Anesthesia-Induced Neuronal Apoptosis in the Developing Retina: A Window of Opportunity.
Anesthesia-Induced Neuronal Apoptosis in the Developing Retina: A Window of Opportunity.
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DOI：
10.1213/ane.0000000000000714
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发表时间：
2015-11
影响因子：
5.7
通讯作者：
Levy RJ
中科院分区：
文献类型：
--
作者：
Cheng Y;He L;Prasad V;Wang S;Levy RJ
麻醉剂在发育中的大脑中引起广泛的细胞凋亡，导致神经认知异常。然而，目前尚不清楚麻醉诱导的神经毒性是否发生在人体中，因为目前还没有评估体内神经元凋亡的方法。视网膜的独特之处在于它是中枢神经系统中唯一可以无创直接可视化的部分。因此，我们的目的是确定是否异氟烷诱导细胞凋亡的发展中的视网膜。CD-1雄性小鼠幼仔暴露于异氟烷（2%）或空气中1小时。暴露后，定量视网膜中活化的半胱天冬酶-3、半胱天冬酶-9和半胱天冬酶-8，评估视网膜线粒体释放的细胞色素c，并鉴定发生细胞凋亡的细胞的数量和类型。视网膜摄取和荧光标记的膜联蛋白V的能力，以结合到经历自然细胞死亡和麻醉诱导的细胞凋亡的视网膜后，确定全身注射。异氟烷激活内核层（INL）的内源性凋亡途径，并激活神经节细胞层的内源性和外源性途径。免疫荧光显示，双极和无长突神经元内的INL进行生理性细胞死亡，在两个队列和无长突细胞的异氟烷诱导的细胞凋亡的可能目标。注射后，荧光标记的膜联蛋白V在空气暴露和异氟烷暴露小鼠的INL中很容易检测到，并与活化的caspase-3阳性细胞共定位。这些研究结果表明，异氟醚诱导的神经元凋亡发生在发育中的视网膜，并奠定了基础的非侵入性成像技术的发展，以检测麻醉诱导的神经毒性的婴儿和儿童。
Anesthetics cause widespread apoptosis in the developing brain, resulting in neurocognitive abnormalities. However, it is unknown whether anesthesia-induced neurotoxicity occurs in humans because there is currently no modality to assess for neuronal apoptosis in vivo. The retina is unique in that it is the only portion of the central nervous system that can be directly visualized noninvasively. Thus, we aimed to determine whether isoflurane induces apoptosis in the developing retina. CD-1 male mouse pups underwent 1-hour exposure to isoflurane (2%) or air. After exposure, activated caspase-3, caspase-9, and caspase-8 were quantified in the retina, cytochrome c release from retinal mitochondria was assessed, and the number and types of cells undergoing apoptosis were identified. Retinal uptake and the ability of fluorescent-labeled annexin V to bind to cells undergoing natural cell death and anesthesia-induced apoptosis in the retina were determined after systemic injection. Isoflurane activated the intrinsic apoptosis pathway in the inner nuclear layer (INL) and activated both the intrinsic and extrinsic pathways in the ganglion cell layer. Immunofluorescence demonstrated that bipolar and amacrine neurons within the INL underwent physiologic cell death in both cohorts and that amacrine cells were the likely targets of isoflurane-induced apoptosis. After injection, fluorescent-labeled annexin V was readily detected in the INL of both air-exposed and isoflurane-exposed mice and colocalized with activated caspase-3-positive cells. These findings indicate that isoflurane-induced neuronal apoptosis occurs in the developing retina and lays the groundwork for development of a noninvasive imaging technique to detect anesthesia-induced neurotoxicity in infants and children.