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JoVE Journal
Neuroscience
Sub-acute Cerebral Microhemorrhages Induced by Lipopolysaccharide Injection in Rats
Sub-acute Cerebral Microhemorrhages Induced by Lipopolysaccharide Injection in Rats
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Sub-acute Cerebral Microhemorrhages Induced by Lipopolysaccharide Injection in Rats

Sub-acute Cerebral Microhemorrhages Induced by Lipopolysaccharide Injection in Rats

Full Text
6,935 Views
06:39 min
October 17, 2018

DOI: 10.3791/58423-v

Dandan Li*1,2, Hóngyi Zhào*2,3, Wei Wei2, Nan Liu2, Yonghua Dr. Huang2

1Department of Neurology,Second Hospital of Shanxi Medical University, 2Department of Neurology,PLA Army General Hospital, 3Department of Neurology,NO 261 Hospital of PLA

Overview

This study presents a protocol for inducing and detecting cerebral micro-hemorrhages (CMHs) in Sprague-Dawley rats through LPS injection. The work contributes to understanding the pathogenesis of CMHs and may inform future research into various brain disorders.

Key Study Components

Area of Science

  • Neuroscience
  • Pathophysiology
  • Animal Models

Background

  • Cerebral micro-hemorrhages are associated with various neurological disorders.
  • Inflammation plays a critical role in the development of CMHs.
  • LPS is commonly used to induce inflammation in experimental models.
  • The technique may extend to studying other brain disorders like Parkinson's and Alzheimer's disease.

Purpose of Study

  • To develop a reliable method for inducing CMHs in a laboratory setting.
  • To explore the inflammatory processes involved in CMH pathogenesis.
  • To establish a basis for future research on brain disorders.

Methods Used

  • LPS is administered via intraperitoneal injection at a dose of 1 mg/kg.
  • The animal model used is 10-week-old male Sprague-Dawley rats.
  • Cardiac puncture and Evans blue dye are used for assessing blood-brain barrier integrity.
  • The protocol includes multiple injections and specific tissue preparation steps for imaging.
  • Various staining techniques are applied for analyzing brain tissue.

Main Results

  • The LPS injection reliably induces CMHs, observable through various imaging techniques.
  • Methods allow the visualization of red blood cells outside blood vessels via H&E staining, indicating CMH presence.
  • Evans blue staining indicates compromised blood-brain barrier function and highlights CMH distribution.

Conclusions

  • This study demonstrates a straightforward method for investigating CMHs and their underlying mechanisms.
  • Insights gained could enhance understanding of CMHs and related neurological disorders.
  • The protocol also enables researchers to examine inflammation's role in cerebral injury.

Frequently Asked Questions

What are the advantages of using LPS injections in this model?
LPS injections induce stable inflammation, making them an effective method for studying CMHs and related processes in the brain.
How is the biological model implemented for studying CMHs?
The model involves administering LPS to Sprague-Dawley rats, followed by specific imaging and staining techniques to assess CMH presence.
What types of data are obtained from this method?
Key outcomes include visualization of CMHs, assessment of blood-brain barrier integrity, and molecular readouts from tissue staining.
In what ways can this method be adapted for other studies?
While focused on CMHs, the technique can also be applied to research on other brain disorders like depression and Alzheimer’s disease.
Are there any limitations or considerations when using this protocol?
Mortality rates can vary based on the age and health of the rats; thus, maintaining a clean environment is crucial to minimize risk.

We present a protocol to induce and detect CMHs caused by LPS injection in Sprague-Dawley rats, which may be utilized in future research investigations on the pathogenesis of CMHs.

This method can help answer key questions in cerebral micro-hemorrhage field such as the etiology, distribution, and detection of cerebral micro-hemorrhage and so on. The main advantage of this technique is that LPS injections stably induce inflammation. In addition, LPS injection is quite simple, economical, and cost-effective.

Though this method can provide insight into cerebral hemorrhage induction, it can also be applied to other brain disorders such as depression, schizophrenia, Parkinson's disease, Alzheimer's disease, and prion disease. Administer LPS at a dose of one milligram per kilogram of body weight to 10-week-old male Sprague Dawley rats by intraperitoneal injection, then return each rat to the home cage. Repeat the LPS injection after six hours and 16 hours.

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Cerebral MicrohemorrhagesLipopolysaccharide (LPS) InjectionInflammationBrain DisordersSprague Dawley RatsEvans BlueBlood-brain BarrierCardiac PerfusionParaformaldehydeCryostatDAPI

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