A Method for Patterning Proteins and Its Application to Study the Guidance of Neurite Outgrowth

Perez Arnaud, Enrique (1994) A Method for Patterning Proteins and Its Application to Study the Guidance of Neurite Outgrowth. PhD thesis, University of Glasgow.

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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

Item Type: Thesis (PhD)
Qualification Level: Doctoral
Additional Information: Adviser: Geoffrey Moores
Keywords: Neurosciences
Date of Award: 1994
Depositing User: Enlighten Team
Unique ID: glathesis:1994-76340
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 19 Nov 2019 15:24
Last Modified: 19 Nov 2019 15:24
URI: https://theses.gla.ac.uk/id/eprint/76340

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