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middle cerebral artery occlusion spontaneously hypertensive rat

Spontaneously hypertensive rats develop larger and less variable infarcts after MCA occlusion, and the stroke-prone cohort is used to study hypertension-related stroke.

Outcome measures

存活脑组织染为红色、梗死区呈白色;以 2 mm 连续冠状切片逐孔染色成像后,测量患侧与对侧半球的梗死面积及总面积,计算患侧相对对侧的梗死面积百分比。

Experimental animals

脑切片 TTC 染色与梗死面积测定
Age
成年(来源未报告具体周龄)
Tissue
脑组织(2 mm 冠状切片;梗死区与存活区)
脑切片 TTC 染色与梗死面积测定
Age
成年(来源未报告具体周龄)
Tissue
脑组织(2 mm 冠状切片;梗死区与存活区)

Experimental procedures

TTC Staining TTC Staining The brain is removed from the skull and placed into a brain slicer mold placed in ice. Razor blades are positioned in the mold to obtain 2mm thick sections. Sections are placed in individual wells of a 24 well tissue culture dish with 500 μl of 2% TTC in 0.1M PBS. The plate is wrapped in foil and placed on a 37°C slide warmer for 30 minutes. TTC is removed and 500ul of fixative is added to each well for storage. Images of the sections are captured by Olympus S2H10 research stereo- microscope and Adobe Photoshop software. The infarct area and total area of the contralateral and ipsilateral hemispheres are measured with Scion image software (Scion Corporation). The percent infarct area is calculated as a percentage of the ipsilateral hemisphere relative to the contralateral hemisphere as previously described. [F2.JPG]]()

Model identity

Model name
middle cerebral artery occlusion spontaneously hypertensive rat
Species
rat
Strain
spontaneously hypertensive rat

Biological characteristics

Phenotype stage
Acute infarct is measured after MCAO; hypertension-related injury depends on age and blood pressure.
Research questions
Hypertension effects on infarct size and variability; small-vessel and lacunar stroke.
Limitations
SHR and SHRSP are not interchangeable with normotensive Wistar or Sprague-Dawley MCAO; blood pressure and colony affect lesion size.

Experimental methods

Experimental protocols

  • 脑切片 TTC 染色与梗死面积测定

    Fresh brains are cut to 2-mm coronal sections, stained per well with 2% TTC in 0.1M PBS at 37°C for 30 min protected from light, stored in fixative, imaged on a stereomicroscope, and infarct percentage computed ipsilateral-versus-contralateral.

  • 脑切片 TTC 染色与梗死面积测定

    Fresh brains are cut to 2-mm coronal sections, stained per well with 2% TTC in 0.1M PBS at 37°C for 30 min protected from light, stored in fixative, imaged on a stereomicroscope, and infarct percentage computed ipsilateral-versus-contralateral.

Reagents and materials

References

  1. Liu F, McCullough LD. Middle cerebral artery occlusion model in rodents: methods and potential pitfalls.. Journal of biomedicine & biotechnology. 2011. PMID:21331357.
  2. Ian Simpson. Induce Stroke via Hypoxia-Ischemia. protocols.io, v2, 2019. DOI: 10.17504/protocols.io.8e5htg6
  3. Hu Y, Yang Z-H, Yan F, et al. CCA repair or ECA ligation - Which middle cerebral artery occlusion is better in the reperfusion mouse model? Ibrain. 2023;9(3):258-269. DOI:10.1002/ibra.12128; PMID:37786756.
  4. Bederson JB, Pitts LH, Germano SM, Nishimura MC, Davis RL, Bartkowski HM. Evaluation of 2,3,5-triphenyltetrazolium chloride as a stain for detection and quantification of experimental cerebral infarction in rats. Stroke. 1986;17(6):1304-1308. DOI:10.1161/01.STR.17.6.1304; PMID:2433817. 原文以成年大鼠 MCA 闭塞后 TTC 与 H&E 对比,24 小时 TTC 梗死面积 21±2% 的冠状切片面积,r=0.91,支持 TTC 用于大鼠脑梗死检测与定量。
  5. Clark WM, Lessov N, Lauten JD, Hazel K. Doxycycline treatment reduces ischemic brain damage in transient middle cerebral artery occlusion in the rat. J Mol Neurosci. 1997;9(2):103-108. DOI:10.1007/BF02736854; PMID:9407391. 原文在成年 Sprague-Dawley 大鼠以线栓行 2 小时可逆 MCAO,48 小时以梗死体积为终点(对照 158±44 mm3),支持 SD 大鼠 MCAO 梗死体积检测背景。
  6. ARRIVE 2.0 guidelines