Dumon, Valerie M-C.; (1997) Cell surface molecules involved in the survival and growth of Purkinje cells. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

During the development of the central nervous system (CNS), cell surface molecules play an important role in determining the fate of neurones. The expression of these cell surface molecules changes throughout development. The changes are indicative of their involvement in events such as proliferation, differentiation, synaptic plasticity and neurite outgrowth. As a neurone matures and gains a differentiated character, synaptic plasticity remains but neurite outgrowth in the mature CNS is limited. The reasons for this limitation are unclear. This study investigates some of the events which determine the survival and the development of mature cerebellar Purkinje cells in vitro. It was found that the yield of live Purkinje cells is greatly increased (6-fold) when sucrose and pyruvate are present in the buffer used for cell isolation. The enzyme pronase is also found to be of benefit in preserving the Purkinje cell's dendritic tree. The increase of cell adhesion to the culture plate via Thy-1 or membrane-bound glutamate acid decarboxylase (GAD) support the survival of mature neurones in vitro, Purkinje cells' morphological and molecular characteristics were preserved and these cells survived up to one month. Soluble antibodies to Thy-1, added to the medium of the culture were found to promote significant neurite outgrowth from mature Purkinje cells and their growth is greatly enhanced when NGF is also added. The neurite outgrowth pattern induced by anti-Thy-1 antibodies treatment differs from those induced by NGF. Anti-Thy-1 antibodies promote a single-branched type of elongation while NGF enhances mainly arborisation as well as a neurite elongation. This study concludes that modulation of cell surface molecules can cause neurones to elongate neurites de novo and further differentiate even after they have reached maturity.

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