Expression Profiling of Ascorbic Acid–Related Transporters in Human and Mouse Eyes
| Autor: | Hong Yan, Jie Huang, Belinda Dana, Nan Ma, Margaret Liu, Andrew J.W. Huang, Miyuki Kubota, David C. Beebe, Carla J. Siegfried, Ying Liu, Ying-Bo Shui |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
transporters In situ hybridization 03 medical and health sciences Ciliary processes chemistry.chemical_compound SVCT2 0302 clinical medicine medicine Laser capture microdissection biology Glucose transporter Ascorbic acid Molecular biology Epithelium 030104 developmental biology medicine.anatomical_structure Biochemistry chemistry 030221 ophthalmology & optometry biology.protein ascorbic acid GLUT1 Dehydroascorbic acid Anatomy and Pathology/Oncology sense organs |
| Zdroj: | Investigative Ophthalmology & Visual Science |
| ISSN: | 1552-5783 |
| DOI: | 10.1167/iovs.16-19162 |
| Popis: | Purpose Ascorbic acid (AsA) is an important antioxidant in the eye. Ascorbic acid is usually transported by sodium-dependent AsA transporters (SVCTs), and dehydroascorbic acid (DHA) by glucose transporters (GLUTs). This study investigates these AsA-related transporters in human compared with mouse eyes. Methods Five pairs of human donor eyes and 15 pairs of mouse eyes were collected. Immunofluorescence and in situ hybridization were performed to detect SVCTs and GLUTs expression in the ciliary epithelium, retina, and lens epithelial cells (LECs). These tissues were isolated with laser microdissection followed by extraction of total RNA. Quantitative PCR (qPCR) was performed to examine the mRNA level of SVCTs and GLUTs in human and mouse ocular tissues. Results Immunofluorescence and in situ hybridization showed SVCT2 and GLUT1 expression in human ciliary epithelium with varied distributions. Sodium-dependent AsA transporter 2 is expressed only in the pigmented epithelium (PE), and GLUT1 is predominately expressed in the nonpigmented epithelium (NPE). However, SVCT2 was not identified in mouse ciliary epithelium, whereas GLUT1 expressed in both PE and NPE. Laser microdissection and qPCR revealed high levels of SVCT2 mRNA in human RPE cells and murine neural retina. Sodium-dependent AsA transporter 1 mRNA could be detected only in human and murine LECs. Glucose transporter 3 and GLUT4 mRNA could not be detected in either the human or mouse ciliary processes or in the lens epithelium. Conclusions These fundamental findings indicate AsA transporter expression in eyes of humans is significantly different compared with mice. This may explain why human aqueous and vitreous humors contain higher AsA levels compared with other animals. |
| Databáze: | OpenAIRE |
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