Expansion microscopy of zebrafish for neuroscience and developmental biology studies

被引：70

作者：

Freifeld, Limor ^{[1
]}

Odstrcil, Iris ^{[2
]}

Foerster, Dominique ^{[3
]}

Ramirez, Alyson ^{[2
]}

Gagnon, James A. ^{[2
]}

Randlett, Owen ^{[2
]}

Costa, Emma K. ^{[4
]}

Asano, Shoh ^{[1
]}

Celiker, Orhan T. ^{[5
]}

Gao, Ruixuan ^{[1
,6
]}

Martin-Alarcon, Daniel A. ^{[7
]}

Reginato, Paul ^{[7
,8
]}

Dick, Cortni ^{[1
]}

Chen, Linlin ^{[1
,9
]}

Schoppik, David ^{[10
,11
,12
]}

Engert, Florian ^{[2
]}

Baier, Herwig ^{[3
]}

Boyden, Edward S. ^{[1
,4
,5
,6
,13
]}

机构：

[1] MIT, Media Lab, Cambridge, MA 02139 USA

[2] Harvard Univ, Dept Mol & Cellular Biol, Cambridge, MA 02138 USA

[3] Max Planck Inst Neurobiol, Dept Genes Circuits Behav, D-82152 Martinsried, Germany

[4] MIT, Dept Brain & Cognit Sci, E25-618, Cambridge, MA 02139 USA

[5] MIT, Dept Elect Engn & Comp Sci, Cambridge, MA 02139 USA

[6] MIT, McGovern Inst Brain Res, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[7] MIT, Dept Biol Engn, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[8] Harvard Med Sch, Dept Genet, Cambridge, MA 02138 USA

[9] Wellesley Coll, Neurosci Program, Wellesley, MA 02481 USA

[10] NYU, Dept Otolaryngol, Sch Med, 550 1St Ave, New York, NY 10016 USA

[11] NYU, Sch Med, Dept Neurosci & Physiol, New York, NY 10016 USA

[12] NYU, Sch Med, Inst Neurosci, New York, NY 10016 USA

[13] MIT, Ctr Neurobiol Engn, 77 Massachusetts Ave, Cambridge, MA 02139 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2017年 / 114卷 / 50期

关键词：

zebrafish; superresolution; microscopy; brain; STRUCTURED ILLUMINATION MICROSCOPY; NUCLEAR-ENVELOPE BREAKDOWN; IN-VIVO; MAUTHNER-CELL; NUCLEOPLASMIC RETICULUM; FLUORESCENT PROTEINS; OPTICAL MICROSCOPY; INTERPHASE NUCLEI; MAMMALIAN-CELLS; NEURAL CIRCUIT;

D O I：

10.1073/pnas.1706281114

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：

Expansion microscopy (ExM) allows scalable imaging of preserved 3D biological specimens with nanoscale resolution on fast diffraction-limited microscopes. Here, we explore the utility of ExM in the larval and embryonic zebrafish, an important model organism for the study of neuroscience and development. Regarding neuroscience, we found that ExM enabled the tracing of fine processes of radial glia, which are not resolvable with diffraction-limited microscopy. ExM further resolved putative synaptic connections, as well as molecular differences between densely packed synapses. Finally, ExM could resolve subsynaptic protein organization, such as ring-like structures composed of glycine receptors. Regarding development, we used ExM to characterize the shapes of nuclear invaginations and channels, and to visualize cytoskeletal proteins nearby. We detected nuclear invagination channels at late prophase and telophase, potentially suggesting roles for such channels in cell division. Thus, ExM of the larval and embryonic zebrafish may enable systematic studies of how molecular components are configured in multiple contexts of interest to neuroscience and developmental biology.

引用

页码：E10799 / E10808

页数：10

共 113 条

[1] Myocyte differentiation generates nuclear invaginations traversed by myofibrils associating with sarcomeric protein mRNAs [J].