Abstract
Stopping opioid medications can result in a debilitating withdrawal syndrome in chronic users. Opioid withdrawal can occur at all ages, but mechanistic understanding of this condition is predominantly derived from adult studies. Here, we examined whether there are age-dependent differences in the behavioural phenotype and cellular indices of opioid withdrawal. We tested this by assessing the behavioural and cFos response (a surrogate marker for neuronal activation) to morphine withdrawal in C57BL/6J mice across key developmental stages—neonatal, adolescent, and adulthood. Mice in all age groups received escalating doses of morphine (10–50 mg/kg) over 5 days and withdrawal was precipitated by a single injection of the opioid receptor antagonist naloxone (2 mg/kg) two hours after the last morphine dose. In adult and adolescent mice, withdrawal behaviours were robust, with age-related differences in autonomic and somatic signs. In both groups, cFos expression was increased in spinally projecting neurons within the Periaqueductal Grey (PAG), Rostro-ventromedial Medulla (RVM), and Locus Coeruleus. Neonatal animals displayed both a distinct behavioural withdrawal and cFos expression profile. Notably, in young animals cFos expression was increased within the PAG and LC, but decreased in the RVM. In summary, naloxone challenge precipitated robust opioid withdrawal behaviours across all developmental stages with neonatal animals displaying differences in withdrawal behaviours and unique neuronal activation patterns within key brainstem regions.
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Datasets and materials generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We would like to acknowledge Dr. Churmy Fan for her assistance with illustrations.
Funding
This work was supported by the Vi Riddell Program for Pediatric Pain, and grants from the Natural Sciences and Engineering Research Council of Canada (RGPIN06289-2019) and Canadian Institutes of Health Research (PJT-162271; PJ8-169697) awarded to T.T. N.v.d.H. is supported by a Harley Hotchkiss-Samuel Weiss Postdoctoral Fellowship and C.H.T.K. by a Postdoctoral Fellowship from the Canadian Institutes of Health Research.
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by NvdH and CHTK. The first draft of the manuscript was written by NvdH and CHTK and all authors commented on previous versions of the manuscript. TT supervised the experiments. All authors read and approved the final manuscript.
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All experiments were approved by the University of Calgary Animal Care Committees and all animals were under veterinary supervision in accordance with the Canadian Council on Animal Care. Health Canada exemption for use of a controlled substance and biosafety permits were obtained.
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van den Hoogen, N.J., Kwok, C.H.T. & Trang, T. Identifying the Neurodevelopmental Differences of Opioid Withdrawal. Cell Mol Neurobiol 41, 1145–1155 (2021). https://doi.org/10.1007/s10571-020-01035-6
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DOI: https://doi.org/10.1007/s10571-020-01035-6