Injury Response of Resected Human Brain Tissue In Vitro

Brain injury affects a significant number of people each year. Organotypic cultures from resected normal neocortical tissue provide unique opportunities to study the cellular and neuropathological consequences of severe injury of adult human brain tissue in vitro. The in vitro injuries caused by resection (interruption of the circulation) and aggravated by the preparation of slices (severed neuronal and glial processes and blood vessels) reflect the reaction of human brain tissue to severe injury. We investigated this process using immunocytochemical markers, reverse transcriptase quantitative polymerase chain reaction and Western blot analysis. Essential features were rapid shrinkage of neurons, loss of neuronal marker expression and proliferation of reactive cells that expressed Nestin and Vimentin. Also, microglia generally responded strongly, whereas the response of glial fibrillary acidic protein‐positive astrocytes appeared to be more variable. Importantly, some reactive cells also expressed both microglia and astrocytic markers, thus confounding their origin. Comparison with post‐mortem human brain tissue obtained at rapid autopsies suggested that the reactive process is not a consequence of epilepsy.

[1]  P. London.  Injury , 1969, Definitions.

[2]  A. Moorman,et al.  Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data , 2009, Nucleic acids research.

[3]  J. C. Baayen,et al.  Mature astrocytes in the adult human neocortex express the early neuronal marker doublecortin. , 2007, Brain : a journal of neurology.

[4]  R. Verwer,et al.  Neural stem cells improve neuronal survival in cultured postmortem brain tissue from aged and Alzheimer patients , 2007, Journal of cellular and molecular medicine.

[5]  M. Sofroniew,et al.  Astrocytes: biology and pathology , 2009, Acta Neuropathologica.

[6]  Robert Leech,et al.  White matter damage and cognitive impairment after traumatic brain injury , 2010, Brain : a journal of neurology.

[7]  Q. Lu,et al.  The Basic Helix-Loop-Helix Transcription Factor Olig2 Is Critical for Reactive Astrocyte Proliferation after Cortical Injury , 2008, The Journal of Neuroscience.

[8]  F. Helmchen,et al.  Resting Microglial Cells Are Highly Dynamic Surveillants of Brain Parenchyma in Vivo , 2005, Science.

[9]  M. Risling,et al.  Rapid, widespread, and longlasting induction of nestin contributes to the generation of glial scar tissue after CNS injury , 1995, The Journal of cell biology.

[10]  A. von Deimling,et al.  Decreased expression of glutamate transporters in astrocytes after human traumatic brain injury. , 2006, Journal of neurotrauma.

[11]  M. Götz,et al.  Progeny of Olig2-Expressing Progenitors in the Gray and White Matter of the Adult Mouse Cerebral Cortex , 2008, The Journal of Neuroscience.

[12]  Johan Pallud,et al.  NG2+/Olig2+ Cells are the Major Cycle‐Related Cell Population of the Adult Human Normal Brain , 2010, Brain pathology.

[13]  Christian E. Elger,et al.  Synaptic Plasticity in the Human Dentate Gyrus , 2000, The Journal of Neuroscience.

[14]  D. Kooy,et al.  Separate blood and brain origins of proliferating cells during gliosis in adult brains , 1990, Brain Research.

[15]  Magdalena Götz,et al.  The stem cell potential of glia: lessons from reactive gliosis , 2011, Nature Reviews Neuroscience.

[16]  F. Speleman,et al.  Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes , 2002, Genome Biology.

[17]  Timothy J. Mitchison,et al.  A chemical method for fast and sensitive detection of DNA synthesis in vivo , 2008, Proceedings of the National Academy of Sciences.

[18]  David A Ramsay,et al.  The cellular inflammatory response in human spinal cords after injury. , 2006, Brain : a journal of neurology.

[19]  W. Richardson,et al.  Cell cycle dynamics of NG2 cells in the postnatal and ageing brain. , 2009, Neuron glia biology.

[20]  R. Folkerth,et al.  Proliferative Potential of Human Astrocytes , 2005, Journal of neuropathology and experimental neurology.

[21]  R. Verwer,et al.  Post-mortem brain tissue cultures from elderly control subjects and patients with a neurodegenerative disease , 2003, Experimental Gerontology.

[22]  J. A. Connor,et al.  Spontaneous pyramidal cell death in organotypic slice cultures from rat hippocampus is prevented by glutamate receptor antagonists , 1994, Neuroscience.

[23]  M. Götz,et al.  Progenitors in the adult cerebral cortex: Cell cycle properties and regulation by physiological stimuli and injury , 2011, Glia.

[24]  D. C. Sterio.  The unbiased estimation of number and sizes of arbitrary particles using the disector , 1984, Journal of microscopy.

[25]  M. Sofroniew,et al.  Essential protective roles of reactive astrocytes in traumatic brain injury. , 2006, Brain : a journal of neurology.

[26]  R. Verwer,et al.  Cells in human postmortem brain tissue slices remain alive for several weeks in culture , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[27]  C. Delphin,et al.  S100B expression defines a state in which GFAP‐expressing cells lose their neural stem cell potential and acquire a more mature developmental stage , 2007, Glia.

[28]  R. Vink,et al.  Traumatic brain injury: Therapeutic challenges and new directions , 2011, Neurotherapeutics.

[29]  S. Sisodiya,et al.  A spatiotemporal study of gliosis in relation to depth electrode tracks in drug-resistant epilepsy , 2014, The European journal of neuroscience.

[30]  D. Rowitch,et al.  Expression pattern of the transcription factor Olig2 in response to brain injuries: implications for neuronal repair. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[31]  J. Baudier,et al.  Nuclear expression of S100B in oligodendrocyte progenitor cells correlates with differentiation toward the oligodendroglial lineage and modulates oligodendrocytes maturation , 2004, Molecular and Cellular Neuroscience.

[32]  H. Yamada,et al.  Nestin-positive microglia in adult rat cerebral cortex , 2009, Brain Research.

[33]  Fabian J Theis,et al.  Live imaging of astrocyte responses to acute injury reveals selective juxtavascular proliferation , 2013, Nature Neuroscience.

[34]  B. Barres,et al.  Genomic Analysis of Reactive Astrogliosis , 2012, The Journal of Neuroscience.

[35]  B. Zlokovic,et al.  Central nervous system pericytes in health and disease , 2011, Nature Neuroscience.

[36]  R. McKay,et al.  Re-expression of the intermediate filament nestin in reactive astrocytes , 1995, Neurobiology of Disease.

[37]  C. D. de Kock,et al.  Frontiers in Synaptic Neuroscience Synaptic Neuroscience , 2022 .

[38]  David W. McNeal,et al.  Prolonged microgliosis in the rhesus monkey central nervous system after traumatic brain injury. , 2007, Journal of neurotrauma.

[39]  Y. Benjamini,et al.  False Discovery Rate–Adjusted Multiple Confidence Intervals for Selected Parameters , 2005 .

[40]  M. Mattson,et al.  Evidence that nucleocytoplasmic Olig2 translocation mediates brain‐injury‐induced differentiation of glial precursors to astrocytes , 2007, Journal of neuroscience research.

[41]  Elizabeth Roberts-Pedersen,et al.  Damage , 2014 .

[42]  K. Janeczko.  Spatiotemporal patterns of the astroglial proliferation in rat brain injured at the postmitotic stage of postnatal development: a combined immunocytochemical and autoradiographic study , 1989, Brain Research.

[43]  M. Graeber,et al.  The microglial cytoskeleton: vimentin is localized within activated cellsin situ , 1988, Journal of neurocytology.

[44]  Kevin J. Staley,et al.  Interictal spikes, seizures and ictal cell death are not necessary for post-traumatic epileptogenesis in vitro , 2012, Neurobiology of Disease.

[45]  H. Romijn,et al.  A procedure for culturing rat neocortex explants in a serum-free nutrient medium , 1988, Journal of Neuroscience Methods.

[46]  Karl Herrup,et al.  Cell cycle regulation in the postmitotic neuron: oxymoron or new biology? , 2007, Nature Reviews Neuroscience.