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Is tenascin-C an autocrine/paracrine factor for neural stem cell development?

Reference	BB/C007425/1
Principal Investigator / Supervisor	Professor Charles ffrench-Constant
Co-Investigators / Co-Supervisors
Institution	University of Cambridge
Department	Pathology
Funding type	Research
Value (£)	230,972
Status	Completed
Type	Research Grant
Start date	04/04/2005
End date	03/04/2008
Duration	36 months

Abstract

Neural stem cells exhibit three fundamental properties during the formation of the CNS; maintenance (by which the stem cells self-renew with each division throughout life), differentiation (with daughter cells becoming neurones or glia as appropriate for location and developmental stage) and development (with the stem cells changing their morphological and biochemical properties as well as their division rate as growth proceeds whilst retaining their stem cell identity). Here we will examine the role of the extracellular matrix glycoprotein tenascin-c in the third or these properties, stem cell development so as to test the hypothesis generated by our previous studies that tenascin-C acts as an autocrine paracrine factor to promote neural stem cell development. First, we will determine which cell type produces the tenascin-C present around the stem cells in the intact CNS at all stages of growth, using immunolabelling, in situ hybridisation and beta-galactosidase localisation in mice heterozygous for a transgenic allele in which the tenascin-C gene has been interrupted by a lac-Z cassette. Second, we will examine stem cell development specifically by looking at radial glial cells (the stem cell of the late embryonic and early postnatal CNS) and asking whether the sequence of changes normally observed in the expression of the markers RC2, GLAST and BLBP occur at the same schedule in tenascin-C knockout mice homozygous for the lac-Z allele. Third, we will analyse the differentiation of individual radial glial cells either isolated by FACS and then grown at low density in co-culture with rat cells so as to allow the mouse cells to be distinguished and phenotyped at the end of the assay, or grown in cultures infected with low titres of retroviruses expressing GFP so that well-spaced single clones derived from one stem cell are labelled and can be individually phenotyped. Finally, we will examine the subventricular zone of the adult CNS, in which the identity and three dimensional architecture of the stem cells and their neighbours has been well characterised, in the knock-out mice to see how stem cell numbers and architecture are affected by the absence of tenascin-C. We will also use the previously described techniques of precursor cell depletion by infusion of the anti-mitotic drug ara-C to determine whether reconstitution by stem cells occurs normally in the absence of tenascin-C, so examining the role of tenascin-C in a regenerative response of the stem cells within the intact CNS.

Summary

unavailable

Committee	Closed Committee - Genes & Developmental Biology (GDB)
Research Topics	Neuroscience and Behaviour, Regenerative Biology, Stem Cells
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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