Name: Evaluating Cell Death Signaling by Immunofluorescence in a Rat Model of Ischemic Stroke (Video) | JoVE Journal
Uploaded: 2025-01-03T15:00:00.000+00:00
Duration: 11 min 32 s
Description: Learn immunofluorescence techniques for evaluating cell death signaling in a rat model of ischemic stroke using MCAO. This method helps researchers identify molecular targets like cathepsins for therapeutic development.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

Simiao Wu was supported by the Science &#38; Technology Department of Sichuan Province (2024YFHZ0330) and the West China Hospital of Sichuan University. Weihong He was supported by the Science &#38; Technology Department of Sichuan Province (2023YFS0297) and Sichuan University. We thank Yi Zhang and Yue Li at the Research Core Facility, West China Hospital of Sichuan University, for their technical support on TUNEL imaging and analysis.

This protocol and the experiments reported here were approved by the Sichuan University Medical Ethics Committee. The care and use of animals were in compliance with local and international ethical criteria.
NOTE: Adult male Sprague-Dawley rats weighing 250-280 g were used in this study. The rats were housed in a standard environmental condition (20-22 &#176;C temperature, 12 h light/dark cycle), fed with a standard rat formula diet and high-purity water. The surgical procedure was performed under sterile conditions.
1. Rat model of MCAO
<ol>
	<li>Induce initial anesthesia using 4% isoflurane with oxygen at 1 L/min. Monitor depth of anesthesia by foot pinch. While anesthesia is achieved, administer an ocular ointment to prevent dryness. Inject rat with 1 mL of saline subcutaneously as volume replenishment and with 5 mg/kg carprofen and 0.1 mg/kg buprenorphine for preoperative analgesia. Reduce isoflurane to 3.5% initially and then to 2% gradually throughout the surgery.</li>
	<li>Place the rat in a supine position and maintain body temperature at 36.5 &#177; 0.1 &#176;C on a regulated heat mat with a rectal temperature monitor probe. Shave and disinfect the skin in the neck with an iodophor disinfecting solution.</li>
	<li>Make a 2 cm midline incision in the neck and retract the soft tissues to expose the carotid vessels. Use forceps to isolate the right common carotid artery (CCA), external carotid artery (ECA), and internal carotid artery (ICA).</li>
	<li>Use sutures to ligate the proximal right CCA and clamp the right ICA and ECA with a micro-clip at their origin.</li>
	<li>Insert a nylon monofilament coated with a round silicone tip into the lumen of the CCA through a small incision and tie a knot in the CCA to prevent bleeding and dislodging of the nylon monofilament.</li>
	<li>Open the right ICA micro-clip and insert the nylon monofilament into the right middle cerebral artery (MCA) through the ICA stump until the silicone tip occludes the origin of MCA (~18-20 mm).</li>
	<li>Cut off the exposed part of the nylon monofilament and close the neck midline incision.</li>
	<li>After 2 h occlusion of MCA, open the neck incision and untie the knot in the CCA. Then withdraw the nylon monofilament to reperfuse the MCA. Ligate the distal right CCA, and close the neck incision with either continuous or interrupted sutures.</li>
	<li>Post-operatively, give buprenorphine by subcutaneous injection every 8 h, at a dose of 0.03 mg/kg. Observe rats throughout the recovery from the anesthesia in a heated cage with a temperature-regulated heating mat. To encourage eating, place a Petri dish with mashed chow in the cage. House one rat per cage after the surgery.</li>
	<li>Subject the sham rats to the same procedure without the monofilament insertion. 
	NOTE: A total of 9 animals are used in each group for measurement of infarct size (n = 3), immunofluorescence analysis (n = 3), and apoptosis detection(n = 3).</li>
	<li>Evaluate neurological function at 2 h after MCAO and 22 h after reperfusion using Zea-Longa behavioral rating scale with a 0-5 point scale14. Exclude rates with a score of 0 or 4.</li>
</ol>
2. Measurement of infarct size (Figure 1)
<ol>
	<li>Sacrifice rats under deep anesthesia (4% isoflurane) by guillotine and carefully isolate the whole brain from the skull.</li>
	<li>Freeze the brain at -20 &#176;C for 20 min and cut the frozen brain into six coronal slices with a 2 mm distance between slices. Stain the brain slices with 2% TTC for 15 min at 37 &#176;C in the dark.</li>
	<li>Fix brain slices with 4% paraformaldehyde for 24 h at room temperature (RT). Photograph the stained slices with a digital camera, with auto mode setting for close-up objects.</li>
	<li>Transfer the pictures to a computer and measure the infarct size using ImageJ. Present the infarct size as the ratio of the sum of the infarct area in 6 slices to the sum of the whole brain area in the same slices. Express data as mean &#177; standard error of the mean (SEM).</li>
</ol>
3. Immunofluorescence analysis on brain cryosection (Figure 2)
<ol>
	<li>Induce anesthesia with 4% isoflurane and 1 L/min oxygen. Incise the skin and subcutaneous tissue to expose the heart.</li>
	<li>Remove the pericardium and make a small cut in the right atrium. Perfuse 200 mL of saline through the left ventricle, followed by 200 mL of 4% paraformaldehyde.</li>
	<li>Open the skull and remove the intact brain.</li>
	<li>Fix the brain in 4% paraformaldehyde at RT for over 24 h. Then, dehydrate it in a series of sucrose solution gradients of 10%, 15%, and 30% concentrations until the tissue sinks, approximately 6-8 hours for each concentration.</li>
	<li>Embed the brain in optimal cutting temperature (OCT) compound and remove air bubbles if there are any. Then, snap-freeze the brain in liquid nitrogen and store it in the -80 &#176;C freezer.</li>
	<li>Section the brain using a cryostat at a thickness of 5 &#181;m. Throughout sectioning, ensure the cabinet temperature remains between -18 &#176;C and -20 &#176;C. Label the sectioning location based on the model site and experimental conditions. Store the sections in a freezer.</li>
	<li>Allow the frozen sections to equilibrate at RT for ~30 min to perform immunofluorescence staining. Then, soak the sections with sterile PBS 3x for 5 min each.</li>
	<li>Outline the tissue with an immunohistochemistry pen. Incubate with 20 &#181;g/mL Proteinase K (RNase-free) and 0.3% Triton X-100 at RT for 10 min.</li>
	<li>Soak the sections with sterile PBS 3x for 5 min each. Apply 3% BSA to the outlined tissue and incubate at RT for 1 h for blocking.</li>
	<li>After blocking, remove the solution, add the appropriate primary antibody (Rabbit recombinant monoclonal cathepsin-B antibody, 1:200), and incubate the sections overnight in the dark at 4 &#176;C.</li>
	<li>Remove the primary antibody and wash the sections with sterile PBS containing 0.1% Tween-20 3x for 5 min each.</li>
	<li>Add the appropriate secondary antibody (Goat Anti-Rat IgG [H+L] [FITC conjugated], 1:200) with the required fluorophore, and incubate the sections in a humidified chamber at RT for 1 h, protecting them from light.</li>
	<li>After secondary antibody incubation, wash the sections with sterile PBS containing 0.1% Tween-20 3x for 5 min each. Stain the sections with a mounting medium containing DAPI and cover them with a coverslip.</li>
	<li>Capture images using a fluorescence microscope with the following settings: excitation wavelength of 488 nm, emission wavelength of 520 nm, and exposure time of 500 ms. Store the sections in the dark at 4 &#176;C for long-term preservation.</li>
</ol>
4. Detection of apoptosis by TUNEL staining (Figure 3)
NOTE: A commercial TUNEL assay kit is used for TUNEL staining. This TUNEL kit includes a DNase I Buffer working solution, deoxynucleotidyl transferase (TdT) equilibration buffer, labeling working solution, and DAPI working solution.
<ol>
	<li>Perfuse and isolate the brain using the same procedure as above (steps 3.2-3.3). Immerse the brain in 4% paraformaldehyde for 3-4 h for fixation.</li>
	<li>Cut the brain into coronal slices approximately 2 mm thick, starting from the rostral end to the caudal end, and immerse in 4% paraformaldehyde for an additional 12 h. Following fixation, wash the brain slices in running water for 1 h.</li>
	<li>Immerse the brain slices in ethanol solutions of 50%, 70%, 80%, 90%, 95%, and 100% concentrations sequentially for 35-50 min each for dehydration.</li>
	<li>Treat the brain slices with TO-type transparent biological agent I for 35-50 min, followed by clearing agent II for 35-50 min, until the tissue is fully transparent.</li>
	<li>Immerse the brain slices sequentially in embedding medium I for 60 min, embedding medium II for 60 min, embedding medium III for 60 min, and embedding medium IV for 60 min.</li>
	<li>Place the brain slices into an embedding cassette. Seal the cassette, label it clearly, and place it in a cold storage room for solidification.</li>
	<li>Set the microtome to an initial thickness of 10 &#181;m to trim the tissue block. Then, adjust the thickness to 5 &#181;m for slicing.</li>
	<li>Heat the water bath to 45 &#176;C, float the tissue sections on the water surface, and pick them up onto slides. After approximately 30 s, remove the slides, dry them at RT, and store them in a drying box.</li>
	<li>Deparaffinize the stored brain sections by immersing them in a clearing agent for 2 cycles, 10 min each.</li>
	<li>To rehydrate the sections, soak them in anhydrous ethanol for 5 min, repeating 3 times. Transfer the sections to 90% ethanol for 3 min. Further, soak the sections in 80% ethanol for 3 min. Finally, immerse the sections in 70% ethanol for 3 min.</li>
	<li>Rinse the samples in PBS 3x for 5 min each. Gently remove excess moisture using filter paper.</li>
	<li>Administer 100 &#181;L of 1x proteinase K working solution to each sample and incubate at 37 &#176;C for 20 min. Following permeabilization, rinse the samples in PBS 3x for 5 min each.</li>
	<li>Prepare samples for positive control.
	<ol>
		<li>Add 100 &#181;L of 1x DNase I Buffer working solution (TUNEL assay kit) to the permeabilized sample and allow it to equilibrate at RT for 5 min. Carefully remove any excess liquid from the sample using absorbent paper.</li>
		<li>Add 100 &#181;L of diluted DNase I working solution (200 U/mL) to the sample and incubate at 37 &#176;C for 10-30 min. Rinse the samples in PBS 3x for 5 min each.</li>
	</ol>
	</li>
	<li>Prepare samples for negative control.
	<ol>
		<li>Apply 100 &#181;L of 1x DNase I Buffer working solution to the permeabilized sample and equilibrate at RT for 5 min. Incubate the negative control sample with DNase I Buffer at 37 &#176;C for 10-30 min, ensuring no DNase I enzyme is added. Rinse the samples in PBS 3x for 5 min each.</li>
	</ol>
	</li>
	<li>Apply 100 &#181;L of terminal deoxynucleotidyl transferase (TdT) equilibration buffer (TUNEL assay kit) to each sample and incubate in a humidified chamber at 37 &#176;C for 10-30 min. Then, remove the TdT Equilibration Buffer with absorbent paper.</li>
	<li>Add 50 &#181;L of labeling working solution (TUNEL assay kit) to each sample, then incubate them at 37 &#176;C in a humidified, dark chamber for 60 min. If the signal intensity is low, consider extending the DNA labeling reaction time. Then, rinse the samples in PBS 3x for 5 min each.</li>
	<li>After blotting excess moisture, apply the DAPI working solution (TUNEL assay kit) and incubate at RT in the dark for 5 min to counterstain the nuclei. Then, rinse the samples in PBS 4x for 5 min each. Remove excess liquid with absorbent paper, and seal the slides using an anti-fade mounting medium.</li>
	<li>Scan the stained slides using the Vectra Polaris multispectral imaging system and analyze data with the system software.</li>
</ol>
5. Statistical analysis
<ol>
	<li>Present the experimental results as means &#177; SEM and analyze data with a Student&#39;s t-test for two group comparisons using GraphPad Prism 10 software (GraphPad Software, San Diego, CA).</li>
	<li>Set a p-value less than 0.05 as a statistical difference.</li>
</ol>

Immunofluorescence Staining to Study Cell Death in Rat MCAO Stroke Models

The authors have nothing to disclose.

Occlusion of a cerebral artery leads to deprivation of oxygen and nutrition, followed by the activation of reactive oxygen species, intracellular calcium overload, glutamate release, and induction of inflammatory responses15. This series of events results in cell death and irreversible tissue damage in the infarcted brain. The penumbra is potentially salvageable through medical intervention within an appropriate therapeutic window16. The only approved treatment for ischemic...

Cerebral stroke is a leading cause of death and disability worldwide1. The high mortality and morbidity of stroke result in immense public healthcare burden and serious socio-economic consequences. Although the rate of stroke incidence and mortality remain stable, the number of stroke patients and stroke-related deaths are increasing over decades2,3. Strokes can be categorized as ischemic or hemorrhagic. The majority of stroke cases are ischemic strokes caused by the occlusion of a main cerebral artery4. To date, tissue plasminogen activator (tPA) is the only FDA-approved drug for ischemic stroke, but its application is highly limited by the narrow therapeutic window, complications, and contraindications5,6,7. Thus, it is urgent to develop new treatment options with a wider therapeutic window to mitigate the effects of stroke.
Experimental animal models are useful tools to study the pathophysiology of ischemic stroke. In most human patients, ischemic stroke is caused by the blockage of the middle cerebral artery (MCA)8. Therefore, rodent models of middle cerebral artery occlusion (MCAO) have been developed to resemble human cerebral infarction. While there are several MCAO approaches that are used in small animal studies, the method most widely used is the intraluminal suture model, which involves inserting a nylon filament into the middle cerebral artery from either the external or internal carotid arteries, resulting in transient or permanent occlusion of the blood flow4,9. This model leads to a large volume of cerebral infarction and allows examination of cell death signaling in the infarcted brain tissue with immunofluorescent techniques.
The area surrounding the infarcted core, referred to as penumbra, is the target for potential stroke therapies10,11. Since blood flow in the penumbra is partially maintained, injured neurons and non-neuronal cells in this area may be salvaged by inhibiting the activation of cell death signaling. Targeting cell death pathways may be a promising strategy for neuroprotection after stroke12. Therefore, evaluating cell death signaling is crucial for experimental stroke research. Recently, cathepsin-B, a lysosomal protease, has been shown to play a role in mediating programmed cell death after stroke, and it has gained substantial attention in the neurological field13. Cathepsin-B represents a potential target for neuroprotection that merits further investigation.
This article demonstrates how to induce MCAO using the intraluminal suture approach in rats. We also show how to perform 2,3,5-triphenyltetrazalium chloride (TTC) staining to determine infarct size and detect apoptotic cells using Terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining.

<table><tbody><tr><td>Akoya Vectra Polaris</td><td>Akoya Biosciences</td><td>N/A</td><td>Vectra Polaris multispectral imaging system&amp;nbsp;</td></tr><tr><td>Anhydrous ethanol</td><td>Chengdu Jinshan Chemical Reagent Co.</td><td>N/A</td><td></td></tr><tr><td>Buprenorphine</td><td>West China Hospital</td><td>N/A</td><td></td></tr><tr><td>Carprofen</td><td>West China Hospital</td><td>N/A</td><td></td></tr><tr><td>Cathepsin-B antibody</td><td>Abcam</td><td>Ab214428</td><td></td></tr><tr><td>Cryostat</td><td>LEICA&amp;nbsp;</td><td>LEICA CM 1950</td><td></td></tr><tr><td>Digital camera</td><td>OLYMPUS</td><td>TG-7</td><td></td></tr><tr><td>Goat Anti-Rat IgG (H+L) (FITC conjugated)</td><td>Elabscience</td><td>&amp;nbsp;E-AB-1021</td><td></td></tr><tr><td>Goat serum</td><td>Solarbio</td><td>SL038</td><td></td></tr><tr><td>GraphPad Prism 10 software</td><td>N/A</td><td>https://www.graphpad-prism.cn/?c=i&amp;amp;a=prismdownload_cn</td><td></td></tr><tr><td>ImageJ</td><td>N/A</td><td>https://imagej.net/ij/</td><td></td></tr><tr><td>Immunohistochemistry pen</td><td>Biosharp</td><td>http://www.biosharp.cn/index/product/details/language/en/product_id/2828.html</td><td></td></tr><tr><td>InForm software</td><td>Akoya Biosciences</td><td>Version 2.6.0</td><td>system software for the multispectral imaging system&amp;nbsp;</td></tr><tr><td>Iodophor disinfecting solution</td><td>Huatian Technology Industry Co.</td><td>N/A</td><td></td></tr><tr><td>Isoflurane</td><td>RWD Life Science Co.</td><td>https://www.rwdstco.com/inhalation-anesthesia-solutions/</td><td></td></tr><tr><td>Mounting medium containing DAPI</td><td>Solarbio</td><td>S2110</td><td></td></tr><tr><td>Nylon monofilament</td><td>RWD Life Science Co.</td><td>https://www.rwdstco.com/</td><td></td></tr><tr><td>Optimal cutting temperature (OCT) compound</td><td>Epredia</td><td>N/A</td><td></td></tr><tr><td>Paraformaldehyde</td><td>Biosharp</td><td>N/A</td><td></td></tr><tr><td>Phosphate buffer solution (PBS)</td><td>Solarbio</td><td>N/A</td><td></td></tr><tr><td>Prism</td><td>GraphPad&amp;nbsp;</td><td>Version 10</td><td></td></tr><tr><td>Proteinase K</td><td>Tiangen</td><td>RT403-02</td><td></td></tr><tr><td>Saline</td><td>Kelun Co.</td><td>N/A</td><td></td></tr><tr><td>Sprague-Dawley rats</td><td>Huafukang Co.</td><td>N/A</td><td></td></tr><tr><td>Sucrose</td><td>BioFroxx</td><td>1245GR500</td><td></td></tr><tr><td>Surgical instruments</td><td>RWD Life Science Co.</td><td>N/A</td><td></td></tr><tr><td>TO-type transparent biological agent&amp;nbsp;&amp;nbsp;</td><td>Beijing Solarbio Science &amp;amp; Tecnology Co., Ltd.</td><td>http://www.solarbio.net/</td><td></td></tr><tr><td>Triphenyltetrazolium chloride (TTC) solution</td><td>Solarbio</td><td>G3005</td><td></td></tr><tr><td>Triton X-100</td><td>BioFroxx</td><td>1139ML100</td><td></td></tr><tr><td>TUNEL kit</td><td>Elabscience</td><td>&amp;nbsp;E-CK-A321</td><td></td></tr></tbody></table>

evaluating cell death signaling by immunofluorescence in a rat model of ischemic stroke

Evaluating Cell Death Signaling by Immunofluorescence in a Rat Model of Ischemic Stroke

Stroke is a leading cause of death and disability worldwide. Most cases of stroke are ischemic and result from the occlusion of the middle cerebral artery (MCA). Current pharmacological approaches for the treatment of ischemic stroke are limited; therefore, novel therapies providing effective neuroprotection against ischemic injury following stroke are urgently needed. In the brain tissue following ischemic stroke, the area of the ischemic penumbra is salvageable but at risk of progressing to irreversible damage. The penumbral area surrounding the infarcted core is a conceptual target for neuroprotection. Since the blood flow is partially maintained in the penumbral area, neuronal and non-neuronal cells in this area transiently survive after stroke, and these cells that are still viable may be rescued by appropriate medical interventions. Understanding the pathophysiology of the penumbra is important in the development of neuroprotective therapies because the cell death pathways activated in the ischemic penumbra may indicate therapeutic targets, such as RUNX1 and cathepsins. These protein targets functioning as mediators of programmed cell death can be further exploited in translational research. With moderate size and similarity to the human brain, the in vivo rat model of MCA occlusion (MCAO) mimics the human ischemic stroke and offers an applicable tool to investigate the penumbral pathology, examine the cell death signaling, and evaluate the effects of potential targets in the context of MCAO. Here, we describe how to induce MCAO in rats and how to perform immunofluorescence staining for the detection of cell death signaling in the rat brain following MCAO.

Rats are subjected to 120 min ischemia during MCAO followed by 22 h reperfusion. The mortality rate was minimal during MCAO surgery and was around 25% within the period of reperfusion. Significant brain damage was observed in the MCAO group, whereas no infarct was observed in the cerebrum of the Sham group (24.87 ± 1.21% vs. 0% sum of infarct area to the whole brain area; MCAO [n = 3] vs. Sham [n = 3], P &lt; 0.001; Figure 1A,B). In the MCAO group, immunoreactivitie...

Rat in vivo models are indispensable tools to investigate the pathological mechanisms and therapeutic targets for ischemic stroke. This work will describe performing immunofluorescence staining in infarcted brain slices following middle cerebral artery occlusion (MCAO) in rats.

Stroke is a leading cause of death and disability worldwide. Most cases of stroke are ischemic and result from the occlusion of the middle cerebral artery ...

evaluating-cell-death-signaling-immunofluorescence-rat-model-ischemic

Research

JoVE Journal

Neuroscience

Surgical induction of ischemic brain damage in the rat is a widely used model for stroke research. Here we demonstrate the induction of focal cerebral ischemia by occlusion of the middle cerebral artery. Visualization of the resulting infarct by histological staining and magnetic resonance imaging is also shown.

Focal Cerebral Ischemia Model by Endovascular Suture Occlusion of the Middle Cerebral Artery in the Rat

Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats.  In addition, important considerations, advantages, and shortcomings of this model are discussed.

Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat

Bilateral carotid occlusion coupled with systemic hypotension produces global brain ischemia in the rat, resulting in damage to the hippocampus with reproducible severity. Animal subjects are impaired with predictable patterns of brain damage, they recover expediently, and mortality rates are comparatively low.

2-Vessel Occlusion/Hypotension: A Rat Model of Global Brain Ischemia

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

We present a new fluorescence technique for selective in situ labeling of active phagocytic cells, which clear off cell corpses in stroke. The approach is important for assessing brain reaction to ischemia because only a small proportion of phagocytes present in ischemic brain participate in clearance of cell death.

Assessing Phagocytic Clearance of Cell Death in Experimental Stroke by Ligatable Fluorescent Probes

Brain damage resulting from cerebral ischemia may be non-invasively imaged and studied in rats using pre-clinical positron emission tomography coupled with the injectable radioactive probe, 18F-fluorodeoxyglucose. Further, the use of modern software tools that include volume of interest (VOI) brain templates dramatically increase the quantitative information gleaned from these studies.

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

A method for delivering neural stem cells, adaptable for injecting solutions or suspensions, through the common carotid artery (mouse) or external carotid artery (rat) after ischemic stroke is reported. Injected cells are distributed broadly throughout the brain parenchyma and can be detected up to 30 d after delivery.

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia

Here, we present a protocol to assess immune cell function following ischemic stroke using flow cytometric analysis in a mini-stroke model. This method enables detailed examination of neuroimmune interactions and can be adapted for studying other neurodegenerative diseases, enhancing our understanding of immune responses in stroke pathology.

Isolation and Flow Cytometric Assessment of Neuroimmune Interactions in a Mini-Stroke Murine Model

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors.  Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

A General Method for Evaluating Incubation of Sucrose Craving in Rats

Learn immunofluorescence techniques for evaluating cell death signaling in a rat model of ischemic stroke using MCAO. This method helps researchers identify molecular targets like cathepsins for therapeutic development.

Evaluating Cell Death Signaling by Immunofluorescence in a Rat Model of Ischemic Stroke

Explore More Videos