Variations of Neurotrophic Factors and It's Importances in Spinal Cord Injured Rats and Beagle Dogs

In-Soo Song; Jun-Young Yang; June-Kyu Lee; Yong-Bum Joo; Soo-Min Cha

doi:10.4184/jkss.2011.18.1.1

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Song, Yang, Lee, Joo, and Cha: Variations of Neurotrophic Factors and It's Importances in Spinal Cord Injured Rats and Beagle Dogs

Original Article

Journal of Korean Spine Surg 2011;18(1):1-12.

Published online: 21 March 2011

DOI: https://doi.org/10.4184/jkss.2011.18.1.1

Variations of Neurotrophic Factors and It's Importances in Spinal Cord Injured Rats and Beagle Dogs

In-Soo Song, M.D.^∗, Jun-Young Yang, M.D., June-Kyu Lee, M.D., Yong-Bum Joo, M.D., Soo-Min Cha, M.D.

Department of Orthopaedic Surgery, Chungnam National University School of Medicine, Research Institute of Medical Science, Daejeon, Korea

^∗Department of Orthopaedic Surgery, Sun General Hospital, Daejeon, Korea

Corresponding author: Jun-Young Yang, M.D. Department of Orthopedic Surgery, Chungnam National University School of Medicine, 640, Daesa-Dong, Jung-Gu, Daejeon, Korea TEL: 82-42-280-7351, FAX: 82-42-252-7098 E-mail: jyyang@cnu.ac.kr

Received 27 October 2010 Revised 18 February 2011 Accepted 21 February 2011

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Study Design

Experimental, prospective study

Objectives

To examine the changes in the variable factors after an acute spinal cord injury(SCI) in rats and dogs simultaneously.

Summary of Literature Review

No study has examined the variations of several factors in a SCI model in different species.

Materials and Methods

In rats, a laminectomy was performed at the T10 level and the injured spinal cord was extracted. In Beagle dogs, the laminectomy level was T10 and T11. The motor function was evaluated using a modified Tarlov's scale. A RT2 profiler PCR array was used to examine each factor (inflammatory cytokines, factors-related with apoptosis, neurotrophic factors, factors-related with extraceullar matrix).

Results

IL-2, TNF, TNFRSF11B increased with time and showed no statistical difference between two species, but TNFSF13B showed a significant difference. BDNF decreased with time in both species, and GDNF was significantly lower in dogs. NGFβ, CTNF and its receptors showed no significant changes in the two species. MMP1 increased in both species but MMP7 decreased in rats and increased in dogs with time, and showed a significant difference between species.

Conclusion

The change in inflammatory cytokines and extracellular matrix correlates with each factor in the combined patterns. Moreover, during the first week after SCI, inflammatory cytokines, apoptosis, neutrophic factors, and extracellular matrix factors may show a partial difference between experimental animals, which means that an animal model can be selected according to the particular experimental plan.

Keywords: Spinal cord injury, Inflammatory cytokines, Factors-related with apoptosis, Neurotrophic factors, Factors-related with extraceullar matrix

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Figures and Tables%

Fig. 1.

EMG recorded after spinal cord inury in dog: Noted severe training wave when spinal cord impacted. *SCI, spinal cord injury.

Fig. 2.

This photograph shows the procedure and instrument of extraction RNA.

Fig. 3.

Quality control of total RNA. All experimental material was well controlled.

Fig. 4.

(A) Motor function after spinal cord injury at 1 day, 7 days by Tarlov's scale. (B) The count of motor function; There was no statistical difference between two groups. (C) Slides from Extracted spinal cord, vesicle and infiltration of inflammatory cells, edema was shown(H&E stain, ×20).

Fig. 5.

This graphs show the acute inflammatory cytokines changes after spinal cord injury. Between two species, there was no significant difference(p>0.05).

Fig. 6.

This graphs show the factors related with apoptosis. Also, there was no significant differences after spinal cord injury(p>0.05).

Fig. 7.

This graphs show the changes of several neurotrophic factors. (A) GDNF was reduced more, statistically in dogs(p<0.05). (B),(C) These factors were no statistically differences(p>0.05).

Fig. 8.

This graphs show the changes of factors related with extracellular matrix. (A), (B) These factors were all increased after spinal cord injury and there was no statistical differences(p>0.05). (C) MMP7 was reduced in rat, but increased in dog, also there was significant difference in changes(<0.05).

Table 1.

Modified Tarlov's scale for motor function evaluation

Scale	Degree of Paraplegia	Symptoms
0	Flaccid	Minimal or no leg movements; variable tone; loss of bowel/bladder control
1	Spastic	Moderate or vigorous purposeless legmovements; leg spastic and extended; no sitting; loss of bowel/bladder control
2	Severe	Only moderate spasticity in legs;vigorous, coordinated movements suggesting walking; can sit and stand; loss of bowel/bladder control
3	Moderate	Sitting, standing, and walking are shown; legs and hips obviously unstable; leg lagging; loss of bowel/bladder control
4	Mild	Walking with some leg and hip instability; bowel/bladder functional
5	Normal	Hip instability seen only with jumping or running

Table 2.

Prepare of RT Cocktail

RT Cocktail	1 reaction	2 reaction	4 reaction
BC3(5X Rt Buffer 3)	4µl	8µl	16µl
P2(Primer & External Control Mix)	1µl	2µl	4µl
RE3(RT Enzyme Mix3)	2µl	4µl	8µl
H₂ O	3µl	6µl	12µl
Final Volume	10µl	20µl	40µl

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