Name: induction of complete transection-type spinal cord injury in mice
Uploaded: 2020-05-06T14:00:00
Description: Watch this Scientific Journal Video about Induction of Complete Transection-Type Spinal Cord Injury in Mice at JoVE.com

This work was supported by a Griffith University International Student (PhD) stipend to RR, a Perry Cross Foundation Grant to JE and JSJ, a Clem Jones Foundation Grant to JSJ and JE, and a Motor Accident Insurance Commission of Queensland grant to JSJ and JE.

All procedures were carried out with the approval of the Griffith University Animal Ethics Committee (ESK/04/16 AEC and MSC/04/18 AEC) under the guidelines of the National Health and Medical Research Council of Australia.
1. Animal set-up procedure for the surgery
<ol>
	<li>Anesthetize and stabilize the animal.
	<ol>
		<li>Use 8&#8211;10-week old female C57BL/6 mice. Use 5% isoflurane in 1 L/min oxygen for induction of anesthesia. For maintenance of anesthesia, use 1.5&#8211;2% isoflurane in...

<ol>
	<li>Sharif-Alhoseini, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+spinal+cord+injury:+a+systematic+review.">Animal models of spinal cord injury: a systematic review.</a> Spinal Cord. 55 (8), 714-721 (2017).</li><li>Lee, D. H., Lee, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+axon+regeneration+after+spinal+cord+injury.">Animal models of axon regeneration after spinal cord injury.</a> Neuroscience Bulletin. 29 (4), 436-444 (2013).</li><li>Sharif-Alhoseini, M., Rahimi-Movaghar, V., Dionyssiotis, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Topics in Paraplegia. , (2014).</li><li>Talac, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+spinal+cord+injury+for+evaluation+of+tissue+engineering+treatment+strategies.">Animal models of spinal cord injury for evaluation of tissue engineering treatment strategies.</a> Biomaterials. 25 (9), 1505-1510 (2004).</li><li>Nakae, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+animal+model+of+spinal+cord+injury+as+an+experimental+pain+model.">The animal model of spinal cord injury as an experimental pain model.</a> Journal of Biomedicine &amp; Biotechnology. 2011, 939023 (2011).</li><li>Kwon, B., Oxland, T., Tetzlaff, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+used+in+spinal+cord+regeneration+research.">Animal models used in spinal cord regeneration research.</a> Spine. 27, 1504-1510 (2002).</li><li>Kundi, S., Bicknell, R., Ahmed, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+injury:+current+mammalian+models.">Spinal cord injury: current mammalian models.</a> American Journal of Neuroscience. (4), 1-12 (2013).</li><li>Harrison, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vertebral+landmarks+for+the+identification+of+spinal+cord+segments+in+the+mouse.">Vertebral landmarks for the identification of spinal cord segments in the mouse.</a> Neuroimage. 68, 22-29 (2013).</li><li>Basso, D. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Basso+Mouse+Scale+for+locomotion+detects+differences+in+recovery+after+spinal+cord+injury+in+five+common+mouse+strains.">Basso Mouse Scale for locomotion detects differences in recovery after spinal cord injury in five common mouse strains.</a> Journal of Neurotrauma. 23 (5), 635-659 (2006).</li><li>Seitz, A., Aglow, E., Heber-Katz, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recovery+from+spinal+cord+injury:+a+new+transection+model+in+the+C57Bl/6+mouse.">Recovery from spinal cord injury: a new transection model in the C57Bl/6 mouse.</a> Journal of Neuroscience Research. 67 (3), 337-345 (2002).</li></ol>

This method induces a complete transection type injury at the T10 vertebral level in mice, which results in complete paraplegia of the animal, below the level of injury. Overall, this method results in minimal bleeding, negligible collateral damage and a stable, reproducible injury. As compared to previously published methods of transection without laminectomy10, this method offers the benefits in terms of direct visualization without manipulating the curvature of the spine, better control over co...

Spinal cord injury (SCI) is a complex medical problem that results in drastic changes in health and lifestyle. There is no cure for SCI, and the pathophysiology of SCI is not thoroughly understood. Animal SCI models, in particular rodent models, offer an invaluable tool for trialing new treatments, and have been used to explore SCI for decades. To date, over 72% of pre-clinical SCI studies have employed rat models, as compared to a mere 16% that have used mice1. Although rats, due to their larger size and tendency to form cavities akin to human SCIs, have traditionally been the preferred model animals for studying novel therapeutic approaches, mice (including many transgenic mouse models) are now being used more frequently to study cellular and molecular mechanisms of the SCI2. The mouse model offers additional benefits in terms of easier handling, faster reproductive rates and lower costs than rats; mice also exhibit a high degree of genomic similarity with humans1,2,3. The major disadvantage of the mouse model has been identified as the significantly smaller size that creates challenges for surgical interventions for creating and treating spinal cord injuries4,5.
There is a gap in the existing literature that highlights the need for a robust and reproducible surgical protocol to induce stable SCI in the mouse model. Therefore, we provide a novel and precise surgical approach in this protocol to overcome these limitations. This protocol provides in-depth guidelines to induce a transection-type injury in mice, as this injury type has been recognized to be the most appropriate to study regenerative and degenerative changes following an injury6, as well as neuroplasticity, neural circuitry and tissue engineering approaches7. We have chosen to induce the injury in the lower thoracic region, since thoracic level SCI is used most commonly in the literature1.

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			<td height="21" style="height:21px;width:449px;">Baytril injectable 50 mg/mL, 50 mL</td>
			<td style="width:243px;">Provet</td>
			<td style="width:161px;">BAYT I</td>
			<td style="width:184px;">Post-operative care drug</td>
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			<td height="21" style="height:21px;width:449px;">Betadine 500 mL</td>
			<td style="width:243px;">Provet</td>
			<td style="width:161px;">BETA AS</td>
			<td>Consumable</td>
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			<td height="21" style="height:21px;width:449px;">Castroviejo needle holder, locking</td>
			<td style="width:243px;">ProSciTech</td>
			<td style="width:161px;">T149C</td>
			<td>Reusable</td>
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			<td height="20" style="height:21px;width:449px;">Ceramic zirconia blade, round with sharp sides, single edge, angled</td>
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			<td>Reusable</td>
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			<td height="21" style="height:21px;width:449px;">Cotton swabs (5pcs)</td>
			<td style="width:243px;">Multigate</td>
			<td style="width:161px;">21-893</td>
			<td>Consumable</td>
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			<td height="21" style="height:21px;width:449px;">Dremel Micro</td>
			<td style="width:243px;">DREMEL</td>
			<td style="width:161px;">8050-N/18</td>
			<td>Cordless rotary tool</td>
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			<td height="20" style="height:21px;width:449px;">Dressing forceps fine</td>
			<td style="width:243px;">Multigate</td>
			<td style="width:161px;">06-306</td>
			<td>Single use disposable</td>
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			<td height="21" style="height:21px;width:449px;">Drill bits</td>
			<td style="width:243px;">Kemmer Pr&#228;zision</td>
			<td style="width:161px;">SM 32 M 0550 070</td>
			<td>Reusable</td>
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			<td height="21" style="height:21px;width:449px;">Dumont #7b forceps</td>
			<td style="width:243px;">Fine Science Tools</td>
			<td style="width:161px;">11270-20</td>
			<td>Reusable</td>
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			<td height="21" style="height:21px;width:449px;">Dumont tweezers, style 5</td>
			<td style="width:243px;">ProSciTech</td>
			<td style="width:161px;">T05-822</td>
			<td>Reusable</td>
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			<td height="21" style="height:21px;width:449px;">Fur trimmer</td>
			<td style="width:243px;">WAHL</td>
			<td style="width:161px;">WA9884-312</td>
			<td>Zero Overlap Hair Trimmer</td>
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			<td height="21" style="height:21px;width:449px;">Iris scissors, Ti, sharp tips, straight, 90mm</td>
			<td style="width:243px;">ProSciTech</td>
			<td style="width:161px;">TY-3032</td>
			<td>Reusable</td>
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			<td height="21" style="height:21px;width:449px;">Isoflurane isothesia NXT 250</td>
			<td style="width:243px;">Provet</td>
			<td style="width:161px;">ISOF 00 HS</td>
			<td>Anaesthetic agent</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:449px;">Colibri Retractor - 4cm</td>
			<td style="width:243px;">Fine Science Tools</td>
			<td>17000-04</td>
			<td>Reusable</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:449px;">Scalpel handle</td>
			<td style="width:243px;">ProSciTech</td>
			<td style="width:161px;">T133</td>
			<td>Reusable</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:449px;">Signature latex surgical gloves size 7.5</td>
			<td style="width:243px;">Medline</td>
			<td style="width:161px;">MSG5475</td>
			<td>Consumable</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:449px;">Sodium Chloride 0.9%</td>
			<td style="width:243px;">STS</td>
			<td>PHA19042005</td>
			<td>Consumable</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:449px;">Sterile Dressing Pack</td>
			<td style="width:243px;">Multigate</td>
			<td style="width:161px;">08-709</td>
			<td>Single use disposable</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:449px;">Sterile Fluid Impervious Drape 60x60 cm</td>
			<td style="width:243px;">Multigate</td>
			<td style="width:161px;">29-220</td>
			<td>Single use disposable</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:449px;">Surgical spirit 100 mL</td>
			<td style="width:243px;">Provet</td>
			<td style="width:161px;"># SURG SP</td>
			<td>Consumable</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:449px;">Suture Material - SILK BLK 45CM 5/0 FS-2</td>
			<td style="width:243px;">Johnson &#38; Johnson Medical</td>
			<td style="width:161px;">682G</td>
			<td>Silk Suture</td>
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			<td height="21" style="height:21px;width:449px;">Suture Material - Vicryl 70CM 5-0 S/A FS-2</td>
			<td style="width:243px;">Johnson &#38; Johnson Medical</td>
			<td style="width:161px;">VCP421H</td>
			<td>Vicryl Suture</td>
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		<tr height="22">
			<td height="22" style="height:22px;width:449px;">Temgesic 0.3 mg in 1 mL, x 5 ampoules (class S8 drug)</td>
			<td style="width:243px;">Provet</td>
			<td style="width:161px;">TEMG I</td>
			<td>Post-operative care drug</td>
		</tr>
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induction of complete transection-type spinal cord injury in mice

Spinal cord injury (SCI) largely leads to irreversible and permanent loss of function, most commonly as a result of trauma. Several treatment options, such as cell transplantation methods, are being researched to overcome the debilitating disabilities arising from SCI. Most pre-clinical animal trials are conducted in rodent models of SCI. While rat models of SCI have been widely used, mouse models have received less attention, even though mouse models can have significant advantages over rat models. The small size of mice equates to lower animal maintenance costs than for rats, and the availability of numerous transgenic mouse models is advantageous for many types of studies. Inducing repeatable and precise injury in the animals is the primary challenge for SCI research, which in small rodents requires high-precision surgery. The transection-type injury model has been a commonly used injury model over the last decade for transplantation-based therapeutic research, however a standardized method for inducing a complete transection-type injury in mice does not exist. We have developed a surgical protocol for inducing a complete transection type injury in C57BL/6 mice at thoracic vertebral level 10 (T10). The procedure uses a small tip drill instead of rongeurs to precisely remove the lamina, after which a thin blade with rounded cutting edge is used to induce the spinal cord transection. This method leads to reproducible transection-type injury in small rodents with minimal collateral muscle and bone damage and therefore minimizes confounding factors, specifically where behavioral functional outcomes are analyzed.

The resulting method as depicted in Figure 1, involves adequate stabilization of the mouse (Figure 1A) and good visualization of the spine and paraspinous tissue (Figure 1B). Spinous process and laminae can be clearly visualized with minimal muscle dissection and blood loss (Figure 1C, highlighted zone). The fine tip drilling is performed as shown in Figure 1D, to create a ...

Watch this Scientific Journal Video about Induction of Complete Transection-Type Spinal Cord Injury in Mice at JoVE.com

Induction of Complete Transection-Type Spinal Cord Injury in Mice

This protocol describes how to create a precise laminectomy for induction of stable transection-type spinal cord injury in the mouse model, with minimal collateral damage for spinal cord injury research.

Spinal cord injury (SCI) largely leads to irreversible and permanent loss of function, most commonly as a result of trauma. Several treatment options, ...

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