The Endolysosomal Transporter DMT1 is Required for Morphine Regulation of Neuronal Ferritin Heavy Chain.
The Endolysosomal Transporter DMT1 is Required for Morphine Regulation of Neuronal Ferritin Heavy Chain.
复制标题
DOI：
10.1007/s11481-023-10082-x
复制
发表时间：
2023-09
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
NeuroHIV和其他神经系统疾病表现为中枢神经系统神经元铁代谢的改变。许多艾滋病毒感染者还使用阿片类药物，这可能会通过进一步失调神经元铁代谢来恶化神经艾滋病毒症状。我们以前的工作表明，μ-阿片激动剂吗啡导致神经元内溶酶体释放其铁储存，神经元通过上调铁蛋白重链（FHC）（一种与neuroHIV认知障碍相关的铁储存蛋白）作出反应。在这里，我们研究了这个过程是否需要二价金属转运蛋白1（DMT 1），一个众所周知的铁转运蛋白表达的内溶酶体。我们首先优化条件以使用在HEK-293细胞中表达的荧光标记的大鼠DMT 1构建体检测DMT 1同种型（DMT 11B ±铁响应元件）。我们还表达了这些结构在原代大鼠皮层神经元比较其表达和亚细胞分布与内源性DMT 1亚型。我们在细胞质中发现了与溶酶体相关蛋白1（LAMP 1）共定位的内源性DMT 1亚型，这是一种内溶酶体的标志物。接下来，我们使用一种有效的DMT 1铁转运药理学抑制剂ebselen阻断内源性DMT 1亚型。依布硒啉预处理阻断了吗啡上调FHC蛋白的能力，表明该途径需要从内溶酶体转运DMT 1铁。这一点在皮质神经元中使用DMT 1 ±IRE的病毒介导的遗传沉默得到了进一步验证，这也阻断了吗啡存在下FHC的上调。总之，我们的工作表明，μ-阿片激动剂吗啡利用内溶酶体铁转运蛋白DMT 1来调节神经元细胞铁代谢，上调FHC蛋白，并导致neuroHIV的认知下降。吗啡需要DMT 1来上调神经元FHC。用吗啡处理的皮质神经元将其内溶酶体铁储存释放到细胞质并上调FHC，FHC是一种与神经HIV中树突棘缺陷和认知障碍相关的铁储存蛋白。该途径需要内溶酶体铁转运蛋白DMT 1，因为转运蛋白的药理学和遗传抑制剂完全阻断吗啡上调FHC的能力。使用BioRender.com创建。
NeuroHIV and other neurologic disorders present with altered iron metabolism in central nervous system neurons. Many people with HIV also use opioids, which can worsen neuroHIV symptoms by further dysregulating neuronal iron metabolism. Our previous work demonstrated that the μ-opioid agonist morphine causes neuronal endolysosomes to release their iron stores, and neurons respond by upregulating ferritin heavy chain (FHC), an iron storage protein associated with cognitive impairment in neuroHIV. Here, we investigated if this process required divalent metal transporter 1 (DMT1), a well-known iron transporter expressed on endolysosomes. We first optimized conditions to detect DMT1 isoforms (DMT1 1B ± iron responsive element) using fluorescently labeled rat DMT1 constructs expressed in HEK-293 cells. We also expressed these constructs in primary rat cortical neurons to compare their expression and subcellular distribution with endogenous DMT1 isoforms. We found endogenous DMT1 isoforms in the cytoplasm that colocalized with lysosomal-associated protein 1 (LAMP1), a marker of endolysosomes. Next, we blocked endogenous DMT1 isoforms using ebselen, a potent pharmacological inhibitor of DMT1 iron transport. Ebselen pre-treatment blocked morphine’s ability to upregulate FHC protein, suggesting this pathway requires DMT1 iron transport from endolysosomes. This was further validated using viral-mediated genetic silencing of DMT1±IRE in cortical neurons, which also blocked FHC upregulation in the presence of morphine. Overall, our work demonstrates that the μ-opioid agonist morphine utilizes the endolysosomal iron transporter DMT1 to modulate neuronal cellular iron metabolism, upregulate FHC protein, and contribute to cognitive decline in neuroHIV. Morphine requires DMT1 to upregulate neuronal FHC. Cortical neurons treated with morphine release their endolysosomal iron stores to the cytoplasm and upregulate FHC, an iron storage protein associated with dendritic spine deficits and cognitive impairment in neuroHIV. This pathway requires the endolysosomal iron transporter DMT1, as pharmacological and genetic inhibitors of the transporter completely block morphine’s ability to upregulate FHC. Created with BioRender.com.
影响因子：
6
作者：
通讯作者：
--