Abstract
Genetic tools are enabling the molecular dissection of the functions and mechanisms of many biological processes. Transgenic manipulations provide powerful tools with which to test hypotheses regarding functions of specific cell types and molecules in vivo in combination with different types of experimental models. Various techniques are available that can target genetic manipulations specifically to astrocytes and that are enabling the molecular dissection of astrocyte biology in vivo. This article summarizes procedures and experience from our laboratory using transgenic strategies that enable either the ablation of proliferating astrocytes and related cells, or the deletion of specific molecules selectively from astrocytes, to study the functions of astrocytes and related cell types in health and disease in vivo using different experimental mouse models.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Sofroniew, M. V. (2009) Molecular dissection of reactive astrogliosis and glial scar formation, Trends Neurosci. 32, 638–647.
Sofroniew, M. V., and Vinters, H. V. (2010) Astrocytes: biology and pathology, Acta Neuropathol 119, 7–35.
Borrelli, E., Heyman, R., Hsi, M., and Evans, R. M. (1988) Targetting of an inducible toxic phenotype in animal cells, Proc. Natl. Acad. Sci. USA 85, 7572–7576.
Bush, T. G., N., P., Horner, C. H., Polito, A., Ostenfeld, T., Svendsen, C. N., Mucke, L., Johnson, M. H., and Sofroniew, M. V. (1999) Leukocyte infiltration, neuronal degeneration and neurite outgrowth after ablation of scar-forming, reactive astrocytes in adult transgenic mice, Neuron 23, 297–308.
Bush, T. G., Savidge, T. C., Freeman, T. C., Cox, H. J., Campbell, E. A., Mucke, L., Johnson, M. H., and Sofroniew, M. V. (1998) Fulminant jejuno-ileitis following ablation of enteric glia in adult transgenic mice, Cell 93, 189–201.
Borrelli, E., Heyman, R. A., Arias, C., Sawchenko, P. E., and Evans, R. M. (1989) Transgenic mice with inducible dwarfism, Nature 339, 538–541.
Fischer, U., Steffens, S., Frank, S., Rainov, N. G., Schulze-Osthoff, K., and Kramm, C. M. (2005) Mechanisms of thymidine kinase/ganciclovir and cytosine deaminase/ 5-fluorocytosine suicide gene therapy-induced cell death in glioma cells, Oncogene 24, 1231–1243.
Sauer, B. (1994) Site-specific recombination: developments and applications, Curr. Opin. Biotech. 5, 521–527.
Soriano, P. (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain, Nature Genetics 21, 70–71.
Johnson, W. B., Ruppe, M. D., Rockenstein, E. M., Price, J., Sarthy, V. P., Vederber, L. C., and Mucke, L. (1995) Indicator expression directed by regulatory sequences of the glial fibrillary acidic protein (GFAP) gene: in vitro comparison of distinct GFAP-lacZ transgenes, Glia 13, 174–184.
Bunting, M., Bernstein, K., Greer, J., Capecchi, M., and Thomas, K. (1999) Targeting genes for self-excision in the germ line, Genes Dev. 13, 1524–1528.
Garcia, A. D. R., Doan, N. B., Imura, T., Bush, T. G., and Sofroniew, M. V. (2004) GFAP-expressing progenitors are the principle source of constitutive neurogenesis in adult mouse forebrain, Nature Neurosci. 7, 1233–1241.
Faulkner, J. R., Herrmann, J. E., Woo, M. J., Tansey, K. E., Doan, N. B., and Sofroniew, M. V. (2004) Reactive astrocytes protect tissue and preserve function after spinal cord injury, J. Neurosci. 24, 2143–2155.
Al-Shawi, R., Burke, J., Jones, C. T., Simons, J. P., and Bishop, J. O. (1988) A Mup promoter-thymidine kinase reporter gene shows relaxed tissue-specific expression and confers male sterility upon transgenic mice, Mol. Cell. Biol. 8, 4821–4828.
Herrmann, J. E., Imura, T., Song, B., Qi, J., Ao, Y., Nguyen, T. K., Korsak, R. A., Takeda, K., Akira, S., and Sofroniew, M. V. (2008) STAT3 is a critical regulator of astrogliosis and scar formation after spinal cord injury, J. Neurosci. 28, 7231–7243.
Forni, P. E., Scuoppo, C., Imayoshi, I., Taulli, R., Dastru, W., Sala, V., Betz, U. A., Muzzi, P., Martinuzzi, D., Vercelli, A. E., Kageyama, R., and Ponzetto, C. (2006) High levels of Cre expression in neuronal progenitors cause defects in brain development leading to microencephaly and hydrocephaly, J Neurosci 26, 9593–9602.
Myer, D. J., Gurkoff, G. G., Lee, S. M., Hovda, D. A., and Sofroniew, M. V. (2006) Essential protective roles of reactive astrocytes in traumatic brain injury, Brain 129, 2761–2772.
Voskuhl, R. R., Peterson, R. S., Song, B., Ao, Y., Morales, L. B., Tiwari-Woodruff, S., and Sofroniew, M. V. (2009) Reactive astrocytes form scar-like perivascular barriers to leukocytes during adaptive immune inflammation of the CNS, J. Neurosci. 29, 11511–11522.
Savidge, T. C., Newman, P., Pothoulakis, C., Ruhl, A., Neunlist, M., Bourreille, A., Hurst, R., and Sofroniew, M. V. (2007) Enteric glia regulate intestinal barrier function and inflammation via release of S-nitrosoglutathione, Gastroenterology 132, 1344–1358.
Saxe, M., Battaglia, F., Wang, J. W., Malleret, G., David, D. J., Monckton, J. E., Garcia, A. D. R., Sofroniew, M. V., Kandel, E. R., Santarelli, L., Hen, R., and Drew, M. R. (2006) Ablation of hippocampal neurogenesis impairs contextual fear conditioning and synaptic plasticity in the dentate gyrus, Proc. Natl. Acad. Sci. USA 103, 17501–17506.
Saxe, M. D., Malleret, G., Vronskaya, S., Mendez, I., Garcia, A. D., Sofroniew, M. V., Kandel, E. R., and Hen, R. (2007) Paradoxical influence of hippocampal neurogenesis on working memory, Proc. Natl. Acad. Sci. USA 104, 4642–4646.
Hatten, M. E., Liem, R. K. H., Shelanski, M. L., and Mason, C. A. (1991) Astroglia in CNS injury, Glia 4, 233–243.
Amat, J. A., Ishiguro, H., Nakamura, K., and Norton, W. T. (1996) Phenotypic diversity and kinetics of proliferating microglia and astrocytes following cortical stab wounds, Glia 16, 368–382.
Doetsch, F., Caille, I., Lim, D. A., Garcia-Verdugo, J. M., and Alvarez-Buylla, A. (1999) Subventricular zone astrocytes are neural stem cells in the adult mammalian brain, Cell 97, 703–716.
Noctor, S. C., Flint, A. C., Weissman, T. A., Dammerman, R. S., and Kriegstein, A. R. (2001) Neurons derived from radial glial cells establish radial units in neocortex, Nature 409, 714–720.
Imura, T., Kornblum, H. I., and Sofroniew, M. V. (2003) The predominant neural stem cell isolated from postnatal and adult forebrain but not from early embryonic forebrain expresses GFAP, J. Neurosci. 23, 2824–2832.
Imura, T., Nakano, I., Kornblum, H. I., and Sofroniew, M. V. (2006) Phenotypic and functional heterogeneity of GFAP-expressing cells in vitro: Differential expression of LeX/CD15 by GFAP-expressing multipotent neural stem cells and non-neurogenic astrocytes, Glia 53, 277–293.
Gregorian, C., Nakashima, J., Le Belle, J., Ohab, J., Kim, R., Liu, A., Smith, K. B., Groszer, M., Garcia, A. D., Sofroniew, M. V., Carmichael, S. T., Kornblum, H. I., Liu, X., and Wu, H. (2009) Pten deletion in adult neural stem/progenitor cells enhances constitutive neurogenesis, J. Neurosci. 29, 1874–1886.
Lutz, S. E., Zhao, Y., Gulinello, M., Lee, S. C., Raine, C. S., and Brosnan, C. F. (2009) Deletion of astrocyte connexins 43 and 30 leads to a dysmyelinating phenotype and hippocampal CA1 vacuolation, J. Neurosci. 29, 7743–7752.
Sahni, V., Mukhopadhyay, A., Tysseling, V., Hebert, A., Birch, D., McGuire, T. L., Stupp, S. I., and Kessler, J. A. (2010) BMPR1a and BMPR1b signaling exert opposing effects on gliosis after spinal cord injury, J Neurosci 30, 1839–1855.
Gregorian, C., Nakashima, J., Dry, S. M., Nghiemphu, P. L., Smith, K. B., Ao, Y., Dang, J., Lawson, G., Mellinghoff, I. K., Mischel, P. S., Phelps, M., Parada, L. F., Liu, X., Sofroniew, M. V., Eilber, F. C., and Wu, H. (2009) PTEN dosage is essential for neurofibroma development and malignant transformation, Proc. Natl. Acad. Sci. USA 106, 19479–19484.
Ohab, J. J., Fleming, S., Blesch, A., and Carmichael, S. T. (2006) A neurovascular niche for neurogenesis after stroke, J. Neurosci. 26, 13007–13016.
Jessen, K. R., and Mirsky, R. (1980) Glial cells in the enteric nervous system contain glial fibrillary acidic protein, Nature 286, 736–737.
Jessen, K. R., and Mirsky, R. (1992) Schwann cells: Early lineage, regulation of proliferation and control of myelin formation, Curr. Opin. Neurobiol. 2, 575–581.
Madison, R. D., Sofroniew, M. V., and Robinson, G. A. (2009) Schwann cell influence on motor neuron regeneration accuracy, Neuroscience 163, 213–221.
Feng, G., Mellor, R., Bernstein, M., Keller-Peck, C., Nguyen, Q., Wallace, M., Nerbonne, J., Lichtman, J., and Sanes, J. (2000) Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP, Neuron 28, 41–51.
Zhuo, L., Theis, M., Alvarez-Maya, I., Brenner, M., Willecke, K., and Messing, A. (2001) hGFAP-cre transgenic mice for manipulation of glial and neuronal function in vivo, Genesis 31, 85–94.
Malatesta, P., Hack, M. A., Hartfuss, E., Kettenmann, H., Klinkert, W., Kirchhoff, F., and Götz, M. (2003) Neuronal or glial progeny: Regional differences in radial glia fate, Neuron 37, 751–764.
Barres, B. A. (2008) The mystery and magic of glia: a perspective on their roles in health and disease, Neuron 60, 430–440.
Ganat, Y. M., Silbereis, J., Cave, C., Ngu, H., Anderson, G. M., Ohkubo, Y., Ment, L. R., and Vaccarino, F. M. (2006) Early postnatal astroglial cells produce multilineage precursors and neural stem cells in vivo, J Neurosci 26, 8609–8621.
Casper, K. B., Jones, K., and McCarthy, K. D. (2007) Characterization of astrocyte-specific conditional knockouts, Genesis 45, 292–299.
Slezak, M., Goritz, C., Niemiec, A., Frisen, J., Chambon, P., Metzger, D., and Pfrieger, F. W. (2007) Transgenic mice for conditional gene manipulation in astroglial cells, Glia 55, 1565–1576.
Barreto, G., Santos-Galindo, M., Diz-Chaves, Y., Pernia, O., Carrero, P., Azcoitia, I., and Garcia-Segura, L. M. (2009) Selective estrogen receptor modulators decrease reactive astrogliosis in the injured brain: effects of aging and prolonged depletion of ovarian hormones, Endocrinology 150, 5010–5015.
Acknowledgments
The author’s work is supported by grants from the National Institutes of Health (NINDS) NS057624, Wings for Life, Multiple Sclerosis Society, and Adelson Medical Research Foundation.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Sofroniew, M.V. (2012). Transgenic Techniques for Cell Ablation or Molecular Deletion to Investigate Functions of Astrocytes and Other GFAP-Expressing Cell Types. In: Milner, R. (eds) Astrocytes. Methods in Molecular Biology, vol 814. Humana Press. https://doi.org/10.1007/978-1-61779-452-0_35
Download citation
DOI: https://doi.org/10.1007/978-1-61779-452-0_35
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-451-3
Online ISBN: 978-1-61779-452-0
eBook Packages: Springer Protocols