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Comparison of surgical methods of transient middle cerebral artery occlusion between rats and mice.

Lee S, Hong Y, Park S, Lee SR, Chang KT, Hong Y - J. Vet. Med. Sci. (2014)

Bottom Line: In rodent MCAo models, has to be considered body temperature during the operative period, as well as the need for the use of a standardized tip in terms of the outer diameter of probes.Our methods could induce stable moderate-severity ischemic brain injury models and histological alteration at 24 hr after MCAo surgery.Finally, we described and compared major parameters between rats and mice, including probe size, thread insert length, operation and occlusion periods, and differences in the procedures.

View Article: PubMed Central - PubMed

Affiliation: Biohealth Products Research Center (BPRC), Inje University, Gimhae, Korea.

ABSTRACT
Rodent models of focal cerebral ischemia that do not require craniotomy have been developed by intraluminal suture middle cerebral artery occlusion (MCAo). Mouse MCAo models have been widely used and extended to genetic studies of cell death or recovery mechanisms. Therefore, we compared surgery-related parameters and techniques between such rats and mice. In rodent MCAo models, has to be considered body temperature during the operative period, as well as the need for the use of a standardized tip in terms of the outer diameter of probes. Induction of focal cerebral ischemia was measured by neurological dysfunction parameters. Our methods could induce stable moderate-severity ischemic brain injury models and histological alteration at 24 hr after MCAo surgery. Moreover approximately 80% (rats) and 85% (mice) survival ratios were shown indicating with model engineering success. Finally, we described and compared major parameters between rats and mice, including probe size, thread insert length, operation and occlusion periods, and differences in the procedures.

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Structural diagram of the MCAo surgery region and histological results followed by anocclusion period in rats and mice. (A) Monofilament was inserted into MCA via ECAstump. H&E- and Nissl-stained brains of (B) MCAo mice and (C) rats. Brain sliceswere cut at 1-mm (mice) and 2-mm (rats) intervals rostral and caudal to the bregma(0.00 mm). The yellow (H&E staining) and black (Nissl staining) dotted linesindicate the marginal penumbra lesion. The black arrow indicates the pale violet colorof the ipsilateral lesion of the corpus callosum. The 90 min MCAo models not onlyshowed severe infarct lesion but also failure to confirmed corpus callosum. CCA:common carotid artery; ECA: external carotid artery; ICA: internal carotid artery; OA:occipital artery; PPA: pterygopalatine artery; MCA: middle cerebral artery.
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fig_001: Structural diagram of the MCAo surgery region and histological results followed by anocclusion period in rats and mice. (A) Monofilament was inserted into MCA via ECAstump. H&E- and Nissl-stained brains of (B) MCAo mice and (C) rats. Brain sliceswere cut at 1-mm (mice) and 2-mm (rats) intervals rostral and caudal to the bregma(0.00 mm). The yellow (H&E staining) and black (Nissl staining) dotted linesindicate the marginal penumbra lesion. The black arrow indicates the pale violet colorof the ipsilateral lesion of the corpus callosum. The 90 min MCAo models not onlyshowed severe infarct lesion but also failure to confirmed corpus callosum. CCA:common carotid artery; ECA: external carotid artery; ICA: internal carotid artery; OA:occipital artery; PPA: pterygopalatine artery; MCA: middle cerebral artery.

Mentions: Microsurgical procedures of middle cerebral artery occlusion: All animalswere anesthetized with 40 mg/kg tiletamine/zolazepam cocktail (Zoletil) and 10 mg/kgxylazine (Rompun) via intraperitoneal injection. After anesthesia, the animals’ bodytemperature was maintained at 36.5°C to 37.0°C using a heating pad on the surgical table.The animals were placed in the supine position and fixed to the surgical table usingadhesive tape. The incision region was disinfected with povidone-iodine solution. Themidline neck skin was incised, and the common carotid artery (CCA), external carotid artery(ECA) and internal carotid artery (ICA) were carefully separated from the vagus nerve. TheICA bifurcates into the middle cerebral artery (MCA) and the pterygopalatine artery (PPA)from the CCA. The occipital artery (OA) originates from the bifurcation point of the ECA andthe ICA. A transient knot was placed on the distal portion of the CCA, near the manubrium ofthe sternum. A microvascular clamp was transiently placed on the ICA proximal to the CCAjunction. Two closely spaced permanent knots were then placed on the distal portion of theECA, below the suprathyroid artery, to prevent the backflow of blood. The tied section ofthe ECA was dissected using microscissors to insert the probe, which reached the ICA throughthe CCA junction. The microvascular clamp that had been placed on the ICA was then removedto allow probe insertion. The probe was carefully inserted into the MCA from the CCAjunction (up to 18–20 mm in rats and 9–11 mm in mice). After confirmation of MCA blockage(mild bending of the ICA), the rat model exhibited blood supply from the CCA, but the mousemodel only exhibited blood supply after the occlusion period (dual occlusion by CCAligation). After 60 to 90 min, the probe was carefully withdrawn until the tip was near thearteriotomy for reperfusion. After confirmation of blood flow reperfusion, topical lidocainegel was applied onto the surgical region to relieve pain and discomfort in the postoperativeperiod. Each animal also received 1.0 ml of normal saline subcutaneouslyfor volume replenishment after the surgery. All surgical procedures were finished within 15min, excluding the occlusion and reperfusion periods (Fig. 1AFig. 1.


Comparison of surgical methods of transient middle cerebral artery occlusion between rats and mice.

Lee S, Hong Y, Park S, Lee SR, Chang KT, Hong Y - J. Vet. Med. Sci. (2014)

Structural diagram of the MCAo surgery region and histological results followed by anocclusion period in rats and mice. (A) Monofilament was inserted into MCA via ECAstump. H&E- and Nissl-stained brains of (B) MCAo mice and (C) rats. Brain sliceswere cut at 1-mm (mice) and 2-mm (rats) intervals rostral and caudal to the bregma(0.00 mm). The yellow (H&E staining) and black (Nissl staining) dotted linesindicate the marginal penumbra lesion. The black arrow indicates the pale violet colorof the ipsilateral lesion of the corpus callosum. The 90 min MCAo models not onlyshowed severe infarct lesion but also failure to confirmed corpus callosum. CCA:common carotid artery; ECA: external carotid artery; ICA: internal carotid artery; OA:occipital artery; PPA: pterygopalatine artery; MCA: middle cerebral artery.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4300368&req=5

fig_001: Structural diagram of the MCAo surgery region and histological results followed by anocclusion period in rats and mice. (A) Monofilament was inserted into MCA via ECAstump. H&E- and Nissl-stained brains of (B) MCAo mice and (C) rats. Brain sliceswere cut at 1-mm (mice) and 2-mm (rats) intervals rostral and caudal to the bregma(0.00 mm). The yellow (H&E staining) and black (Nissl staining) dotted linesindicate the marginal penumbra lesion. The black arrow indicates the pale violet colorof the ipsilateral lesion of the corpus callosum. The 90 min MCAo models not onlyshowed severe infarct lesion but also failure to confirmed corpus callosum. CCA:common carotid artery; ECA: external carotid artery; ICA: internal carotid artery; OA:occipital artery; PPA: pterygopalatine artery; MCA: middle cerebral artery.
Mentions: Microsurgical procedures of middle cerebral artery occlusion: All animalswere anesthetized with 40 mg/kg tiletamine/zolazepam cocktail (Zoletil) and 10 mg/kgxylazine (Rompun) via intraperitoneal injection. After anesthesia, the animals’ bodytemperature was maintained at 36.5°C to 37.0°C using a heating pad on the surgical table.The animals were placed in the supine position and fixed to the surgical table usingadhesive tape. The incision region was disinfected with povidone-iodine solution. Themidline neck skin was incised, and the common carotid artery (CCA), external carotid artery(ECA) and internal carotid artery (ICA) were carefully separated from the vagus nerve. TheICA bifurcates into the middle cerebral artery (MCA) and the pterygopalatine artery (PPA)from the CCA. The occipital artery (OA) originates from the bifurcation point of the ECA andthe ICA. A transient knot was placed on the distal portion of the CCA, near the manubrium ofthe sternum. A microvascular clamp was transiently placed on the ICA proximal to the CCAjunction. Two closely spaced permanent knots were then placed on the distal portion of theECA, below the suprathyroid artery, to prevent the backflow of blood. The tied section ofthe ECA was dissected using microscissors to insert the probe, which reached the ICA throughthe CCA junction. The microvascular clamp that had been placed on the ICA was then removedto allow probe insertion. The probe was carefully inserted into the MCA from the CCAjunction (up to 18–20 mm in rats and 9–11 mm in mice). After confirmation of MCA blockage(mild bending of the ICA), the rat model exhibited blood supply from the CCA, but the mousemodel only exhibited blood supply after the occlusion period (dual occlusion by CCAligation). After 60 to 90 min, the probe was carefully withdrawn until the tip was near thearteriotomy for reperfusion. After confirmation of blood flow reperfusion, topical lidocainegel was applied onto the surgical region to relieve pain and discomfort in the postoperativeperiod. Each animal also received 1.0 ml of normal saline subcutaneouslyfor volume replenishment after the surgery. All surgical procedures were finished within 15min, excluding the occlusion and reperfusion periods (Fig. 1AFig. 1.

Bottom Line: In rodent MCAo models, has to be considered body temperature during the operative period, as well as the need for the use of a standardized tip in terms of the outer diameter of probes.Our methods could induce stable moderate-severity ischemic brain injury models and histological alteration at 24 hr after MCAo surgery.Finally, we described and compared major parameters between rats and mice, including probe size, thread insert length, operation and occlusion periods, and differences in the procedures.

View Article: PubMed Central - PubMed

Affiliation: Biohealth Products Research Center (BPRC), Inje University, Gimhae, Korea.

ABSTRACT
Rodent models of focal cerebral ischemia that do not require craniotomy have been developed by intraluminal suture middle cerebral artery occlusion (MCAo). Mouse MCAo models have been widely used and extended to genetic studies of cell death or recovery mechanisms. Therefore, we compared surgery-related parameters and techniques between such rats and mice. In rodent MCAo models, has to be considered body temperature during the operative period, as well as the need for the use of a standardized tip in terms of the outer diameter of probes. Induction of focal cerebral ischemia was measured by neurological dysfunction parameters. Our methods could induce stable moderate-severity ischemic brain injury models and histological alteration at 24 hr after MCAo surgery. Moreover approximately 80% (rats) and 85% (mice) survival ratios were shown indicating with model engineering success. Finally, we described and compared major parameters between rats and mice, including probe size, thread insert length, operation and occlusion periods, and differences in the procedures.

Show MeSH
Related in: MedlinePlus