Skip to main content
SpringerLink
Log in

In Vivo Analysis of Cell Proliferation and Apoptosis in the CNS

  • Chapter
Cellular and Molecular Methods in Neuroscience Research

Abstract

The balance between cell proliferation and death is fundamental in several morphogenetic processes and ultimately determines the mass, shape, and function of the various tissues and organs that form the animal body. Apoptosis is a gene-regulated process of programmed cell death (PCD) that plays fundamental roles in several normal and pathological conditions (56,126). This form of “cell suicide” is most often detected during embryonic development, but is also found in normal cell and tissue turnover (26,77,81,133). Although the nervous tissue is traditionally regarded as being fundamentally constituted by postmitotic nonproliferating cells, analysis of cell proliferation and apoptosis in vivo has recently gained an increasing importance mainly considering that: (i) proliferative and/or apoptotic events have been extensively characterized not only during embryonic development but also in several areas of the postnatal and adult brain (9,10, 46,63–65,79,97,101); (ii) trophic factor deprivation often results in apoptotic cell death of target neurons (25,82,131); and (iii) links have been hypothesized between apoptosis and signal transduction (31,60).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Alnemri, E.S., J.N. Livingston, D.W. Nicholson, G. Salvesen, N.A. Thornberry, W.W. Wong, and J. Yuan. 1996. Human ICE/CED-3 protease nomenclature. Cell 87:171–181.

    Article  PubMed  CAS  Google Scholar 

  2. Altman, J. 1966. Autoradiographic and histological studies of postnatal neurogenesis. A longitudinal investigation of the kinetics, migration and transformation of cells incorporating tritiated thymidine in infant rat, with special reference to postnatal neurogenesis in some brain regions. J. Comp. Neurol. 128:431–474.

    Article  Google Scholar 

  3. Altman, J. 1969. Autoradiographic and histological studies of postnatal neurogenesis. Cell proliferation and migration in the anterior forebrain, with special reference to persisting neurogenesis in the olfactory bulb. J. Comp. Neurol. 137:433–458.

    Article  PubMed  CAS  Google Scholar 

  4. Altman, J. 1972a. Postnatal development of the cerebellar cortex in the rat. I. The external germinal layer and the transitional molecular layer. J. Comp. Neurol. 145:353–398.

    Article  PubMed  CAS  Google Scholar 

  5. Altman, J. 1972b. Postnatal development of the cerebellar cortex in the rat. II. Phases in the maturation of the Purkinje cells and of the molecular layer. J. Comp. Neurol. 145:399–464.

    Article  PubMed  CAS  Google Scholar 

  6. Altman, J. 1982. Morphological development of the rat cerebellum and some of its mechanism, p. 8–46. In S.L. Palay and V. Chan-Palay (Eds.), The Cerebellum—New Vistas. Springer-Verlag, New York.

    Google Scholar 

  7. Altman, J. 1992. Programmed cell death: the paths to suicide. Trends Neurol. Sci. 15:278–280.

    Article  CAS  Google Scholar 

  8. Altman, J. and S.A. Bayer. 1978. Prenatal development of the cerebellar system in the rat. I. Cytogenesis and histogenesis of the deep nuclei and the cortex of the cerebellum. J. Comp. Neurol. 179:23–48.

    Article  PubMed  CAS  Google Scholar 

  9. Altman, J. and S.A. Bayer. 1997. Development of the Cerebellar System in Relation to Its Evolution, Structure and Functions. CRC Press, Boca Raton.

    Google Scholar 

  10. Alvarez-Buylla, A. 1997. Neurogenesis in the adult brain: prospects for brain repair, p. 86–100. In E.H. Gage and Y. Christen (Eds.), Isolation, Characterization and Utilization of CNS Stem Cells. Springer-Verlag, Berlin.

    Google Scholar 

  11. Ankarcrona, M., J.M. Dypbukt, E. Bonfoco, B. Zhivotovsky, S. Orrenius, S.A. Lipton, and P. Nicotera. 1995. Glutamate-induced neuronal death: a succession of necrosis or apoptosis depending on mitochondrial function. Neuron 15:961–973.

    Article  PubMed  CAS  Google Scholar 

  12. Banki, K., E. Hutter, E. Colombo, N.J. Gonchoroff, and A. Perl. 1996. Glutathione levels and sensitivity to apoptosis are regulated by changes in transaldolase expression. J. Biol. Chem. 271:32994–33001.

    Article  PubMed  CAS  Google Scholar 

  13. Barbiero, G., F. Duranti, G. Bonelli, J.S. Amenta, and F.M. Baccino. 1997. Intracellular ionic variations in the apoptotic death of L cells by inhibitors of cell cycle progression. Exp. Cell Res. 217:410–418.

    Article  Google Scholar 

  14. Barres, B.A., I.K. Hart, H.S.R. Coles, J.F. Burne, J.T. Voyvodic, W.D. Richardson, and M.C. Raff. 1992. Cell death and control of cell survival in the oligodendrocyte lineage. Cell 70:31–46.

    Article  PubMed  CAS  Google Scholar 

  15. Baum, H.-P., J. Reichrath, A. Theobald, and G. Schock. 1994. Fixation requirements for the immunohistochemical reactivity of PCNA antibody PC10 on cryostat sections. Histochem. J. 26:929–933.

    PubMed  CAS  Google Scholar 

  16. Bayer, S.A. 1983. 3H-Thymidine-radiographic studies of neurogenesis in the rat olfactory bulb. Exp. Brain Res. 50:329–340.

    Article  PubMed  CAS  Google Scholar 

  17. Belecky-Adams, T., B. Cook, and R. Adler. 1996. Correlation between terminal mitosis and differentiated fate of retinal precursor cells in vivo and in vitro: analysis with the „window-labeling„ technique. Dev. Biol. 178:304–315.

    Article  PubMed  CAS  Google Scholar 

  18. Beppu, T., Y. Ishida, H. Arai, T. Wada, N. Uesugi, and K. Sasaki. 1994. Identification of S-phase cells with PC10 antibody to proliferating cell nuclear antigen (PCNA) by flow cytometric analysis. J. Histochem. Cytochem. 42:1177–1182.

    CAS  Google Scholar 

  19. Bernier, P.J. and A. Parent. 1998. Bcl-2 protein as a marker of neuronal immaturity in postnatal primate brain. J. Neurosci. 18:2486–2497.

    PubMed  CAS  Google Scholar 

  20. Bonatz, G., J. Luttges, J. Hedderich, D. Inform, W. Jonat, P. Rudolph, and R. Parwaresch. 1999. Prognostic significance of a novel proliferation marker, anti-repp 86, for endometrial carcinoma: a multivariate study. Hum. Pathol. 30:949–956.

    Article  PubMed  CAS  Google Scholar 

  21. Bonfanti, L., P. Peretto, A. Merighi, and A. Fasolo. 1997. Newly generated cells from the rostral migratory stream in the accessory olfactory bulb of the adult rat. Neuroscience 81:489–502.

    Article  PubMed  CAS  Google Scholar 

  22. Bonfanti, L. and D.T. Theodosis. 1994. Expression of polysialylated neural cell adhesion molecule by proliferating cells in the subependymal layer of the adult rat, in its rostral extension and in the olfactory bulb. Neuroscience 62:291–305.

    Article  PubMed  CAS  Google Scholar 

  23. Bonfoco, E., D. Krainic, M. Ankarcrona, P. Nicotera, and S. Lipton. 1995. Apoptosis and necrosis: two distinct events induced, respectively, by mild and intense insults with N-methyl-D-aspartate or nitric oxide/superoxide in cortical cell cultures. Proc. Natl. Acad. Sci. USA 92:7162–7166.

    Article  PubMed  CAS  Google Scholar 

  24. Bravo, R. and H. Macdonald-Bravo. 1987. Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J. Cell Biol. 105:1549–1554.

    Article  PubMed  CAS  Google Scholar 

  25. Bredesen, D.E. 1995. Neural apoptosis. Ann. Neurol. 38:839–851.

    Article  PubMed  CAS  Google Scholar 

  26. Carrio, R., M. Lopez-Hoyos, J. Jimeno, M.A. Benedict, R. Merino, A. Benito, J.L. Fernandez-Luna, G. Nunez, J.A. Garcia-Porrero, and J. Merino. 1996. A demonstrates restricted tissue distribution during embryonic development and functions to protect against cell death. Am. J. Pathol. 149:2133–2142.

    PubMed  CAS  Google Scholar 

  27. Celis, J.E., R. Bravo, P.M. Larsen, and S.J. Fey. 1984. Cyclin: a nuclear protein whose level correlates directly with the proliferative state of normal as well as transformed cells. Leukemia Res. 8:143–157.

    Article  CAS  Google Scholar 

  28. Chang, L.M.S. and F.J. Bollum. 1986. Molecular biology of terminal transferase. Crit. Rev. Biochem. 21:27–52.

    Article  CAS  Google Scholar 

  29. Chun, J. 1998. Apoptotic DNA fragmentation detection using ligation-mediated PCR, p. 23–33. In L. Zhu and J. Chun (Eds.), Apoptosis Detection and Assay Methods. Eton Publishing, Natick, MA.

    Google Scholar 

  30. Clarke, P.G.H. 1990. Developmental cell death: morphological diversity and multiple mechanisms. Anat. Embryol. 181:195–213.

    Article  PubMed  CAS  Google Scholar 

  31. Cunningham, T.J. 1982. Naturally occurring neuron death and its regulation by developing neural pathways. Int. Rev. Cytol. 74:163–186.

    PubMed  CAS  Google Scholar 

  32. Dachary-Prigent, J., J.M. Freyssinet, J.M. Pasquet, J.C. Carron, and A.T. Nurden. 1993. Annexin V as a probe of aminophospolipid exposure and platelet membrane vesiculation: a flow cytometry study showing a role for free sulfhydryl groups. Blood 81:2554–2565.

    PubMed  CAS  Google Scholar 

  33. Daly, J.M., C.B. Jannot, R.R. Beerli, D. Grasus-Porta, F.G. Maurer, and N.E. Hynes. 1997. Neu differentiation factor induces ErbB2 down-regulation and apoptosis of ErbB2-overexpressing breast tumor cells. Cancer Res. 57:3804–3811.

    PubMed  CAS  Google Scholar 

  34. Darzynkiewicz, Z., X. Li, and J. Gong. 1994. Assays of cell viability: discrimination of cells dying by apoptosis. Methods Cell Biol. 41:15–38.

    Article  PubMed  CAS  Google Scholar 

  35. De Bilbao, E., E. Guarin, P. Nef, P. Vallet, P. Giannakopoulos, and M. Dubois-Dauphin. 1999. Postnatal distribution of cpp32/caspase 3 mRNA in the mouse central nervous system: an in situ hybridization study. J. Comp. Neurol. 409:339–357.

    Article  PubMed  Google Scholar 

  36. DelRio, J.A. and E. Soriano. 1989. Immunocytochemical detection of 5’-bromodeoxyuridine incorporation in the central nervous system of the mouse. Dev. Brain Res. 49:311–317.

    Article  CAS  Google Scholar 

  37. Didenko, V.V. and P.J. Hornsby. 1996. Presence of double-strand breaks with single-base 3’ overhangs in cells undergoing apoptosis but not necrosis. J. Cell Biol. 135:1369–1376.

    Article  PubMed  CAS  Google Scholar 

  38. Duchrow, C. Schluter, G. Key, M.H. Kubbutat, C. Wohlenberg, H.D. Flad, and J. Gerdes. 1995. Cell proliferation-associated nuclear antigen defined by antibody Ki-67: a new kind of cell cycle-maintaining proteins. Arch. Immunol. Ther. Exp. 43:117–121.

    CAS  Google Scholar 

  39. Fadok, V.A., D. Voelker, P.A. Campbell, J.J. Cohen, D.L. Bratton, and P.M. Henson. 1992. Exposure of phosphatidylserine on the surface of apoptotic lypmphocytes triggers specific recognition and removal by macrophages. J. Immunol. 148:2207–2216.

    PubMed  CAS  Google Scholar 

  40. Fernandez-Alnemri, T., G. Litwack, and E.S. Alnemri. 1994. CPP32, a novel human apoptotic protein with homology to Caenorhabditis elegans cell death protein Ced-3 and mammalian interleukin-1-beta-converting enzyme. J. Biol. Chem. 269:30761–30764.

    Google Scholar 

  41. Fernandez, A., J. Kiefer, L. Fosdick, and D.J. McConkey. 1995. Oxygen radical production and thiol depletion are required for Ca(2+)-mediated endogenous endonuclease activation in apoptotic thymocytes. J. Immunol. 155:5133–5139.

    PubMed  CAS  Google Scholar 

  42. Fesus, L., Z. Nemes, L. Piredda, A. Madi, M. di Rao, and M. Picentini. 1987. Induction and activation of tissue transglutaminase during programmed cell death. FEBS Lett. 224:104–108.

    Article  PubMed  CAS  Google Scholar 

  43. Frade, J.-M., P. Bovolenta, J.R. Martinez-Morales, A. Arribas, J.A. Barbas, and A. Rodriguez-Tébar. 1997. Control of early cell death by BDNF in the chick retina. Development 124:3313–3320.

    PubMed  CAS  Google Scholar 

  44. France-Lanord, V., B. Brugg, P.P. Michel, Y. Agid, and M. Ruberg. 1997. Mitochondrial free radical signal in ceramide-dependent apoptosis: a putative mechanism for neuronal death in Parkinson’s disease. J. Neurochem. 69:1612–1621.

    PubMed  CAS  Google Scholar 

  45. Frankfurt, O.S. 1998. Detection of apoptotic cells with monoclonal antibodies to single-stranded DNA, p. 47–62. In L. Zhu and J. Chun (Eds.), Apoptosis Detection and Assay Methods. Eaton Publishing, Natick, MA.

    Google Scholar 

  46. Gage, F.H. 1998. Stem cells of the central nervous system. Curr. Opin. Neurobiol. 8:671–676.

    Article  PubMed  CAS  Google Scholar 

  47. Galli-Resta, L. and M. Ensini. 1996. An intrinsic time limit between genesis and death of individual neurons in the developing retinal ganglion cell layer. J. Neurosci. 16:2318–2324.

    PubMed  CAS  Google Scholar 

  48. Gates, M.A., L.B. Thomas, E.M. Howard, E.D. Laywell, B. Sajin, A. Faissner, B. Gotz, J. Silver, and D.A. Steindler. 1995. Cell and molecular analysis of the developing and adult mouse subventricular zone of the cerebral hemispheres. J. Comp. Neurol. 361:249–266.

    Article  PubMed  CAS  Google Scholar 

  49. Gavrieli, Y., Y. Sherman, and S.A. Ben-Sasson. 1992. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119:493–501.

    Article  PubMed  CAS  Google Scholar 

  50. Gerdes, J., U. Schwab, H. Lemke, and H. Stein. 1983. Production of a mouse monoclonal antibody reactive with a human antigen associated with cell proliferation. Int. J. Cancer 31:13–20.

    Article  PubMed  CAS  Google Scholar 

  51. Gold, R., M. Schmidt, G. Rothe, H. Zischler, H. Breitschopf, H. Wekerle, and H. Lassmann. 1993. Detection of DNA fragmentation in apoptosis: application of in situ nick translation to cell culture systems and tissue sections. J. Histochem. Cytochem. 41:1023–1030.

    PubMed  CAS  Google Scholar 

  52. Hall, P.A., P.J. Coates, R.A. Goodlad, I.R. Hart, and D.P. Lane. 1994. Proliferating cell nuclear antigen expression in non-cycling cells may be induced by growth factors in vivo. Br. J. Cancer 70:244–247.

    PubMed  CAS  Google Scholar 

  53. Hines, M.D. and B.L. Allen-Hoffmann. 1996. Keratinocyte growth factor inhibits cross-linked envelope formation and nucleosomal fragmentation in cultured human keratinocytes. J. Biol. Chem. 271:6245–6251.

    Article  PubMed  CAS  Google Scholar 

  54. Holt, S.E. and J.V. Shay. 1999. Role of telomerase in cellular proliferation and cancer. J. Cell. Physiol. 180:10–18.

    Article  PubMed  CAS  Google Scholar 

  55. Jin, K., J. Chen, T. Nagayama, M. Chen, J. Sinclair, S.H. Graham, and R.P. Simon. 1999. In situ detection of neuronal DNA strand breaks using the Klenow fragment of DNA polymerase I reveals different mechanisms of neuron death after global cerebral ischemia. J. Neurochem. 72:1204–1214.

    Article  PubMed  CAS  Google Scholar 

  56. Kerr, J.F., A.H. Wyllie, and A.R. Currie. 1972. Apoptosis: a basic biological phenomewith wide-ranging implications in tissue kinetics. Br. J. Cancer 26:239–257.

    PubMed  CAS  Google Scholar 

  57. Kondo, K. and T. Makita. 1996. Electron microscopic immunogold labeling of bromodeoxyuridine (BrdU) in routine electron microscopy. Acta Histochem. Cytochem. 29:115–119.

    CAS  Google Scholar 

  58. Korhonen, L., S. Hamner, P.A. Olsson, and D. Lindholm. 1997. Bcl-2 regulates the levels of the cysteine proteases ICH and CPP32/Yama in human neuronal precursor cells. Eur. J. Neurosci. 9:2489–2496.

    Article  PubMed  CAS  Google Scholar 

  59. Krueger, B.K., J.E Burne, and M.C. Raff. 1995. Evidence for large-scale astrocyte death in the developing cerebellum. J. Neurosci. 15:3366–3374.

    PubMed  CAS  Google Scholar 

  60. Lee, S., S. Christakos, and M.B. Small. 1993. Apoptosis and signal transduction: clues to a molecular mechanism. Curr. Opin. Cell Biol. 5:286–291.

    Article  PubMed  CAS  Google Scholar 

  61. Lois, C., J.-M. Garcia-Verdugo, and A. Alvarez-Buylla. 1996. Chain migration of neuronal precursors. Science 271:978–981.

    Article  PubMed  CAS  Google Scholar 

  62. Lossi, L., S. Mioletti, and A. Merighi. 2000. Undifferentiated progenitors and granule cell precursors undergo apoptosis within a few hours after their generation, but represent an endogenous source of brainderived neurotrophic factor for the post-natal cerebellar cortex. Soc. Neurosci. Abstr. 26:598.

    Google Scholar 

  63. Lossi, L., D. Zagzag, M.A. Greco, and A. Merighi. 1998. Apoptosis of undifferentiated progenitors and granule cell precursors in the post-natal human cerebellar cortex correlates with expression of BCL-2, ICE and CPP-32 proteins. J. Comp. Neurol. 399:359–372.

    Article  PubMed  CAS  Google Scholar 

  64. Luskin, M.B. 1993. Restricted proliferation and migration of postnatally generated neurons derived from the forebrain subventricular zone. Neuron 11:173–189.

    Article  PubMed  CAS  Google Scholar 

  65. Luskin, M.B. 1997. Characterization of neural progenitor cells of the neonatal forebrain, p. 67–86. In F.H. Gage and Y. Christen (Eds.), Isolation, Characterization and Utilization of CNS Stem Cells. Springer-Verlag, Berlin.

    Google Scholar 

  66. Meisenholder, G.W., S.G. Martin, D.R. Green, J. Nordber, B.M. Babior, and R.A. Gottlieb. 1996. Events in apoptosis. Acidification is downstream of protease activation and bcl-2 protection. J. Biol. Chem 271:16260–12262.

    Article  PubMed  CAS  Google Scholar 

  67. Migheli, A., A. Attanasio, and D. Schiffer. 1995. Ultrastructural detection of DNA strand beaks in apoptotic neural cell by in situ end-labelling techniques. J. Pathol. 176:27–35.

    Article  PubMed  CAS  Google Scholar 

  68. Miller, M.A., C.M. Mazewski, Y. Sheikh, L.M. White, G.A. Yanik, D.M. Hyams, B.C. Lampkin, and A. Raza. 1991. Simultaneous immunohistochemical detection of IUdR and BrdU infused intravenously to cancer patients. J. Histochem. Cytochem. 39:407–412.

    PubMed  CAS  Google Scholar 

  69. Miller, M.W. and R.S. Novakowski. 1988. Use of bromodeoxyuridine-immunohistochemistry to examine the proliferation, migration and time of origin of cells in the central nervous system. Brain Res. 457:44–52.

    Article  PubMed  CAS  Google Scholar 

  70. Mitev, V., A. Pauloin, and L.M. Houdebine. 1994. Purification and characterization of two casein kinase type II isozymes from bovine gray matter. J. Neurochem. 63:717–726.

    Article  PubMed  CAS  Google Scholar 

  71. Muto, M., M. Utsuyama, T. Horiguchi, E. Kubo, T. Sado, and K. Hirokawa. 1995. The characterization of the monoclonal antibody Th-10a, specific for a nuclear protein appearing in the S phase of the cell cycle in normal thymocytes and its unregulated expression in lymphoma cell lines. Cell Prolif. 28:645–657.

    PubMed  CAS  Google Scholar 

  72. Nakajima, T., K. Kagawa, T. Deguchi, H. Hikita, T. Okanoue, K. Kashima, E. Konishi, and T. Ashihara. 1994. Biological role of proliferating cell nuclear antigen (PCNA) expression during rat liver regeneration. Acta Histochem. Cytochem. 27:135–140.

    Google Scholar 

  73. Naot, D., R.V. Sionov, and D. Ish-Shalom. 1997. CD44: structure, function, and association with the malignant process. Adv. Cancer Res. 71:241–319.

    Google Scholar 

  74. Nicholson, D.W., A. Ali, N.A. Thornberry, J.P. Vaillancourt, C.K. Ding, M. Gallant, Y. Gareau, and P.R. Griffin. 1995. Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature 376:37–43.

    Article  PubMed  CAS  Google Scholar 

  75. Nicotera, P. and A.D. Rossi. 1994. Nuclear Ca2+: physiological regulation and role in apoptosis. Mol. Cell Biochem. 135:89–98.

    Article  PubMed  CAS  Google Scholar 

  76. Nishikawa, S. and F. Sasaki. 1995. DNA localization in nuclear fragments of apoptotic ameloblasts using anti-DNA immunoelectron microscopy: programmed cell death of ameloblasts. Histochem. Cell. Biol. 104:151–159.

    Article  PubMed  CAS  Google Scholar 

  77. Oppenheim, R.W. 1985. Naturally occurring cell death during neural development. Trends Neurosci. 8:487–493.

    Article  Google Scholar 

  78. Peretto, P., A. Merighi, A. Fasolo, and L. Bonfanti. 1996. Glial tubes in the rostral migratory stream of the adult rat. Brain Res. Bull. 42:9–21.

    Article  Google Scholar 

  79. Peretto, P., A. Merighi, A. Fasolo, and L. Bonfanti. 1999. The subependymal layer in rodents: a site of structural plasticity and cell migration in the adult mammalian brain. Brain Res. Bull. 49:221–243.

    Article  PubMed  CAS  Google Scholar 

  80. Piacentini, M., L. Fesus, M.G. Farrace, L. Ghibelli, L. Piredda, and G. Melino. 1991. The expression of „tissue„ transglutaminase in two human cancer cell lines is related with programmed cell death (apoptosis). Eur. J. Cell Biol. 54:246–254.

    PubMed  CAS  Google Scholar 

  81. Polakowska, R.R., M. Piacentini, R. Batlett, L.A. Goldsmith, and A.R. Haake. 1994. Apoptosis in human skin development: morphogenesis, periderm and stem cells. Dev. Dynamics 199:176–188.

    CAS  Google Scholar 

  82. Raff, M.C., B.A. Barres, J.F. Burne, H.S.R. Coles, Y. Ishizaki, and M.D. Jacobson. 1994. Programmed cell death and the control of cell survival. Philos. Trans. R. Soc. Lond. [Biol] 345:265–268.

    Article  CAS  Google Scholar 

  83. Rakic, P. 1996. Radial versus tangential migration of neuronal clones in the developing cerebral cortex. Proc. Natl. Acad. Sci. USA 92:11323–11327.

    Article  Google Scholar 

  84. Ray, S., T.J. Kelley, S. Campion, A.P. Seve, and S. Basu. 1991. Developmental expression of embryonic chicken brain DNA polymerase alpha and its binding with monoclonal antibodies against human KB cell DNA polymerase alpha. Cell Growth Differ. 2:567–573.

    PubMed  CAS  Google Scholar 

  85. Reynolds, J.E. and A. Eastman. 1996. Intracellular calcium stores are not required for Bcl-2-mediated protection from apoptosis. J. Biol. Chem. 271:27739–27743.

    Article  PubMed  CAS  Google Scholar 

  86. Rubini, M., C. D’Ambrosio, S. Carturan, G. Yumet, E. Catalano, S. Shan, Z. Huang, M. Criscuolo, M. Pifferi, and R. Baserga. 1999. Characterization of an antibody that can detect an activated IGF-I receptor in human cancers. Exp. Cell Res. 251:22–32.

    Article  PubMed  CAS  Google Scholar 

  87. Rudolph, P., R. Knuchel, E. Endl, H.J. Heidebrecht, F. Hofstader. and R. Parwaresch. 1998. The immunohistochemical marker Ki-S2: cell cycle kinetics and tissue distribution of a novel proliferation-specific antigen. Mod. Pathol. 11:450–456.

    PubMed  CAS  Google Scholar 

  88. Ryder, E.E. and C.L. Cepko. 1994. Migration patterns of clonally related granule cells and their progenitors in the developing chick cerebellum. Neuron 12:1011–1029.

    Article  PubMed  CAS  Google Scholar 

  89. Sadi, M.V. and E.R. Barrack. 1991. Determination of growth fraction in advanced prostate cancer by Ki-67 immunostaining and its relationship to the time of tumor progression after hormonal therapy. Cancer 67:3065–3071.

    Article  PubMed  CAS  Google Scholar 

  90. Sallinen, P., H. Haapasalo, T. Kerttula, I. Rantala, H. Kalimo, Y. Collan, J. Isola, and H. Helin. 1994. Sources of variation in the assesment of cell proliferation using proliferating cell nuclear antigen histochemistry. Anal. Quant. Cytol. and Histol. 16:261–268.

    CAS  Google Scholar 

  91. Santisteban, M.S. and G. Brugal. 1994. Image analysis of in situ cell cycle related changes of PCNA and Ki-67 proliferating antigen expression. Cell Prolif. 27:435–453.

    CAS  Google Scholar 

  92. Sasaki, K., T. Murakami, M. Kawasaki, and M. Takahashi. 1987. The cell cycle associated change of the Ki-67 reactive nuclear antigen expression. J. Cell. Physiol. 133:579–584.

    Article  PubMed  CAS  Google Scholar 

  93. Sato, Y., T. Ito, A. Nozawa, and M. Kanisawa. 1995. Bromodeoxyuridine and iododeoxyuridine double immunostaining for epoxy resin sections. Biotech. Hischem. 70:159–174.

    Google Scholar 

  94. Schlegel, J., I. Peter, S. Orrenius, D.K. Miller, N.A. Thornberry, T.T. Yamin, and D.W. Nicholson. 1996. CPP32/apopain is a key interleukin 1 beta converting-like protease involved in Fas-mediated apoptosis. J. Biol. Chem. 271:1841–1844.

    Article  PubMed  CAS  Google Scholar 

  95. Schluter, C., M. Duchrow, C. Vohlenberg, M.H. Becker, G. Key, H.D. Flad, and J. Gerdes. 1993. The cell proliferation-associated antigen of antibody Ki-67: a very large, ubiquitous nuclear protein with numerous repeated elements, representing a new kind of cell cycle-maintaining proteins. J. Cell Biol. 123:513–522.

    Article  PubMed  CAS  Google Scholar 

  96. Schwartz, L.M., S.W. Smith, M.E. Jones, and B.A. Osborne. 1993. Do all programmed cell death occur via apoptosis? Proc. Natl. Acad. Sci. USA 90:980–984.

    Article  PubMed  CAS  Google Scholar 

  97. Simonati, A., C. Tosati, T. Rosso, E. Piazzola, and N. Izzuto. 1999. Cell proliferation and death: morphological evidence during corticogenesis in the developing human brain. Microsc. Res. Tech. 45:341–352.

    Article  PubMed  CAS  Google Scholar 

  98. Sirri, V., P. Roussel, and D. Hernandez-Verdun. 2000. The AgNOR proteins: qualitative and quantitative changes during the cell cycle. Micron 31:121–126.

    Article  PubMed  CAS  Google Scholar 

  99. Soriano, E., J.A. Del Rio, and C. Auladell. 1993. Characterization of the phenotype and birthdates of pyknotic dead cells in the nervous system by a combination of DNA staining and immunocytochemistry for 5’-bromodeoxyuridine and neural antigens. J. Histochem. Cytochem. 41:819–827.

    PubMed  CAS  Google Scholar 

  100. Soriano, E., J.A. Del Rio, F.J. Martinez-Guijarro, I. Ferrer, and C. Lopez. 1991. Immunocytochemical detection of 5’-bromodeoxyuridine in fluoro-gold-labeled neurons: a simple technique to combine retrograde axonal. J. Histochem. Cytochem. 39:1565–1570.

    PubMed  CAS  Google Scholar 

  101. Spreafico, R., P. Arcelli, C. Frassoni, P. Canetti, G. Giaccone, T. Rizzuti, M. Mastrangelo, and M. Bentivoglio. 1999. Development of layer I of the human cerebral cortex after midgestation: architectonic findings, immunocytochemical identification of neurons and glia, and in situ labeling of apoptotic cells. J. Comp. Neurol. 410:126–142.

    Article  PubMed  CAS  Google Scholar 

  102. Srinivasan, A., K.A. Roth, R.O. Sayers, K.S. Schindler, A.M. Wong, L.C. Fritz, and K.J. Tomaselli. 1998. In situ immunodetection of activated caspase-3 in apoptotic neurons in the developing nervous system. Cell Death Differ. 5:1004–1016.

    Article  PubMed  CAS  Google Scholar 

  103. Takahashi, T. and V.S. Caviness, Jr. 1993. PCNA-binding to DNA at the G1/S transition in proliferating cells of the developing cerebral wall. J. Neurocytol. 22:1096–1102.

    Article  PubMed  CAS  Google Scholar 

  104. Tamatani, R., Y. Taniguchi, and Y. Kawarai. 1995. Ultrastructural study of proliferating cells with an improved immunocytochemical detection of DNA-incorporated bromodeoxyuridine. J. Histochem. Cytochem. 43:21–29.

    PubMed  CAS  Google Scholar 

  105. Tampanaru-Sarmesiu, A., L. Stefaneau, K. Thapar, G. Kontogeorgos, T. Sumi, and K. Kovacs. 1998. Transferrin and transferrin receptor in human hypophysis and pituitary adenomas. Am. J. Pathol. 152:413–422.

    PubMed  CAS  Google Scholar 

  106. Tanaka, M. and T. Marunouchi. 1998. Immunohistochemical analysis of developmental stage of external granular layer neurons which undergo apoptosis in postnatal rat cerebellum. Neurosci. Lett. 242:85–88.

    Article  PubMed  CAS  Google Scholar 

  107. Thiry, M. 1992. Highly sensitive immunodetection of DNA on sections with exogenous terminal deoxynucleotidyl transferase and non-isotopic nucleotide analogues. J. Histochem. Cytochem. 40:411–419.

    PubMed  CAS  Google Scholar 

  108. Thiry, M. 1993. Immunodetection of RNA on ultrathin sections incubated with polydenylated nucleotidyl tranferase. J. Histochem. Cytochem. 41:657–665.

    PubMed  CAS  Google Scholar 

  109. Thiry, M. 1995. Nucleic acid compartmentalization within the cell nucleus by in situ transferase-immunogold techniques. Microsc. Res. Tech. 31:4–21.

    Article  PubMed  CAS  Google Scholar 

  110. Thiry, M. and F. Puvion Dutilleul. 1995. Differential distribution of single-stranded DNA, double-stranded DNA, and RNA in adenovirus-induced intranuclear regions of HeLa cells. J. Histochem. Cytochem. 43:749–759.

    PubMed  CAS  Google Scholar 

  111. Torunsciolo, D.R.Z., R.E. Schimdt, and K.A. Roth. 1995. Simultaneous detection of TdT-mediated dUTP-biotyn nick end-labeling (TUNEL) positive cells and multiple immunohistochemical markers in single tissue sections. BioTechniques 19:800–805.

    Google Scholar 

  112. Trere, D. 2000. AgNOR staining and quantification. Micron 31:127–131.

    Article  PubMed  CAS  Google Scholar 

  113. Tsanaclis, A.M., S.S. Brem, S. Gately, H.M. Shipper, and R. Wang. 1991. Statin immunolocalization in human brain tumors. Detection of noncycling cells using a novel marker of cell quiescence. Cancer 68:786–792.

    Article  PubMed  CAS  Google Scholar 

  114. Tsurusawa, M., M. Ito, Z. Zha, S. Kawai, Y. Takasaki, and T. Fujimoto. 1992. Cell-cycle-associated expressions of proliferating cell nuclear antigen and Ki-67 reactive antigen of bone marrow blast cells in childhood acute leukemia. Leukemia 6:669–674.

    PubMed  CAS  Google Scholar 

  115. Tsutsui, K., S. Okada, M. Watanabe, T. Shohmori, S. Seki, and Y. Inoue. 1993. Molecular cloning of partial cDNAs for rat DNA topoisomerase II isoforms and their differential expression in brain development. J. Biol. Chem. 268:19076–19083.

    PubMed  CAS  Google Scholar 

  116. Ulaner, G.A. and L.C. Giudice. 1997. Developmental regulation of telomerase activity in human fetal tissues during gestation. Mol. Hum. Reprod. 3:769–773.

    Article  PubMed  CAS  Google Scholar 

  117. van Dierendonck, J.H., R. Verheijen, H.J. Kuijpers, R. van Driel, J.L. Beck, G.J. Brakenhoff, and F.C. Ramaekers. 1989. Ki-67 detects a nuclear matrix-associated proliferation-related antigen. II. Locaflization in mitotic cells and association with chromosomes. J. Cell Sci. 92:531–540.

    PubMed  Google Scholar 

  118. van Dierendonck, J.H., J.H. Wijsman, R. Keijzer, C.J. van de Velde, and C.J. Cornelisse. 1991. Cell-cycle-related staining patterns of anti-proliferating cell nuclear antigen monoclonal antibodies. Comparison with BrdUrd labeling and Ki-67 staining. Am. J. Pathol. 138:1165–1172.

    PubMed  Google Scholar 

  119. Villalba, M., J. Boeckart, and L. Journot. 1997. Pituitary adenylate cyclase-activating polypeptide (PACAP-38) protects cerebellar granule neurons from apoptosis by activating the mitogen-activated protein kinase (MAP Kinase) pathway. J. Neurosci. 17:83–90.

    PubMed  CAS  Google Scholar 

  120. Wang, X., N.G. Zelenski, Y. Yang, J. Sakai, M.S. Brown, and J.L. Goldstein. 1996. Cleavage of sterol regulatory element binding proteins (SREBPs) by CPP32 during apoptosis. EMBO J. 15:1012–1020.

    PubMed  CAS  Google Scholar 

  121. Waseem, N.H. and D.P. Lane. 1990. Monoclonal antibody analysis of the proliferating cell nuclear antigen (PCNA). Structural conversation and the detection of a nucleolar form. J. Cell Sci. 96:121–129.

    PubMed  CAS  Google Scholar 

  122. Watanabe, M., K. Tsutsui, K. Tstusui, and Y. Inoue. 1994. Differential expression of the topoisomerase II alpha and II beta mRNAs in developing rat brain. Neurosci. Res. 19:51–57.

    Article  PubMed  CAS  Google Scholar 

  123. Wood, K.A., B. Dipasquale, and R.J. Youle. 1993. In situ labeling of granule cells for apoptosis-associated DNA fragmentation reveals different mechanisms of cell loss in developing cerebellum. Neuron 11:621–632.

    Article  PubMed  CAS  Google Scholar 

  124. Wood, K.A. and R.J. Youle. 1995. The role of free radicals and p53 in neuron apoptosis in vivo. J. Neurosci. 15:5851–5857.

    PubMed  CAS  Google Scholar 

  125. Wright, W.E., M.A. Piatyszek, W.E. Rainey, W. Byrd, and J.W. Shay. 1996. Telomerase activity in human germline and embryonic tissues and cells. Dev. Genet. 18:173–179.

    Article  PubMed  CAS  Google Scholar 

  126. Wyllie, A.H. 1980. Glucocorticoid-induced tymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284:555–556.

    Article  PubMed  CAS  Google Scholar 

  127. Wyllie, A.H., R.G. Morris, A.L. Smith, and D. Dunlop. 1984. Chromatin cleavage in apoptosis: association with condensed chromatin morphology and dependence on macromolecular synthesis. J. Pathol. 142:67–77.

    Article  PubMed  CAS  Google Scholar 

  128. Xiong, Y., H. Zhang, and D. Beach. 1992. D type cyclins associate with multiple protein kinases and the DNA replication and repair factor PCNA. Cell 71:505–514.

    Article  PubMed  CAS  Google Scholar 

  129. Yamaguchi, Y., K.S.E. Nozawa, N. Hayakawa, Y. Nimura, and S. Yoshida. 1998. Change in telomerase activity of rat organs during growth and aging. Exp. Cell Res. 242:120–127.

    Article  PubMed  CAS  Google Scholar 

  130. Yanik, G.A., N. Yousuf, M.A. Miller, S.H. Swerdlow, B.C. Lampkin, and A. Raza. 1992. In vivo determination of cell cicle kinetics of non-Hodgkin’s lymphomas using iododeoxyuridine and bromodeoxyuridine. J. Histochem. Cytochem. 40:723–728.

    PubMed  CAS  Google Scholar 

  131. Yao, R. and G.M. Cooper. 1995. Requirement for phosphatidylinositol-3 kinase in the prevention of apoptosis by nerve growth factor. Science 267:2003–2006.

    Article  PubMed  CAS  Google Scholar 

  132. Yuan, J. 1995. Molecular control of life and death. Curr. Opin. Cell Biol. 7:211–214.

    Article  PubMed  CAS  Google Scholar 

  133. Zakeri, Z. and R.A. Lockshin. 1994. Physiological cell death during development and its relationship to aging. Ann. NY Acad. Sci. 719:212–229.

    Article  PubMed  CAS  Google Scholar 

  134. Zandvilit, D.W., A.M. Hanby, C.A. Austin, K.L. Marsch, I.B. Clak, N.A. Wright, and R. Poulsom. 1996. Analysis of foetal expression sites of human type II DNA topoisomerase alpha and beta mRNAs by in situ hybridization. Biochim. Biophys. Acta 1307:239–247.

    Google Scholar 

  135. Zhang, C. 1998. Monoclonal antibody as a probe for characterization and separation of apoptotic cells, p. 63–73. In L. Zhu and J. Chun (Eds.), Apoptosis Detection and Assay Methods. Eaton Publishing, Natick, MA.

    Google Scholar 

  136. Zhang, G., V. Gurtu, J.-T. Ma, and S.R. Kain. 1998. Sensitive detection of apoptosis using enhanced color variants of Annexin V conjugates, p. 1–6. In L. Zhu and J. Chun (Eds.), Apoptosis Detection and Assay Methods. Eaton Publishing, Natick, MA.

    Google Scholar 

  137. Zhang, G., V. Gurtu, C. Spencer, J.-T. Ma, and S.R. Kain. 1998. Detection of caspase activity associated with apoptosis using fluorimetric and colorimetric methods, p. 7–14. In L. Zhu and J. Chun (Eds.), Apoptosis Detection and Assay Methods. Eaton Publishing, Natick, MA.

    Google Scholar 

  138. Zhang, L. and J.E. Goldman. 1996. Generation of cerebellar interneurons from dividing progenitors in the white matter. Neuron 16:47–54.

    Article  PubMed  Google Scholar 

  139. Zurita, E., R. Himenez, R. Diaz de la Guardia, and M. Burgos. 1999. The relative rDNA content of a NOR determines its level of expression and its probability of becoming active. A sequential silver staining and in-situ hybridization study. Chromosome Res. 7:563–570.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Veterinary Morphophysiology, Università degli Studi di Torino, Torino, Italy, EU

    Laura Lossi, Silvia Mioletti, Patrizia Aimar, Renato Bruno & Adalberto Merighi

  2. Neuroscience Research Group, Università degli Studi di Torino, Torino, Italy, EU

    Laura Lossi, Patrizia Aimar & Adalberto Merighi

  3. Rita Levi Montalcini Center for Brain Repair, Università degli Studi di Torino, Torino, Italy, EU

    Adalberto Merighi

Authors
  1. Laura Lossi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Silvia Mioletti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Patrizia Aimar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Renato Bruno
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Adalberto Merighi
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Department of Veterinary Morphophysiology Rita Levi Montalcini Center Brain Repair, University of Torino, Gruglasco, I-10055, Italy

    Adalberto Merighi

  2. Department of Experimental and Biomedical Sciences CNR Center for the Study of Biomembranes, University of Padova, Padova, I-35121, Italy

    Giorgio Carmignoto

Rights and permissions

Reprints and permissions

Copyright information

© 2002 Springer-Verlag New York, Inc.

About this chapter

Cite this chapter

Lossi, L., Mioletti, S., Aimar, P., Bruno, R., Merighi, A. (2002). In Vivo Analysis of Cell Proliferation and Apoptosis in the CNS. In: Merighi, A., Carmignoto, G. (eds) Cellular and Molecular Methods in Neuroscience Research. Springer, New York, NY. https://doi.org/10.1007/978-0-387-22460-2_14

Download citation

  • DOI: https://doi.org/10.1007/978-0-387-22460-2_14

  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-0-387-95386-1

  • Online ISBN: 978-0-387-22460-2

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation