This study was partially supported by NIH grant RO1NS053407 to J.H. Zhang. We would like to thank Mr. Damon Klebe for his valuable contributions.

All procedures were conducted in compliance with the NIH Guide for the Care and Use of Laboratory Animals and approved by the Animal Care and Use Committee at Loma Linda University.
1. Presurgical Preparations
Aseptic techniques are recommended for all surgical procedures. Disinfect the stereotactic apparatus and prepare sterile surgical tools prior to surgery. Wear personal protective equipment (PPE) during all animal handling. Use a heating pad during surge...

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Animal models of intracerebral hemorrhage (ICH) contribute greatly to an advanced understanding of the disease's pathophysiology, and are commonly used to develop and evaluate novel therapeutic strategies in a preclinical setting. Intraparenchymal injections of autologous blood or bacterial collagenase are well-established methods to generate ICH in rodents. Both methods were initially developed in the rat; however, due to the rapidly increasing availability of transgenic and knockout strains, mice became indispensable t...

<table border="1">
 <tbody>
 <tr>
 <td>Material</td>
 <td>Company</td>
 <td>Catalogue 				Number</td>
 <td>Comment</td>
 </tr>
 <tr>
 <td>Stereotactic Head Frame</td>
 <td>Stoelting Co.</td>
 <td>51600</td>
 <td></td>
 </tr>
 <tr>
 <td>Nanomite Syringe Pump</td>
 <td>Harvard Apparatus</td>
 <td>PY2 70-2217</td>
 <td></td>
 </tr>
 <tr>
 <td>Hamilton Syringe</td>
 <td>Hamilton Company</td>
 <td>1725RN 				(250 &mu;l) 
1701 RN (10 &mu;l)</td>
 <td>26 Gauge needle for 				250 &mu;l and 10 &mu;l syringes.</td>
 </tr>
 <tr>
 <td>Microdrill</td>
 <td>Fine Science Tools</td>
 <td>18000-17</td>
 <td></td>
 </tr>
 <tr>
 <td>Microdrill burr</td>
 <td>Fine Science Tools</td>
 <td>19007-09</td>
 <td>0.9 mm diameter</td>
 </tr>
 <tr>
 <td>Collagenase Type VII-S</td>
 <td>Sigma-Aldrich</td>
 <td>C2399</td>
 <td></td>
 </tr>
 <tr>
 <td>Microhematocrit 				Capillary Tubes</td>
 <td>Fisher Scientific</td>
 <td>22-362-574</td>
 <td>unheparinized</td>
 </tr>
 <tr>
 <td>Bone Wax </td>
 <td>Ethicon</td>
 <td>W31</td>
 <td></td>
 </tr>
 <tr>
 <td>Suture</td>
 <td>Ethicon</td>
 <td>1676G</td>
 <td></td>
 </tr>
 <tr>
 <td>Ketamine</td>
 <td>JHP Pharmaceuticals</td>
 <td>42023-115-10</td>
 <td>Ketalar</td>
 </tr>
 <tr>
 <td>Xylazine </td>
 <td>LLOYD Laboratories</td>
 <td>139-236</td>
 <td>AnaSed</td>
 </tr>
 </tbody>
 </table>

modeling intracerebral hemorrhage in mice: injection of autologous blood or bacterial collagenase

Spontaneous intracerebral hemorrhage (ICH) defines a potentially life-threatening neurological malady that accounts for 10-15% of all stroke-related hospitalizations and for which no effective treatments are available to date1,2. Because of the heterogeneity of ICH in humans, various preclinical models are needed to thoroughly explore prospective therapeutic strategies3. Experimental ICH is commonly induced in rodents by intraparenchymal injection of either autologous blood or bacterial collagenase4. The appropriate model is selected based on the pathophysiology of hemorrhage induction and injury progression. The blood injection model mimics a rapidly progressing hemorrhage. Alternatively, bacterial collagenase enzymatically disrupts the basal lamina of brain capillaries, causing an active bleed that generally evolves over several hours5. Resultant perihematomal edema and neurofunctional deficits can be quantified from both models. In this study, we described and evaluated a modified double injection model of autologous whole blood6 as well as an ICH injection model of bacterial collagenase7, both of which target the basal ganglia (corpus striatum) of male CD-1 mice. We assessed neurofunctional deficits and brain edema at 24 and 72 hr after ICH induction. Intrastriatal injection of autologous blood (30 &mu;l) or bacterial collagenase (0.075U) caused reproducible neurofunctional deficits in mice and significantly increased brain edema at 24 and 72 hr after surgery (p&lt;0.05). In conclusion, both models yield consistent hemorrhagic infarcts and represent basic methods for preclinical ICH research.

Watch this Scientific Journal Video about Modeling Intracerebral Hemorrhage in Mice: Injection of Autologous Blood or Bacterial Collagenase at JoVE.com

Modeling Intracerebral Hemorrhage in Mice: Injection of Autologous Blood or Bacterial Collagenase

Clinically relevant animal models of intracerebral hemorrhage (ICH) are needed to extend our knowledge of hemorrhagic stroke and to examine novel therapeutic strategies. In this study, we describe and evaluate two ICH models that implement unilateral injections of either autologous whole blood or bacterial collagenase into the basal ganglia (corpus striatum) of mice.

Spontaneous intracerebral hemorrhage (ICH) defines a potentially life-threatening neurological malady that accounts for 10-15% of all stroke-related ...

modeling-intracerebral-hemorrhage-mice-injection-autologous-blood-or

Research

JoVE Journal

Medicine

23.9K visualizações.  Loma Linda University School of Medicine. Modelos animais clinicamente relevantes de hemorragia intracerebral (ICH) são necessários para ampliar nosso conhecimento sobre AVC hemorrágico e para analisar novas estratégias terapêuticas. Neste estudo, descrever e avaliar dois modelos ICH que implementam injeções unilaterais de tanto sangue autólogo todo ou colagenase bacteriana no gânglio basal (corpo estriado) de camundongos.