Mutation of wrb, a Component of the Guided Entry of Tail-Anchored Protein Pathway, Disrupts Photoreceptor Synapse Structure and Function

被引:26
作者
Daniele, Lauren L. [1 ]
Emran, Farida [2 ]
Lobo, Glenn P. [1 ]
Gaivin, Robert J. [1 ]
Perkins, Brian D. [1 ]
机构
[1] Cleveland Clin, Cole Eye Inst, Dept Ophthalm Res, 9500 Euclid Ave, Cleveland, OH 44195 USA
[2] McGill Univ, Ctr Res Neurosci, Montreal, PQ, Canada
关键词
zebrafish; ribbon synapse; optokinetic response; photoreceptors; ENDOPLASMIC-RETICULUM MEMBRANE; ZEBRAFISH NRC MUTANT; CONE PHOTORECEPTOR; ER MEMBRANE; HAIR-CELLS; RETINAL PHOTORECEPTORS; RIBBON FORMATION; GANGLION-CELLS; INSERTION; IDENTIFICATION;
D O I
10.1167/iovs.15-18996
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
PURPOSE. Tail-anchored (TA) proteins contain a single hydrophobic domain at the C-terminus and are posttranslationally inserted into the ER membrane via the GET (guided entry of tail-anchored proteins) pathway. The role of the GET pathway in photoreceptors is unexplored. The goal of this study was to characterize the zebrafish pinball wizard mutant, which disrupts Wrb, a core component of the GET pathway. METHODS. Electroretinography, optokinetic response measurements (OKR), immunohistochemistry, and electron microscopy analyses were employed to assess ribbon synapse function, protein expression, and ultrastructure in 5-day-old zebrafish larvae. Expression of wrb was investigated with real-time qRT-PCR and in situ hybridization. RESULTS. Mutation of wrb abolished the OKR and greatly diminished the ERG b-wave, but not the a-wave. Ribeye and SV2 were partially mislocalized in both photoreceptors and hair cells of wrb mutants. Fewer contacts were seen between photoreceptors and bipolar cells in wrb(-/-) mutants. Expression of wrb was observed throughout the nervous system and Wrb localized to the ER and synaptic region of photoreceptors. Morpholino knockdown of the cytosolic ATPase trc40, which targets TA proteins to the ER, also diminished the OKR. Overexpression of wrb fully restored contrast sensitivity in mutants, while overexpression of mutant wrb(R73A), which cannot bind Trc40, did not. CONCLUSIONS. Proteins Wrb and Trc40 are required for synaptic transmission between photoreceptors and bipolar cells, indicating that TA protein insertion by the TRC pathway is a critical step in ribbon synapse assembly and function.
引用
收藏
页码:2942 / 2954
页数:13
相关论文
共 69 条
  • [1] Signal recognition particle mediates post-translational targeting in eukaryotes
    Abell, BM
    Pool, MR
    Schlenker, O
    Sinning, I
    High, S
    [J]. EMBO JOURNAL, 2004, 23 (14) : 2755 - 2764
  • [2] Synapse formation is arrested in retinal photoreceptors of the zebrafish nrc mutant
    Allwardt, BA
    Lall, AB
    Brockerhoff, SE
    Dowling, JE
    [J]. JOURNAL OF NEUROSCIENCE, 2001, 21 (07) : 2330 - 2342
  • [3] Identification of 315 genes essential for early zebrafish development
    Amsterdam, A
    Nissen, RM
    Sun, ZX
    Swindell, EC
    Farrington, S
    Hopkins, N
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (35) : 12792 - 12797
  • [4] A large-scale insertional mutagenesis screen in zebrafish
    Amsterdam, A
    Burgess, S
    Golling, G
    Chen, WB
    Sun, ZX
    Townsend, K
    Farrington, S
    Haldi, M
    Hopkins, N
    [J]. GENES & DEVELOPMENT, 1999, 13 (20) : 2713 - 2724
  • [5] Rod contributions to the electroretinogram of the dark-adapted developing zebrafish
    Bilotta, J
    Saszik, S
    Sutherland, SE
    [J]. DEVELOPMENTAL DYNAMICS, 2001, 222 (04) : 564 - 570
  • [6] Targeting of a tail-anchored protein to endoplasmic reticulum and mitochondrial outer membrane by independent but competing pathways
    Borgese, N
    Gazzoni, I
    Barberi, M
    Colombo, S
    Pedrazzini, E
    [J]. MOLECULAR BIOLOGY OF THE CELL, 2001, 12 (08) : 2482 - 2496
  • [7] How tails guide tail-anchored proteins to their destinations
    Borgese, Nica
    Brambillasca, Silvia
    Colombo, Sara
    [J]. CURRENT OPINION IN CELL BIOLOGY, 2007, 19 (04) : 368 - 375
  • [8] A BEHAVIORAL SCREEN FOR ISOLATING ZEBRAFISH MUTANTS WITH VISUAL-SYSTEM DEFECTS
    BROCKERHOFF, SE
    HURLEY, JB
    JANSSENBIENHOLD, U
    NEUHAUSS, SCF
    DRIEVER, W
    DOWLING, JE
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (23) : 10545 - 10549
  • [9] Retina-Specific GTPase Accelerator RGS11/Gβ5S/R9AP Is a Constitutive Heterotrimer Selectively Targeted to mGluR6 in ON-Bipolar Neurons
    Cao, Yan
    Masuho, Ikuo
    Okawa, Haruhisa
    Xie, Keqiang
    Asami, Junko
    Kammermeier, Paul J.
    Maddox, Dennis M.
    Furukawa, Takahisa
    Inoue, Takayoshi
    Sampath, Alapakkam P.
    Martemyanov, Kirill A.
    [J]. JOURNAL OF NEUROSCIENCE, 2009, 29 (29) : 9301 - 9313
  • [10] Uncoordinated (UNC)119: Coordinating the trafficking of myristoylated proteins
    Constantine, Ryan
    Zhang, Houbin
    Gerstner, Cecilia D.
    Frederick, Jeanne M.
    Baehr, Wolfgang
    [J]. VISION RESEARCH, 2012, 75 : 26 - 32