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Title: Control of neurite outgrowth from avian sensory ganglia
Author: Bryce, Graeme J.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1993
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Methods used for studying neurite outgrowth in culture range from attachment of dissociated neurons or explants to coated substrata, to embedding cells or tissues in three-dimensional matrices. Here I describe a new technique in which isolated chick embryo dorsal root ganglia (DRG) can be cultured floating on the surface of serum-free culture medium. The utility of the preparation for studies of neurite growth at the cellular level, and at the level of pattern formation during development is demonstrated. 1. The quantitative effects of neurotrophic factors in regulating neurite outgrowth can be measured using this new preparation. The extent of outgrowth from floating DRG was compared with that from ganglia adhered to laminin, or to polylysine. This preparation provides a substrate-independent assay for neurotrophic factors which offers a number of advantages over existing bioassays. It is also potentially useful in the search for novel neurotrophic molecules. 2. The movement of neuronal growth cones is not well understood. Cytoskeletal dynamics and cell-substratum adhesion are involved. Since the growth of neurites from floating ganglia occurs in the absence of a conventional substratum it was thought that this preparation might provide new insight into the mechanism of growth cone movement and neurite elongation. Experiments were carried out using the drugs colcemid and nocodazole which affect microtubules and cytochalasin B which affects actin filaments. Colcemid and nocodazole inhibited neurite outgrowth from both floating and adhered ganglia. In contrast cytochalasin B did not inhibit outgrowth from floating ganglia although it completely inhibited outgrowth from ganglia adhered to laminin. 3. During the development of sensory innervation, segmental differences arise in the rates of growth of axons invading limb and non-limb regions. Growth rates of axons invading developing limb are faster. It is not known how these different axonal growth rates are controlled.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available