Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2019 Aug 2;27(3):1067–1085. doi: 10.1038/s41418-019-0398-2

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s), under exclusive licence to ADMC Associazione Differenziamento e Morte Cellulare 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 7 — Cones respond to MCU overexpression by decreasing MICU3 transcript and selectively transporting abnormal mitochondria away from the ellipsoid. a qRT-PCR quantification of relative mRNA of MICU proteins (relative to reference gene Ef1α, see “Methods”) between WT and MCU OE retinas at 3 months of age (n = 3). The mean is reported and bars = standard error. **p < 0.01 and ns = not significant using Welch’s t-test. b Cone mitochondrial clusters in live larvae expressing gnat2:mito-GCaMP3. WT mitochondria were imaged with higher laser power to show localization. In MCU OE models, mitochondrial clusters were found near the synapse and nuclear layer (white arrows), which was not observed in WT siblings. Left: cone schematic with OS = outer segment, M = mitochondrial cluster, and N = nucleus. Scale bar = 5 µm. c 8 h timelapse of a migrating mitochondrial cluster (green, white arrow) in live MCU OE gnat2:mito-GCaMP3 larvae. Cone cell bodies express cytosolic RFP (magenta). Scale bar = 5 µm. d Quantification of cone mitochondria in WT sibling and MCU OE fish from EM images of whole zebrafish larval eyes (single slice at optic nerve) at 6 days of age. Fraction of swollen mitochondria was determined by counting swollen mitochondria relative to total cone mitochondria either in the ellipsoid or mislocalized. n = 3 larvae for both WT and MCU OE fish. The mean is reported and bars = standard error. e EM image from MCU OE larvae at 14 days of age. A single cone photoreceptor can contain both healthy mitochondria (black arrows) in the ellipsoid region and swollen mitochondria (white arrows) near the synapse. Cone cell membrane outlined in red overlay to aid visualization. Scale bar = 2 µm. OS = outer segment, N = nucleus. f 3D reconstruction of an MCU OE cone from a 6-day-old fish using serial block-face EM (synapse not shown). Electron-lucent mitochondria (M⁻) displace the nucleus (N) of the cone to move toward the synapse region. OS = outer segment. M⁻ = electron-lucent mitochondria. M⁺ = healthy mitochondria. N = nucleus. Outline (yellow) = cell body. g Quantification of mislocalized mitochondria using the mito-GCaMP3 probe in 6-day-old MCU OE fish under a normal light cycle or subjected to constant darkness at 5 days of age. Quantification was performed across a whole eye at a total depth of 50 µm. Scale bar = 10 µm. ***p < 0.001 using Welch’s t-test (n = 16 fish each). h Quantification of mislocalized mitochondria in Richardsons’-stained sections of 6-day-old fish under a normal light cycle or subjected to constant darkness at 5 days of age. Quantification was performed in 2–3 slices per eye at/near the optic nerve and the average # of mitochondria for each eye is reported. Scale bar = 25 µm. **p < 0.01 using Welch’s t-test (n = 10 eyes in light, 12 eyes in dark)