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A retrograde adeno-associated virus for collecting ribosome-bound mRNA from anatomically defined projection neurons

Overview of attention for article published in Frontiers in Molecular Neuroscience, September 2015
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Title
A retrograde adeno-associated virus for collecting ribosome-bound mRNA from anatomically defined projection neurons
Published in
Frontiers in Molecular Neuroscience, September 2015
DOI 10.3389/fnmol.2015.00056
Pubmed ID
Authors

Denise R. Cook-Snyder, Alexander Jones, Leon G. Reijmers

Abstract

The brain contains a large variety of projection neurons with different functional properties. The functional properties of projection neurons arise from their connectivity with other neurons and their molecular composition. We describe a novel tool for obtaining the gene expression profiles of projection neurons that are anatomically defined by the location of their soma and axon terminals. Our tool utilizes adeno-associated virus serotype 9 (AAV9), which we found to retrogradely transduce projection neurons after injection at the site of the axon terminals. We used AAV9 to express Enhanced Green Fluorescent Protein (EGFP)-tagged ribosomal protein L10a (EGFP-L10a), which enables the immunoprecipitation of EGFP-tagged ribosomes and associated mRNA with a method known as Translating Ribosome Affinity Purification (TRAP). To achieve high expression of the EGFP-L10a protein in projection neurons, we placed its expression under control of a 1.3 kb alpha-calcium/calmodulin-dependent protein kinase II (Camk2a) promoter. We injected the AAV9-Camk2a-TRAP virus in either the hippocampus or the bed nucleus of the stria terminalis (BNST) of the mouse brain. In both brain regions the 1.3 kb Camk2a promoter did not confer complete cell-type specificity around the site of injection, as EGFP-L10a expression was observed in Camk2a-expressing neurons as well as in neuronal and non-neuronal cells that did not express Camk2a. In contrast, cell-type specific expression was observed in Camk2a-positive projection neurons that were retrogradely transduced by AAV9-Camk2a-TRAP. Injection of AAV9-Camk2a-TRAP into the BNST enabled the use of TRAP to collect ribosome-bound mRNA from basal amygdala projection neurons that innervate the BNST. AAV9-Camk2a-TRAP provides a single-virus system that can be used for the molecular profiling of anatomically defined projection neurons in mice and other mammalian model organisms. In addition, AAV9-Camk2a-TRAP may enable the discovery of protein synthesis events that support information storage in projection neurons.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 58 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Germany 1 2%
Unknown 57 98%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 16 28%
Researcher 13 22%
Student > Bachelor 7 12%
Student > Master 5 9%
Student > Doctoral Student 3 5%
Other 7 12%
Unknown 7 12%
Readers by discipline Count As %
Neuroscience 23 40%
Agricultural and Biological Sciences 18 31%
Medicine and Dentistry 3 5%
Biochemistry, Genetics and Molecular Biology 2 3%
Pharmacology, Toxicology and Pharmaceutical Science 1 2%
Other 2 3%
Unknown 9 16%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 2. 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 12 November 2015.
All research outputs
#14,698,309
of 22,829,083 outputs
Outputs from Frontiers in Molecular Neuroscience
#1,642
of 2,878 outputs
Outputs of similar age
#149,569
of 274,665 outputs
Outputs of similar age from Frontiers in Molecular Neuroscience
#15
of 22 outputs
Altmetric has tracked 22,829,083 research outputs across all sources so far. This one is in the 35th percentile – i.e., 35% of other outputs scored the same or lower than it.
So far Altmetric has tracked 2,878 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 5.7. This one is in the 42nd percentile – i.e., 42% of its peers scored the same or lower than it.
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 274,665 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 44th percentile – i.e., 44% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 22 others from the same source and published within six weeks on either side of this one. This one is in the 31st percentile – i.e., 31% of its contemporaries scored the same or lower than it.