A Neuronal Apoptosis Model induced by Spinal Cord Compression in Rat

Summary

Here, we present a protocol to generate a rat spinal cord compression model, assess its behavioral score, and observe the compressed spinal cord region. The behavioral assessments showed decreased monitor motor disability. Hematoxylin and eosin staining and immunostaining revealed considerable neuronal apoptosis in the compressed region of the spinal cord.

Abstract

As a severe progressive degenerative disease, cervical spondylotic myelopathy (CSM) has a poor prognosis and is associated with physical pain, stiffness, motor or sensory dysfunction, and a high risk of spinal cord injury and acroparalysis. Thus, therapeutic strategies that promote efficient spinal cord regeneration in this chronic and progressive disease are urgently needed. Effective and reproducible animal spinal cord compression models are required to understand the complex biological mechanism underlying CSM. Most spinal cord injury models reflect acute and structural destructive conditions, whereas animal models of CSM present a chronic compression in the spinal cord. This paper presents a protocol to generate a rat spinal cord compression model, which was further evaluated by assessing the behavioral score and observing the compressed spinal cord region. The behavioral assessments showed decreased monitor motor disability, including joint movements, stepping ability, coordination, trunk stability, and limb muscle strength. Hematoxylin and eosin (H&E) staining and immunostaining revealed considerable neuronal apoptosis in the compressed region of the spinal cord.

Introduction

As a common progressive degenerative disease, CSM accounts for 5-10% of all cervical spondylosis¹. If patients suffering from CSM ignore their symptoms and fail to treat them in a timely and effective manner, this could lead to severe complications, such as spinal cord injury and limb paralysis, which would deteriorate with aging, posing a substantial economic and mental burden to patients and their families^2,3. The pathogenesis of CSM is complex, involving static and dynamic factors, the hypoxia-ischemia theory, endothelial cell injury, the blood spinal cord barrier destruction theory,....

Protocol

The following procedure was performed with approval from the Institutional Animal Care and Use Committee (IACUC), Shanghai University of Traditional Chinese Medicine. All survival surgeries were performed under sterile conditions as outlined by the NIH guidelines. Pain and risk of infections were managed with appropriate analgesics and antibiotics to ensure a successful outcome. This surgical procedure is optimized for Sprague-Dawley (SD) outbred male rats at 12 weeks of age and 400 g weight.

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Discussion

The goal of this surgical procedure was to generate reproducible, prolonged, neural apoptosis in the rat spinal cord. A key advantage of this model is that the expandable hydrogel implants provide a prolonged compression on the spinal cord, thereby leading to a progressive neural apoptotic response (Figure 2C), which is consistent with the pathological process of CSM. In the current study, the mortality from spinal cord injury was extremely low (~2 in 50), whereas the repeatability of t.......

Materials

Name	Company	Catalog Number	Comments
Antibiotic ointment			Prevent wound infection
Buprenorphine-SR			Pain relief
Isoflurane	Veteasy		Anesthesia
Inhalant anesthesia equipment			Anesthesia
Micro ophthalmic forceps	Mingren medical equipment	Length: 11 cm, Head diameter: 0.3 mm	Clip the muscle
Ophthalmic forceps	Shanghai Medical Devices (Group) Co., Ltd. Surgical Instruments Factory	JD1050	Clip the skin
Ophthalmic scissors (10 cm)	Shanghai Medical Devices (Group) Co., Ltd. Surgical Instruments Factory	Y00030	Skin incision
SD male rats	Shanghai SLAC Laboratory Animal Co., Ltd	SCXK2018-0004	Animal model
Sterile surgical blades (22#)	Shanghai Pudong Jinhuan Medical Products Co., Ltd.	35T0707	Muscle incision
Small animal trimmer			Hair removal
Veet hair removal cream	RECKITT BENCKISER (India) Ltd		Hair removal
Venus shears	Mingren medical equipment	Length: 12.5 cm	Muscle incision