RRC ID 49671
Author Kamezaki A, Sato F, Aoki K, Asakawa K, Kawakami K, Matsuzaki F, Sehara-Fujisawa A.
Title Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo.
Journal Sci Rep
Abstract Neuregulin1 (NRG1) plays diverse developmental roles and is likely involved in several neurological disorders including schizophrenia. The transmembrane NRG1 protein is proteolytically cleaved and released as a soluble ligand for ErbB receptors. Such post-translational processing, referred to as 'ectodomain shedding', is thought to be crucial for NRG1 function. However, little is known regarding the regulatory mechanism of NRG1 cleavage in vivo. Here, we developed a fluorescent probe, NRG1 Cleavage Indicating SenSOR (N-CISSOR), by fusing mCherry and GFP to the extracellular and intracellular domains of NRG1, respectively. N-CISSOR mimicked the subcellular localization and biochemical properties of NRG1 including cleavage dynamics and ErbB phosphorylation in cultured cells. mCherry/GFP ratio imaging of phorbol-12-myristate-13-acetate-stimulated N-CISSOR-expressing HEK293T cells enabled to monitor rapid ectodomain shedding of NRG1 at the subcellular level. Utilizing N-CISSOR in zebrafish embryos revealed preferential axonal NRG1 ectodomain shedding in developing motor neurons, demonstrating that NRG1 ectodomain shedding is spatially regulated at the subcellular level. Thus, N-CISSOR will be a valuable tool for elucidating the spatiotemporal regulation of NRG1 ectodomain shedding, both in vitro and in vivo.
Volume 6
Pages 28873
Published 2016-7-1
DOI 10.1038/srep28873
PII srep28873
PMID 27364328
PMC PMC4929465
MeSH Animals Animals, Genetically Modified Binding Sites / genetics Cell Line Embryo, Nonmammalian / metabolism Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism* HEK293 Cells Humans Luminescent Proteins / genetics Luminescent Proteins / metabolism* Mice Microscopy, Fluorescence Neuregulin-1 / genetics Neuregulin-1 / metabolism* Phorbol Esters / pharmacology Protein Processing, Post-Translational* Proteolysis / drug effects Time-Lapse Imaging / methods* Zebrafish / embryology Zebrafish / genetics Zebrafish / metabolism
IF 3.998
Times Cited 7
Zebrafish Tg(HuC:GVP)