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In vivo development of retinal ON-bipolar cell axonal terminals visualized in nyx::MYFP transgenic zebrafish

Published online by Cambridge University Press:  04 October 2006

ERIC H. SCHROETER ,
RACHEL O.L. WONG  and
RONALD G. GREGG
ERIC H. SCHROETER
Affiliation:
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri
RACHEL O.L. WONG
Affiliation:
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri
RONALD G. GREGG
Affiliation:
Department of Biochemistry and Molecular Biology, and Center for Genetics and Molecular Medicine, University of Louisville, Louisville, Kentucky
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Abstract

Axonal differentiation of retinal bipolar cells has largely been studied by comparing the morphology of these interneurons in fixed tissue at different ages. To better understand how bipolar axonal terminals develop in vivo, we imaged fluorescently labeled cells in the zebrafish retina using time-lapse confocal and two photon microscopy. Using the upstream regulatory sequences from the nyx gene that encodes nyctalopin, we constructed a transgenic fish in which a subset of retinal bipolar cells express membrane targeted yellow fluorescent protein (MYFP). Axonal terminals of these YFP-labeled bipolar cells laminated primarily in the inner half of the inner plexiform layer, suggesting that they are likely to be ON-bipolar cells. Transient expression of MYFP in isolated bipolar cells indicates that two or more subsets of bipolar cells, with one or two terminal boutons, are labeled. Live imaging of YFP-expressing bipolar cells in the nyx::MYFP transgenic fish at different ages showed that initially, filopodial-like structures extend and retract from their primary axonal process throughout the inner plexiform layer (IPL). Over time, filopodial exploration becomes concentrated at discrete foci prior to the establishment of large terminal boutons, characteristic of the mature form. This sequence of axonal differentiation suggests that synaptic targeting by bipolar cell axons may involve an early process of trial and error, rather than a process of directed outgrowth and contact. Our observations represent the first in vivo visualization of axonal development of bipolar cells in a vertebrate retina.

Keywords

ZebrafishBipolar cellNyctalopinAxonal development
Type
Research Article
Information
Visual Neuroscience , Volume 23 , Issue 5 , September 2006 , pp. 833 - 843
Copyright
2006 Cambridge University Press

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Supplemental movie 1

High magnification serial confocal sections (0.5 Μm steps) through the IPL of a 6 dpf nyx::MYFP fish.
Axon terminals display many filopodia but contain a core within which YFP is absent. Scale bar = 5Μm.

Download Supplemental movie 1(Video)
Video 1.1 MB

Supplemental movie 2

Time-lapse series of an individual axon terminal from 7 dpf nyx::MYFP fish (15 minute intervals), showing rapid extension and retraction of filopodia in an immature terminal. Scale bar = 5 Μm.

Download Supplemental movie 2(Video)
Video 60.4 KB