Korean J Pain.  2013 Jul;26(3):255-264. 10.3344/kjp.2013.26.3.255.

The Effects of Pre-emptive Administration of Ketamine and norBNI on Pain Behavior, c-Fos, and Prodynorphin Protein Expression in the Rat Spinal Cord after Formalin-induced Pain Is Modulated by the DREAM Protein

• Long I
• Suppian R
• Ismail Z

• KMID: 2074026
• DOI: http://doi.org/10.3344/kjp.2013.26.3.255

Abstract

BACKGROUND
We investigated the effects of pre-emptive administration of ketamine and norBNI on pain behavior and the expression of DREAM, c-Fos, and prodynorphin proteins on the ipsilateral side of the rat spinal cord at 2 and 4 hours after formalin injection.
METHODS
Eighty-four male Sprague Dawley rats were divided into 4 major groups consisting of control rats (C) (n = 12), rats given only formalin injections (F) (n = 24), and rats treated with pre-emptive administration of either ketamine (K+F) (n = 24) or norBNI (N+F) (n = 24). The non-control groups were further divided into subgroups consisting of rats that were sacrificed at 2 and 4 hours (n = 12 for each group) after formalin injection. Pain behavior was recorded for 1 hour. After 2 and 4 hours, the rats were sacrificed and the spinal cords (L4-L5 sections) were removed for immunohistochemistry and Western blot analysis.
RESULTS
The pain behavior response was reduced in the K+F group compared to the other groups during the second phase of the formalin pain response. We detected an increase in the nuclear DREAM protein level in the K+F group at 2 and 4 hours and a transient decrease in the N+F group at 2 hours; however, it increased at 4 hours after injection. Fos-like immunoreactivity (FLI) and Prodynorphin-like immunoreactivity (PLI) neurons decreased in the K+F group but increased in the N+F group at 2 hours after injection. While FLI decreased, PLI increased in all groups at 4 hours after injection.
CONCLUSIONS
We suggest that NMDA and kappa opioid receptors can modulate DREAM protein expression, which can affect pain behavior and protein transcriptional processes at 2 hours and bring about either harmful or protective effects at 4 hours after formalin injection.

Keyword

DREAM protein; formalin test; kappa opioid receptor; NMDA receptor; rat spinal cord

MeSH Terms

Animals
Blotting, Western
Enkephalins
Formaldehyde
Humans
Immunohistochemistry
Ketamine
Male
N-Methylaspartate
Neurons
Pain Measurement
Protein Precursors
Proteins
Rats
Rats, Sprague-Dawley
Receptors, Opioid, kappa
Spinal Cord
Enkephalins
Formaldehyde
Ketamine
N-Methylaspartate
Protein Precursors
Proteins
Receptors, Opioid, kappa

Figure

• 

  Fig. 1 (A) Pain behavior response for all groups in 1-hour periods. Control group (C), formalin injected group (F), ketamine and formalin injected group (K+F), norBNI and formalin injected group (N+F). Values are the means ± S.E.M. n = 6 for all groups. (B) Pain behavior response during phase 1. *P < 0.001 compared to C group (C) Pain behavior response during phase 2. *P < 0.001 compared to K+F group.

• 

  Fig. 2 (A) A representative example of Western blot results for nuclear extracts on the ipsilateral side between all groups with quantification analysis of the integrated density value. (B) Columns represent the mean relative DREAM protein level ± S.E.M for 6 separate experiments. The mean relative DREAM protein level (fold change) represents comparative levels of the DREAM protein in the experimental groups (formalin injected group, F, ketamine and formalin injected group, K+F, norBNI and formalin injected group, N+F) over the calibrator group (control group, C) after normalization by its loading control (housekeeping protein, β-actin protein) at 2 and 4 hours after formalin injection. n = 6 for each group. *P < 0.01 compared between F and N+F groups, compared between K+F and N+F groups.

• 

  Fig. 3 Photomicrographs (original magnification 40× objective lens) showing Fos-like immunoreactivity (FLI) expression on the ipsilateral side of spinal cord sections of control group (C) (A) and formalin injected group (F) (B). Arrow indicates the dark staining FLI.

• 

  Fig. 4 Photomicrographs (original magnification 40× objective lens) showing Prodynorphin-like immunoreactivity (PLI) expression on the ipsilateral side of spinal cord sections of the control group (C) (A) and formalin injected group (F) (B). Arrow indicates the dark staining PLI.

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