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J Neurosci. 2015 Feb 18;35(7):2860-70. doi: 10.1523/JNEUROSCI.3199-14.2015.

Interleukin 1 type 1 receptor restore: a genetic mouse model for studying interleukin 1 receptor-mediated effects in specific cell types.

Author information

1
Division of Biosciences, The Ohio State University, Columbus, Ohio 43210, Institute for Behavioral Medicine Research, The Ohio State University Wexner Medical Center, Columbus, Ohio 43210.
2
Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105.
3
Institute for Behavioral Medicine Research, The Ohio State University Wexner Medical Center, Columbus, Ohio 43210, Department of Neuroscience, The Ohio State University, Columbus, Ohio 43210, and.
4
Department of MVIMG, Wexner Medical Center Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210.
5
Institute for Behavioral Medicine Research, The Ohio State University Wexner Medical Center, Columbus, Ohio 43210.
6
Division of Biosciences, The Ohio State University, Columbus, Ohio 43210, Institute for Behavioral Medicine Research, The Ohio State University Wexner Medical Center, Columbus, Ohio 43210, quan.14@osu.edu.

Abstract

Interleukin-1 (IL-1) mediates diverse neurophysiological and neuropathological effects in the CNS through type I IL-1 receptor (IL-1R1). However, identification of IL-1R1-expressing cell types and cell-type-specific functions of IL-1R1 remains challenging. In this study, we created a novel genetic mouse model in which IL-1R1 gene expression is disrupted by an intronic insertion of a loxP flanked disruptive sequence that can be deleted by Cre recombinase, resulting in restored IL-1R1 gene expression under its endogenous promoters. A second mutation was introduced at stop codon of the IL-1R1 gene to allow tracking of the restored IL-1R1 protein by a 3HA tag and IL-1R1 mRNA by tdTomato fluorescence. These animals were designated as IL-1R1(r/r) and exhibited an IL-1R1 knock-out phenotype. We used IL-1R1 globally restored mice (IL-1R1(GR/GR)) as an IL-1R1 reporter and observed concordant labeling of IL-1R1 mRNA and protein in brain endothelial cells. Two cell-type-specific IL-1R1 restore lines were generated: Tie2Cre-IL-1R1(r/r) and LysMCre-IL-1R1(r/r). Brain endothelial COX-2 expression, CNS leukocyte infiltration, and global microglia activation induced by intracerebroventricular injection of IL-1β were not observed in IL-1R1(r/r) or LysMCre-IL-1R1(r/r) mice, but were restored in Tie2Cre-IL-1R1(r/r) mice. These results reveal IL-1R1 expression in endothelial cells alone is sufficient to mediate these central IL-1-induced responses. In addition, ex vivo IL-1β stimulation increased IL-1β expression in bone marrow cells in wild-type, Tie2Cre-IL-1R1(r/r), and LysMCre-IL-1R1(r/r), but not IL-1R1(r/r) mice. These results demonstrate this IL-1R1 restore model is a valuable tool for studying cell-type-specific functions of IL-1R1.

KEYWORDS:

blood–brain barrier; endothelial cells; interleukin 1; knock-in; neuroinflammation; restore

PMID:
25698726
PMCID:
PMC4331620
DOI:
10.1523/JNEUROSCI.3199-14.2015
[Indexed for MEDLINE]
Free PMC Article

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