Information about the developmental regulation of the extracellular matrix in the CNS is incomplete and the function of proteins such as the tissue inhibitor of matrix metalloproteinases (TIMPS) is poorly understood. We recently characterized the expression of the four known TIMPs during CNS development in vivo. The expression of TIMP-2 by post-mitotic neurons correlates with neuronal differentiation. We demonstrate herein that TIMP-2 induces cell cycle arrest and neuronal differentiation in a cell autonomous manner. Strikingly, TIMP-2 's effect was independent of its MMP-inhibitory activity. The punctate labeling of TIMP-2 on the cell surface combined with the interaction of TIMP-2 with alpha3 beta1 integrin suggests that TIMP-2 may exert its MMP-independent activities via integrins. The persistence of nestin-positive progenitors in the neocortical ventricular zone and the reduced neurite length of Timp-2 -null neurons suggest that neuronal differentiation is delayed in the absence of TIMP-2. This is the first report to detail the mechanism of TIMP-2 action in neuronal ceils and demonstrates a novel function for TIMP- 2 in neurons independent of MMP inhibition. Primary cultured Timp-2 -/- and wild-type cerebral cortical neurons will be used to test the hypothesis that TIMP- 2 plays a role in neuronal differentiation independent of MMP-inhibitory activity via interaction with integrins.
In aim 1, the number, location and affinity of TIMP-2 receptors will be determined and the Timp-2 null neurite length phenotype will be rescued by exogenous application of TIMP-2 protein with and without MMP-inhibitory activity.
In aim 2, the integrin(s) to which TIMP-2 binds will be identified and competitive binding assays will be performed to block the phenotypic rescue by TIMP-2.
Aim 3 will determine whether Timp-2 deletion alters cortical progenitor cell cycle parameters or laminar fate in vivo using two BrdU injection paradigms. Unlike somatic cells, neurons irreversibly withdraw from the cell cycle and permanently remain quiescent. The mechanisms responsible for the maintenance of neuronal quiescence are poorly understood. The identification of molecules responsible for neuronal quiescence has significant implications for the ability of the adult brain to generate new neurons in response injury as well as in neurological deficits such as Alzheimer's and Parkinson's disease.

National Institute of Health (NIH)
National Institute of Neurological Disorders and Stroke (NINDS)
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Neurogenesis and Cell Fate Study Section (NCF)
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Owens, David F
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University of Vermont & St Agric College
Anatomy/Cell Biology
Schools of Medicine
United States
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Long, Patrick M; Tighe, Scott W; Driscoll, Heather E et al. (2015) Acetate supplementation as a means of inducing glioblastoma stem-like cell growth arrest. J Cell Physiol 230:1929-43
Tsen, Andrew R; Long, Patrick M; Driscoll, Heather E et al. (2014) Triacetin-based acetate supplementation as a chemotherapeutic adjuvant therapy in glioma. Int J Cancer 134:1300-10
Long, Patrick M; Tighe, Scott W; Driscoll, Heather E et al. (2013) Acetate supplementation induces growth arrest of NG2/PDGFR?-positive oligodendroglioma-derived tumor-initiating cells. PLoS One 8:e80714
Long, Patrick M; Moffett, John R; Namboodiri, Aryan M A et al. (2013) N-acetylaspartate (NAA) and N-acetylaspartylglutamate (NAAG) promote growth and inhibit differentiation of glioma stem-like cells. J Biol Chem 288:26188-200
Stradecki, H M; Jaworski, D M (2011) Hyperphagia and leptin resistance in tissue inhibitor of metalloproteinase-2 deficient mice. J Neuroendocrinol 23:269-81
Long, Patrick M; Stradecki, Holly M; Minturn, Jane E et al. (2011) Differential aminoacylase expression in neuroblastoma. Int J Cancer 129:1322-30
Jaworski, Diane M; Sideleva, Olga; Stradecki, Holly M et al. (2011) Sexually dimorphic diet-induced insulin resistance in obese tissue inhibitor of metalloproteinase-2 (TIMP-2)-deficient mice. Endocrinology 152:1300-13
Rivera, Santiago; Khrestchatisky, Michel; Kaczmarek, Leszek et al. (2010) Metzincin proteases and their inhibitors: foes or friends in nervous system physiology? J Neurosci 30:15337-57
Lluri, Gentian; Langlois, Garret D; Soloway, Paul D et al. (2008) Tissue inhibitor of metalloproteinase-2 (TIMP-2) regulates myogenesis and beta1 integrin expression in vitro. Exp Cell Res 314:11-24
Jaworski, Diane M; Beem-Miller, Micah; Lluri, Gentian et al. (2007) Potential regulatory relationship between the nested gene DDC8 and its host gene tissue inhibitor of metalloproteinase-2. Physiol Genomics 28:168-78

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