Publication Date
2-2-2016
Document Type
Article
Organizational Units
College of Natural Science and Mathematics, Biological Sciences, Physics and Astronomy
Keywords
Cellularization, Drosophila melanogaster, Furrow formation, Rab8
Abstract
One of the most fundamental changes in cell morphology is the ingression of a plasma membrane furrow. The Drosophila embryo undergoes several cycles of rapid furrow ingression during early development that culminate in the formation of an epithelial sheet. Previous studies have demonstrated the requirement for intracellular trafficking pathways in furrow ingression; however, the pathways that link compartmental behaviors with cortical furrow ingression events are unclear. Here, we show that Rab8 has striking dynamic behaviors in vivo. As furrows ingress, cytoplasmic Rab8 puncta are depleted and Rab8 accumulates at the plasma membrane in a location that coincides with known regions of directed membrane addition. We additionally use CRISPR/Cas9 technology to N-terminally tag Rab8, which is then used to address endogenous localization and function. Endogenous Rab8 displays partial coincidence with Rab11 and the Golgi, and this colocalization is enriched during the fast phase of cellularization. When Rab8 function is disrupted, furrow formation in the early embryo is completely abolished. We also demonstrate that Rab8 behaviors require the function of the exocyst complex subunit Sec5 as well as the recycling endosome protein Rab11. Active, GTP-locked Rab8 is primarily associated with dynamic membrane compartments and the plasma membrane, whereas GDP-locked Rab8 forms large cytoplasmic aggregates. These studies suggest a model in which active Rab8 populations direct furrow ingression by guiding the targeted delivery of cytoplasmic membrane stores to the cell surface through interactions with the exocyst tethering complex.
Copyright Date
2-2-2016
Rights Holder
The Authors
Provenance
Received from author
Language
English (eng)
Extent
12 pgs
Publication Statement
Copyright is held by the authors. Published by The Company of Biologists Ltd. User is responsible for all copyright compliance.
Publication Title
Development
Volume
143
Issue
5
First Page
892
Last Page
903
ISSN
1477-9129
PubMed ID
26839362
Recommended Citation
Mavor, L. M., Miao, H., Zuo, Z., Holly, R. M., Xie, Y., Loerke, D., & Blankenship, J. T. (2016). Rab8 directs furrow ingression and membrane addition during epithelial formation in Drosophila melanogaster. Development, 143(5), 892–903. DOI: 10.1242/dev.128876