scholarly journals Immunocytochemical Study of an Early Microglial Activation in Ischemia

1992 ◽  
Vol 12 (2) ◽  
pp. 257-269 ◽  
Author(s):  
Jochen Gehrmann ◽  
Petra Bonnekoh ◽  
Takahito Miyazawa ◽  
Konstantin-Alexander Hossmann ◽  
Georg W. Kreutzberg
Keyword(s):  
Monoclonal Antibodies ◽  
Distribution Pattern ◽  
Rat Brain ◽  
Mhc Class Ii ◽  
Neuronal Damage ◽  
Dorsal Hippocampus ◽  
Neuronal Cell ◽  
Microglial Cells ◽  
Vessel Occlusion ◽  
Temporal And Spatial Distribution

Transient arrest of the cerebral blood circulation results in neuronal cell death in selectively vulnerable regions of the rat brain. To elucidate further the involvement of glial cells in this pathology, we have studied the temporal and spatial distribution pattern of activated microglial cells in several regions of the ischemic rat brain. Transient global ischemia was produced in rats by 30 min of a four-vessel occlusion. Survival times were 1, 3, and 7 days after the ischemic injury. The microglial reaction was studied immunocytochemically using several monoclonal antibodies, e.g., against CR3 complement receptor and major histocompatibility complex (MHC) antigens. Two recently produced monoclonal antibodies against rat microglial cells, designated MUC 101 and 102, were also used to identify microglial cells. Following ischemia, the microglial reaction was correlated with the development of neuronal damage. The earliest presence of activated microglial cells was observed in the dorsolateral striatum, the CA1 area, and the dentate hilus of the dorsal hippocampus. However, the microglial reaction was not confined to areas showing selective neuronal damage, but also occurred in regions that are rather resistant to ischemia, such as the CA3 area. Particularly in the frontoparietal cortex, the appearance of MHC class II–positive microglial cells provided an early indication of the subsequent distribution pattern of neuronal damage. The microglial reaction would thus seem to be an early, sensitive, and reliable marker for the occurrence of neuronal damage in ischemia.

scholarly journals The Microglial Reaction in the Rat Dorsal Hippocampus following Transient Forebrain Ischemia

1991 ◽  
Vol 11 (6) ◽  
pp. 966-973 ◽  
Author(s):  
Takato Morioka ◽  
Audrey N. Kalehua ◽  
Wolfgang J. Streit
Keyword(s):  
Time Course ◽  
Neuronal Damage ◽  
Dorsal Hippocampus ◽  
Microglial Cells ◽  
Forebrain Ischemia ◽  
Dentate Granule Cell ◽  
Vessel Occlusion ◽  
Reactive Microglia ◽  
Transient Forebrain Ischemia ◽  
Dentate Hilus

We have examined the distribution and time course of the microglial reaction in the rat dorsal hippocampus after 25-min transient forebrain ischemia (four-vessel occlusion model). Microglial cells were visualized in brain sections using lectin staining with the Griffonia simplicifolia B4-isolectin following intervals of reperfusion ranging from 20 min to 4 weeks. Increased staining of microglial cells was detected in the dentate hilus and area CA1 as early as 20 min after reperfusion. These same regions demonstrated more intense microglial staining after 24 h. The strongest microglial reaction was observed 4–6 days after reperfusion when reactive microglia were abundant throughout all laminae of CA1 and the dentate hilus. Following longer reperfusion intervals, the microglial reaction became less intense; however, it remained above normal levels until the end of the fourth week. At all time points examined, microglial reactivity in the CA3 pyramidal and dentate granule cell layers was considerably lower than that observed in CA1 and dentate hilus. Our results are consistent with the known serial pathological changes associated with cerebral ischemia, but, in addition, show that the examination of the microglial reaction provides an extremely sensitive indicator of subtle and morphologically nonapparent neuronal damage during the early stages of injury.

scholarly journals Lipid Peroxidation in Chronic Cerebral HypoperfusionInduced Neurodegeneration in Rats

2020 ◽  
Vol 10 (2) ◽  
Author(s):  
Anil Kumar S ◽  
Saif SA ◽  
Oothuman P ◽  
Mustafa MIA
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Lipid Peroxidation ◽  
Neurodegenerative Disorders ◽  
Neuronal Damage ◽  
Neuronal Cell ◽  
Chronic Cerebral Hypoperfusion ◽  
Cerebral Hypoperfusion ◽  
Control Group ◽  
Vessel Occlusion

Introduction: Reduced cerebral blood fl ow is associated with neurodegenerative disorders and dementia, in particular. Experimental evidence has demonstrated the initiating role of chronic cerebral hypoperfusion in neuronal damage to the hippocampus, the cerebral cortex, the white matter areas and the visual system. Permanent, bilateral occlusion of the common carotid arteries of rats (two vessel occlusion - 2VO) has been introduced for the reproduction of chronic cerebral hypoperfusion as it occurs in Alzheimer’s disease and human aging. Increased generation of free radicals through lipid peroxidation can damage neuronal cell membrane. Markers of lipid peroxidation have been found to be elevated in brain tissues and body fl uids in neurodegenerative diseases, including Alzheimer’s disease, Parkinson disease and amyotrophic lateral sclerosis. Materials and Methods: Malondialdehyde (MDA), final product of lipid peroxidation, was estimated by thiobarbituric acid-reactive substances (TBARS) assay kit at eight weeks after induction of 2VO in the rats and control group. Results: Our study revealed a highly signifi cant (p<0.001) increase in the mean MDA concentration (12.296 ± 1.113 μM) in 2VO rats as compared to the control group (5.286 ± 0.363 μM) rats. Conclusion: Therapeutic strategies to modulate lipid peroxidation early throughout the course of the disease may be promising in slowing or possibly preventing neurodegenerative disorders.

Alterations in τ Immunostaining in the Rat Hippocampus following Transient Cerebral Ischemia

1994 ◽  
Vol 14 (4) ◽  
pp. 554-564 ◽  
Author(s):  
James W. Geddes ◽  
Claudia Schwab ◽  
Susan Craddock ◽  
Janice L. Wilson ◽  
L. Creed Pettigrew
Keyword(s):  
Neuronal Damage ◽  
Hippocampal Formation ◽  
Molecular Layer ◽  
Neuronal Cell ◽  
Direct Result ◽  
Ischemic Insult ◽  
Vessel Occlusion ◽  
Neuronal Populations ◽  
Neuronal Cell Bodies ◽  
Hilar Neurons

Previous studies in gerbils have shown that cytoskeletal disruption and a loss of the dendritic microtubule-associated protein, MAP2, may occur after short periods of transient global ischemia. τ, a predominantly axonal microtubule-associated protein, has not been examined following ischemia. We compared neuronal damage with alterations in MAP2, τ, and 72-kD heat shock protein (HSP72) immunostaining at various reperfusion times following 20 min of ischemia in the rat four-vessel occlusion model. τ accumulated in neuronal cell bodies throughout the hippocampal formation 30 min to 2 h after the ischemic insult. Perikaryal τ immunostaining was transient in most regions, but persisted in polymorphic hilar neurons. This was accompanied by a loss of immunostaining in the target of many hilar neurons, the inner molecular layer of the dentate gyrus. The same neuronal populations that exhibited increased τ immunostaining of perikarya later displayed an induction of HSP72 immunoreactivity. In contrast, loss of MAP2 immunostaining was not consistently observed before neuronal death and did not correspond to HSP72 induction. The altered τ immunostaining is not the direct result of excitotoxic insult, as intrahippocampal injection of kainic acid did not cause the somal accumulation of τ, but did cause disruption of MAP2 immunostaining. Taken together, the results suggest that the somal accumulation of τ is an early, sensitive, and selective marker of ischemic insult.

Changes in Extracellular Glutamate Release on Repetitive Transient Occlusion in Global Ischemia Model

2009 ◽  
Author(s):  
Gija Lee ◽  
Seokkeun Choi ◽  
Sungwook Kang ◽  
Samjin Choi ◽  
Jeonghoon Park ◽  
...  
Keyword(s):  
Neuronal Damage ◽  
Glutamate Release ◽  
Short Interval ◽  
Neuronal Cell ◽  
Release Time ◽  
Protective Mechanism ◽  
Time Interval ◽  
Extracellular Glutamate ◽  
Vessel Occlusion ◽  
Doppler Flowmetry

During the operation, surgeons in neurosurgical area usually performed the multiple temporary occlusions of parental artery which may induce the neuronal damage. It is generally thought that neuronal damage by cerebral ischemia is associated with extracellular concentrations of the excitatory amino acids. In this experiment, we measured the dynamics of extracellular glutamate release in 11 vessel occlusion (VO) model during repeated within short interval. Changes in cerebral blood flow were monitored by laser-Doppler flowmetry simultaneously with cortical glutamate level measured by amperometric biosensor. During ischemia, the peak level of glutamate release was gradually decreased as 112.38±26.21 μM in first period, 82.63±18.50 μM in second period, and 48.58±11.89 μM in third period. The time interval between the ischemia induction and the beginning of glutamate release was increased as 106.7 ± 10.89 (sec) at first attack, 139.11 ± 3.87 (sec) in second attack, 169.00 ± 14.56 (sec) in third ischemic period. From the results of real-time monitoring about glutamate release in 11-VO model during repetitive ischemic episode, it was demonstrated that repetitive ischemia induced less glutamate release from neuronal cell than single ischemia due to endogeneous protective mechanism which delayed glutamate release time in later ischemic injury.

scholarly journals Direct Evidence for Acute and Massive Norepinephrine Release in the Hippocampus during Transient Ischemia

1989 ◽  
Vol 9 (6) ◽  
pp. 892-896 ◽  
Author(s):  
Mordecai Y.-T. Globus ◽  
Raul Busto ◽  
W. Dalton Dietrich ◽  
Elena Martinez ◽  
Isabel Valdés ◽  
...  
Keyword(s):  
Direct Evidence ◽  
Neuronal Damage ◽  
Neuronal Cell Death ◽  
Neuronal Cell ◽  
Fold Increase ◽  
Norepinephrine Release ◽  
Transient Ischemia ◽  
Vessel Occlusion ◽  
Extracellular Release ◽  
Induced Changes

Recent studies suggest the norepinephrine (NE) may play a regulatory role in neuronal cell death in the hippocampus after transient ischemia. However, ischemia-induced changes in extracellular NE release have not been demonstrated. In the present study, we utilized the microdialysis technique to measure extracellular NE levels in the hippocampus before, during, and after 20 min of global ischemia induced by two-vessel occlusion combined with systemic hypotension in the rat. Stable basal concentrations of extracellular NE were detected in three consecutive samples collected prior to ischemia (1.86 ± 1.21 pmol/ml of perfusate mean ± SEM). During ischemia, NE levels increased to 30.1 ± 5.5 pmol/ml, representing an 18-fold increase. The levels gradually returned to baseline by 40 min of reperfusion. These results are the first to demonstrate that acute and massive extracellular release of NE occurs in the hippocampus during ischemia and early recirculation. These results support the hypothesis that the activation of the noradrenergic system may play a significant role in modulating the development of ischemic neuronal damage.

Correlation between lactate and neuronal cell damage in the rat brain after focal ischemia: An in vivo 1H magnetic resonance spectroscopic (1H-MRS) study

2010 ◽  
Vol 51 (3) ◽  
pp. 344-350 ◽  
Author(s):  
Chul-Woong Woo ◽  
Byong Sop Lee ◽  
Sang Tae Kim ◽  
Ki-Soo Kim
Keyword(s):  
Magnetic Resonance ◽  
Rat Brain ◽  
Neuronal Damage ◽  
Cell Damage ◽  
Infarct Volume ◽  
Neuronal Cell ◽  
Focal Ischemia ◽  
Tunel Staining ◽  
Resonance Spectroscopy ◽  
Lipid Levels

Background: Increased levels of lactate are observed by 1H magnetic resonance spectroscopy (1H-MRS) in rat brains after stroke. However, it is not known whether the changes in lactate levels are predictive of the degree of neuronal damage. Purpose: To investigate the correlation between changes in lactate and lipid levels measured by 1H-MRS and neuronal cell damage in the rat brain. Material and Methods: A middle cerebral artery occlusion (MCAO) model was used to evaluate focal ischemia in rats (n=36). After MCAO for 90 min T2-weighted images (T2WIs), diffusion-weighted images (DWIs), and 1H-MRS data were obtained from brains immediately, 6 hours, 9 hours, 12 hours, 18 hours, 24 hours, 3 days, and 7 days after reperfusion. Infarct volumes were measured in T2WIs obtained 4 weeks after reperfusion. The degree of neuronal damage was measured by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) staining in three rats from each group at the same time as brain images were collected. Results: Creatine (Cr)-normalized lactate + lipid levels ([Lac+Lip]/Cr) were negatively correlated with Cr-normalized N-acetyl-L-aspartate levels (NAA/Cr) and positively correlated with TUNEL-positive cell numbers up to 24 hours after reperfusion. (Lac+Lip)/Cr at 6 hours and 9 hours was significantly correlated with NAA/Cr at 7 days, but there was no significant correlation between (Lac+Lip)/Cr during the first 24 hours and infarct volume at 4 weeks. Conclusion: Up to 24 hours after reperfusion, (Lac+Lip)/Cr was strongly negatively correlated with NAA/Cr, and was a good predictor of neuronal damage at 7 days; however, it was not predictive of final infarct volume at 4 weeks.

Human Cortical Neuronal Cell Line (Hcn-1): Further in Vitro Characterization and Suitability for Brain Transplantation

1992 ◽  
Vol 1 (1) ◽  
pp. 3-15 ◽  
Author(s):  
Maciej Poltorak ◽  
Mitsuo Isono ◽  
William J. Freed ◽  
Gabriele V. Ronnett ◽  
Solomon H. Snyder
Keyword(s):  
Rat Brain ◽  
Cell Line ◽  
Mhc Class I ◽  
Mhc Class Ii ◽  
Neuronal Cell ◽  
Antigen Expression ◽  
Interferon Γ ◽  
Class I ◽  
Intracerebral Transplantation

The human neuronal cell-1 (HCN-1) line has recently been established. Under favorable conditions, these cells differentiate into mature neuronal phenotypes. Here we report on further characterization of these cells. Cultured HCN-1 cells express fibronectin immunoreactivity and grow well on fibronectin substrate but do not respond to human bFGF. In the undifferentiated state, some HCN-1 cells show MHC class I antigen expression. After differentiation, HCN-1 cells and their processes are MHC class I negative. On the other hand, interferon-γ stimulation enhances MHC class I expression but does not induce MHC class II immunoreactivity. Our in vitro data indicate that HCN-1 cells express mixed characteristics, including both neuronal and mesenchymal markers, and are consistent with the suggestion that the HCN-1 cell line resembles an immature neuroepithelial cell precursor with a complex origin. One possible application of the use of the HCN-1 cells includes intracerebral transplantation. We also examined the survival of dissociated HCN-1 cells implanted into rat brain parenchyma. The host animals were not immunosuppressed. Despite expression of MHC class I antigens, small clusters of HCN-1 cells survived in the rat brain. These xenografts did not induce distinct immunological responses within the host brain tissue. Surviving HCN-1 cells demonstrated similar features to those observed in culture. Our preliminary results suggest that the HCN-1 cell line would be suitable for intracerebral transplantation in primates or humans. However, it may be that short-term host immunosuppression or addition of HCN-1 cell differentiation factors would be beneficial for enhanced cell survival.

Antisense knockdown of the glial glutamate transporter GLT-1 exacerbates hippocampal neuronal damage following traumatic injury to rat brain

2001 ◽  
Vol 13 (1) ◽  
pp. 119-128 ◽  
Author(s):  
Vemuganti L. Raghavendra Rao ◽  
Aclan Dogan ◽  
Kellie K. Bowen ◽  
Kathryn G. Todd ◽  
Robert J. Dempsey
Keyword(s):  
Rat Brain ◽  
Neuronal Damage ◽  
Traumatic Injury ◽  
Glutamate Transporter ◽  
Glial Glutamate Transporter ◽  
Antisense Knockdown

Use of Both Fluoro-Jade B and Hematoxylin and Eosin to Detect Cell Death in the Juvenile Rat Brain Exposed to NMDA-Receptor Antagonists or GABA-Receptor Agonists in Safety Assessment

2021 ◽  
pp. 019262332110077
Author(s):  
Catherine A. Picut ◽  
Odete R. Mendes ◽  
David S. Weil ◽  
Sarah Davis ◽  
Cynthia Swanson
Keyword(s):  
Cell Death ◽  
Nmda Receptor ◽  
Rat Brain ◽  
Neuronal Cell Death ◽  
Neuronal Cell ◽  
Nmda Receptor Antagonists ◽  
Neonatal Rats ◽  
Receptor Antagonists ◽  
Fluoro Jade B ◽  
Hematoxylin And Eosin

Administration of pediatric anesthetics with N-methyl D-aspartate (NMDA)-receptor antagonist and/or γ-aminobutyric acid (GABA) agonist activities may result in neuronal degeneration and/or neuronal cell death in neonatal rats. Evaluating pediatric drug candidates for this potential neurotoxicity is often part of overall preclinical new drug development strategy. This specialized assessment may require dosing neonatal rats at postnatal day 7 at the peak of the brain growth spurt and evaluating brain tissue 24 to 48 hours following dosing. The need to identify methods to aid in the accurate and reproducible detection of lesions associated with this type of neurotoxic profile is paramount for meeting the changing needs of neuropathology assessment and addressing emerging challenges in the neuroscience field. We document the use of Fluoro-Jade B (FJB) staining, to be used in conjunction with standard hematoxylin and eosin staining, to detect acute neurodegeneration and neuronal cell death that can be caused by some NMDA-receptor antagonists and/or GABA agonists in the neonatal rat brain. The FJB staining is simple, specific, and sensitive and can be performed on brain specimens from the same cohort of animals utilized for standard neurotoxicity assessment, thus satisfying animal welfare recommendations with no effect on achievement of scientific and regulatory goals.

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