Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo

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 ‘ec...

Full description

Bibliographic Details
Main Authors:	Kamezaki, Aosa, Sato, Fuminori, Aoki, Kazuhiro, Asakawa, Kazuhide, Kawakami, Koichi, Matsuzaki, Fumio, Sehara-Fujisawa, Atsuko
Format:	Online
Language:	English
Published:	Nature Publishing Group 2016
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4929465/

id	pubmed-4929465
recordtype	oai_dc
spelling	pubmed-49294652016-07-06 Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo Kamezaki, Aosa Sato, Fuminori Aoki, Kazuhiro Asakawa, Kazuhide Kawakami, Koichi Matsuzaki, Fumio Sehara-Fujisawa, Atsuko Article 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. Nature Publishing Group 2016-07-01 /pmc/articles/PMC4929465/ /pubmed/27364328 http://dx.doi.org/10.1038/srep28873 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Kamezaki, Aosa Sato, Fuminori Aoki, Kazuhiro Asakawa, Kazuhide Kawakami, Koichi Matsuzaki, Fumio Sehara-Fujisawa, Atsuko
spellingShingle	Kamezaki, Aosa Sato, Fuminori Aoki, Kazuhiro Asakawa, Kazuhide Kawakami, Koichi Matsuzaki, Fumio Sehara-Fujisawa, Atsuko Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo
author_facet	Kamezaki, Aosa Sato, Fuminori Aoki, Kazuhiro Asakawa, Kazuhide Kawakami, Koichi Matsuzaki, Fumio Sehara-Fujisawa, Atsuko
author_sort	Kamezaki, Aosa
title	Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo
title_short	Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo
title_full	Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo
title_fullStr	Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo
title_full_unstemmed	Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo
title_sort	visualization of neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo
description	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.
publisher	Nature Publishing Group
publishDate	2016
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4929465/
_version_	1613602868200210432

Visualization of Neuregulin 1 ectodomain shedding reveals its local processing in vitro and in vivo

Similar Items