title: A mouse model of yellow fluorescent protein (YFP) expression in hematopoietic cells to assess leukocyte-endothelial interactions in the microcirculation.
abstract: In this study, we describe the use of intravital microscopy in a transgenic mouse model expressing yellow fluorescent protein (YFP) under the control of a monocyte specific promoter c-fms (CD115) to track and quantify specific leukocyte subsets. Flow cytometry on peripheral and bone marrow leukocytes revealed that YFP was predominantly expressed by CD11a(+), CD11b(+), and CD14(+) monocytes. In the bone marrow, 67+/-4% of Ly6C(high) F4/80(+) cells were YFP(high) while 55+/-1% of Ly6C(low) F4/80(+) cells were YFP(low) supporting the use of c-fms(YFP) expression as a marker of monocyte lineage. 70+/-7% of CD11b(+) F4/80(+) Ly6C(+) ("triple positive") cells expressed YFP. To assess leukocyte-endothelial interactions in YFP(+) cells in c-fms(YFP+) mice, we evaluated leukocyte adhesion, rolling and local shear stress responses in the cremasteric endothelium 4 h following administration of TNFalpha. TNFalpha resulted in a five-fold increase in adhesion of YFP(+) cells to the endothelium and provided superior discriminative ability in assessing rolling and adhesion events when compared with bright field microscopy. Additionally, when compared with Rhodamine-6G labeled leukocytes or GFP(+) cells in mice transplanted with green fluorescent protein (GFP) positive bone marrow, the level of detail observed in the c-fms(YFP+) was greater, with both GFP(+) and YFP(+) cells demonstrating superior signal to noise compared to bright field microscopy. A weak positive linear correlation between wall shear stress and YFP(+) cell adhesion (r(2)=0.20, p<0.05) was seen in the cremasteric microcirculation. Taken together, these data demonstrate the use of c-fms(YFP+) mice in identifying distinct monocyte subsets and highlight the potential of this model for real-time monocyte-endothelial interactions using intravital microscopy.
authors.affiliation: Davis Heart and Lung Research Institute, The Ohio State University College of Medicine, 473 W. 12th Avenue, Room 110, Columbus, OH 43210, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Kampfrath
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Deiuliis
authors.forename: Susan D
authors.initials: SD
authors.lastname: Moffatt-Bruce
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Anderson
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Sun
authors.forename: Karen
authors.initials: K
authors.lastname: Wood
authors.forename: Michael C
authors.initials: MC
authors.lastname: Ostrowski
authors.forename: Sanjay
authors.initials: S
authors.lastname: Rajagopalan
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D016026:Bone Marrow Transplantation; D002448:Cell Adhesion / Q000502:physiology; D002450:Cell Communication; D019070:Cell Lineage; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology; D005434:Flow Cytometry; D006410:Hematopoiesis / Q000502:physiology; D036904:Leukocyte Rolling / Q000502:physiology; D008164:Luminescent Proteins / Q000378:metabolism*; D016131:Lymphocyte Subsets / Q000166:cytology* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008833:Microcirculation / Q000502:physiology; D018715:Microscopy, Video; D023421:Models, Animal; D018482:Muscle, Skeletal / Q000098:blood supply
chemical_list: D015415:Biomarkers; D008164:Luminescent Proteins
references.citation: Am J Physiol. 1993 May;264(5 Pt 2):H1504-8
references.pmid: 7684574
references.citation: Am Heart J. 1999 Nov;138(5 Pt 2):S419-20
references.pmid: 10539839
references.citation: J Immunol. 2004 Apr 1;172(7):4410-7
references.pmid: 15034056
references.citation: Am J Physiol Gastrointest Liver Physiol. 2003 Jan;284(1):G123-9
references.pmid: 12388188
references.citation: Int J Microcirc Clin Exp. 1995 Mar-Apr;15(2):85-91
references.pmid: 8655257
references.citation: J Clin Invest. 2008 Feb;118(2):710-21
references.pmid: 18202748
references.citation: Immunol Cell Biol. 2008 Jul;86(5):398-408
references.pmid: 18392044
references.citation: J Clin Invest. 2007 Jan;117(1):185-94
references.pmid: 17200718
references.citation: J Exp Med. 1999 Jan 4;189(1):37-50
references.pmid: 9874562
references.citation: J Clin Invest. 2007 Jun;117(6):1718-26
references.pmid: 17549259
references.citation: J Immunol Methods. 2000 May 26;239(1-2):109-19
references.pmid: 10821952
references.citation: Blood. 2003 Feb 1;101(3):1155-63
references.pmid: 12393599
references.citation: J Immunol. 2004 Apr 1;172(7):4418-24
references.pmid: 15034057
references.citation: Microcirculation. 2004 Jan-Feb;11(1):55-67
references.pmid: 15280097
references.citation: J Clin Invest. 2007 Apr;117(4):902-9
references.pmid: 17364026
references.citation: J Leukoc Biol. 1994 May;55(5):662-75
references.pmid: 8182345
references.citation: Immunobiology. 2006;211(6-8):609-18
references.pmid: 16920499
references.citation: Immunity. 2003 Jul;19(1):71-82
references.pmid: 12871640
references.citation: J Clin Invest. 2007 Jan;117(1):195-205
references.pmid: 17200719
references.citation: J Exp Med. 2009 Mar 16;206(3):595-606
references.pmid: 19273628
references.citation: Am J Physiol Lung Cell Mol Physiol. 2002 May;282(5):L959-67
references.pmid: 11943660
references.citation: Br J Haematol. 2007 Sep;138(6):792-801
references.pmid: 17672885
references.citation: Science. 2007 Aug 3;317(5838):666-70
references.pmid: 17673663
references.citation: J Pharmacol Toxicol Methods. 2004 Jan-Feb;49(1):1-14
references.pmid: 14670689