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Contenido archivado el 2022-12-23

Lateral organization of membrane proteins on surfaces of developing nerve cells exposed to effects of neuroplasticity and cytoskeletal disruption

Objetivo



Molecular mechanisms of neural development and network formation, which provide the basis of brain activity and a clue to brain dysfunctions, recruit neural cell adhesion molecules (N-CAM) and excitatory amino acid receptors that are arranged on cell surfaces. This arrangement is believed to depend upon the cytoskeleton and alters with neural developmental and plastic changes, thus contributing to mechanisms of intra- and extra-cellular signalling. Subtle relationships between the cell plasma membrane organization, the state of the cytoskeleton, and neural development and plasticity have to be highlighted.

A quantitative examination, at the electron-microscopic (EM) level, will be undertaken of the (re-)arrangement of N-CAM and/or glutamate receptors (Glu-R) labelled with specific immunogold on surfaces of developing nerve cells in: an in vivo model of neural plasticity, a one-trial passive avoidance task in one-day chicks and in adult rats; and an in vitro model, a monolayer culture of hippocampus cells that allows disruption of neuron cytoskeletal elements in different developmental stages. Computerized microscopy, recently developed stereological and image analysis techniques, as well as stochastic approaches, will be employed to quantify and compare statistically the surface expression and topographies of N-CAM (in both models) and Glu-R (in vitro) with regard to: identified forebrain regions from control and trained chicks and rats; two stages of culture development; control cultured nerve cells and those with disrupted microtubules, actin- or neurofilamens; cell somata, processes, growth cones.

The results will allow determination of subtle relationships between the lateral organization of the two proteins in neuronal membranes, the state of the cytoskeleton, and the phenomena of development and/or plasticity. While the in vitro studies will elucidate data about cellular mechanisms of the cytoskeleton -membrane interactions and the age-dependent plasmalemma organization, the in vivo model will enable us to determine how these mechanisms can contribute to the actual process of memorization. Such results will provide insights into molecular mechanisms involved in neural circuitry within the central nervous system.

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Coordinador

Open University
Aportación de la UE
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Dirección
Walton Hall
MK7 6AA Milton Keynes
Reino Unido

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Participantes (5)