Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan.
Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan
Mol Pharmacol. 2022 May;101(5):371-380. doi: 10.1124/molpharm.121.000377. Epub 2022 Mar 2.
The synthetic cannabinoid WIN55,212-2 (WIN) is widely used as a pharmacological tool to study the biologic activity of cannabinoid receptors. In contrast to many other cannabinoid agonists, however, WIN also causes broad effects outside of neurons, such as reducing inflammatory responses, causing cell cycle arrest, and reducing general protein expression. How exactly WIN causes these broad effects is not known. Here we show that WIN partially disrupts the Golgi apparatus at nanomolar concentrations and fully disperses the Golgi apparatus in neuronal and non-neuronal cells at micromolar concentrations. WIN55,212-3, the enantiomer of WIN; JWH-018, a related alkylindole; or 2-arachidonoylglycerol, an endocannabinoid, did not cause Golgi disruption, suggesting that the effect was specific to the chirality of WIN. WIN treatment also perturbed the microtubule network. Importantly, WIN disrupted the Golgi in primary cortical neurons derived from mice where cannabinoid receptor-1 (CB1) was genetically knocked out, indicating that the effects were independent of CB1 signaling. The Golgi dispersion could not be explained by WIN's action on peroxisome proliferator-activated receptors. Our results show that WIN can disrupt the Golgi apparatus independent of CB1 in cultured cells. These effects could contribute to the unique physiologic effects that WIN exhibits in neuronal behavior, as well as its role as an antiproliferative and anti-inflammatory agent. SIGNIFICANCE STATEMENT: The synthetic cannabinoid WIN55,212-2 (WIN), widely used to investigate the cannabinoid system, also shows unique broader effects at cellular and organismal levels compared to endogenous cannabinoids. Our study shows that WIN can disrupt the Golgi apparatus and the microtubule network in multiple cell types, independent of cannabinoid receptors. These results could explain how WIN reduces surface levels of proteins and contributes to the unique physiological effects observed with WIN.
合成大麻素 WIN55,212-2（WIN）被广泛用作研究大麻素受体生物活性的药理学工具。然而，与许多其他大麻素激动剂不同，WIN 还会引起神经元以外的广泛作用，例如减少炎症反应、引起细胞周期停滞和减少总蛋白表达。WIN 如何引起这些广泛的作用尚不清楚。在这里，我们发现 WIN 在纳摩尔浓度下部分破坏高尔基体，在微摩尔浓度下完全分散神经元和非神经元细胞中的高尔基体。WIN 的对映异构体 WIN55,212-3、相关的烷基吲哚 JWH-018 或内源性大麻素 2-花生四烯酸甘油酯均不会引起高尔基体解体，表明这种作用是 WIN 手性特异性的。WIN 处理还扰乱了微管网络。重要的是，WIN 在从基因敲除大麻素受体 1（CB1）的小鼠中分离的原代皮质神经元中破坏了高尔基体，表明这些作用独立于 CB1 信号。WIN 对过氧化物酶体增殖物激活受体的作用不能解释高尔基体的分散。我们的结果表明，WIN 可以在培养细胞中独立于 CB1 破坏高尔基体。这些影响可能有助于 WIN 在神经元行为中表现出的独特生理效应，以及其作为抗增殖和抗炎剂的作用。
合成大麻素 WIN55,212-2（WIN）被广泛用于研究大麻素系统，与内源性大麻素相比，它在细胞和机体水平上也表现出独特的更广泛的作用。我们的研究表明，WIN 可以在多种细胞类型中破坏高尔基体和微管网络，而不依赖于大麻素受体。这些结果可以解释 WIN 如何降低蛋白质的表面水平，并有助于解释与 WIN 相关的独特生理效应。
