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| Status |
Public on Aug 03, 2009 |
| Title |
Microarray analysis of splenic reservoir monocytes and their blood counterparts |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
A current paradigm states that monocytes circulate freely and patrol blood vessels, but differentiate irreversibly into dendritic cells or macrophages upon tissue entry. Here we show that bona fide undifferentiated monocytes reside in the spleen and outnumber their equivalents in circulation. The reservoir monocytes are relatively immotile, assemble in clusters in the cords of the subcapsular red pulp, and are distinct from macrophages and dendritic cells. In response to ischemic myocardial injury, splenic monocytes increase their motility, exit the spleen en masse, accumulate in injured tissue and participate in wound healing. These observations uncover a role for the spleen as a site for storage and rapid deployment of monocytes, and identify the splenic monocyte reservoir as a resource that the body exploits to regulate inflammation.
The goal of this gene expression study was to compare the gene expression of Ly-6C hi inflammatory monocytes residing in the spleen and their circulating counterparts in the blood.
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| Overall design |
Monocyte subsets of a group of four mice (C57BL/6, 8-12 weeks) were isolated by fluorescence activated cell sorting (FACS Aria, BD biosciences) as CD11bhi (CD90/B220/CD49b/NK1.1/Ly-6G)lo (F4/80/I-Ab/CD11c)lo Ly-6Chi (all antibodies BD biosciences) cells. Samples of 1,000 Ly-6Chi blood and Ly-6Chi splenic monocytes of each mouse were collected directly into 20 µl lysis buffer of the PicoPure RNA isolation kit (Arcturus). RNA extraction was subsequently performed according to the manufacturer’s instructions (Arcturus). RNA quality was assessed using RNA pico lab chips on the Agilent Bioanalyzer. For all samples a RIN above 8 could be achieved. All further steps were performed at the UCSF Shared Microarray Core Facilities according to standard protocols (http://www.arrays.ucsf.edu and http://www.agilent.com).
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| Contributor(s) |
Swirski FK, Nahrendorf M, Etzrodt M, Wildgruber M, Cortez-Retamozo V, Panizzi P, Figueiredo JL, Kohler R, Chudnovskiy A, Waterman P, Aikawa E, Mempel T, Libby P, Weissleder R, Pittet MJ |
| Citation(s) |
19644120 |
| Submission date |
Feb 16, 2009 |
| Last update date |
Jan 12, 2017 |
| Contact name |
Martin Etzrodt |
| E-mail(s) |
ETZRODT.MARTIN@MGH.HARVARD.EDU
|
| Phone |
(617) 643-0500
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| Fax |
(617) 643-6133
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| URL |
http://csb.mgh.harvard.edu/pittet
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| Organization name |
Massachusetts General Hospital
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| Department |
Center for Systems Biology
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| Lab |
Pittet Lab
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| Street address |
185 Cambridge Street
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| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02114 |
| Country |
USA |
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| Platforms (1) |
| GPL7202 |
Agilent-014868 Whole Mouse Genome Microarray 4x44K G4122F (Probe Name version) |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA112129 |
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