Confocal and Super-Resolution Imaging of Polarized Intracellular Trafficking and Secretion of Basement Membrane Proteins during Drosophila Oogenesis

被引:2
作者
Shah, Hemin P. [1 ]
Devergne, Olivier [1 ]
机构
[1] Northern Illinois Univ, Dept Biol Sci, De Kalb, IL 60115 USA
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2022年 / 183期
基金
美国国家科学基金会;
关键词
IV COLLAGEN; FLUORESCENCE MICROSCOPY; LYSYL HYDROXYLASE; CELL; MORPHOGENESIS; LOCALIZATION; DEPOSITION; EPITHELIUM; INSIGHTS; LAMININ;
D O I
10.3791/63778
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The basement membrane (BM) - a specialized sheet of extracellular matrix present at the basal side of epithelial cells - is critical for the establishment and maintenance of epithelial tissue morphology and organ morphogenesis. Moreover, the BM is essential for tissue modeling, serving as a signaling platform, and providing external forces to shape tissues and organs. Despite the many important roles that the BM plays during normal development and pathological conditions, the biological pathways controlling the intracellular trafficking of BM-containing vesicles and how basal secretion leads to the polarized deposition of BM proteins are poorly understood. The follicular epithelium of the Drosophila ovary is an excellent model system to study the basal deposition of BM membrane proteins, as it produces and secretes all major components of the BM. Confocal and super-resolution imaging combined with image processing in fixed tissues allows for the identification and characterization of cellular factors specifically involved in the intracellular trafficking and deposition of BM proteins. This article presents a detailed protocol for staining and imaging BM-containing vesicles and deposited BM using endogenously tagged proteins in the follicular epithelium of the Drosophila ovary. This protocol can be applied to address both qualitative and quantitative questions and it was developed to accommodate high-throughput screening, allowing for the rapid and efficient identification of factors involved in the polarized intracellular trafficking and secretion of vesicles during epithelial tissue development.
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页数:29
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