Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.797017
Title: A method for patterning proteins and its application to study the guidance of neurite outgrowth
Author: Perez Arnaud, Enrique
Awarding Body: University of Glasgow
Current Institution: University of Glasgow
Date of Award: 1994
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Abstract:
The work described in this thesis has consisted of two well differentiated parts. In the first part, a method for patterning proteins at a micrometer level over a glass surface was developed and assessed. This method was based upon the use of amino groups attached to certain parts of the glass surface, to define the areas to which proteins could be bound, and of a non-specific protein attached to the remaining parts of the surface, to define the areas that would not bind the specific protein. It was assessed with a variety of protein detection techniques, among them silver staining, detection of radiolabelled proteins, and enzymatic assays, and in the end it proved to be very effective. In the second part, patterns of laminin were produced with this method, and these were used to culture nerve cells over them. The purpose of this was to see whether they would guide the processes produced by the nerve cells, and they actually proved to be able to guide them. Then, this patterning method was used to study the geometrical conditions that would have to be fulfilled by a laminin pattern that would guide this processes efficiently. The geometrical characteristics that were investigated were A): The maximum distance between separate laminin areas that the processes could ignore, B): The minimum width of laminin pathways that could be recognised by the processes, and C): The angles of the turns in the pathways that the processes could follow. The results of these investigations yielded a number of insights into the mechanisms by which nerve growth cones find their way towards their targets, that can be sumarised as follows: A) In the case of the cell type studied here, i.e. chicken embryo dorsal root ganglion cells, the maximum width of a non-adsorbant stretch than can be crossed by its growth cones is around 20 mum. This figure correlates well with the length of the filopodia in those growth cones. B) The minimum width of a laminin line that a growth cone can recognize and follow is smaller than 1 mum. C) growth cones following a thin laminin line seem to travel faster than growth cones over unpattemed laminin. D) Growth cones can follow laminin pathways with turns as sharp as 30° E). However, the sharper the angles are, the longer the time that growth cones take in turning through them.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.797017  DOI: Not available
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