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A method for isolation of cone photoreceptors from adult zebrafish retinae

Overview of attention for article published in BMC Neuroscience, November 2016
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  • In the top 25% of all research outputs scored by Altmetric
  • Good Attention Score compared to outputs of the same age (75th percentile)
  • Good Attention Score compared to outputs of the same age and source (75th percentile)

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Title
A method for isolation of cone photoreceptors from adult zebrafish retinae
Published in
BMC Neuroscience, November 2016
DOI 10.1186/s12868-016-0307-2
Pubmed ID
Authors

Antonino Glaviano, Andrew J. Smith, Alfonso Blanco, Sarah McLoughlin, Maria L. Cederlund, Theresa Heffernan, Beata Sapetto-Rebow, Yolanda Alvarez, Jun Yin, Breandán N. Kennedy

Abstract

Cone photoreceptors are specialised sensory retinal neurons responsible for photopic vision, colour perception and visual acuity. Retinal degenerative diseases are a heterogeneous group of eye diseases in which the most severe vision loss typically arises from cone photoreceptor dysfunction or degeneration. Establishing a method to purify cone photoreceptors from retinal tissue can accelerate the identification of key molecular determinants that underlie cone photoreceptor development, survival and function. The work herein describes a new method to purify enhanced green fluorescent protein (EGFP)-labelled cone photoreceptors from adult retina of Tg(3.2gnat2:EGFP) zebrafish. Methods for dissecting adult zebrafish retinae, cell dissociation, cell sorting, RNA isolation and RNA quality control were optimised. The dissociation protocol, carried out with ~30 retinae from adult zebrafish, yielded approximately 6 × 10(6) cells. Flow cytometry cell sorting subsequently distinguished 1 × 10(6) EGFP(+) cells and 4 × 10(6) EGFP(-) cells. Electropherograms confirmed downstream isolation of high-quality RNA with RNA integrity number (RIN) >7.6 and RNA concentration >5.7 ng/µl obtained from both populations. Reverse Transcriptase-PCR confirmed that the EGFP-positive cell populations express known genetic markers of cone photoreceptors that were not expressed in the EGFP-negative cell population whereas a rod opsin amplicon was only detected in the EGFP-negative retinal cell population. This work describes a valuable adult zebrafish cone photoreceptor isolation methodology enabling future identification of cone photoreceptor-enriched genes, proteins and signalling networks responsible for their development, survival and function. In addition, this advancement facilitates the identification of novel candidate genes for inherited human blindness.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 30 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 30 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 5 17%
Researcher 5 17%
Student > Bachelor 3 10%
Professor > Associate Professor 3 10%
Student > Master 3 10%
Other 4 13%
Unknown 7 23%
Readers by discipline Count As %
Agricultural and Biological Sciences 7 23%
Biochemistry, Genetics and Molecular Biology 6 20%
Neuroscience 4 13%
Medicine and Dentistry 3 10%
Social Sciences 1 3%
Other 1 3%
Unknown 8 27%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 7. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 15 February 2017.
All research outputs
#4,851,599
of 23,881,329 outputs
Outputs from BMC Neuroscience
#218
of 1,265 outputs
Outputs of similar age
#78,625
of 315,006 outputs
Outputs of similar age from BMC Neuroscience
#6
of 20 outputs
Altmetric has tracked 23,881,329 research outputs across all sources so far. Compared to these this one has done well and is in the 79th percentile: it's in the top 25% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 1,265 research outputs from this source. They receive a mean Attention Score of 4.5. This one has done well, scoring higher than 83% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 315,006 tracked outputs that were published within six weeks on either side of this one in any source. This one has done well, scoring higher than 75% of its contemporaries.
We're also able to compare this research output to 20 others from the same source and published within six weeks on either side of this one. This one has done well, scoring higher than 75% of its contemporaries.