Elsevier

Neuropharmacology

Volume 46, Issue 6, May 2004, Pages 824-835
Neuropharmacology

Naloxone-induced morphine withdrawal increases the number and degranulation of mast cells in the thalamus of the mouse

https://doi.org/10.1016/j.neuropharm.2003.11.022Get rights and content

Abstract

Naloxone-induced jumping in morphine-dependent mice is inhibited by cromolyn, a mast cell stabilizer, suggesting that this characteristic withdrawal behavior results from degranulation of mast cells. Because withdrawal is considered as a central phenomenon, degranulation of mast cells located within the CNS may influence aspects of opioid withdrawal. The present study evaluates histologically whether naloxone, injected into opioid dependent mice, induces degranulation of mast cells. Seventy-two hours after the s.c. implantation of a 75 mg morphine pellet, the number and degranulation of thalamic mast cells did not differ from those in placebo-implanted controls. However, two injections of 50 mg/kg of naloxone, 30 and 60 min before tissue collection, increased the number of degranulated mast cells compared to those in mice injected with saline. Analysis throughout the entire thalamus (90 40-μ sections) revealed increases in the total number of mast cells as well as the number that were degranulated, especially in sections 52–60, corresponding to Bregma −2.18 to 2.54. Here, mast cells were clustered in the IGL and VPL/VPM nuclei, and redistributed from the ventromedial to the dorsolateral aspects of the Po and PF nuclei during withdrawal. Degranulation was also greater throughout the LD, LP nuclei during withdrawal. These data reveal a novel neuroimmune reaction to opioid withdrawal in the CNS.

Introduction

Mast cells are derived from pluripotent progenitor stem cells in the bone marrow (Kitamura et al., 1979). They are present in the peripheral and central nervous system (CNS) of several species, including rodents (Dropp, 1972). In the brain, mast cells are predominantly located in the thalamus in the rat (Florenzano and Bentivoglio, 2000) and mouse (Yang et al., 1999). Here, and elsewhere in the body, they are closely apposed to neurons (Manning et al., 1994) and blood vessels (Dropp, 1972, Dropp, 1976, Yang et al., 1999, Florenzano and Bentivoglio, 2000), raising the possibility that they influence vascular and neuronal function.

Intracranial mast cell populations fluctuate under normal physiological and pathological conditions. The number of mast cells in the brain of rodents decreases in response to immobilization stress or handling (Persinger, 1980, Theoharides et al., 1995), but increases in response to chronic subordination stress (Cirulli et al., 1998). In addition, the population of mast cells in the brain can increase under pathological conditions. These include conditions such as autoimmune demyelination (Bo et al., 1991), multiple sclerosis (Theoharides, 1996, Secor et al., 2000), and thiamine deficiency (Ferguson et al., 1999). Thus, mast cells may represent an important effector cell in normal physiology as well as pathophysiology of brain lesions and disorders (Theoharides, 1996, Dines and Powell, 1997).

When mast cells are activated, they release a broad array of substances from their granules in a process called degranulation. Substances liberated include, among others, bioactive amines (histamine and serotonin), nerve growth factor (NGF), cytokines and chemokines, and free radicals (see review by Galli, 1993, Cocchiara et al., 1999). Several stimuli are capable of degranulating mast cells, including opioids (Casale et al., 1984). This action is not due to opioid activity at a traditional mu-, delta- or kappa-type opioid receptor, but perhaps merely due to the strong positive charge, such as that on compound 48/80, substance P and dynorphin, compounds that are also potent degranulators of mast cells.

Degranulation of mast cells can be prevented by “mast cell stabilizers”, such as sodium cromoglycate (cromolyn). Cromolyn is not only used therapeutically for conditions of hypersensitivity and mastocytosis, but also to assess experimentally the role of mast cells in various diseases. For example, the amelioration by cromolyn of experimental allergic encephalomyelitis (EAE), an animal model of multiple sclerosis (Dietsch and Hinrichs, 1989), supports the hypothesis that this condition depends, in part, on mast cell degranulation (Brenner et al., 1994). Used in this fashion, it was discovered that sodium cromoglycate inhibits naloxone-induced withdrawal jumping behavior in morphine-dependent mice (Leza et al., 1992), thereby suggesting that this characteristic symptom of morphine withdrawal depends on mast cell degranulation. Because opioid dependence and withdrawal are thought to be centrally mediated phenomena, this raises the possibility that mast cells in the thalamus play a role in some aspects of opioid withdrawal. We sought to verify the effect of morphine withdrawal on thalamic mast cells by a direct histological analysis of their number and state of degranulation in brain sections from mice during withdrawal. We precipitated withdrawal by injecting a large dose of naloxone in mice that were made highly dependent on morphine by the implantation of a morphine pellet.

Section snippets

Animals

Adult male Swiss-Webster mice weighing 20–30 g (Harlan, Indianapolis, IN) were housed 4 per cage and allowed to acclimate for at least 1 week prior to use. Mice were allowed free access to food and water, and housed in a room with a constant temperature of 23 °C on a 12-h light:12-h dark cycle. Animals were used strictly in accordance with the Guidelines of IASP and the University of Minnesota Animal Care and Use Committee, and those prepared by the Committee on Care and Use of Laboratory

Distribution of mast cells in unmanipulated mice

In naïve mice, many metachromatically stained granulated cells but few degranulated cells were seen, distributed unevenly in the thalamic parenchyma (Fig. 1, Fig. 2). Consistent with the literature, mast cells were typically in close proximity to blood vessels. There were few mast cells in the anterior section of the thalamus (mean±SEM=2.3±1.1, n=15) compared to the more numerous cells located in the middle (27.9±6.9) and posterior regions (25.2±10.0) (Fig. 2A). The distribution of mast cells

Discussion

Naloxone-induced morphine withdrawal has a distinct influence on the number and granulation of mast cells in specific regions of the mouse thalamus. This is consistent with the ability of cromolyn, a mast cell stabilizer, to inhibit naloxone-induced withdrawal jumping in a dose-dependent fashion in mice (Leza et al., 1992). Morphine alone, administered either acutely or chronically, had no influence on the total number of thalamic mast cells and their state of granulation. This allows us to

Acknowledgements

This work was supported by the National Institutes of Health grants DA07234 (O.B.T) from the National Institute on Drug Abuse and NS39740 (A.A.L.) funded by the National Institute of Neurological Disorders and Stroke and the National Institutes on Arthritis and Musculoskeletal and Skin Diseases. The authors wish to thank Christopher Hall and Xin Ge for their excellent technical assistance and Dr. Jian Ding for his helpful editorial comments.

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