Abstract
We analysed the spatial and temporal distribution of apoptosis in human cerebellum development, during embryonic and fetal periods. Cerebella excised from two human embryos (8 weeks old) and eight fetuses (12‐22 weeks old), were paraffin embedded and serially sectioned. Apoptotic cells were identified by propidium iodide staining, and TUNEL. In addition, immunohistochemistry for suicide receptor Fas(APO‐1/CD95) was performed. We determined the distribution and percentage of apoptotic cells as well as Fas(APO‐1/CD95)‐positive cells in different regions and stages of development. Apoptotic cells were seen in both proliferative zones and postmitotic regions along the migratory pathways as well as in the developing cerebellar cortex in all examined stages. The Fas(APO‐1/CD95) immunoreactivity was present in all examined stages in a small population of apoptotic cells: either neuroblasts or differentiated cells in postmitotic zones. These findings suggest that apoptosis drives the selection of the cells which are committed to differentiate during the early stages of cerebellar development. The differences between apoptotic cells distribution and Fas receptor expression suggest that cell selection is driven by different apoptotic pathways.
Keywords: apotosis, human embryo, cerebellum, TUNEL, Fas(APO‐1/CD95)
References
- 1. Oppenheim R.W., Cell death during development of nervous system, Ann. Rev. Neurosci., 14: 453–501, 1991. [DOI] [PubMed] [Google Scholar]
- 2. Jacobson M.D., Programmed cell death and the control of the cell survival: lessons from the nervous system, Science, 262: 695–700, 1993. [DOI] [PubMed] [Google Scholar]
- 3. Saunders J.W., Death in embryonic system, Science, 154: 604–612, 1966. [DOI] [PubMed] [Google Scholar]
- 4. Caviness V.S.J., Takahaski T., Nowakowski R.S., Time and neocortical neurogenesis: a general development and evolutionary model, Trends. Neurosci., 18: 379–383, 1995. [DOI] [PubMed] [Google Scholar]
- 5. Cowan W.M., Fawcett J.W., Regressive events in neurogenesis, Science, 225: 1258–1265, 1984. [DOI] [PubMed] [Google Scholar]
- 6. Boulder Commitee , Embryonic vertebrate central nervous system: revised terminology, Anat Rec, 166: 257–261, 1970. [DOI] [PubMed] [Google Scholar]
- 7. Blaschke A.J., Stanley K., Chun J., Widespread programmed cell death in proliferative and postmitotic regions of the fetal cerebral cortex, Development, 122: 1165–1174, 1996. [DOI] [PubMed] [Google Scholar]
- 8. Blaschke A.J., Weiner J.A., Chung J., Programmed cell death is a universal feature of embryonic and postnatal neuroproliferative regions throughout the central nervous system, J. Comp. Neurol., 396:1 39–50, 1998. [DOI] [PubMed] [Google Scholar]
- 9. Clarke P.G., Posada A., Primi M.G., Castagne V., Neuronal death in the central nervous system during development, Biomed. Pharmacother., 52(9): 356–62, 1998. [DOI] [PubMed] [Google Scholar]
- 10. Felderhoff‐Mueser U., Taylor L., Greenwoo K., Kozma M., Stibenz D., Joashi U.C., Ewards A.D., Mehmet H., Fas/CD95/APO‐1 can function as a death receptor for neuronal cells in vitro and in vivo and is upregulated following cerebral hypoxicischemic injury to the developing rat brain, Brain Pathol. 10:17–29, 2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11. Popescu B.O., Popescu L.M., Neuronal apoptosis triggered by anti‐Fas (CD95, APO‐1) antibody is enhanced by dexamethasone, J.Med.Biochem., 4:1–14, 2000. [Google Scholar]
- 12. Becher B., Barker P.A., Owens T., Antel J., CD 95‐CD95L: can the brain learn from the immune system?, Trends. Neurosci., 21: 114–117, 1998. [DOI] [PubMed] [Google Scholar]
- 13. Gorman A.M., Orrenius S., Ceccatelli S., Apoptosis in neuronal cells: role of caspases, Neuroreport, 9:R49–R55, 1998. [DOI] [PubMed] [Google Scholar]
- 14. Martin‐Villalba A., Herr I., Jeremias I., Hahne M., Brandt R., Vogel J., Schenkel J., Herdegen T., Debatin K.M., CD95 ligand (Fas‐L/APO1‐L) and tumor necrosis factor‐related apoptosis‐inducing ligand mediate ischemia‐induced apoptosis in neurons, J. Neurosci., 19: 3809–3817, 1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15. Larsen W.J., Human embriology, Churchill Livingstone, New York , 1997. [Google Scholar]
- 16. Gavrieli Y., Sherman Y., Ben‐Sasson S.A., Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation, J. Cell. Biol., 119: 493–501, 1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 17. Wijsman J.H., Jonker R.R., Keijer R., van de Velde C.J., Cornelisse C.J. and van Dierendonck J.H., A new method to detect apoptosis in paraffin sections: in situ end labeling of fragmented DNA, J. Histochem. Cytochem., 41: 7–12, 1993. [DOI] [PubMed] [Google Scholar]
- 18. Simonati A., Rosso T., Rizzuto N., DNA fragmentation in normal development of the human central nervous system, Neuropathol. Appl. Neurobiol., 23:3,203–11, 1997. [PubMed] [Google Scholar]
- 19. Angevine J.B., Mancall E.L., Yakovlev P.I., The human cerebellum, Boston , Little, Brown, 1961. [Google Scholar]
- 20. Fields W.D., Willis W.D. Jr., The cerebellum in health and disease, St. Louis , Green, 1970. [Google Scholar]
- 21. Ghez C., The cerebellum In Kandel E.R., Schwartz J.H., and Jessell T.M., (eds.): Principles of neural science, 3rd ed., Elsevier, New York , 1991. [Google Scholar]
- 22. Llinas R.R., Walton K.D., Cerebellum In Shepherd G.M., (ed): The synaptic organization of the brain, 3rd ed., New York , Oxford University Press, 1990. [Google Scholar]
- 23. Llinas R.R., The cortex of the cerebellum, Sci. Am. 232:56, 1975. [DOI] [PubMed] [Google Scholar]
- 24. Norman M.G., Central nervous system In Developmental Pathology of the Embryo and Fetus (Eds Dimmick J.E. and Kalousek D.K.), Philadelphia , Lippincott, 1992. [Google Scholar]
- 25. Palay S.L., Chan‐Palay V., Cerebellar cortex: cytology and organization, Springer‐Verlag, Heidelberg , 1974. [Google Scholar]
- 26. Perry D.G., Hanson V., Benuck M., Puszkin S., Neuronal protein NP185 in avian and murine cerebellum: expression during development and evidence for its presence in nerve endings, Histochem. Cytochem., 39:1461–1470, 1991. [DOI] [PubMed] [Google Scholar]
