Open AccessAn in vitro assay reveals a role for the diaphragm protein PV-1 in endothelial fenestra morphogenesis
Author(s) -
Sofia Ioannidou,
Katrin Deinhardt,
Jadwiga M. Miotla,
John R. Bradley,
Eunice Cheung,
Steven J. Samuelsson,
Y. Jack Ng,
David T. Shima
Publication year - 2006
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.0603501103
Subject(s) - microbiology and biotechnology , biology , microfilament , morphogenesis , in vitro , diaphragm (acoustics) , endothelium , ultrastructure , anatomy , cytoskeleton , cell , biochemistry , endocrinology , physics , acoustics , loudspeaker , gene
Fenestrae are small pores in the endothelium of renal glomerular, gastrointestinal, and endocrine gland capillaries and are involved in the bidirectional exchange of molecules between blood and tissues. Although decades of studies have characterized fenestrae at the ultrastructural level, little is known on the mechanisms by which fenestrae form. We present the development of anin vitro assay in which rapid and abundant fenestra induction enables a detailed study of their biogenesis. Through the use of agents that stabilize or disassemble actin microfilaments, we show that actin microfilament remodeling is part of fenestra biogenesis in this model. Furthermore, by using a loss-of-function approach, we show that the diaphragm protein PV-1 is necessary for fenestral pore architecture and the ordered arrangement of fenestrae in sieve plates. Together, these data provide insight into the cell biology of fenestra formation and open up the future study of the fenestra to a combined morphological and biochemical analysis.