• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

利用鼠角膜活检进行实时评估线粒体呼吸的方法。

A Method for Real-Time Assessment of Mitochondrial Respiration Using Murine Corneal Biopsy.

机构信息

Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States.

Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, North Carolina, United States.

出版信息

Invest Ophthalmol Vis Sci. 2023 Aug 1;64(11):33. doi: 10.1167/iovs.64.11.33.

DOI:10.1167/iovs.64.11.33
PMID:37642632
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10476441/
Abstract

PURPOSE

To develop and optimize a method to monitor real-time mitochondrial function by measuring the oxygen consumption rate (OCR) in murine corneal biopsy punches with a Seahorse extracellular flux analyzer.

METHODS

Murine corneal biopsies were obtained using a biopsy punch immediately after euthanasia. The corneal metabolic profile was assessed using a Seahorse XFe96 pro analyzer, and mitochondrial respiration was analyzed with specific settings.

RESULTS

Real-time adenosine triphosphate rate assay showed that mitochondrial oxidative phosphorylation is a major source of adenosine triphosphate production in ex vivo live murine corneal biopsies. Euthanasia methods (carbon dioxide asphyxiation vs. overdosing on anesthetic drugs) did not affect corneal OCR values. Mouse corneal biopsy punches in 1.5-mm diameter generated higher and more reproducible OCR values than those in 1.0-mm diameter. The biopsy punches from the central and off-central cornea did not show significant differences in OCR values. There was no difference in OCR reading by the tissue orientations (the epithelium side up vs. the endothelium side up). No significant differences were found in corneal OCR levels between sexes, strains (C57BL/6J vs. BALB/cJ), or ages (4, 8, and 32 weeks). Using this method, we showed that the wound healing process in the mouse cornea affected mitochondrial activity.

CONCLUSIONS

The present study validated a new strategy to measure real-time mitochondrial function in fresh mouse corneal tissues. This procedure should be helpful for studies of the ex vivo live corneal metabolism in response to genetic manipulations, disease conditions, or pharmacological treatments in mouse models.

摘要

目的

开发和优化一种通过 Seahorse 细胞外通量分析仪测量鼠角膜活检穿孔的耗氧量（OCR）来实时监测线粒体功能的方法。

方法

在安乐死后立即使用活检穿孔器获取鼠角膜活检。使用 Seahorse XFe96 pro 分析仪评估角膜代谢谱，并采用特定设置分析线粒体呼吸。

结果

实时三磷酸腺苷率测定表明，线粒体氧化磷酸化是体外活体鼠角膜活检中三磷酸腺苷产生的主要来源。安乐死方法（二氧化碳窒息与麻醉药物过量）不影响角膜 OCR 值。直径为 1.5 毫米的鼠角膜活检穿孔产生的 OCR 值更高且更具重现性，而直径为 1.0 毫米的穿孔则较低且重现性较差。中央和非中央角膜的活检穿孔在 OCR 值方面没有显著差异。组织方向（上皮面朝上与内皮面朝上）对 OCR 读数没有影响。性别、品系（C57BL/6J 与 BALB/cJ）或年龄（4、8 和 32 周）之间的角膜 OCR 水平无差异。使用该方法，我们表明小鼠角膜的愈合过程会影响线粒体活性。

结论

本研究验证了一种测量新鲜鼠角膜组织实时线粒体功能的新策略。该程序对于研究体外活体角膜代谢对遗传操作、疾病状况或药物治疗的反应在小鼠模型中的作用应该是有帮助的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/410c1682e9d5/iovs-64-11-33-f007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/b2f8b2a7e59a/iovs-64-11-33-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/8ec29a94b05b/iovs-64-11-33-f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/7bf395141b44/iovs-64-11-33-f003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/6a46f5b3f5d2/iovs-64-11-33-f004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/428c064e4222/iovs-64-11-33-f005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/de67703d6ba9/iovs-64-11-33-f006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/410c1682e9d5/iovs-64-11-33-f007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/b2f8b2a7e59a/iovs-64-11-33-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/8ec29a94b05b/iovs-64-11-33-f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/7bf395141b44/iovs-64-11-33-f003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/6a46f5b3f5d2/iovs-64-11-33-f004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/428c064e4222/iovs-64-11-33-f005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/de67703d6ba9/iovs-64-11-33-f006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cf9/10476441/410c1682e9d5/iovs-64-11-33-f007.jpg

相似文献

1
A Method for Real-Time Assessment of Mitochondrial Respiration Using Murine Corneal Biopsy.利用鼠角膜活检进行实时评估线粒体呼吸的方法。
Invest Ophthalmol Vis Sci. 2023 Aug 1;64(11):33. doi: 10.1167/iovs.64.11.33.
2
Peroxisome proliferator-activated receptor-α (PPARα) regulates wound healing and mitochondrial metabolism in the cornea.过氧化物酶体增殖物激活受体-α（PPARα）调节角膜的伤口愈合和线粒体代谢。
Proc Natl Acad Sci U S A. 2023 Mar 28;120(13):e2217576120. doi: 10.1073/pnas.2217576120. Epub 2023 Mar 21.
3
Monitoring Mitochondrial Respiration in Mouse Cerebellar Granule Neurons.监测小鼠小脑颗粒神经元中的线粒体呼吸。
Methods Mol Biol. 2022;2515:1-15. doi: 10.1007/978-1-0716-2409-8_1.
4
Real-Time Assessment of Mitochondrial Function in Cytotrophoblast and Syncytialized Trophoblast Cells Using the Seahorse XFe24 Extracellular Flux Analyzer.使用 Seahorse XFe24 细胞外通量分析仪实时评估滋养细胞和合体滋养层细胞中的线粒体功能。
Methods Mol Biol. 2024;2728:137-147. doi: 10.1007/978-1-0716-3495-0_12.
5
Quantification of Oxygen Consumption in Retina Ex Vivo Demonstrates Limited Reserve Capacity of Photoreceptor Mitochondria.视网膜离体耗氧量的定量分析表明光感受器线粒体的储备能力有限。
Invest Ophthalmol Vis Sci. 2015 Dec;56(13):8428-36. doi: 10.1167/iovs.15-17901.
6
Short Communication: Beta-adrenergic agonists alter oxidative phosphorylation in primary myoblasts.短篇通讯：β-肾上腺素激动剂改变原代成肌细胞的氧化磷酸化。
J Anim Sci. 2022 Aug 1;100(8). doi: 10.1093/jas/skac208.
7
A novel high-throughput assay for respiration in isolated brain microvessels reveals impaired mitochondrial function in the aged mice.一种新型高通量检测脑微血管细胞耗氧率的方法揭示了衰老小鼠中线粒体功能受损。
Geroscience. 2018 Aug;40(4):365-375. doi: 10.1007/s11357-018-0037-8. Epub 2018 Aug 3.
8
Real-Time Assessment of Mitochondrial Toxicity in HepG2 Cells Using the Seahorse Extracellular Flux Analyzer.使用 Seahorse 细胞外通量分析仪实时评估 HepG2 细胞中的线粒体毒性。
Curr Protoc. 2021 Mar;1(3):e75. doi: 10.1002/cpz1.75.
9
Live Metabolic Profile Analysis of Zebrafish Embryos Using a Seahorse XF 24 Extracellular Flux Analyzer.使用海马XF 24细胞外通量分析仪对斑马鱼胚胎进行实时代谢谱分析。
Methods Mol Biol. 2018;1797:393-401. doi: 10.1007/978-1-4939-7883-0_21.
10
Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism.测量培养细胞中的氧消耗率（OCR）和细胞外酸化率（ECAR）以评估能量代谢。
Bio Protoc. 2018 May 20;8(10):e2850. doi: 10.21769/BioProtoc.2850.

引用本文的文献

1
Exploring the Therapeutic Potential of Salivary Exosomes in Corneal Epithelial Wound Healing.探索唾液外泌体在角膜上皮伤口愈合中的治疗潜力。
Invest Ophthalmol Vis Sci. 2025 Aug 1;66(11):8. doi: 10.1167/iovs.66.11.8.
2
Corneal epithelial cells upregulate macropinocytosis to engulf metabolically active axonal mitochondria released by injured axons.角膜上皮细胞上调巨胞饮作用，以吞噬受损轴突释放的具有代谢活性的轴突线粒体。
Ocul Surf. 2025 Jul;37:173-188. doi: 10.1016/j.jtos.2025.03.007. Epub 2025 Apr 1.
3
Mitochondria Transplantation Promotes Corneal Epithelial Wound Healing.

本文引用的文献

1
Peroxisome proliferator-activated receptor-α (PPARα) regulates wound healing and mitochondrial metabolism in the cornea.过氧化物酶体增殖物激活受体-α（PPARα）调节角膜的伤口愈合和线粒体代谢。
Proc Natl Acad Sci U S A. 2023 Mar 28;120(13):e2217576120. doi: 10.1073/pnas.2217576120. Epub 2023 Mar 21.
2
Monocarboxylate Transporters: Role and Regulation in Corneal Diabetes.单羧酸转运蛋白：在角膜糖尿病中的作用和调控。
Anal Cell Pathol (Amst). 2022 Oct 26;2022:6718566. doi: 10.1155/2022/6718566. eCollection 2022.
3
Prolactin-Induced Protein facilitates corneal wound healing.
线粒体移植促进角膜上皮伤口愈合。
Invest Ophthalmol Vis Sci. 2024 Jun 3;65(6):14. doi: 10.1167/iovs.65.6.14.
催乳素诱导蛋白促进角膜伤口愈合。
Exp Eye Res. 2022 Dec;225:109300. doi: 10.1016/j.exer.2022.109300. Epub 2022 Oct 31.
4
Pathogenic Role of Diabetes-Induced Overexpression of Kallistatin in Corneal Wound Healing Deficiency Through Inhibition of Canonical Wnt Signaling.糖尿病诱导 kallistatin 过表达通过抑制经典 Wnt 信号在角膜愈合缺陷中的致病作用。
Diabetes. 2022 Apr 1;71(4):747-761. doi: 10.2337/db21-0740.
5
Cisd2 plays an essential role in corneal epithelial regeneration.Cisd2 在角膜上皮再生中发挥重要作用。
EBioMedicine. 2021 Nov;73:103654. doi: 10.1016/j.ebiom.2021.103654. Epub 2021 Oct 29.
6
Pathogenic mitochondrial dysfunction and metabolic abnormalities.致病的线粒体功能障碍和代谢异常。
Biochem Pharmacol. 2021 Nov;193:114809. doi: 10.1016/j.bcp.2021.114809. Epub 2021 Oct 19.
7
Mitochondrial Dysfunction and Mitophagy in Fuchs Endothelial Corneal Dystrophy.线粒体功能障碍与 Fuchs 内皮角膜营养不良中的自噬
Cells. 2021 Jul 26;10(8):1888. doi: 10.3390/cells10081888.
8
Measurement of mitochondrial respiration in the murine retina using a Seahorse extracellular flux analyzer.使用 Seahorse 细胞外通量分析仪测量鼠视网膜中的线粒体呼吸。
STAR Protoc. 2021 May 12;2(2):100533. doi: 10.1016/j.xpro.2021.100533. eCollection 2021 Jun 18.
9
Impairment of corneal epithelial wound healing is association with increased neutrophil infiltration and reactive oxygen species activation in tenascin X-deficient mice.角膜上皮伤口愈合受损与 tenascin X 缺陷小鼠中性粒细胞浸润和活性氧物种激活增加有关。
Lab Invest. 2021 Jun;101(6):690-700. doi: 10.1038/s41374-021-00576-8. Epub 2021 Mar 29.
10
Mitochondrial DNA lesions and copy number are strain dependent in endurance-trained mice.线粒体 DNA 损伤和拷贝数在经过耐力训练的小鼠中依赖于品系。
Physiol Rep. 2020 Nov;8(21):e14605. doi: 10.14814/phy2.14605.