Biomedical subjects

Brigitte van Zundert

Publications and source records attributed to Brigitte van Zundert.

5 recordsLinked to original sources

PubMed2005

Glycinergic and GABAergic synaptic transmission are differentially affected by gephyrin in spinal neurons.

In the present study, we have examined the physiological properties of synaptic currents mediated by GlyRs and GABAARs after culturing spinal neurons with a gephyrin antisense oligonucleotide. Application of gephyrin antisense, but not the sense, reduced the glycinergic mIPSC amplitude ( approximately 50%) and frequency ( approximately 85%), indicating the importance of gephyrin for GlyR anchoring at postsynaptic sites. On the other hand, the glycine-evoked current amplitude was unchanged indicating that functional GlyRs were still located in the extrasynaptic membrane. The analysis of the GABAergic transmission in the same neurons revealed approximately 70% reduction in the frequency of the GABAergic mIPSCs, without changes in the amplitude. Interestingly, the modulation of remaining GABAAR-mediated synaptic events by zinc and diazepam was significantly altered by the antisense. These results indicate that gephyrin is required for the membrane insertion/stabilization of the GABAAR gamma2 subunit as well as for its subsequent localization in the postsynaptic membrane.

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PubMed2004

Changes on the properties of glycine receptors during neuronal development.

Glycine receptors (GlyRs) play a major role in the excitability of spinal cord and brain stem neurons. During development, several properties of these receptors undergo significant changes resulting in major modifications of their physiological functions. For example, the receptor structure switches from a monomeric alpha or heteromeric alpha 2 beta in immature neurons to an alpha 1 beta receptor type in mature neurons. Together with these changes in receptor subunits, the postsynaptic cluster size increases with development. Parallel to these modifications, the apparent receptor affinity to glycine and strychnine, as well as that of Zn(2+) and ethanol increases with time. The mature receptor is characterized by a slow desensitizing current and high sensitivity to modulation by protein kinase C. Also, the high level of glycinergic transmission in immature spinal neurons modulates neuronal excitability causing membrane depolarization and changes in intracellular calcium. Due to these properties, chronic inhibition of glycinergic transmission affects neurite outgrowth and produces changes in the level of synaptic transmission induced by GABA(A) and AMPA receptors. Finally, the high level of plasticity found in immature GlyRs is likely associated to changes in cytoskeleton dynamics.

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PubMed2004

Receptor compartmentalization and trafficking at glutamate synapses: a developmental proposal.

This article focuses on NMDA receptor subunit changes that occur in the forebrain and midbrain during development, namely the switch from predominance of NMDA receptors rich in NR2B subunits to that of NMDA receptors rich in NR2A subunits. We review the potential roles in brain plasticity of two membrane-associated guanylate kinases (MAGUKs), SAP102 and PSD95, which form a scaffold for the ion-passing glutamate receptors at the postsynaptic density, and we consider the known functional significance of these molecules in subunit switching. In addition, based on recent analyses of the synaptic location of glutamate receptors, activity-dependent changes in developing visual neurons, and extensive data on MAGUKs, we propose a model of glutamatergic synaptic differentiation. In this model, different NMDA receptor scaffolding and signaling complexes effect the trafficking and synaptic localization of NR2A-rich and NR2B-rich receptors, leading to tangential compartmentalization of these receptors and their movement between synaptic and extrasynaptic compartments.

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PubMed2003

Developmental-dependent action of microtubule depolymerization on the function and structure of synaptic glycine receptor clusters in spinal neurons.

Microtubules have been proposed to interact with gephyrin/glycine receptors (GlyRs) in synaptic aggregates. However, the consequence of microtubule disruption on the structure of postsynaptic GlyR/gephyrin clusters is controversial and possible alterations in function are largely unknown. In this study, we have examined the physiological and morphological properties of GlyR/gephyrin clusters after colchicine treatment in cultured spinal neurons during development. In immature neurons (5-7 DIV), disruption of microtubules resulted in a 33 +/- 4% decrease in the peak amplitude and a 72 +/- 15% reduction in the frequency of spontaneous glycinergic miniature postsynaptic currents (mIPSCs) recorded in whole cell mode. However, similar colchicine treatments resulted in smaller effects on 10-12 DIV neurons and no effect on mature neurons (15-17 DIV). The decrease in glycinergic mIPSC amplitude and frequency reflects postsynaptic actions of colchicine, since postsynaptic stabilization of microtubules with GTP prevented both actions and similar reductions in mIPSC frequency were obtained by modifying the Cl(-) driving force to obtain parallel reductions in mIPSC amplitude. Confocal microscopy revealed that colchicine reduced the average length and immunofluorescence intensity of synaptic gephyrin/GlyR clusters in immature (approximately 30%) and intermediate (approximately 15%) neurons, but not in mature clusters. Thus the structural and functional changes of postsynaptic gephyrin/GlyR clusters after colchicine treatment were tightly correlated. Finally, RT-PCR, kinetic analysis and picrotoxin blockade of glycinergic mIPSCs indicated a reorganization of the postsynaptic region from containing both alpha2beta and alpha1beta GlyRs in immature neurons to only alpha1beta GlyRs in mature neurons. Microtubule disruption preferentially affected postsynaptic sites containing alpha2beta-containing synaptic receptors.

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PubMed2002

Glycine receptors involved in synaptic transmission are selectively regulated by the cytoskeleton in mouse spinal neurons.

Using whole cell patch-clamp recordings, we examined the effect of colchicine, a microtubule disrupter, on the properties of glycine receptors (GlyRs) in cultured spinal cord neurons. Confocal microscopy revealed that colchicine treatment effectively altered microtubule bundles and neuronal morphology. Application of colchicine via the culture media or the patch-pipette, however, did not affect the whole cell current rundown (73 +/- 6% of control after 1 h), the sensitivity of the GlyR to glycine (EC(50) = 29 +/- 1 microM), or strychnine inhibition (47 +/- 5% of control after 100 nM strychnine). On the other hand, colchicine dialyzed for 25 min via the patch pipette selectively reduced the quantal amplitude of spontaneous glycinergic miniature inhibitory postsynaptic currents (mIPSCs) to 68 +/- 5% of control. This effect was specific for GlyRs since synaptic events mediated by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and GABA(A) receptors were unchanged. In conclusion, this study indicates that microtubules can regulate the function of GlyRs involved in inhibitory synaptic transmission.

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