• 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

用于完整大脑中PTEN动力学荧光寿命成像的基因编码生物传感器。

Genetically encoded biosensor for fluorescence lifetime imaging of PTEN dynamics in the intact brain.

作者信息

Kagan Tomer, Gabay Matan, Meenakshisundaram Aasha, Levi Yossi, Eid Sharbel, Malchenko Nikol, Maman Maya, Nitzan Anat, Ravotto Luca, Zaidel-Bar Ronen, Eickholt Britta Johanna, Gal Maayan, Laviv Tal

机构信息

Department of Physiology and Pharmacology, Faculty of Medical and Health Sciences, Tel Aviv University, Tel Aviv, Israel.

Department of Oral Biology, The Goldschleger School of Dental Medicine, Faculty of Medical and Health Sciences, Tel Aviv University, Tel Aviv, Israel.

出版信息

Nat Methods. 2025 Apr;22(4):764-777. doi: 10.1038/s41592-025-02610-9. Epub 2025 Feb 20.

DOI:10.1038/s41592-025-02610-9
PMID:39979596
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11978514/
Abstract

The phosphatase and tensin homolog (PTEN) is a vital protein that maintains an inhibitory brake for cellular proliferation and growth. Accordingly, PTEN loss-of-function mutations are associated with a broad spectrum of human pathologies. Despite its importance, there is currently no method to directly monitor PTEN activity with cellular specificity within intact biological systems. Here we describe the development of a FRET-based biosensor using PTEN conformation as a proxy for the PTEN activity state, for two-photon fluorescence lifetime imaging microscopy. We identify a point mutation that allows the monitoring of PTEN activity with minimal interference to endogenous PTEN signaling. We demonstrate imaging of PTEN activity in cell lines, intact Caenorhabditis elegans and in the mouse brain. Finally, we develop a red-shifted sensor variant that allows us to identify cell-type-specific PTEN activity in excitatory and inhibitory cortical cells. In summary, our approach enables dynamic imaging of PTEN activity in vivo with unprecedented spatial and temporal resolution.

摘要

磷酸酶和张力蛋白同源物（PTEN）是一种重要的蛋白质，它对细胞增殖和生长起着抑制作用。因此，PTEN功能丧失突变与多种人类疾病相关。尽管其很重要，但目前尚无方法在完整的生物系统中以细胞特异性直接监测PTEN活性。在此，我们描述了一种基于荧光共振能量转移（FRET）的生物传感器的开发，该传感器利用PTEN构象作为PTEN活性状态的替代指标，用于双光子荧光寿命成像显微镜。我们鉴定出一个点突变，可在对内源性PTEN信号干扰最小的情况下监测PTEN活性。我们展示了在细胞系、完整的秀丽隐杆线虫和小鼠大脑中对PTEN活性的成像。最后，我们开发了一种红移传感器变体，使我们能够在兴奋性和抑制性皮质细胞中识别细胞类型特异性的PTEN活性。总之，我们的方法能够以前所未有的空间和时间分辨率在体内动态成像PTEN活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/008272734737/41592_2025_2610_Fig16_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/0fe82edb1eed/41592_2025_2610_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/ff23954c3f7d/41592_2025_2610_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/78832ca3d3e2/41592_2025_2610_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/9d6dc34f5765/41592_2025_2610_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/d3d3b3e81386/41592_2025_2610_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/4ad5d5688619/41592_2025_2610_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/35b143a679a7/41592_2025_2610_Fig7_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/98f461d50358/41592_2025_2610_Fig8_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/35913bbfac1d/41592_2025_2610_Fig9_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/a8c02ac5bc53/41592_2025_2610_Fig10_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/08c7bdf26d16/41592_2025_2610_Fig11_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/8f435843085d/41592_2025_2610_Fig12_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/f230e696cf55/41592_2025_2610_Fig13_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/2c66fa735a66/41592_2025_2610_Fig14_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/e10ed2f89746/41592_2025_2610_Fig15_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/008272734737/41592_2025_2610_Fig16_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/0fe82edb1eed/41592_2025_2610_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/ff23954c3f7d/41592_2025_2610_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/78832ca3d3e2/41592_2025_2610_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/9d6dc34f5765/41592_2025_2610_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/d3d3b3e81386/41592_2025_2610_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/4ad5d5688619/41592_2025_2610_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/35b143a679a7/41592_2025_2610_Fig7_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/98f461d50358/41592_2025_2610_Fig8_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/35913bbfac1d/41592_2025_2610_Fig9_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/a8c02ac5bc53/41592_2025_2610_Fig10_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/08c7bdf26d16/41592_2025_2610_Fig11_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/8f435843085d/41592_2025_2610_Fig12_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/f230e696cf55/41592_2025_2610_Fig13_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/2c66fa735a66/41592_2025_2610_Fig14_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/e10ed2f89746/41592_2025_2610_Fig15_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ff4/11978514/008272734737/41592_2025_2610_Fig16_ESM.jpg

相似文献

1
Genetically encoded biosensor for fluorescence lifetime imaging of PTEN dynamics in the intact brain.用于完整大脑中PTEN动力学荧光寿命成像的基因编码生物传感器。
Nat Methods. 2025 Apr;22(4):764-777. doi: 10.1038/s41592-025-02610-9. Epub 2025 Feb 20.
2
The ketone body β-hydroxybutyrate ameliorates neurodevelopmental deficits in the GABAergic system of mutants.酮体 β-羟丁酸改善 突变体 GABA 能系统的神经发育缺陷。
Elife. 2024 Oct 18;13:RP94520. doi: 10.7554/eLife.94520.
3
A biosensor to monitor dynamic regulation and function of tumour suppressor PTEN in living cells.一种用于监测活细胞中肿瘤抑制因子 PTEN 动态调节和功能的生物传感器。
Nat Commun. 2014 Jul 16;5:4431. doi: 10.1038/ncomms5431.
4
Imaging Cellular Inorganic Phosphate in Caenorhabditis elegans Using a Genetically Encoded FRET-Based Biosensor.使用基于荧光共振能量转移（FRET）的基因编码生物传感器对秀丽隐杆线虫中的细胞无机磷酸盐进行成像。
PLoS One. 2015 Oct 20;10(10):e0141128. doi: 10.1371/journal.pone.0141128. eCollection 2015.
5
Automated screening of AURKA activity based on a genetically encoded FRET biosensor using fluorescence lifetime imaging microscopy.基于荧光寿命成像显微镜的遗传编码 FRET 生物传感器对 AURKA 活性的自动筛选。
Methods Appl Fluoresc. 2020 Feb 20;8(2):024006. doi: 10.1088/2050-6120/ab73f5.
6
Nuclear Excluded Autism-Associated Phosphatase and Tensin Homolog Mutations Dysregulate Neuronal Growth.核排除自闭症相关磷酸酶和张力蛋白同源物突变扰乱神经元生长。
Biol Psychiatry. 2018 Aug 15;84(4):265-277. doi: 10.1016/j.biopsych.2017.11.025. Epub 2017 Dec 2.
7
Insulin activates the insulin receptor to downregulate the PTEN tumour suppressor.胰岛素激活胰岛素受体以下调 PTEN 肿瘤抑制因子。
Oncogene. 2014 Jul 17;33(29):3878-85. doi: 10.1038/onc.2013.347. Epub 2013 Sep 2.
8
A PTEN variant uncouples longevity from impaired fitness in Caenorhabditis elegans with reduced insulin/IGF-1 signaling.PTEN 变异体通过降低胰岛素/IGF-1 信号传导使秀丽隐杆线虫的寿命与健康受损脱耦。
Nat Commun. 2021 Sep 24;12(1):5631. doi: 10.1038/s41467-021-25920-w.
9
In vivo fluorescent adenosine 5'-triphosphate (ATP) imaging of Drosophila melanogaster and Caenorhabditis elegans by using a genetically encoded fluorescent ATP biosensor optimized for low temperatures.利用低温优化的遗传编码荧光 ATP 生物传感器对果蝇和秀丽隐杆线虫进行体内荧光腺苷 5'-三磷酸 (ATP) 成像。
Anal Chem. 2013 Aug 20;85(16):7889-96. doi: 10.1021/ac4015325. Epub 2013 Aug 8.
10
Balancing Proliferation and Connectivity in PTEN-associated Autism Spectrum Disorder.在与PTEN相关的自闭症谱系障碍中平衡增殖与连接性
Neurotherapeutics. 2015 Jul;12(3):609-19. doi: 10.1007/s13311-015-0356-8.

引用本文的文献

1
Genetically encoded biosensors of metabolic function for the study of neurodegeneration, a review and perspective.用于神经退行性疾病研究的代谢功能基因编码生物传感器：综述与展望
Neurophotonics. 2025 Jun;12(Suppl 2):S22805. doi: 10.1117/1.NPh.12.S2.S22805. Epub 2025 Sep 4.
2
Abundance-biased codon diversification prevents recombination in AAV production and ensures robust in vivo expression of functional FRET sensors.丰度偏向性密码子多样化可防止腺相关病毒生产过程中的重组，并确保功能性荧光共振能量转移传感器在体内的稳定表达。
Commun Biol. 2025 Aug 19;8(1):1244. doi: 10.1038/s42003-025-08677-6.
3
Expanding Horizons in Advancements of FRET Biosensing Technologies.

本文引用的文献

1
Local autocrine plasticity signaling in single dendritic spines by insulin-like growth factors.胰岛素样生长因子在单个树突棘中的局部自分泌可塑性信号。
Sci Adv. 2023 Aug 2;9(31):eadg0666. doi: 10.1126/sciadv.adg0666.
2
Monitoring AKT activity and targeting in live tissue and disease contexts using a real-time Akt-FRET biosensor mouse.使用实时 Akt-FRET 生物传感器小鼠监测活组织和疾病环境中的 AKT 活性和靶向。
Sci Adv. 2023 Apr 28;9(17):eadf9063. doi: 10.1126/sciadv.adf9063. Epub 2023 Apr 26.
3
Fast and sensitive GCaMP calcium indicators for imaging neural populations.
荧光共振能量转移生物传感技术进展中的拓展视野
Biosensors (Basel). 2025 Jul 14;15(7):452. doi: 10.3390/bios15070452.
4
PTEN in somatostatin neurons regulates fear and anxiety and is required for inhibitory synaptic connectivity within central amygdala.生长抑素神经元中的PTEN调节恐惧和焦虑，并且是中央杏仁核内抑制性突触连接所必需的。
Front Cell Neurosci. 2025 Jun 26;19:1597131. doi: 10.3389/fncel.2025.1597131. eCollection 2025.
快速灵敏的 GCaMP 钙指示剂用于神经群体成像。
Nature. 2023 Mar;615(7954):884-891. doi: 10.1038/s41586-023-05828-9. Epub 2023 Mar 15.
4
Phosphatase and tensin homolog (PTEN) variants and epilepsy: A multicenter case series.磷酸酶和张力蛋白同源物（PTEN）变体与癫痫：一项多中心病例系列研究。
Seizure. 2022 Aug;100:82-86. doi: 10.1016/j.seizure.2022.06.013. Epub 2022 Jun 24.
5
haploinsufficiency causes desynchronized growth of brain areas involved in sensory processing.单倍剂量不足会导致参与感觉处理的脑区生长不同步。
iScience. 2022 Jan 19;25(2):103796. doi: 10.1016/j.isci.2022.103796. eCollection 2022 Feb 18.
6
Hypothalamic dopamine neurons motivate mating through persistent cAMP signalling.下丘脑多巴胺神经元通过持续的 cAMP 信号来激发交配行为。
Nature. 2021 Sep;597(7875):245-249. doi: 10.1038/s41586-021-03845-0. Epub 2021 Aug 25.
7
AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions.AIMTOR，一种用于活细胞成像的 BRET 生物传感器，揭示了细胞内 mTOR 信号转导及其功能障碍。
BMC Biol. 2020 Jul 3;18(1):81. doi: 10.1186/s12915-020-00790-8.
8
The Role of PTEN in Neurodevelopment.PTEN在神经发育中的作用。
Mol Neuropsychiatry. 2020 Apr;5(Suppl 1):60-71. doi: 10.1159/000504782. Epub 2020 Jan 21.
9
Multi-model functionalization of disease-associated PTEN missense mutations identifies multiple molecular mechanisms underlying protein dysfunction.多模型功能化与疾病相关的 PTEN 错义突变，鉴定了蛋白功能障碍的多种潜在分子机制。
Nat Commun. 2020 Apr 29;11(1):2073. doi: 10.1038/s41467-020-15943-0.
10
In Vivo Imaging of the Coupling between Neuronal and CREB Activity in the Mouse Brain.在体成像研究小鼠大脑中神经元和 CREB 活性的偶联
Neuron. 2020 Mar 4;105(5):799-812.e5. doi: 10.1016/j.neuron.2019.11.028. Epub 2019 Dec 26.