Korean J Pain.  2022 Jan;35(1):22-32. 10.3344/kjp.2022.35.1.22.

The role of basolateral amygdala orexin 1 receptors on the modulation of pain and psychosocial deficits in nitroglycerin-induced migraine model in adult male rats

• Askari-Zahabi K
• Abbasnejad M
• Kooshki R
• Raoof M
• Esmaeili-Mahani S
• Pourrahimi AM
• Zamyad M

• KMID: 2524643
• DOI: http://doi.org/10.3344/kjp.2022.35.1.22

Abstract

Background
Migraine headaches have been associated with sensory hyperactivity and anomalies in social/emotional responses. The main objective of this study was to evaluate the potential involvement of orexin 1 receptors (Orx1R) within the basolateral amygdala (BLA) in the modulation of pain and psychosocial dysfunction in a nitroglycerin (NTG)-induced rat model of migraine.
Methods
Adult male Wistar rats were injected with NTG (5 mg/kg, intraperitoneal) every second day over nine days to induce migraine. The experiments were done in the following six groups (6 rats per group): untreated control, NTG, NTG plus vehicle, and NTG groups that were post-treated with intra-BLA microinjection of Orx1R antagonist SB-334867 (10, 20, and 40 nM). Thermal hyperalgesia was assessed using the hot plate and tail-flick tests. Moreover, the elevated plus maze (EPM) and open field (OF) tests were used to assess anxiety-like behaviors. The animals’ sociability was evaluated using the three-chamber social task. The NTG-induced photophobia was assessed using a light-dark box.
Results
We observed no change in NTG-induced thermal hyperalgesia following administration of SB-334867 (10, 20, and 40 nM). However, SB-334867 (20 and 40 nM) aggravated the NTG-induced anxiogenic responses in both the EPM and OF tasks. The NTG-induced social impairment was overpowered by SB-334867 at all doses. Time spent in the dark chamber of light-dark box was significantly increased in rats treated with SB-334867 (20 and 40 nM/rat).
Conclusions
The findings suggest a role for Orx1R within the BLA in control comorbid affective complaints with migraine in rats.

Keyword

Amygdala; Anxiety; Basolateral Nuclear Complex; Hyperalgesia; Migraine Disorders; Nitroglycerin; Orexins; Orexin Receptors; Orexin Receptor Antagonists; Rats

Figure

• 

  Fig. 1 Stereotaxic surgical procedure for guide cannula implantation in the basolateral amygdala (BLA) (A-C), a typical section of BLA taken from the Paxinos and Watson rat brain atlas and a representative photographs exhibiting appropriate cannula implantation into the BLA region (D).

• 

  Fig. 2 The flow chart of study recruitment. BLA: basolateral amygdala, NTG: nitroglycerin, DMSO: dimethylsulfoxide, EPM: elevated plus maze, OF: open field.

• 

  Fig. 3 The hot plate (A) and tail flick (B) latencies among the different experimental groups (n = 6 per group). Graphs represent the mean ± standard error of the mean. NTG: nitroglycerin, SB: SB-334867. **P < 0.01 vs. control group, ***P < 0.001.

• 

  Fig. 4 The time spent (A) and the number of entries (B) into the open arms of the elevated plus-maze during a 5 minutes test in nitroglycerin (NTG)-treated rats and NTG groups received intra-basolateral amygdala infusion of SB-334867 (10, 20, and 40 nM/rat) (n = 6 per group). Graphs represent the mean ± standard error of the mean. *P < 0.05, **P < 0.01, ***P < 0.001 vs. control group, #P < 0.05 vs. NTG.

• 

  Fig. 5 The time spent (A) and the number of entries (B) in the central zone of the open filed during a 5 minutes test in nitroglycerin (NTG)-treated rats and NTG groups that received intra-basolateral amygdala infusion of SB-334867 (10, 20, and 40 nM/rat). n = 6 per experimental group. Graphs represent the mean ± standard error of the mean. *P < 0.05, **P < 0.01 vs. control group, #P < 0.05 vs. NTG.

• 

  Fig. 6 Assessment of light aversion behavior in the light dark box test. The data in each graph are represented as mean ± standard error of the mean values (n = 6). NTG: nitroglycerin, DMSO: dimethylsulfoxide, SB: SB-334867. ***P < 0.001 vs. control group, ###P < 0.001 vs. NTG group.

• 

  Fig. 7 Sociability in the three-chambered task. The time spent in the chamber containing stranger 1 (A) and stranger 2 (B), and the time spent in close proximity with the cage containing stranger 1 (C). Social discrimination (D). n = 6 per experimental group. Bar graphs represent mean ± standard error of the mean. NTG: nitroglycerin, SB: SB-334867. *P < 0.05, **P < 0.01, and ***P < 0.001 vs. control, #P < 0.05 NTG and NTG plus vehicle.

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