title: Interleukin 1 type 1 receptor restore: a genetic mouse model for studying interleukin 1 receptor-mediated effects in specific cell types.
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.
authors.affiliation: 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.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105.
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Yamashita
authors.affiliation: 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.
authors.forename: Qun
authors.initials: Q
authors.lastname: Chen
authors.affiliation: 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.
authors.forename: Natalya
authors.initials: N
authors.lastname: Belevych
authors.affiliation: 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.
authors.forename: Daniel B
authors.identifier: http://orcid.org/0000-0001-8402-4154
authors.initials: DB
authors.lastname: Mckim
authors.affiliation: 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.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Tarr
authors.affiliation: Department of MVIMG, Wexner Medical Center Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210.
authors.forename: Vincenzo
authors.identifier: http://orcid.org/0000-0001-6163-1779
authors.initials: V
authors.lastname: Coppola
authors.affiliation: Institute for Behavioral Medicine Research, The Ohio State University Wexner Medical Center, Columbus, Ohio 43210.
authors.forename: Nikitaa
authors.initials: N
authors.lastname: Nath
authors.affiliation: Institute for Behavioral Medicine Research, The Ohio State University Wexner Medical Center, Columbus, Ohio 43210.
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Nemeth
authors.affiliation: Institute for Behavioral Medicine Research, The Ohio State University Wexner Medical Center, Columbus, Ohio 43210.
authors.forename: Zunera W
authors.initials: ZW
authors.lastname: Syed
authors.affiliation: 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.
authors.forename: John F
authors.initials: JF
authors.lastname: Sheridan
authors.affiliation: 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.
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Godbout
authors.affiliation: Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105.
authors.forename: Jian
authors.initials: J
authors.lastname: Zuo
authors.affiliation: 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.
authors.forename: Ning
authors.initials: N
authors.lastname: Quan
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism; D001921:Brain / Q000166:cytology*; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism; D004622:Embryo, Mammalian; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics*; D005838:Genotype; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D053583:Interleukin-1beta / Q000494:pharmacology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D053573:Receptors, Interleukin-1 Type I / Q000235:genetics* / Q000378:metabolism*; D013997:Time Factors; D000099266:Engrailed 2 Protein
chemical_list: D002135:Calcium-Binding Proteins; D051546:Cyclooxygenase 2; D018398:Homeodomain Proteins; D053583:Interleukin-1beta; D008840:Microfilament Proteins; D009419:Nerve Tissue Proteins; D053573:Receptors, Interleukin-1 Type I; D000099266:Engrailed 2 Protein; C491049:Aif1 protein, mouse; C583355:IL1R1 protein, mouse; C077623:Ptgs2 protein, mouse
keywords: blood–brain barrier; endothelial cells; interleukin 1; knock-in; neuroinflammation; restore