Ablation of a Neuronal Population Using a Two-photon Laser and Its Assessment Using Calcium Imaging and Behavioral Recording in Zebrafish Larvae

被引:8
作者
Muto, Akira [1 ]
Kawakami, Koichi [1 ]
机构
[1] SOKENDAI Grad Univ Adv Studies, Natl Inst Genet, Dept Genet, Div Mol & Dev Biol, Hayama, Japan
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2018年 / 136期
关键词
Neuroscience; Issue; 136; Laser-ablation; zebrafish larvae; calcium imaging; prey capture; feeding; pretectum; hypothalamus; two-photon microscopy; vision; neuron; GCaMP; GFP; SPINAL PROJECTION NEURONS; VISUALIZATION; PERCEPTION; ACTIVATION; CIRCUITS; TECTUM; GENE;
D O I
10.3791/57485
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
To identify the role of a subpopulation of neurons in behavior, it is essential to test the consequences of blocking its activity in living animals. Laser ablation of neurons is an effective method for this purpose when neurons are selectively labeled with fluorescent probes. In the present study, protocols for laser ablating a subpopulation of neurons using a two-photon microscope and testing of its functional and behavioral consequences are described. In this study, prey capture behavior in zebrafish larvae is used as a study model. The pretecto-hypothalamic circuit is known to underlie this visually-driven prey catching behavior. Zebrafish pretectum were laser-ablated, and neuronal activity in the inferior lobe of the hypothalamus (ILH; the target of the pretectal projection) was examined. Prey capture behavior after pretectal ablation was also tested.
引用
收藏
页数:11
相关论文
共 27 条
[11]  
Lorent K, 2001, DEVELOPMENT, V128, P2131
[12]   BrdU-, neuroD (nrd)- and Hu-studies reveal unusual non-ventricular neurogenesis in the postembryonic zebrafish forebrain [J].
Mueller, T ;
Wullimann, MF .
MECHANISMS OF DEVELOPMENT, 2002, 117 (1-2) :123-135
[13]   Forward genetic analysis of visual behavior in zebrafish [J].
Muto, A ;
Orger, MB ;
Wehman, AM ;
Smear, MC ;
Kay, JN ;
Page-McCaw, PS ;
Gahtan, E ;
Xiao, T ;
Nevin, LM ;
Gosse, NJ ;
Staub, W ;
Finger-Baier, K ;
Baier, H .
PLOS GENETICS, 2005, 1 (05) :575-588
[14]  
Muto A, 2016, METHODS MOL BIOL, P333, DOI 10.1007/978-1-4939-3771-4_23
[15]   Activation of the hypothalamic feeding centre upon visual prey detection [J].
Muto, Akira ;
Lal, Pradeep ;
Ailani, Deepak ;
Abe, Gembu ;
Itoh, Mari ;
Kawakami, Koichi .
NATURE COMMUNICATIONS, 2017, 8
[16]   Real-Time Visualization of Neuronal Activity during Perception [J].
Muto, Akira ;
Ohkura, Masamichi ;
Abe, Gembu ;
Nakai, Junichi ;
Kawakami, Koichi .
CURRENT BIOLOGY, 2013, 23 (04) :307-311
[17]   Genetic visualization with an improved GCaMP calcium indicator reveals spatiotemporal activation of the spinal motor neurons in zebrafish [J].
Muto, Akira ;
Ohkura, Masamichi ;
Kotani, Tomoya ;
Higashijima, Shin-ichi ;
Nakai, Junichi ;
Kawakami, Koichi .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2011, 108 (13) :5425-5430
[18]   A high signal-to-noise Ca2+ probe composed of a single green fluorescent protein [J].
Nakai, J ;
Ohkura, M ;
Imoto, K .
NATURE BIOTECHNOLOGY, 2001, 19 (02) :137-141
[19]   Control of visually guided behavior by distinct populations of spinal projection neurons [J].
Orger, Michael B. ;
Kampff, Adam R. ;
Severi, Kristen E. ;
Bollmann, Johann H. ;
Engert, Florian .
NATURE NEUROSCIENCE, 2008, 11 (03) :327-333
[20]   Classification of Object Size in Retinotectal Microcircuits [J].
Preuss, Stephanie J. ;
Trivedi, Chintan A. ;
vom Berg-Maurer, Colette M. ;
Ryu, Soojin ;
Bollmann, Johann H. .
CURRENT BIOLOGY, 2014, 24 (20) :2376-2385