دورية أكاديمية
Development of a selective novel fluorescent substrate for sodium-dependent transporters.
| العنوان: | Development of a selective novel fluorescent substrate for sodium-dependent transporters. |
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| المؤلفون: | Agnes RS; Department of Radiology, University Hospitals of Cleveland and Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA. Electronic address: Richard.Agnes@case.edu., Traughber BJ; Department of Radiation Oncology, Mayo Clinic, Rochester, MN 55905, USA. Electronic address: Traughber.Bryan@mayo.edu., Muzic RF Jr; Department of Radiology, University Hospitals of Cleveland and Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA. Electronic address: Raymond.Muzic@case.edu. |
| المصدر: | Life sciences [Life Sci] 2024 Aug 15; Vol. 351, pp. 122847. Date of Electronic Publication: 2024 Jun 14. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 0375521 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-0631 (Electronic) Linking ISSN: 00243205 NLM ISO Abbreviation: Life Sci Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: <2008->: Amsterdam : Elsevier Original Publication: Oxford; Elmsford, N. Y. [etc.] Pergamon Press. |
| مواضيع طبية MeSH: | Fluorescent Dyes*/metabolism , Sodium-Glucose Transporter 1*/metabolism, Humans ; HEK293 Cells ; Animals ; Swine ; Sodium-Glucose Transporter 2/metabolism ; Glucose/metabolism ; LLC-PK1 Cells ; Biological Transport ; Sodium/metabolism ; Carbocyanines/chemistry ; Carbocyanines/metabolism |
| مستخلص: | Aim: To synthesize, characterize, and validate 6FGA, a fluorescent glucose modified with a Cyanine5.5 at carbon-6 position, for probing the function of sodium-dependent glucose transporters, SGLT1 and SGLT2. Main Methods: The synthesis of fluorescent glucose analogue was achieved through "click chemistry" of Cyanine5.5-alkyne and 6-azido-6-deoxy-d-glucose. Cell system studies were conducted to characterize the in vivo transport properties. Key Findings: Optical analyses revealed that 6FGA displayed similar spectral profiles to Cyanine5.5 in DMSO, allowing for concentration determination, thus supporting its utility in quantitative kinetic studies within biological assays. Uptake studies in cell system SGLT models, LLC-PK1 and HEK293 cells, exhibited concentration and time-dependent behavior, indicating saturation at specific concentrations and durations which are hallmarks of transported-mediated uptake. The results of cytotoxicity assays suggested cell viability at micromolar concentrations, enabling usage in assays for at least 1 h without significant toxicity. The dependence of 6FGA uptake on sodium, the co-transported cation, was demonstrated in LLC-PK1 and HEK293 cells. Fluorescence microscopy confirmed intracellular localization of 6FGA, particularly near the nucleus. Competition studies revealed that glucose tends to weakly reduce 6FGA uptake, although the effect did not achieve statistical significance. Assessments using standard SGLT and GLUT inhibitors highlighted 6FGA's sensitivity for probing SGLT-mediated transport. Significance: 6FGA is a new fluorescent glucose analog offering advantages over existing probes due to its improved photophysical properties, greater sensitivity, enabling subcellular resolution and efficient tissue penetration in near-infrared imaging. 6FGA presents practicality and cost-effectiveness, making it a promising tool for nonradioactive, microplate-based assays at investigating SGLT-mediated glucose transport mechanisms. Competing Interests: Declaration of competing interest The authors report that they have no competing interest to declare. (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Fluorescence; Glucose; Molecular imaging; SGLT; Transport |
| المشرفين على المادة: | 0 (Fluorescent Dyes) 0 (Sodium-Glucose Transporter 1) 0 (Sodium-Glucose Transporter 2) IY9XDZ35W2 (Glucose) 0 (SLC5A1 protein, human) 9NEZ333N27 (Sodium) 0 (Carbocyanines) |
| تواريخ الأحداث: | Date Created: 20240616 Date Completed: 20240701 Latest Revision: 20240701 |
| رمز التحديث: | 20240702 |
| DOI: | 10.1016/j.lfs.2024.122847 |
| PMID: | 38880166 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1879-0631 |
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| DOI: | 10.1016/j.lfs.2024.122847 |