Mouse Model of Intraluminal MCAO: Cerebral Infarct Evaluation by Cresyl Violet Staining

Neonatal hypoxic-ischemic brain injury is commonly studied by means of the Vannucci procedure in mice or rats (unilateral common carotid artery occlusion followed by hypoxia). Previously, we modified the postnatal day 7 (P7) rat procedure for use in mice, and later demonstrated that genetic strain strongly influences the degree of brain injury in the P7 mouse model of hypoxia-ischemia (HI). Recently, the P9 or P10 mouse brain was recognized as the developmental equivalent of a term neonatal human brain, rather than P7. Consequently, the Vannucci procedure has again been modified, and a commonly used protocol employs 10% oxygen for 50 min in C57Bl/6 mice. Strain differences have yet to be described for the P9/P10 mouse model. In order to determine if the strain differences we previously reported in the P7 mouse model are present in the P9 model, we compared 2 commonly used strains, CD1 and C57Bl/6J, in both the P7 (carotid ligation [in this case, right] followed by exposure to 8% oxygen for 30 min) and P9 (carotid ligation [in this case left] followed by exposure to 10% oxygen) models of HI. Experiments using the P7 model were performed in 2001–2012 and those using the P9 model were performed in 2012–2016. Five to seven days after the HI procedure, mice were perfused with 4% paraformaldehyde, their brains were sectioned on a Vibratome (50 µm) and alternate sections were stained with Perl’s iron stain or cresyl violet. Brain sections were examined microscopically and scored for the degree of injury. Since brains in the P7 group had been scored previously with a slightly different system, they were reanalyzed using our current scoring system which scores injury in 11 regions: the anterior, middle, and posterior cortex; the anterior, middle, and posterior striatum; CA1, CA2, CA3, and the dentate gyrus of the hippocampus and thalamus, on a scale from 0 (none) to 3 (cystic infarct) for a total score of 0–33. Brains in the P9 group were scored with the same system. Given the same insult, the P7 CD1 mice had greater injury than the C57Bl/6J mice, which agrees with our previous findings. The P9 CD1 mice also had greater injury than the C57Bl/6J mice. This study confirms that CD1 mice are more susceptible to injury than C57Bl/6J mice and that strain selection is important when using mouse models of HI.

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Comparison of age‐dependent alterations in thioredoxin 2 and thioredoxin reductase 2 expressions in hippocampi between mice and rats

Laboratory Animal Research ◽

10.1186/s42826-021-00088-y ◽

2021 ◽

Vol 37 (1) ◽

Author(s):

Yeon Ho Yoo ◽

Dae Won Kim ◽

Bai Hui Chen ◽

Hyejin Sim ◽

Bora Kim ◽

...

Keyword(s):

Thioredoxin Reductase ◽

Pyramidal Cells ◽

Brain Regions ◽

Young Age ◽

Cresyl Violet ◽

Western Blots ◽

Protein Levels ◽

Cresyl Violet Staining ◽

Age Dependent ◽

The Brain

Abstract Background Aging is one of major causes triggering neurophysiological changes in many brain substructures, including the hippocampus, which has a major role in learning and memory. Thioredoxin (Trx) is a class of small redox proteins. Among the Trx family, Trx2 plays an important role in the regulation of mitochondrial membrane potential and is controlled by TrxR2. Hitherto, age-dependent alterations in Trx2 and TrxR2 in aged hippocampi have been poorly investigated. Therefore, the aim of this study was to examine changes in Trx2 and TrxR2 in mouse and rat hippocampi by age and to compare their differences between mice and rats. Results Trx2 and TrxR2 levels using Western blots in mice were the highest at young age and gradually reduced with time, showing that no significant differences in the levels were found between the two subfields. In rats, however, their expression levels were the lowest at young age and gradually increased with time. Nevertheless, there were no differences in cellular distribution and morphology in their hippocampi when it was observed by cresyl violet staining. In addition, both Trx2 and TrxR2 immunoreactivities in the CA1-3 fields were mainly shown in pyramidal cells (principal cells), showing that their immunoreactivities were altered like changes in their protein levels. Conclusions Our current findings suggest that Trx2 and TrxR2 expressions in the brain may be different according to brain regions, age and species. Therefore, further studies are needed to examine the reasons of the differences of Trx2 and TrxR2 expressions in the hippocampus between mice and rats.

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GPVI-Fc-PEG improves cerebral infarct volume and cerebral thrombosis in mouse model with cerebral thrombosis

Molecular Medicine Reports ◽

10.3892/mmr.2017.7556 ◽

2017 ◽

Vol 16 (5) ◽

pp. 7561-7568 ◽

Cited By ~ 1

Author(s):

Yimae Wufuer ◽

Xuefeng Shan ◽

Magaoweiya Sailike ◽

Kamile Adilaimu ◽

Songfeng Ma ◽

...

Keyword(s):

Mouse Model ◽

Infarct Volume ◽

Cerebral Infarct ◽

Cerebral Thrombosis

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Evaluation of the effects of a new series of SMTPs in the acetic acid-induced embolic cerebral infarct mouse model

European Journal of Pharmacology ◽

10.1016/j.ejphar.2017.10.055 ◽

2018 ◽

Vol 818 ◽

pp. 221-227 ◽

Cited By ~ 7

Author(s):

Keita Shibata ◽

Terumasa Hashimoto ◽

Keiji Hasumi ◽

Kazuo Honda ◽

Koji Nobe

Keyword(s):

Acetic Acid ◽

Mouse Model ◽

Cerebral Infarct

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YES-10, A Combination of Extracts from Clematis mandshurica RUPR. and Erigeron annuus (L.) PERS., Prevents Ischemic Brain Injury in A Gerbil Model of Transient Forebrain Ischemia

Plants ◽

10.3390/plants9020154 ◽

2020 ◽

Vol 9 (2) ◽

pp. 154 ◽

Cited By ~ 1

Author(s):

Tae-Kyeong Lee ◽

Joon Ha Park ◽

Bora Kim ◽

Young Eun Park ◽

Jae-Chul Lee ◽

...

Keyword(s):

Reperfusion Injury ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Cresyl Violet ◽

Neuroprotective Effects ◽

Traditional Medicines ◽

Neuronal Nuclei ◽

Cresyl Violet Staining ◽

Fluoro Jade B ◽

Erigeron Annuus

Clematis mandshurica RUPR. (CMR) and Erigeron annuus (L.) PERS. (EALP) have pharmacological effects including anti-inflammatory activity and been used in traditional medicines in Asia. However, neuroprotective effects of CMR and/or EALP extracts against brain ischemic insults have never been addressed. Thus, the aim of this study was to examine neuroprotective effects of YES-10, a combination of extracts from CMR and EALP (combination ratio, 1:1), in the hippocampus following ischemia/reperfusion in gerbils. Protection of neurons was investigated by cresyl violet staining, fluoro-jade B histofluorescence staining and immunohistochemistry for neuronal nuclei. In addition, attenuation of gliosis was studied by immunohistochemistry for astrocytic and microglial markers. Treatments with 50 or 100 mg/kg YES-10 failed to protect neurons in the hippocampus after ischemia/reperfusion injury. However, administration of 200 mg/kg YES-10 protected neurons from ischemia/reperfusion injury and attenuated reactive gliosis. These findings strongly suggest that a combination of extracts from CMR and EALP can be used as a prevention approach/drug against brain ischemic damage.

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Correlation between Hypermetabolism and Neuronal Damage during Status Epilepticus Induced by Lithium and Pilocarpine in Immature and Adult Rats

Journal of Cerebral Blood Flow & Metabolism ◽

10.1097/00004647-199902000-00011 ◽

1999 ◽

Vol 19 (2) ◽

pp. 195-209 ◽

Cited By ~ 76

Author(s):

Maria José da Silva Fernandes ◽

Céline Dubé ◽

Sylvette Boyet ◽

Christian Marescaux ◽

Astrid Nehlig

Keyword(s):

Status Epilepticus ◽

Substantia Nigra ◽

Present Model ◽

Neuronal Damage ◽

Cresyl Violet ◽

Adult Rats ◽

Cresyl Violet Staining ◽

Variable Extent ◽

Extent Of Damage ◽

Deoxyglucose Method

The correlation between seizure-induced hypermetabolism and subsequent neuronal damage was studied in 10-day-old (P10), 21-day-old (P21), and adult rats subjected to lithium-pilocarpine status epilepticus (SE). Local CMRglc (LCMRglc) values were measured by the [14C]2-deoxyglucose method for a duration of 45 minutes starting at 60 minutes after the onset of SE, and neuronal damage was assessed by cresyl violet staining at 6 days after SE. In P21 and adult rats, LCMRglc values were increased by 275 to 875% in all thalamic, cortical, forebrain, and hypothalamic regions plus the substantia nigra. In addition, at P21 there were also large increases in LCMRglc in brainstem regions. In P10 rats, metabolic increases were mostly located in cortical and forebrain regions plus the substantia nigra but did not affect hypothalamic, thalamic, or brainstem areas. In adult rats, there was an anatomical correlation between hypermetabolism and neuronal damage. At P21, although hypermetabolism occurred in regions with damage, the extent of damage varied considerably with the animals and ranged from an almost negligible to a very extended degree. Finally, in P10 rats, although quite pronounced hypermetabolism occurred, there was no neuronal damage induced by the seizures. Thus, in the present model of epilepsy, the correlation between marked hypermetabolism and neuronal damage can be shown in adult rats. Conversely, immature rats can sustain major metabolic activations that lead either to a variable extent of damage, as seen at P21, or no damage, as recorded at P10.

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Inflammasome Regulation at the Level of Cellular and Molecular Function affect by Recombinant Tissue Plasminogen Activator and Exosomes Treatment in Ischemic Stroke Animal Model

10.21203/rs.3.rs-375724/v1 ◽

2021 ◽

Author(s):

Mohsen Safakhil ◽

Mina Ramezani ◽

Azadeh Mohamadgholi

Keyword(s):

Animal Model ◽

Cell Death ◽

Combination Therapy ◽

Functional Recovery ◽

Therapy Group ◽

Neuronal Cell ◽

Cresyl Violet ◽

Treatment Groups ◽

Cresyl Violet Staining ◽

All Treatment

Abstract In the current research, neuroprotection and performance improvement have been investigated in MCAO animal model treated with exosomes combined with rt-PA. Middle cerebral artery occlusion (MCAO) was induced in 25 adult male Wistar rats. Rats received rt-PA with a dose of 100 µg/kg in a single dose or drived-exosome from bone marrow MSCs. The Garcia scoring system and elevated body test were employed as behavioral tests for the functional recovery assessment. Cresyl violet staining was applied to evaluate the cell death degree in brain tissues. Immunohistochemical analysis was performed for the detection of GFAP and IBA1-positive cells. Results Cresyl violet staining revealed that the population of dark cells was significantly reduced in all treatment groups. A considerable increase (P ≤ 0.05) in catalase enzyme was observed in the combination therapy group compared whit the MCAO group (P ≤ 0.05). The amount of GPX despite the increase in all treatment groups compared to MCAO group was not statistically significant (P ≥ 0.05). Our conclusion was approved by the Nlrp1 and Nlrp3 downregulation during combination therapy in MCAO model by reduction in cell death rate. The Density of GFAP-positive cells Showed a decrease in the exosome with or without rt-PA experimental groups in comparison with the MCAO group (P < 0.05). Our observation indicated that exosomes, in combination with rt-PA, resulted in the noticeable functional recovery, neuronal regeneration, and reduction of neuronal cell death after a 7-day period of the MCAO induction. This novel therapeutic strategy could provide a better treatment option for those patients suffered from stroke.

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Cresyl Violet Staining

Encyclopedia of Neuroscience ◽

10.1007/978-3-540-29678-2_1319 ◽

2008 ◽

pp. 904-904

Keyword(s):

Cresyl Violet ◽

Cresyl Violet Staining

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Abstract P27: Microglial Activity and Therapeutic Hypothermia Post Cardiac Arrest Resuscitation in Rats

Circulation ◽

10.1161/circ.118.suppl_18.s_1450-d ◽

2008 ◽

Vol 118 (suppl_18) ◽

Author(s):

Xiaofeng Jia ◽

Shinyi Tsai ◽

Gehua Zhen ◽

Adrian Velasquez ◽

Matthew A Koenig ◽

...

Keyword(s):

Cardiac Arrest ◽

Therapeutic Hypothermia ◽

Calcium Binding ◽

Cresyl Violet ◽

Surface Cooling ◽

Behavioral Recovery ◽

Cresyl Violet Staining ◽

Injury Model ◽

Temperature Manipulation ◽

The Impact

Background Therapeutic hypothermia after cardiac arrest (CA) improves survival and functional outcomes, but its neuroprotective mechanism remains unclear. We previously showed that immediate initiation of hypothermia post-CA led to better neuro-electrical and behavioral recovery. In this study, we quantified the impact of immediate versus conventional hypothermia with stereological technique on microglia (using expression of ionized calcium binding adaptor molecule 1 - Iba1), astrocytes (using expression of glial fibrillary astrocyte protein - GFAP), and neurons (by Cresyl-violet staining with histopathological damage scoring - HDS). Methods Forty-eight rats were randomly divided into 3 groups (n=16 per group), based on normothermia (NT, T=37°C), 6 hours of immediate hypothermia (IH, T=33°C, initiated immediately after CA), or 12 hours of conventional hypothermia (CH, T=33°C, initiated one hour after CA) after CA. Each group was then evenly divided to 2 subgroups (n=8 based on 7 or 9 min asphyxial time). Hypothermia was maintained using surface cooling. Neurological recovery was evaluated using serial Neurological Deficit Score (NDS) calculation. Results Better recovery by NDS 72 hours post-CA was noted in rats treated with IH (median, interquartile range: 72, 63–74), compared to CH (66, 43–70), and NT (51, 0 – 65) (p=0.005). There was a significantly different proportion of ischemic neurons by HDS in cortex (IH: 18.9±2.5%, CH: 33.2±4.4%, NT: 36.2±4.7%, p=0.002) and CA1 (IH: 14.4±2.9%, CH: 24.6±5.4%, NT: 49.2±5.3%, p<0.001). There was significant less Iba-1 expression in CH compared to IH (p=0.042) and NT (p=0.012) in cortex and CA1. There was no difference for GFAP expression in cortex or CA1. Conclusions Immediate hypothermia leads to better functional outcome and lower ischemic neuronal injury in rats after CA. Temperature manipulation seems to affect the immune response to CA, with increased microglial activity in the IH and NT groups compared to CH group in this injury model. The longer hypothermia duration may decrease the microglia activation in response of central nervous system to CA than short duration in this injury model. No difference was noted in the astrocytic activity in the different groups.

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Effect of Low Intensity Magnetic Field Stimulation on Calcium-Mediated Cytotoxicity After Mild Spinal Cord Contusion Injury in Rats

Annals of Neurosciences ◽

10.1177/0972753120950072 ◽

2020 ◽

Vol 27 (2) ◽

pp. 49-56

Author(s):

Supti Bhattacharyya ◽

Shivani Sahu ◽

Sajeev Kaur ◽

Suman Jain

Keyword(s):

Magnetic Field ◽

Spinal Cord ◽

Tissue Injury ◽

Bladder Function ◽

Cresyl Violet ◽

Tail Flick ◽

Low Intensity ◽

Secondary Damage ◽

Pain Status ◽

Cresyl Violet Staining

Background: Magnetic field (MF) stimulation has the potential to reduce secondary damage and promote functional recovery after neural tissue injury. The study aimed to observe the effect of very low intensity (17.96µT) MF on general body condition, secondary damage, pain status, and locomotion. Methods: We exposed rats to MF (2 h/day × 3 weeks) after 6.25 mm contusion spinal injury. Locomotor behavior was evaluated by BBB score, pain assessment was done by recording threshold for tail flick, expression of voltage-gated calcium channels and extent of secondary damage in the spinal cord was assessed by immunofluorescence and Cresyl violet staining, respectively. Results: A significant ( p ≤ .001) improvement in bladder function as well as BBB score was observed after MF exposure in comparison with sham and SCI over the observation period of 3 weeks. SCI group showed an increase in the threshold for vocalization after discharge, which decreased following MF exposure. Cresyl violet staining showed significantly higher tissue sparing (73%) at the epicenter after MF exposure when compared to SCI group. This was accompanied with a significant decrease in calcium channel expression in MF group as compared to SCI. Conclusion: The results suggest facilitation of sensory-motor recovery after MF exposure, which could be due to attenuation of secondary damage and calcium-mediated excitotoxicity in a mild contusion rat model of SCI.

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