Name: modified spared nerve injury surgery model of neuropathic pain in mice
Uploaded: 2022-01-25T14:00:00
Description: Watch this Scientific Journal Video about Modified Spared Nerve Injury Surgery Model of Neuropathic Pain in Mice at JoVE.com

Z.G. is supported by NINDS R01NS100801.

Animal experiments were approved by UCSF Institutional Animal Care and Use Committee and were conducted in accordance with the NIH Guide for the Care and Use of Laboratory animals. Adult C57BL/6 mice weighing 20-30 g was used in this study. The von Frey assessment was performed between 1:00 pm and 3:00 pm.
1. Anesthesia and mouse preparation
<ol>
	<li>Place a mouse in a plastic chamber filled with 2% isoflurane in O2 at a flow rate of 1.0 L/min until it is fully a...

<ol>
	<li>Chung, J. M., Kim, H. K., Chung, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Segmental+spinal+nerve+ligation+model+of+neuropathic+pain.">Segmental spinal nerve ligation model of neuropathic pain.</a> Methods in Molecular Medicine. 99, 35-45 (2004).</li><li>Vissers, K., Adriaensen, H., De Coster, R., De Deyne, C., Meert, T. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+chronic-constriction+injury+of+the+sciatic+nerve+reduces+bilaterally+the+responsiveness+to+formalin+in+rats:+a+behavioral+and+hormonal+evaluation.">A chronic-constriction injury of the sciatic nerve reduces bilaterally the responsiveness to formalin in rats: a behavioral and hormonal evaluation.</a> Anesthesia and Analgesia. 97 (2), 520-525 (2003).</li><li>Malmberg, A. B., Basbaum, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+sciatic+nerve+injury+in+the+mouse+as+a+model+of+neuropathic+pain:+behavioral+and+neuroanatomical+correlates.">Partial sciatic nerve injury in the mouse as a model of neuropathic pain: behavioral and neuroanatomical correlates.</a> Pain. 76 (1-2), 215-222 (1998).</li><li>Moore, K. 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=Partial+peripheral+nerve+injury+promotes+a+selective+loss+of+GABAergic+inhibition+in+the+superficial+dorsal+horn+of+the+spinal+cord.">Partial peripheral nerve injury promotes a selective loss of GABAergic inhibition in the superficial dorsal horn of the spinal cord.</a> The Journal of Neuroscience: the official journal of the Society for Neuroscience. 22 (15), 6724-6731 (2002).</li><li>Decosterd, I., Woolf, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spared+nerve+injury:+an+animal+model+of+persistent+peripheral+neuropathic+pain.">Spared nerve injury: an animal model of persistent peripheral neuropathic pain.</a> Pain. 87 (2), 149-158 (2000).</li><li>Cichon, J., Sun, L., Yang, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spared+Nerve+Injury+Model+of+Neuropathic+Pain+in+Mice.">Spared Nerve Injury Model of Neuropathic Pain in Mice.</a> Bio-Protocol. 8 (6), 2777 (2018).</li><li>Shields, S. D., Eckert, W. A., Basbaum, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spared+nerve+injury+model+of+neuropathic+pain+in+the+mouse:+a+behavioral+and+anatomic+analysis.">Spared nerve injury model of neuropathic pain in the mouse: a behavioral and anatomic analysis.</a> The Journal of Pain. 4 (8), 465-470 (2003).</li><li>Guan, Z., 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=Injured+sensory+neuron-derived+CSF1+induces+microglial+proliferation+and+DAP12-dependent+pain.">Injured sensory neuron-derived CSF1 induces microglial proliferation and DAP12-dependent pain.</a> Nature Neuroscience. 19 (1), 94-101 (2016).</li><li>Richner, M., Bjerrum, O. J., Nykjaer, A., Vaegter, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+spared+nerve+injury+(SNI)+model+of+induced+mechanical+allodynia+in+mice.">The spared nerve injury (SNI) model of induced mechanical allodynia in mice.</a> Journal of Visualized Experiments: JoVE. (54), e3092 (2011).</li><li>Challa, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+animal+models+of+neuropathic+pain:+Pros+and+Cons.">Surgical animal models of neuropathic pain: Pros and Cons.</a> The International Journal of Neuroscience. 125 (3), 170-174 (2015).</li><li>Kosaka, Y., 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=Development+and+persistence+of+neuropathic+pain+through+microglial+activation+and+KCC2+decreasing+after+mouse+tibial+nerve+injury.">Development and persistence of neuropathic pain through microglial activation and KCC2 decreasing after mouse tibial nerve injury.</a> Brain Research. 1733, 146718 (2020).</li><li>Parisien, M., Lima, L. V., Dagostino, C., El-Hachem, N., Drury, G. L., Grant, A. V., Huising, J., Verma, V., Meloto, C. B., Silva, J. R., Dutra, G. C. S., Markova, T., Dang, H., Tessier, P. A., Slade, G. D., Nackley, A. G., Ghasemlou, N., Mogil, J. S., Allegri, M., Diatchenko, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+inflammatory+response+via+neutrophil+activation+protects+against+the+development+of+chronic+pain.">Acute inflammatory response via neutrophil activation protects against the development of chronic pain.</a> Sci Transl Med. 14 (644), eabj9954 (2022).</li><li>Chaplan, S. R., Bach, F. W., Pogrel, J. W., Chung, J. M., Yaksh, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+assessment+of+tactile+allodynia+in+the+rat+paw.">Quantitative assessment of tactile allodynia in the rat paw.</a> Journal of Neuroscience Methods. 53 (1), 55-63 (1994).</li><li>Bourquin, A. F., 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=Assessment+and+analysis+of+mechanical+allodynia-like+behavior+induced+by+spared+nerve+injury+(SNI)+in+the+mouse.">Assessment and analysis of mechanical allodynia-like behavior induced by spared nerve injury (SNI) in the mouse.</a> Pain. 122 (1-2), 1-14 (2006).</li><li>Sulaiman, W., Gordon, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurobiology+of+peripheral+nerve+injury,+regeneration,+and+functional+recovery:+from+bench+top+research+to+bedside+application.">Neurobiology of peripheral nerve injury, regeneration, and functional recovery: from bench top research to bedside application.</a> The Ochsner Journal. 13 (1), 100-108 (2013).</li><li>Kochi, T., 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=Characterization+of+the+arterial+anatomy+of+the+murine+hindlimb:+functional+role+in+the+design+and+understanding+of+ischemia+models.">Characterization of the arterial anatomy of the murine hindlimb: functional role in the design and understanding of ischemia models.</a> PLoS One. 8 (12), 84047 (2013).</li><li>Omori, Y., 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=A+mouse+model+of+sural+nerve+injury-induced+neuropathy:+gabapentin+inhibits+pain-related+behaviors+and+the+hyperactivity+of+wide-dynamic+range+neurons+in+the+dorsal+horn.">A mouse model of sural nerve injury-induced neuropathy: gabapentin inhibits pain-related behaviors and the hyperactivity of wide-dynamic range neurons in the dorsal horn.</a> Journal of Pharmacological Sciences. 109 (4), 532-539 (2009).</li><li>Colloca, L., 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=Neuropathic+pain.">Neuropathic pain.</a> Nature Reviews. Disease Primers. 3, 17002 (2017).</li></ol>

Compared to the traditional mouse SNIt method that ligates the common peroneal nerve and the sural nerve separately6,7,8,9, the modified SNIt model has three advantages: (1) it has less risk of contracting or stretching sciatic or tibial nerves; (2) there is no need to remove the distal nerve stumps after nerve-cutting because by ligating the common peroneal nerve and the sural nerve together, ...

Nerve injury-induced neuropathic pain following surgery or trauma has a significant economic burden that impairs quality of life. A host of nerve injury models, including spinal nerve ligation (SNL)1, chronic constriction injury (CCI)2 to the sciatic nerve, partial sciatic nerve ligation (pSNL)3, sciatic nerve transaction (SNT)4 and spared nerve injury (SNI)5,6,7,8, were successfully developed to mimic the cardinal symptoms of peripheral nerve injury in rats and mice for studying the molecular and cellular mechanism of neuropathic pain6,7,8,9,10. However, each surgical model has its benefits and limitations, therefore particular attention should be given to exploring and developing the surgery models10.
The rodent SNI model produces long-term hypersensitivity to mechanical stimulation. However, it is somewhat confusing because there are two different SNI models. The initial SNI model was developed in Woolf&#39;s lab, in which the common peroneal and the tibial nerves were injured, leaving the sural nerve intact5,6. The second SNI model was developed in Basbaum&#39;s lab, in which the common peroneal and the sural nerves were injured, leaving the tibial nerve intact7,8. The initial Woolf&#39;s model is defined as SNIs here because the sural nerve is left intact, and Basbaum&#39;s model is defined as SNIt here because the tibial nerve is left intact. Because the sural nerve is purely sensory whereas the tibial nerve contains both motor and sensory fibers, the SNIt model has much less motor deficit than the SNIs model. However, unlike the SNIs model, mice in the SNIt model do not develop thermal hypersensitivity, but mechanical hypersensitivity develops in both models. Although the SNIt model is a relatively easy procedure, it requires the ligation of the sural and common peroneal nerves separately with the potential risk of stretching the sciatic or tibial nerves6,7,8,9.
The common peroneal, tibial, and sural nerves are three branches of the sciatic nerve and can be clearly identified at the superior edge of the gastrocnemius muscle (Figure 1): the tibial nerve goes under the gastrocnemius muscle, and the common peroneal (cephalad side) and sural nerve (caudal side) are above the gastrocnemius muscle11. Based on its anatomical features, a modified mouse SNIt surgery procedure was developed to ligate the common peroneal and sural nerves together with only one nerve-ligation and one nerve-cutting, which results in shortened procedure-time.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>6-0 suture</td>
			<td>Henry Schein</td>
			<td>9007482</td>
			<td>Nerve ligation and close the muscular layer</td>
		</tr>
		<tr>
			<td>Iris Scissors</td>
			<td>Integra Miltex</td>
			<td>12460598</td>
			<td>Cut muscle and fascia</td>
		</tr>
		<tr>
			<td>Mayo dissecting scissors</td>
			<td>Fisherbrand</td>
			<td>895120</td>
			<td>Cut skin incision</td>
		</tr>
		<tr>
			<td>Micro forcep</td>
			<td>Fisherbrand</td>
			<td>16100110</td>
			<td>Blunt dissection biceps femoris muscle</td>
		</tr>
		<tr>
			<td>Micro Scissors</td>
			<td>Excelta</td>
			<td>17467496</td>
			<td>Cut nerve</td>
		</tr>
		<tr>
			<td>Microdissection Forceps</td>
			<td>Fisherbrand</td>
			<td>16100123</td>
			<td>Separate the common peroneal and the sural nerves from the neighboring tissues</td>
		</tr>
		<tr>
			<td>Needle Holder</td>
			<td>Fisherbrand</td>
			<td>8966</td>
			<td>Hold 6-0 needle</td>
		</tr>
		<tr>
			<td>Prism software</td>
			<td>Graph Pad</td>
			<td>version 8.0</td>
			<td>Statistical analysis software</td>
		</tr>
		<tr>
			<td>Wound clips</td>
			<td>Roboz Surgical</td>
			<td>NC1878744</td>
			<td>Close skin incision</td>
		</tr>
	</tbody>
</table>

modified spared nerve injury surgery model of neuropathic pain in mice

Spared nerve injury (SNI) is an animal model that mimics the cardinal symptoms of peripheral nerve injury for studying the molecular and cellular mechanism of neuropathic pain in mice and rats. Currently, there are two types of SNI model, one to cut and ligate the common peroneal and the tibial nerves with intact sural nerve, which is defined as SNIs in this study, and another to cut and ligate the common peroneal and the sural nerves with intact tibial nerve, which is defined as SNIt in this study. Because the sural nerve is purely sensory whereas the tibial nerve contains both motor and sensory fibers, the SNIt model has much less motor deficit than the SNIs model. In the traditional SNIt mouse model, the common peroneal and the sural nerves are cut and ligated separately. Here a modified SNIt surgery method is described to damage both common peroneal and sural nerves with only one ligation and one cut with a shorter procedure time, which is easier to perform and reduces the potential risk of stretching the sciatic or tibial nerves, and produces similar mechanical hypersensitivity as the traditional SNIt model.

The comparison of procedure time between modified and traditional methods. 
The procedure time from the beginning of cutting the skin to the end of closing skin was recorded in 5 mice with the modified approach and 5 mice with the traditional approach, respectively. A minimal number of animals was used to obtain results with statistical significance. Compared with the control of the traditional approach7,14 to perform SNIt, the modified appr...

Watch this Scientific Journal Video about Modified Spared Nerve Injury Surgery Model of Neuropathic Pain in Mice at JoVE.com

Modified Spared Nerve Injury Surgery Model of Neuropathic Pain in Mice

The modified surgery is a simplified method for mouse or rat spared nerve injury model that requires only one ligation and one cut to injure both common peroneal and sural nerves.

Spared nerve injury (SNI) is an animal model that mimics the cardinal symptoms of peripheral nerve injury for studying the molecular and cellular ...

To cut and ligate a common peroneal and sural nerves with intact tibial nerve, defined as SNIt, has much less motor deficit with a shorter learning curve. The modified mouse SNIt surgery procedure was developed to ligate the common peroneal and sural nerves together with only one nerve ligation and one nerve cutting, which results in shortened procedure time. To begin, place the mouse in a right lateral position.Keep the left leg up with knee flexion, and secure it with adhesive tape. Remove the hair around the thigh and knee area with an electric shaver and hair removal cream. After placing the mouse on a new sterile surgery towel, disinfect the skin with 2%chlorhexidine and 70%alcohol.Secure it with adhesive tape. Cut a one centimeter incision starting at one third of the horizontal line, crossing the knee with an approximately 30 degree angle from the vertical line with a scalpel. Two white lines can be visualized under the biceps femoris muscle after separating skin incision with the medial cephalad thick line above the femur and lateral caudal thin line above sciatic nerve.Blunt dissect the biceps femoris muscle along the caudal white line using curved micro forceps and micro scissors to expose the sciatic nerve. Differentiate the three branches from the sciatic nerve at the superior edge of the gastrocnemius muscle. The tibial nerve has the biggest diameter passing under the gastrocnemius muscle, whereas the sural nerves are lateral and smallest in diameter.The common peroneal nerve runs above the gastrocnemius muscle. Depending on how the biceps femoris muscle is dissected and opened, visualize the common peroneal nerve as lateral or medial to the tibial nerve. Then separate the common peroneal and the sural nerves from the neighboring tissues using curved micro forceps.Ligate the common peroneal and sural nerves together with a 6/0 suture. Both nerves run above the gastrocnemius muscle, but the tibial nerve passes under the gastrocnemius muscle. Observe the limb for contraction following the tight ligature.Cut nerve at a distal part within two to four millimeters of the ligation with a pair of micro scissors. Ensure that the tibial nerve remains untouched during the whole procedure. Close the muscular layer with a 6/0 silk suture and close the skin incision with wound clips.The results show that the modified approach took almost half the amount of procedure time to perform SNlt compared with the control of the traditional approach. No difference in Von Frey assessment was observed in the two groups at baseline. The traditional method and the modified method induced similar mechanical hypersensitivity on the ipsilateral side from postoperative day one to postoperative day 14.The percentage response between traditional and modified methods showed similar mechanical hypersensitivity on the ipsilateral side from baseline. Easy and rapid exposure of the sciatic nerve, accurate identification of the common peroneal, tibial, and sural nerves, cutting and ligation of the common peroneal and sural nerves are the three critical steps in this protocol. The method was successfully developed to mimic peripheral nerve injury cardio symptoms and randomized to study the molecular and cellular mechanism of neuropathic pain.

Research

JoVE Journal

Neuroscience

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and ...

Transplantation of Olfactory Ensheathing Cells to Evaluate Functional Recovery after Peripheral Nerve Injury

Olfactory ensheathing cells (OECs) are neural crest cells which allow growth and regrowth of the primary olfactory neurons. Indeed, the primary olfactory ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

A Novel In Vitro Model of Blast Traumatic Brain Injury

A Novel In Vitro Model of Blast Traumatic Brain Injury

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the ...

Facial Nerve Surgery in the Rat Model to Study Axonal Inhibition and Regeneration

This protocol describes consistent and reproducible methods to study axonal regeneration and inhibition in a rat facial nerve injury model. The facial ...

Purification of Fibroblasts and Schwann Cells from Sensory and Motor Nerves in Vitro

The principal cells in the peripheral nervous system are the Schwann cells (SCs) and the fibroblasts. Both these cells distinctly express the sensory and ...

Burn Injury-Induced Pain and Depression-Like Behavior in Mice

Scalding water is the most common cause of burn injury in both elderly and young populations. It is one of the major clinical challenges because of the ...

Dural Stimulation and Periorbital von Frey Testing in Mice As a Preclinical Model of Headache

The cranial meninges, comprised of the dura mater, arachnoid, and pia mater, are thought to primarily serve structural functions for the nervous system.  ...

Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and ...

A Spinal Cord Injury Contusion Device for Accurate Mouse Models

Automated Impactor for Contusive Spinal Cord Injury Model in Mice

Author Spotlight: Insight Into Innovations in Spinal Cord Injury Research

Spinal cord injury (SCI) due to traumatic injuries such as car accidents and falls is associated with permanent spinal cord dysfunction. Creation of ...