LV-pIN-KDEL: a novel lentiviral vector demonstrates the morphology, dynamics and continuity of the endoplasmic reticulum in live neurones.

BMC Neuroscience
Vicky C JonesOwen T Jones

Abstract

The neuronal endoplasmic reticulum (ER) is an extensive, complex endomembrane system, containing Ca2+ pumps, and Ca2+ channels that permit it to act as a dynamic calcium store. Currently, there is controversy over the continuity of the ER in neurones, how this intersects with calcium signalling and the possibility of physical compartmentalisation. Unfortunately, available probes of ER structure such as vital dyes are limited by their membrane specificity. The introduction of ER-targeted GFP plasmids has been a considerable step forward, but these are difficult to express in neurones through conventional transfection approaches. To circumvent such problems we have engineered a novel ER-targeted GFP construct, termed pIN-KDEL, into a 3rd generation replication-defective, self-inactivating lentiviral vector system capable of mediating gene transduction in diverse dividing and post-mitotic mammalian cells, including neurones. Following its expression in HEK293 (or COS-7) cells, LV-pIN-KDEL yielded a pattern of fluorescence that co-localised exclusively with the ER marker sec61beta but with no other major organelle. We found no evidence for cytotoxicity and only rarely inclusion body formation. To explore the utility of the probe in...Continue Reading

References

Nov 1, 1991·Developmental Biology·M TerasakiC Sardet
Apr 1, 1989·The Journal of Cell Biology·K Goslin, G Banker
Aug 2, 1994·Proceedings of the National Academy of Sciences of the United States of America·M TerasakiT S Reese
Dec 1, 1995·The Journal of Cell Biology·N NakamuraG Warren
Jan 8, 2000·The Journal of Biological Chemistry·D G JugloffO T Jones
May 12, 2000·Nature Cell Biology·S NehlsJ Lippincott-Schwartz
Apr 20, 2001·Trends in Neurosciences·O H PetersenM K Park
Sep 29, 2001·Trends in Neurosciences·M P Blaustein, V A Golovina
Nov 22, 2001·Journal of Neuroscience Methods·E C OhkiP J Price
Feb 19, 2002·Nature Neuroscience·Bagirathy NadarajahJohn G Parnavelas
Feb 21, 2002·Proceedings of the National Academy of Sciences of the United States of America·Alexander PfeiferInder M Verma
Jan 25, 2003·Cell Calcium·Martin D BootmanAlexei Verkhratsky
Sep 2, 2003·Journal of Neuroscience Research·Johan JakobssonCecilia Lundberg
Dec 20, 2003·Current Gene Therapy·Zeger Debyser
Jan 27, 2004·Molecular Therapy : the Journal of the American Society of Gene Therapy·Liang-Fong WongNicholas D Mazarakis
Feb 3, 2004·Journal of Cellular and Molecular Medicine·A Verkhratsky, E C Toescu
Oct 12, 2004·Annual Review of Cell and Developmental Biology·Marcus C S LeeRandy Schekman
Jun 25, 2005·Current Opinion in Cell Biology·Tim Levine, Catherine Rabouille
Aug 4, 2005·Cell Calcium·Denis BurdakovAlexei Verkhratsky
Oct 4, 2005·The European Journal of Neuroscience·Håkan Toresson, Seth G N Grant
Jan 13, 2006·Molecular Therapy : the Journal of the American Society of Gene Therapy·Johan Jakobsson, Cecilia Lundberg
Jun 30, 2006·Current Opinion in Cell Biology·Nica BorgeseErik Snapp
Oct 5, 2006·Cell·Michael A Sutton, Erin M Schuman
Nov 24, 2006·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Yu Mi ChoiMyoung Kyu Park
Jan 9, 2007·Advances in Experimental Medicine and Biology·Yuichiro Shimizu, Linda M Hendershot
Mar 1, 1945·The Journal of Experimental Medicine·K R PorterE F Fullam

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Citations

Mar 20, 2009·Pflügers Archiv : European journal of physiology·Vicky C JonesOwen T Jones
Aug 11, 2010·Proceedings of the National Academy of Sciences of the United States of America·Qing-Jun MengAndrew S I Loudon
Sep 10, 2010·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Ivan GoussakovGrace E Stutzmann
Dec 11, 2013·Translational Stroke Research·K KucharzH Toresson
Oct 5, 2011·Cellular and Molecular Life Sciences : CMLS·Enrico Zampese, Paola Pizzo
Jul 26, 2011·Molecular and Cellular Neurosciences·José Ignacio ValenzuelaAndrés Couve
Aug 4, 2010·Current Opinion in Neurobiology·Benoît Renvoisé, Craig Blackstone
Oct 20, 2009·Cell Calcium·Richard D RainbowJohn G McCarron
Mar 17, 2010·Current Biology : CB·Weiqun LuAndrew S I Loudon
Oct 24, 2018·Breast Cancer Research : BCR·Eleanor BroadberryCharles H Streuli

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Datasets Mentioned

BETA
GM130
AY841887

Methods Mentioned

BETA
glycosylation
protein folding
Fluorescence
ELISA
fluorescence imaging
transfection
pull-down
transfect
transgenic
transfections

Software Mentioned

Image J
LAS AF
NIH Image J
SoftWoRx

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