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

. 2020 Oct 6;14:556700. doi: 10.3389/fnins.2020.556700

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

Copyright © 2020 Wagner, Reinehr, Gammel, Greulich, Hurst, Dick, Schnichels and Joachim.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PMC Copyright notice

Analysis of photoreceptors. (A) Exemplary immunofluorescence pictures of opsin (red) staining for L-cones and rhodopsin (green) staining for rods in photoreceptor outer segments. Nuclei were labeled with 4′,6-diamidino-2-phenylindole (DAPI) (blue). (B) Fewer opsin⁺ cells were found in control compared to native retinas (p = 0.02). The filter and tweezers method showed, compared to native retinas, a better preservation over a cultivation period of 4 days. The number of opsin⁺ cells was significantly higher in tweezers samples (p = 0.04) compared to the control ones, while no changes were noted between filter and control retinas. A severe loss of opsin⁺ cells was discovered in the control compared to native samples after 8 days (p < 0.001). In the novel methods filter and tweezers, the number of opsin-labeled cells was comparable to native retinas. When comparing filter and tweezers samples (both: p < 0.001) to the controls, more opsin⁺ cells could be detected. (C) OPSIN expression was not altered at 4 days. OPSIN mRNA expression in tweezers samples was significantly upregulated compared to control retinas at 8 days (p = 0.002). (D) The rhodopsin signal found in control (p < 0.001) and filter samples (p = 0.01) was significantly less intense at 4 days compared to that in native samples. A significantly higher signal intensity was documented in tweezers (p < 0.001) and filter retinas (p = 0.047) compared to the controls. Moreover, a higher rhodopsin signal intensity was observed in tweezers retinas compared to filter samples (p = 0.04) at 4 days. At 8 days of cultivation, all three methods showed a significantly diminished rhodopsin intensity compared to native samples (all: p < 0.001). (E) RHODOPSIN mRNA expression was not altered at 4 days. Quantitative real-time PCR (RT-qPCR) examination of RHODOPSIN demonstrated an upregulation in tweezers samples compared to control retinas at 8 days (p = 0.02). OS, photoreceptor outer segments; ONL, outer nuclear layer. Scale bar: 20 μm, values are mean ± SEM for immunofluorescence (IF) and median ± quartile + min/max for RT-qPCR. IF: n = 9–10/group; RT-qPCR: n = 5/group. *p < 0.05 and ***p < 0.001 vs. native group; ^#p < 0.05, ^##p < 0.01, and ^###p < 0.001 vs. controls; ^¥p < 0.05 vs. filter group.