Abstract
Cell death in the immature brain can be studied in many ways using morphological and biochemical markers. Essential requirements for cell death and degeneration assessment techniques in the brain include sufficient sensitivity and the ability to differentiate between apoptotic and necrotic cell death, and different forms of DNA damage, and to distinguish between different stages in the cell death process. Detection of cell death by morphology still remains the gold standard. Morphological analyses can also be combined with immunohistochemistry or in situ labeling using different types of specific antibodies or markers to identify cell types undergoing degeneration. This chapter introduces widely used methods to detect different types of cell death in the immature brain after hypoxic-ischemic brain injury by using morphological assessment combined with tissue staining. The procedures, including tissue preparation, staining, and evaluation, are described.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Martin LJ et al (1998) Neurodegeneration in excitotoxicity, global cerebral ischemia, and target deprivation: a perspective on the contributions of apoptosis and necrosis. Brain Res Bull 46(4):281–309
MacManus JP, Linnik MD (1997) Gene expression induced by cerebral ischemia: an apoptotic perspective. J Cereb Blood Flow Metab 17(8):815–832
Majno G, Joris I (1995) Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol 146(1):3–15
Wyllie AH, Beattie GJ, Hargreaves AD (1981) Chromatin changes in apoptosis. Histochem J 13(4):681–692
Kerr JF, Wyllie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26(4):239–257
Oppenheim RW et al (2008) Developing postmitotic mammalian neurons in vivo lacking Apaf-1 undergo programmed cell death by a caspase-independent, nonapoptotic pathway invol-ving autophagy. J Neurosci 28(6):1490–1497
Zhu C et al (2000) Correlation between caspase- 3 activation and three different markers of DNA damage in neonatal cerebral hypoxia-ischemia. J Neurochem 75(2):819–829
Sidhu RS, Tuor UI, Del Bigio MR (1997) Nuclear condensation and fragmentation following cerebral hypoxia-ischemia occurs more frequently in immature than older rats. Neurosci Lett 223(2):129–132
Portera-Cailliau C, Price DL, Martin LJ (1997) Excitotoxic neuronal death in the immature brain is an apoptosis-necrosis morphological continuum. J Comp Neurol 378(1):70–87
Ishimaru MJ et al (1999) Distinguishing excitotoxic from apoptotic neurodegeneration in the developing rat brain. J Comp Neurol 408(4):461–476
Bonfoco E et al (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 U S A 92(16):7162–7166
Stroemer RP, Rothwell NJ (1998) Exacerbation of ischemic brain damage by localized striatal injection of interleukin-1beta in the rat. J Cereb Blood Flow Metab 18(8):833–839
Nakajima W et al (2000) Apoptosis has a prolonged role in the neurodegeneration after hypoxic ischemia in the newborn rat. J Neurosci 20(21):7994–8004
Johnston MV, Nakajima W, Hagberg H (2002) Mechanisms of hypoxic neurodegeneration in the developing brain. Neuroscientist 8(3):212–220
Puka-Sundvall M et al (2000) Impairment of mitochondrial respiration after cerebral hypoxia-ischemia in immature rats: relationship to activation of caspase-3 and neuronal injury. Brain Res Dev Brain Res 125(1–2):43–50
Puka-Sundvall M et al (2000) Subcellular distribution of calcium and ultrastructural changes after cerebral hypoxia-ischemia in immature rats. Brain Res Dev Brain Res 125(1–2):31–41
Gozuacik D, Kimchi A (2004) Autophagy as a cell death and tumor suppressor mechanism. Oncogene 23(16):2891–2906
Zhu C et al (2005) The influence of age on apoptotic and other mechanisms of cell death after cerebral hypoxia-ischemia. Cell Death Differ 12(2):162–176
Blomgren K, Leist M, Groc L (2007) Pathological apoptosis in the developing brain. Apoptosis 12(5):993–1010
Slikker W Jr et al (2007) Ketamine-induced neuronal cell death in the perinatal rhesus monkey. Toxicol Sci 98(1):145–158
Zhu C et al (2003) Involvement of apoptosis-inducing factor in neuronal death after hypoxia-ischemia in the neonatal rat brain. J Neurochem 86(2):306–317
Galluzzi L, Blomgren K, Kroemer G (2009) Mitochondrial membrane permeabilization in neuronal injury. Nat Rev Neurosci 10(7):481–494
Schmued LC, Albertson C, Slikker W Jr (1997) Fluoro-Jade: a novel fluorochrome for the sensitive and reliable histochemical localization of neuronal degeneration. Brain Res 751(1):37–46
Zhu C et al (2006) Different apoptotic mechanisms are activated in male and female brains after neonatal hypoxia-ischaemia. J Neurochem 96(4):1016–1027
Puyal J et al (2009) Postischemic treatment of neonatal cerebral ischemia should target autophagy. Ann Neurol 66(3):378–389
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
Zhu, C., Blomgren, K. (2012). Morphological Assessments of Neonatal Hypoxia–Ischemia: In Situ Cell Degeneration. In: Chen, J., Xu, XM., Xu, Z., Zhang, J. (eds) Animal Models of Acute Neurological Injuries II. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-61779-782-8_19
Download citation
DOI: https://doi.org/10.1007/978-1-61779-782-8_19
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-781-1
Online ISBN: 978-1-61779-782-8
eBook Packages: Springer Protocols