Improved expression of halorhodopsin for light-induced silencing of neuronal activity

被引:132
作者
Zhao, Shengli [1 ]
Cunha, Catarina [1 ,2 ]
Zhang, Feng [3 ]
Liu, Qun [4 ]
Gloss, Bernd [4 ]
Deisseroth, Karl [3 ]
Augustine, George J. [1 ]
Feng, Guoping [1 ,4 ]
机构
[1] Duke Univ, Med Ctr, Dept Neurobiol, Durham, NC 27710 USA
[2] Univ Porto, Fac Ciencias, P-4100 Oporto, Portugal
[3] Stanford Univ, Dept Bioengn, Stanford, CA 94305 USA
[4] Duke Univ, Med Ctr, Duke Neurotransgen Lab, Durham, NC 27710 USA
关键词
D O I
10.1007/s11068-008-9034-7
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The ability to control and manipulate neuronal activity within an intact mammalian brain is of key importance for mapping functional connectivity and for dissecting the neural circuitry underlying behaviors. We have previously generated transgenic mice that express channelrhodopsin-2 for light-induced activation of neurons and mapping of neural circuits. Here we describe transgenic mice that express halorhodopsin (NpHR), a light-driven chloride pump that can be used to silence neuronal activity via light. Using the Thy-1 promoter to target NpHR expression to neurons, we found that neurons in these mice expressed high levels of NpHR-YFP and that illumination of cortical pyramidal neurons expressing NpHR-YFP led to rapid, reversible photoinhibition of action potential firing in these cells. However, NpHR-YFP expression led to the formation of numerous intracellular blebs, which may disrupt neuronal function. Labeling of various subcellular markers indicated that the blebs arise from retention of NpHR-YFP in the endoplasmic reticulum. By improving the signal peptide sequence and adding an ER export signal to NpHR-YFP, we eliminated the formation of blebs and dramatically increased the membrane expression of NpHR-YFP. Thus, the improved version of NpHR should serve as an excellent tool for neuronal silencing in vitro and in vivo.
引用
收藏
页码:141 / 154
页数:14
相关论文
共 39 条
  • [1] Neural substrates of awakening probed with optogenetic control of hypocretin neurons
    Adamantidis, Antoine R.
    Zhang, Feng
    Aravanis, Alexander M.
    Deisseroth, Karl
    De Lecea, Luis
    [J]. NATURE, 2007, 450 (7168) : 420 - U9
  • [2] In vivo light-induced activation of neural circuitry in transgenic mice expressing channelrhodopsin-2
    Arenkiel, Benjamin R.
    Peca, Joao
    Davison, Ian G.
    Feliciano, Catia
    Deisseroth, Karl
    Augustine, George J.
    Ehlers, Michael D.
    Feng, Guoping
    [J]. NEURON, 2007, 54 (02) : 205 - 218
  • [3] Evolving the lock to fit the key to create a family of G protein-coupled receptors potently activated by an inert ligand
    Armbruster, Blaine N.
    Li, Xiang
    Pausch, Mark H.
    Herlitze, Stefan
    Roth, Bryan L.
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (12) : 5163 - 5168
  • [4] Improved prediction of signal peptides: SignalP 3.0
    Bendtsen, JD
    Nielsen, H
    von Heijne, G
    Brunak, S
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 2004, 340 (04) : 783 - 795
  • [5] Ectopic expression of a microbial-type rhodopsin restores visual responses in mice with photoreceptor degeneration
    Bi, AD
    Cui, JJ
    Ma, YP
    Olshevskaya, E
    Pu, ML
    Dizhoor, AM
    Pan, ZH
    [J]. NEURON, 2006, 50 (01) : 23 - 33
  • [6] Millisecond-timescale, genetically targeted optical control of neural activity
    Boyden, ES
    Zhang, F
    Bamberg, E
    Nagel, G
    Deisseroth, K
    [J]. NATURE NEUROSCIENCE, 2005, 8 (09) : 1263 - 1268
  • [7] Overexpression of growth-associated proteins in the neurons of adult transgenic mice
    Caroni, P
    [J]. JOURNAL OF NEUROSCIENCE METHODS, 1997, 71 (01) : 3 - 9
  • [8] Next-generation optical technologies for illuminating genetically targeted brain circuits
    Deisseroth, Karl
    Feng, Guoping
    Majewska, Ania K.
    Miesenbock, Gero
    Ting, Alice
    Schnitzer, Mark J.
    [J]. JOURNAL OF NEUROSCIENCE, 2006, 26 (41) : 10380 - 10386
  • [9] New insights into membrane trafficking and protein sorting
    Derby, Merran C.
    Gleeson, Paul A.
    [J]. INTERNATIONAL REVIEW OF CYTOLOGY - A SURVEY OF CELL BIOLOGY, VOL 261, 2007, 261 : 47 - +
  • [10] Reinsertion or degradation of AMPA receptors determined by activity-dependent endocytic sorting
    Ehlers, MD
    [J]. NEURON, 2000, 28 (02) : 511 - 525