TY - JOUR TI - Endocytic trafficking determines cellular tolerance of presynaptic opioid signaling AU - Jullié, Damien AU - Benitez, Camila AU - Knight, Tracy A AU - Simic, Milos S AU - von Zastrow, Mark A2 - Shen, Kang A2 - Taffe, Michael A A2 - Williams, John T A2 - Banghart, Matthew Ryan A2 - Ingram, Susan L VL - 11 PY - 2022 DA - 2022/11/15 SP - e81298 C1 - eLife 2022;11:e81298 DO - 10.7554/eLife.81298 UR - https://doi.org/10.7554/eLife.81298 AB - Opioid tolerance is well-described physiologically but its mechanistic basis remains incompletely understood. An important site of opioid action in vivo is the presynaptic terminal, where opioids inhibit transmitter release. This response characteristically resists desensitization over minutes yet becomes gradually tolerant over hours, and how this is possible remains unknown. Here, we delineate a cellular mechanism underlying this longer-term form of opioid tolerance in cultured rat medium spiny neurons. Our results support a model in which presynaptic tolerance is mediated by a gradual depletion of cognate receptors from the axon surface through iterative rounds of receptor endocytosis and recycling. For the μ-opioid receptor (MOR), we show that the agonist-induced endocytic process which initiates iterative receptor cycling requires GRK2/3-mediated phosphorylation of the receptor’s cytoplasmic tail, and that partial or biased agonist drugs with reduced ability to drive phosphorylation-dependent endocytosis in terminals produce correspondingly less presynaptic tolerance. We then show that the δ-opioid receptor (DOR) conforms to the same general paradigm except that presynaptic endocytosis of DOR, in contrast to MOR, does not require phosphorylation of the receptor’s cytoplasmic tail. Further, we show that DOR recycles less efficiently than MOR in axons and, consistent with this, that DOR tolerance develops more strongly. Together, these results delineate a cellular basis for the development of presynaptic tolerance to opioids and describe a methodology useful for investigating presynaptic neuromodulation more broadly. KW - GPCR KW - opioid KW - endocytosis KW - axon KW - neuron KW - tolerance JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -