamus, hypothalamus and amygdala. We find that each cadherin is differentially distributed in distinct, but highly overlapping fields that largely correspond to known anatomical boundaries and are often coordinately expressed in interconnected regions. For example, cadherin-6 expression defines CA1 and its principal target, the subiculum; cadherin-10 is differentially expressed in CA1 and CA3 in a manner correlating with the organization of interconnecting Schaffer collateral axons; and cadherin-9 shows a striking concentration in CA3. Some cadherin mRNAs are highly restricted to particular anatomical fields over the entire time course, while others are more broadly expressed and become concentrated within particular domains coincident with the timing of afferent ingrowth. Our data indicate that classic cadherins are sufficiently diverse and differentially distributed to support a role in cell surface recognition and adhesion during the formation of limbic system connectivity.

Amygdala↗
PubMed2001

Stages of synapse development defined by dependence on F-actin.

It has been widely speculated that actin plays a central role in CNS synapse assembly, but such a requirement for actin filaments (F-actin) has not yet been demonstrated experimentally. We used hippocampal neurons grown in culture and the actin depolymerizing agent, latrunculin A, to examine directly the relationship between F-actin and synapse formation and maturation. During the first week in culture, actin depolymerization results in a near complete loss of synapses defined by synaptophysin-labeled vesicle clusters, synaptic vesicle recycling, and ultrastructure. Over the second week in culture, F-actin becomes increasingly stable, but actin depolymerization no longer disrupts basic synaptic structure. There is, however, a reduction in the number and size of synaptophysin-labeled clusters and in the size of vesicle clusters undergoing FM4-64 recycling, suggesting that synaptic vesicle anchoring remains partially dependent on F-actin. By 18 d in culture, synaptophysin clusters and synaptic vesicle recycling are largely resistant to F-actin depolymerization. The decrease in synapse dependence on F-actin correlates well with the acquisition and retention of presynaptic scaffolding proteins such as Bassoon and postsynaptic scaffolding proteins such as those of the postsynaptic density-95 family. Increased activity stabilizes F-actin and its associated proteins at synaptic sites, suggesting a correlation between active synapses, actin stability, and synapse stability. Our findings demonstrate that F-actin is essential for the development and maintenance of young synapses. Because F-actin is also highly regulatable, we propose that F-actin may be a principal target for stabilizing or destabilizing signals that ultimately result in synapse maintenance or elimination.

Actins↗
PubMed2000

Development and molecular organization of dendritic spines and their synapses.

Nearly all excitatory input in the hippocampus impinges on dendritic spines which serve as multifunctional compartments that can, at the very least, selectively isolate and amplify incoming signals. Their importance to normal brain function is highlighted by the severe mental impairment observed in most individuals having poorly developed spines (Purpura, Science 1974;186:1126-1128). Distinct groups of membrane proteins, cytoskeletal elements, scaffolding proteins, and second messenger-related proteins are concentrated particularly in dendritic spines, but their ability to generate, maintain, and coordinately regulate spine structure or function is poorly understood. Here we review the unique molecular composition of dendritic spines along with the factors known to influence dendritic spine development in order to construct a model of dendritic spine development in relation to synaptogenesis.

Actins↗
PubMed2000

Increasing numbers of synaptic puncta during late-phase LTP: N-cadherin is synthesized, recruited to synaptic sites, and required for potentiation.

It is an open question whether new synapses form during hippocampal LTP. Here, we show that late-phase LTP (L-LTP) is associated with a significant increase in numbers of synaptic puncta identified by synaptophysin and N-cadherin, an adhesion protein involved in synapse formation during development. During potentiation, protein levels of N-cadherin are significantly elevated and N-cadherin dimerization is enhanced. The increases in synaptic number and N-cadherin levels are dependent on cAMP-dependent protein kinase (PKA) and protein synthesis, both of which are also required for L-LTP. Blocking N-cadherin adhesion prevents the induction of L-LTP, but not the early-phase of LTP (E-LTP). Our data suggest that N-cadherin is synthesized during the induction of L-LTP and recruited to newly forming synapses. N-cadherin may play a critical role in L-LTP by holding nascent pre-and postsynaptic membranes in apposition, enabling incipient synapses to acquire function and contribute to potentiation.

Animals↗
PubMed2000

Molecular modification of N-cadherin in response to synaptic activity.

The relationship between adhesive interactions across the synaptic cleft and synaptic function has remained elusive. At certain CNS synapses, pre- to postsynaptic adhesion is mediated at least in part by neural (N-) cadherin. Here, we demonstrate that upon depolarization of hippocampal neurons in culture by K+ treatment, or application of NMDA or alpha-latrotoxin, synaptic N-cadherin dimerizes and becomes markedly protease resistant. These properties are indices of strong, stable, enhanced cadherin-mediated intercellular adhesion. N-cadherin retained protease resistance for at least 2 hr after recovery, while other surface molecules, including other cadherins, were completely degraded. The acquisition of protease resistance and dimerization of N-cadherin is not dependent on new protein synthesis, nor is it accompanied by internalization of N-cadherin. By immunocytochemistry, we found that high K+ selectively induces surface dispersion of N-cadherin, which, after recovery, returns to synaptic puncta. N-cadherin dispersion under K+ treatment parallels the rapid expansion of the presynaptic membrane consequent to the massive vesicle fusion that occurs with this type of depolarization. In contrast, with NMDA application, N-cadherin does not disperse but does acquire enhanced protease resistance and dimerizes. Our data strongly suggest that synaptic adhesion is dynamically and locally controlled, and modulated by synaptic activity.

2-Amino-5-phosphonovalerate↗
PubMed2000

Making memories stick: cell-adhesion molecules in synaptic plasticity.

Synapses are adhesive junctions highly specialized for interneuronal signalling in the central nervous system. The strength of the synaptic signal can be modified (synaptic plasticity), a key feature of the cellular changes thought to underlie learning and memory. Cell-adhesion molecules are important constituents of synapses, with well-recognized roles in building and maintaining synaptic structure during brain development. However, growing evidence indicates that cell-adhesion molecules also play important and diverse roles in regulating synaptic plasticity and learning and memory. This review focuses on recent advances in understanding the molecular mechanisms through which adhesion molecules might regulate synaptic plasticity.

Animals↗
PubMed1999

Neural (N)-cadherin at developing thalamocortical synapses provides an adhesion mechanism for the formation of somatopically organized connections.

Thalamic projections from the ventrobasal (VB) nucleus to rodent somatosensory cortex develop highly ordered terminations that form discrete, clustered patches localized to layer IV cellular aggregates that are termed "barrels." The molecular signaling and adhesion events that occur at the synapse as barrel-clustered thalamic connections form are unknown. Here, we show that neural (N)-cadherin, a membrane glycoprotein mediating strong homophilic adhesion, is concentrated at the developing thalamocortical synaptic junctional complex and demarcates these synaptic junctions as they form their characteristic barrel clusters during the first postnatal week. Furthermore, experimentally altering the distribution of thalamocortical axon terminals by peripheral manipulation leads to an identically altered N-cadherin distribution pattern, which is significant in establishing that N-cadherin does not define region-specific patterns of synapse distribution proactively but, rather, conforms to patterning imposed by thalamic axons through instructional cues conveyed through several synaptic relays. At postnatal day 9, levels of N-cadherin expression rapidly decrease, leading to loss of N-cadherin labeling of the barrels and, at adulthood, elimination from VB thalamocortical synapses. However, alphaN- and beta-catenin, which are critical binding partners of the classic cadherins, persist at the adult synapse, suggesting the presence of another classic cadherin as the thalamocortical synapse matures. This is the first evidence linking a synapse adhesion molecule with the establishment of patterned thalamocortical synapse distribution, suggesting strongly that N-cadherin performs a critical role in this process by adhering presynaptic and postsynaptic membranes as ingrowing thalamic axon terminals and postsynaptic thalamorecipient sites link and stabilize into mature synaptic junctional complexes distributed with precise topographic order. It is speculated that the developmental redistribution of N-cadherin may reflect dynamic regulation of synaptic membrane adhesion, which, in turn, might modulate plasticity of thalamocortical synaptic function.

Aging↗
PubMed1998

N-cadherin redistribution during synaptogenesis in hippocampal neurons.

Cadherins are homophilic adhesion molecules that, together with their intracellular binding partners the catenins, mediate adhesion and signaling at a variety of intercellular junctions. This study shows that neural (N)-cadherin and beta-catenin, an intracellular binding partner for the classic cadherins, are present in axons and dendrites before synapse formation and then cluster at developing synapses between hippocampal neurons. N-cadherin is expressed initially at all synaptic sites but rapidly becomes restricted to a subpopulation of excitatory synaptic sites. Sites of GABAergic, inhibitory synapses in mature cultures therefore lack N-cadherin but are associated with clusters of beta-catenin, implying that they contain a different classic cadherin. These findings indicate that N-cadherin adhesion may stabilize early synapses that can then be remodeled to express a different cadherin and that cadherins systematically differentiate between functionally (excitatory and inhibitory) and spatially distinct synaptic sites on single neurons. These results suggest that differential cadherin expression may orchestrate the point-to-point specificity displayed by developing synapses.

Animals↗
PubMed1998

Polarized distribution and cell type-specific localization of telencephalin, an intercellular adhesion molecule.

Telencephalin is an intercellular adhesion molecule (ICAM) restricted to the telencephalon. This study demonstrates that immunolabeled telencephalin is targeted to the somatodendritic domain of cultured hippocampal pyramidal neurons beginning with the first stages of dendritic differentiation. In contrast, it is entirely excluded from all gamma-aminobutyric acid (GABA)ergic inhibitory interneurons at all stages of development. Prior to the stage at which nearly all pyramidal neurons express telencephalin, labeled neurons possess more extensive dendritic arbors than unlabeled pyramidal neurons. More synaptic boutons form with the larger, more elaborate telencephalin-expressing dendritic trees, but bouton number per unit length is similar between neurons with and without telencephalin. These findings suggest that telencephalin identifies pyramidal neurons, it may identify plasma membrane as dendritic, and it may be generally adhesive or stabilize dendritic membranes, but it is probably not specifically synaptic. Such characteristics would be expected to be important for the formation of cell type-specific dendritic arbors.

Animals↗
PubMed1997

Differential subcellular regulation of NMDAR1 protein and mRNA in dendrites of dentate gyrus granule cells after perforant path transection.

Unilateral transection of the excitatory perforant path results in the acute deafferentation of a segregated zone on the distal dendrites of hippocampal dentate gyrus granule cells (i.e., outer molecular layer), followed by sprouting, reactive synaptogenesis, and a return of physiological and behavioral function. To investigate cellular mechanisms underlying NMDA receptor plasticity in response to such extensive synaptic reorganization, we quantitatively evaluated changes in intensity levels of NMDAR1 immunofluorescence and NMDAR1 mRNA hybridization within subcellular compartments of dentate gyrus granule cells 2, 5, and 9 d after perforant path lesions. There were no significant changes in either measure at 2 d postlesion. However, at 5 and 9 d postlesion, during the period of axonal sprouting and synaptogenesis, there was an increase in NMDAR1 immunolabeling that was restricted to the dendritic segments of the denervated outer molecular layer and the granule cell somata. In contrast, NMDAR1 mRNA levels at 5 and 9 d postlesion increased throughout the full extent of the molecular layer, including both denervated and nondenervated segments of granule cell dendrites. These findings reveal that NMDAR1 mRNA is one of a limited population of mRNAs that is transported into dendrites and further suggest that in response to terminal proliferation and sprouting, increased mRNA transport occurs throughout the full dendritic extent, whereas increased local protein synthesis is restricted to denervated regions of the dendrites whose afferent activity is perturbed. These results begin to elucidate the dynamic postsynaptic subcellular regulation of receptor subunits associated with synaptic plasticity after denervation.

Animals↗
PubMed1997

Dendritic compartmentation of NMDA receptor mRNA in cultured hippocampal neurons.

The intracellular localization of mRNA encoding the obligatory subunit for the N-methyl-D-aspartate (NMDA) receptor complex (NMDAR1) was determined in cultured hippocampal neurons by in situ hybridization. From the time at which dendrites begin to develop, labeling for NMDAR1 mRNA extended beyond cell somata into incipient dendritic processes. By 10 days in culture, label was found throughout dendritic arbors in a distribution pattern similar to that for microtublin associated proteins 2 mRNA, but contrasted sharply with that for GluR1 or beta-actin mRNAs, both of which were restricted to cell somata. NMDAR1 mRNA was also expressed in astrocytes, where it was diffusely localized throughout the cytoplasm. These data suggest that transport of NMDAR1 mRNA may serve as a mechanism to target and manipulate local concentrations of NMDA receptors.

Animals↗
PubMed1996

Expression and polarization of VGF in developing hippocampal neurons.

VGF is an NGF-inducible protein whose expression closely correlates with PC12 cell neurite outgrowth. Immunolocalization of VGF in PC12 cells by electron microscopy revealed labeling of a subpopulation of large dense core vesicles. To determine whether VGF expression may be involved in neural and axonal differentiation, we examined VGF immunoreactivity in developing cultured hippocampal neurons. Most VGF appeared to be in vesicles which were present in neurons from the earliest developmental stages and were particularly abundant in a small number of neurons, most of which were GABAergic. VGF became restricted to axons after dendrites had begun to mature, a stage more closely associated with synaptogenesis than axonal polarization. Colocalization studies suggest that VGF-containing and synaptophysin-containing vesicles are distinct and differentially distributed.

Animals↗
PubMed1996

Activity-independent segregation of excitatory and inhibitory synaptic terminals in cultured hippocampal neurons.

Cultured hippocampal neurons were used as a model system to address experimentally the spatial and temporal sequence leading to the appropriate sorting of excitatory and inhibitory synaptic terminals to different cellular target domains and the role of neural activity in this process. By using antibodies against glutamic acid decarboxylase 65 (GAD65) and synaptophysin, we examined the development and segregation of GABAergic and non-GABAergic synaptic terminals on single neurons. Electron microscopy confirmed that GAD65-labeled swellings observed using light microscopy corresponded to synaptic boutons. From the time at which GABAergic terminals first appeared, they developed at a more rapid rate on neuronal somata than non-GABAergic terminals did, such that by 18 d in culture, 60% of the total boutons on somata were GABAergic. By contrast, the majority (70%) of boutons on dendrites were non-GABAergic. These data suggest that inhibitory synaptic terminals are targeted preferentially to or maintained on cell somata at the expense of excitatory terminals. Interestingly, non-GABAergic terminals were not inhibited from forming synapses on cell somata, because in the absence of GABAergic terminals they attained the same total somatic terminal density seen in the presence of GABAergic terminals. Chronic blockade of neuronal activity did not affect the differential targeting of GABAergic and non-GABAergic axons; however, it did reduce the extent of dendritic arborization. Our findings support a two-step model for synaptic segregation whereby the majority of terminals is initially targeted in an activity-independent manner to the appropriate cellular domains, but an additional developmental mechanism serves to further restrict and refine the original synaptic distribution.

Animals↗
PubMed1996

Compartmentation of alpha-internexin and neurofilament triplet proteins in cultured hippocampal neurons.

Intermediate filaments comprise an integral part of the neuronal cytoskeleton. However, little is known about their function, and there remains some uncertainty about their precise subcellular localization. We examined the timing of expression and distribution of alpha-internexin, neurofilament triplet proteins and peripherin using immunocytochemistry in cultured hippocampal neurons. alpha-Internexin immunostaining was present in all neurons at all developmental stages. Immunostaining appeared as long filaments in axons and short fragments in dendrites which extended into dendritic spines. The presence of alpha-internexin in dendritic spines was confirmed in situ by electron microscopy of rat hippocampal tissue sections and suggests that this intermediate filament may serve as a link between cytoskeletal elements in dendritic shafts and spines. In culture, immunostaining using antibodies against individual triplet protein subunits indicated that light (NF-L) and middle (NF-M) subunits were first expressed in cells shortly after the initiation of axonal outgrowth. Expression of the heavy (NF-H) subunit occurred a few days later. Although timing and localization of expression did not correlate with the initiation of axonal or dendritic processes, it was coincident with periods of rapid outgrowth. Triplet proteins were more abundant in axons and appeared to be incorporated into lengthier filaments than in dendrites. Highly phosphorylated NFH/M immunoreactivity was polarized to axons after 6 days in culture. The distribution of one NF-H epitope was restricted to GABAergic neurons in mature cultures, suggesting a cell-type specific modification. Peripherin was not detectable at any time in hippocampal cultures. Our results show that intermediate filaments are integral components of the neuronal cytoskeleton of cultured hippocampal neurons throughout development. Furthermore, the localization of alpha-internexin suggests that it may be involved in the formation or maintenance of dendritic spines.

Animals↗
PubMed1995

Decreased expression of the alpha subunit of Ca2+/ calmodulin-dependent protein kinase type II mRNA in the adult rat CNS following recurrent limbic seizures.

Calcium/calmodulin-dependent protein kinase type II (CamKII) is a ubiquitous brain enzyme implicated in a wide variety of neuronal processes. Understanding CamKII has become increasingly complicated with the recent identification of multiple gene transcripts coding for separate subunits. Previous studies have shown that mRNA for the alpha subunit of CamKII can be increased by reduction of afferent input. In this study we have examined the regulation of alpha CamKII mRNA following increased activity due to seizures. Using in situ hybridization with a cRNA probe against the rat alpha CamKII sequence we found reduced levels of hybridization following limbic seizures induced by lesions of the hilus of the dentate gyrus. Hybridization was most dramatically reduced in the granule cells of the dentate gyrus and the pyramidal cells of hippocampal region CA1. There were also significant reductions in hybridization in the superficial layers of neocortex and piriform cortex. In each of these region hybridization was decreased in the molecular layers which is consistent with the reported dendritic localization of alpha CamKII mRNA. All changes in mRNA content were transient, with maximal reductions at 24 h following lesion placement and a return to control levels by 96 h. These findings demonstrate the negative regulation of alpha CamKII mRNA by seizure activity and raise the possibility that synthesis of this kinase may be regulated by normal physiological activity.

Animals↗
PubMed1994

Characterization of GABAergic neurons in hippocampal cell cultures.

The morphological characteristics of GABAergic neurons and the distribution of GABAergic synaptic terminals were examined in cultures of hippocampal neurons from 4-35 days in vitro. Neurons expressing GABA immunoreactivity represented about 6% of the total number of cultured neurons at all time points. Although the morphological characteristics of GABAergic cells suggested a heterogeneous population, GABAergic cells as a class were notably different from the non-GABAergic, presumably pyramidal cells. Most GABAergic cells had more fusiform or polygonal shaped somata, non-spiny and less tapering dendrites and appeared more phase-dense than nonGABAergic cells. Quantitative analysis revealed that GABAergic cells had fewer primary dendrites, more elongated dendritic arbors, and longer dendritic segments than non-GABAergic neurons-characteristics that are similar to GABAergic cells in situ. Double immunostaining revealed that GAD65-positive varicosities were also immunopositive for synapsin I, suggesting that GAD65-positive varicosities that contacted somata and dendrites represented presynaptic specializations. Confocal microscopy revealed the proportion of the synaptic specializations on the cell soma that were GAD65-positive was greater than on the dendrites, suggesting that somata and dendrites differ in their ability to induce the formation of presynaptic specializations by GABAergic axons. These data indicate that the GABAergic cells that develop in culture exhibit distinctive morphological characteristics and participate in different synaptic interactions that nonGABA cells. Thus many of the features that distinguish GABAergic neurons in culture are reminiscent of the characteristics that distinguish GABAergic neurons in situ.

Animals↗