PMID- 26557053
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20151111
LR  - 20200930
IS  - 1662-5099 (Print)
IS  - 1662-5099 (Electronic)
IS  - 1662-5099 (Linking)
VI  - 8
DP  - 2015
TI  - A retrograde adeno-associated virus for collecting ribosome-bound mRNA from 
      anatomically defined projection neurons.
PG  - 56
LID - 10.3389/fnmol.2015.00056 [doi]
LID - 56
AB  - 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.
FAU - Cook-Snyder, Denise R
AU  - Cook-Snyder DR
AD  - Department of Neuroscience, School of Medicine, Tufts University Boston, MA, USA 
      ; Department of Biology and Neuroscience Program, Carthage College Kenosha, WI, 
      USA.
FAU - Jones, Alexander
AU  - Jones A
AD  - Department of Neuroscience, School of Medicine, Tufts University Boston, MA, USA 
      ; Graduate Program in Neuroscience, Sackler School of Graduate Biomedical 
      Sciences, Tufts University Boston, MA, USA.
FAU - Reijmers, Leon G
AU  - Reijmers LG
AD  - Department of Neuroscience, School of Medicine, Tufts University Boston, MA, USA.
LA  - eng
GR  - DP2 OD006446/OD/NIH HHS/United States
GR  - K12 GM074869/GM/NIGMS NIH HHS/United States
GR  - P30 NS047243/NS/NINDS NIH HHS/United States
GR  - R21 MH099794/MH/NIMH NIH HHS/United States
PT  - Journal Article
DEP - 20150924
PL  - Switzerland
TA  - Front Mol Neurosci
JT  - Frontiers in molecular neuroscience
JID - 101477914
PMC - PMC4617378
OTO - NOTNLM
OT  - AAV9
OT  - Camk2a
OT  - EGFP-L10a
OT  - TRAP
OT  - projection neuron
EDAT- 2015/11/12 06:00
MHDA- 2015/11/12 06:01
PMCR- 2015/01/01
CRDT- 2015/11/12 06:00
PHST- 2015/06/09 00:00 [received]
PHST- 2015/09/07 00:00 [accepted]
PHST- 2015/11/12 06:00 [entrez]
PHST- 2015/11/12 06:00 [pubmed]
PHST- 2015/11/12 06:01 [medline]
PHST- 2015/01/01 00:00 [pmc-release]
AID - 10.3389/fnmol.2015.00056 [doi]
PST - epublish
SO  - Front Mol Neurosci. 2015 Sep 24;8:56. doi: 10.3389/fnmol.2015.00056. eCollection 
      2015.