Advertisement

Astrocytes pp 277-292 | Cite as

Investigating Age-Related Changes in Proliferation and the Cell Division Repertoire of Parenchymal Reactive Astrocytes

  • Gábor Heimann
  • Swetlana SirkoEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1938)

Abstract

Reactive gliosis is a complicated process involving all types of glial cells and is the therapeutic target of efforts to treat several types of neuropathologies. Parenchymal astrocytes continuously survey their microenvironment to identify even tiny abnormalities in the central nervous system (CNS) homeostasis and react rapidly to brain damage, such as following ischemia, trauma, or neurodegenerative diseases, to prevent propagation of tissue damage. Aging can play causal roles in certain astroglial dysfunctions, however, still little is known to what extent the heterogeneous reaction of astrocytes at the injury site might be impaired over the course of aging. Based on our experience with both in vitro and in vivo experimental paradigms, we describe here in detail the analysis of age-related changes in (1) proliferative response of parenchymal astrocytes within the posttraumatic cerebral cortex grey matter (GM), and (2) repertoire of their cell divisions in adherent cell culture prepared from the injured GM of young and old double transgenic GFAP-mRFP1/(FUCCI)-S/G2/M-mAG-hGeminin mice by single cell time-lapse imaging.

Key words

Reactive astrocyte Brain injury Proliferation Cell division Cell cycle Cell imaging 

References

  1. 1.
    Allen NJ, Barres BA (2009) Neuroscience: glia—more than just brain glue. Nature 457(7230):675–677. https://doi.org/10.1038/457675aCrossRefPubMedGoogle Scholar
  2. 2.
    Herculano-Houzel S (2014) The glia/neuron ratio: how it varies uniformly across brain structures and species and what that means for brain physiology and evolution. Glia 62(9):1377–1391. https://doi.org/10.1002/glia.22683CrossRefPubMedGoogle Scholar
  3. 3.
    Sofroniew MV, Vinters HV (2010) Astrocytes: biology and pathology. Acta Neuropathol 119(1):7–35. https://doi.org/10.1007/s00401-009-0619-8CrossRefPubMedGoogle Scholar
  4. 4.
    Götz M, Sirko S (2013) Potential of glial cells. In: Sell S (ed) Stem cells handbook, 2nd edn. Springer, New York, pp 347–361. https://doi.org/10.1007/978-1-4614-7696-2_24CrossRefGoogle Scholar
  5. 5.
    Dimou L, Gotz M (2014) Glial cells as progenitors and stem cells: new roles in the healthy and diseased brain. Physiol Rev 94(3):709–737. https://doi.org/10.1152/physrev.00036.2013CrossRefPubMedGoogle Scholar
  6. 6.
    Sirko S, Neitz A, Mittmann T, Horvat-Brocker A, von Holst A, Eysel UT, Faissner A (2009) Focal laser-lesions activate an endogenous population of neural stem/progenitor cells in the adult visual cortex. Brain 132(Pt 8):2252–2264. https://doi.org/10.1093/brain/awp043CrossRefPubMedGoogle Scholar
  7. 7.
    Sirko S, Behrendt G, Johansson PA, Tripathi P, Costa M, Bek S, Heinrich C, Tiedt S, Colak D, Dichgans M, Fischer IR, Plesnila N, Staufenbiel M, Haass C, Snapyan M, Saghatelyan A, Tsai LH, Fischer A, Grobe K, Dimou L, Gotz M (2013) Reactive glia in the injured brain acquire stem cell properties in response to sonic hedgehog. [corrected]. Cell Stem Cell 12(4):426–439. https://doi.org/10.1016/j.stem.2013.01.019CrossRefPubMedGoogle Scholar
  8. 8.
    Wilhelmsson U, Li L, Pekna M, Berthold CH, Blom S, Eliasson C, Renner O, Bushong E, Ellisman M, Morgan TE, Pekny M (2004) Absence of glial fibrillary acidic protein and vimentin prevents hypertrophy of astrocytic processes and improves post-traumatic regeneration. J Neurosci 24(21):5016–5021. https://doi.org/10.1523/JNEUROSCI.0820-04.2004CrossRefPubMedGoogle Scholar
  9. 9.
    Sirko S, Irmler M, Gascon S, Bek S, Schneider S, Dimou L, Obermann J, De Souza Paiva D, Poirier F, Beckers J, Hauck SM, Barde YA, Gotz M (2015) Astrocyte reactivity after brain injury-: the role of galectins 1 and 3. Glia 63(12):2340–2361. https://doi.org/10.1002/glia.22898CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Zamanian JL, Xu L, Foo LC, Nouri N, Zhou L, Giffard RG, Barres BA (2012) Genomic analysis of reactive astrogliosis. J Neurosci 32(18):6391–6410. https://doi.org/10.1523/JNEUROSCI.6221-11.2012CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Buffo A, Rite I, Tripathi P, Lepier A, Colak D, Horn AP, Mori T, Gotz M (2008) Origin and progeny of reactive gliosis: a source of multipotent cells in the injured brain. Proc Natl Acad Sci U S A 105(9):3581–3586. https://doi.org/10.1073/pnas.0709002105CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Gotz M, Sirko S, Beckers J, Irmler M (2015) Reactive astrocytes as neural stem or progenitor cells: in vivo lineage, in vitro potential, and genome-wide expression analysis. Glia 63(8):1452–1468. https://doi.org/10.1002/glia.22850CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Shimada IS, LeComte MD, Granger JC, Quinlan NJ, Spees JL (2012) Self-renewal and differentiation of reactive astrocyte-derived neural stem/progenitor cells isolated from the cortical peri-infarct area after stroke. J Neurosci 32(23):7926–7940. https://doi.org/10.1523/JNEUROSCI.4303-11.2012CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Bardehle S, Kruger M, Buggenthin F, Schwausch J, Ninkovic J, Clevers H, Snippert HJ, Theis FJ, Meyer-Luehmann M, Bechmann I, Dimou L, Gotz M (2013) Live imaging of astrocyte responses to acute injury reveals selective juxtavascular proliferation. Nat Neurosci 16(5):580–586. https://doi.org/10.1038/nn.3371CrossRefPubMedGoogle Scholar
  15. 15.
    Simon C, Gotz M, Dimou L (2011) Progenitors in the adult cerebral cortex: cell cycle properties and regulation by physiological stimuli and injury. Glia 59(6):869–881. https://doi.org/10.1002/glia.21156CrossRefPubMedGoogle Scholar
  16. 16.
    Heimann G, Canhos LL, Frik J, Jager G, Lepko T, Ninkovic J, Gotz M, Sirko S (2017) Changes in the proliferative program limit astrocyte homeostasis in the aged post-traumatic murine cerebral cortex. Cereb Cortex 27(8):4213–4228. https://doi.org/10.1093/cercor/bhx112CrossRefPubMedGoogle Scholar
  17. 17.
    Frik J, Merl-Pham J, Plesnila N, Mattugini N, Kjell J, Kraska J, Gomez RM, Hauck SM, Sirko S, Gotz M (2018) Cross-talk between monocyte invasion and astrocyte proliferation regulates scarring in brain injury. EMBO Rep 19(5):e45294. https://doi.org/10.15252/embr.201745294CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Burda JE, Sofroniew MV (2014) Reactive gliosis and the multicellular response to CNS damage and disease. Neuron 81(2):229–248. https://doi.org/10.1016/j.neuron.2013.12.034CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Buga AM, Sascau M, Pisoschi C, Herndon JG, Kessler C, Popa-Wagner A (2008) The genomic response of the ipsilateral and contralateral cortex to stroke in aged rats. J Cell Mol Med 12(6B):2731–2753. https://doi.org/10.1111/j.1582-4934.2008.00252.xCrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Rodriguez-Arellano JJ, Parpura V, Zorec R, Verkhratsky A (2015) Astrocytes in physiological aging and Alzheimer's disease. Neuroscience 323:170–182. https://doi.org/10.1016/j.neuroscience.2015.01.007CrossRefPubMedGoogle Scholar
  21. 21.
    Clarke D, Penrose MA, Harvey AR, Rodger J, Bates KA (2017) Low intensity rTMS has sex-dependent effects on the local response of glia following a penetrating cortical stab injury. Exp Neurol 295:233–242. https://doi.org/10.1016/j.expneurol.2017.06.019CrossRefPubMedGoogle Scholar
  22. 22.
    Acaz-Fonseca E, Duran JC, Carrero P, Garcia-Segura LM, Arevalo MA (2015) Sex differences in glia reactivity after cortical brain injury. Glia 63(11):1966–1981. https://doi.org/10.1002/glia.22867CrossRefPubMedGoogle Scholar
  23. 23.
    Tropepe V, Craig CG, Morshead CM, van der Kooy D (1997) Transforming growth factor-alpha null and senescent mice show decreased neural progenitor cell proliferation in the forebrain subependyma. J Neurosci 17(20):7850–7859CrossRefGoogle Scholar
  24. 24.
    Seki T (2002) Expression patterns of immature neuronal markers PSA-NCAM, CRMP-4 and NeuroD in the hippocampus of young adult and aged rodents. J Neurosci Res 70(3):327–334. https://doi.org/10.1002/jnr.10387CrossRefPubMedGoogle Scholar
  25. 25.
    Jin K, Sun Y, Xie L, Batteur S, Mao XO, Smelick C, Logvinova A, Greenberg DA (2003) Neurogenesis and aging: FGF-2 and HB-EGF restore neurogenesis in hippocampus and subventricular zone of aged mice. Aging Cell 2(3):175–183CrossRefGoogle Scholar
  26. 26.
    Garcia A, Steiner B, Kronenberg G, Bick-Sander A, Kempermann G (2004) Age-dependent expression of glucocorticoid- and mineralocorticoid receptors on neural precursor cell populations in the adult murine hippocampus. Aging Cell 3(6):363–371. https://doi.org/10.1111/j.1474-9728.2004.00130.xCrossRefPubMedGoogle Scholar
  27. 27.
    Enwere E, Shingo T, Gregg C, Fujikawa H, Ohta S, Weiss S (2004) Aging results in reduced epidermal growth factor receptor signaling, diminished olfactory neurogenesis, and deficits in fine olfactory discrimination. J Neurosci 24(38):8354–8365. https://doi.org/10.1523/JNEUROSCI.2751-04.2004CrossRefPubMedGoogle Scholar
  28. 28.
    Luo J, Daniels SB, Lennington JB, Notti RQ, Conover JC (2006) The aging neurogenic subventricular zone. Aging Cell 5(2):139–152. https://doi.org/10.1111/j.1474-9726.2006.00197.xCrossRefPubMedGoogle Scholar
  29. 29.
    Tang H, Wang Y, Xie L, Mao X, Won SJ, Galvan V, Jin K (2009) Effect of neural precursor proliferation level on neurogenesis in rat brain during aging and after focal ischemia. Neurobiol Aging 30(2):299–308. https://doi.org/10.1016/j.neurobiolaging.2007.06.004CrossRefPubMedGoogle Scholar
  30. 30.
    Amat JA, Ishiguro H, Nakamura K, Norton WT (1996) Phenotypic diversity and kinetics of proliferating microglia and astrocytes following cortical stab wounds. Glia 16(4):368–382. https://doi.org/10.1002/(SICI)1098-1136(199604)16:4<368::AID-GLIA9>3.0.CO;2-WCrossRefPubMedGoogle Scholar
  31. 31.
    Scholzen T, Gerdes J (2000) The Ki-67 protein: from the known and the unknown. J Cell Physiol 182(3):311–322. https://doi.org/10.1002/(SICI)1097-4652(200003)182:3<311::AID-JCP1>3.0.CO;2-9CrossRefPubMedGoogle Scholar
  32. 32.
    Mahadevan LC, Willis AC, Barratt MJ (1991) Rapid histone H3 phosphorylation in response to growth factors, phorbol esters, okadaic acid, and protein synthesis inhibitors. Cell 65(5):775–783CrossRefGoogle Scholar
  33. 33.
    Bambakidis NC, Petrullis M, Kui X, Rothstein B, Karampelas I, Kuang Y, Selman WR, LaManna JC, Miller RH (2012) Improvement of neurological recovery and stimulation of neural progenitor cell proliferation by intrathecal administration of sonic hedgehog. J Neurosurg 116(5):1114–1120. https://doi.org/10.3171/2012.1.JNS111285CrossRefPubMedGoogle Scholar
  34. 34.
    Sakaue-Sawano A, Kurokawa H, Morimura T, Hanyu A, Hama H, Osawa H, Kashiwagi S, Fukami K, Miyata T, Miyoshi H, Imamura T, Ogawa M, Masai H, Miyawaki A (2008) Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. Cell 132(3):487–498. https://doi.org/10.1016/j.cell.2007.12.033CrossRefPubMedGoogle Scholar
  35. 35.
    Wechsler-Reya RJ, Scott MP (1999) Control of neuronal precursor proliferation in the cerebellum by sonic hedgehog. Neuron 22(1):103–114CrossRefGoogle Scholar
  36. 36.
    Yang R, Wang M, Wang J, Huang X, Yang R, Gao WQ (2015) Cell division mode change mediates the regulation of cerebellar granule neurogenesis controlled by the sonic hedgehog Signaling. Stem Cell Reports 5(5):816–828. https://doi.org/10.1016/j.stemcr.2015.09.019CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Badan I, Buchhold B, Hamm A, Gratz M, Walker LC, Platt D, Kessler C, Popa-Wagner A (2003) Accelerated glial reactivity to stroke in aged rats correlates with reduced functional recovery. J Cereb Blood Flow Metab 23(7):845–854. https://doi.org/10.1097/01.WCB.0000071883.63724.A7CrossRefPubMedGoogle Scholar
  38. 38.
    Copen WA, Schwamm LH, Gonzalez RG, Wu O, Harmath CB, Schaefer PW, Koroshetz WJ, Sorensen AG (2001) Ischemic stroke: effects of etiology and patient age on the time course of the core apparent diffusion coefficient. Radiology 221(1):27–34. https://doi.org/10.1148/radiol.2211001397CrossRefPubMedGoogle Scholar
  39. 39.
    Fonarow GC, Reeves MJ, Zhao X, Olson DM, Smith EE, Saver JL, Schwamm LH, Get With the Guidelines-Stroke Steering C, Investigators (2010) Age-related differences in characteristics, performance measures, treatment trends, and outcomes in patients with ischemic stroke. Circulation 121(7):879–891. https://doi.org/10.1161/CIRCULATIONAHA.109.892497CrossRefPubMedGoogle Scholar
  40. 40.
    Pekna M, Pekny M, Nilsson M (2012) Modulation of neural plasticity as a basis for stroke rehabilitation. Stroke 43(10):2819–2828. https://doi.org/10.1161/STROKEAHA.112.654228CrossRefPubMedGoogle Scholar
  41. 41.
    Hilsenbeck O, Schwarzfischer M, Skylaki S, Schauberger B, Hoppe PS, Loeffler D, Kokkaliaris KD, Hastreiter S, Skylaki E, Filipczyk A, Strasser M, Buggenthin F, Feigelman JS, Krumsiek J, van den Berg AJ, Endele M, Etzrodt M, Marr C, Theis FJ, Schroeder T (2016) Software tools for single-cell tracking and quantification of cellular and molecular properties. Nat Biotechnol 34(7):703–706. https://doi.org/10.1038/nbt.3626CrossRefPubMedGoogle Scholar
  42. 42.
    Nolte C, Matyash M, Pivneva T, Schipke CG, Ohlemeyer C, Hanisch UK, Kirchhoff F, Kettenmann H (2001) GFAP promoter-controlled EGFP-expressing transgenic mice: a tool to visualize astrocytes and astrogliosis in living brain tissue. Glia 33(1):72–86CrossRefGoogle Scholar
  43. 43.
    Hirrlinger PG, Scheller A, Braun C, Quintela-Schneider M, Fuss B, Hirrlinger J, Kirchhoff F (2005) Expression of reef coral fluorescent proteins in the central nervous system of transgenic mice. Mol Cell Neurosci 30(3):291–303. https://doi.org/10.1016/j.mcn.2005.08.011CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Physiological Genomics, Biomedical CenterLudwig-Maximilians-UniversityMunchGermany
  2. 2.Institute of Stem Cell ResearchHelmholtz Center MunichNeuherbergGermany

Personalised recommendations