Nuclear envelope composition determines the ability of neutrophil-type cells to passage through micron-scale constrictions.
Neutrophils are characterized by their distinct nuclear shape, which is thought to facilitate the transit of these cells through pore spaces less than one-fifth of their diameter. We used human promyelocytic leukemia (HL-60) cells as a model system to investigate the effect of nuclear shape in whole cell deformability. We probed neutrophil-differentiated HL-60 cells lacking expression of lamin B receptor, which fail to develop lobulated nuclei during granulopoiesis and present an in vitro model for Pelger-Huët anomaly; despite the circular morphology of their nuclei, the cells passed through micron-scale constrictions on similar timescales as scrambled controls. We then investigated the unique nuclear envelope composition of neutrophil-differentiated HL-60 cells, which may also impact their deformability; although lamin A is typically down-regulated during granulopoiesis, we genetically modified HL-60 cells to generate a subpopulation of cells with well defined levels of ectopic lamin A. The lamin A-overexpressing neutrophil-type cells showed similar functional characteristics as the mock controls, but they had an impaired ability to pass through micron-scale constrictions. Our results suggest that levels of lamin A have a marked effect on the ability of neutrophils to passage through micron-scale constrictions, whereas the unusual multilobed shape of the neutrophil nucleus is less essential.
Department of Integrative Biology and Physiology, UCLA, Los Angeles, California 90095; Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts 02139; Department of Physics and School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts 02138. Electronic address: rowat@ucla.edu.
Amy C
AC
Rowat
Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts 02139.
Diana E
DE
Jaalouk
Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts 02139.
Monika
M
Zwerger
Department of Physics and School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts 02138.
W Lloyd
WL
Ung
Department of Physics and School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts 02138.
Irwin A
IA
Eydelnant
Department of Pharmaceutical Sciences, College of Pharmacy, University of New England, Portland, Maine 04103.
Don E
DE
Olins
Department of Pharmaceutical Sciences, College of Pharmacy, University of New England, Portland, Maine 04103.
Ada L
AL
Olins
Division of Molecular Genetics, German Cancer Research Center, D-69120 Heidelberg, Germany.
Harald
H
Herrmann
Department of Physics and School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts 02138.
David A
DA
Weitz
Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts 02139; Weill Institute for Cell and Molecular Biology, Department of Biomedical Engineering, Cornell University, Ithaca, New York 14853.
Jan
J
Lammerding
D002465:Cell Movement; D002467:Cell Nucleus / Q000378:metabolism / Q000502:physiology; D053144:Cell Nucleus Shape; D015870:Gene Expression; D018922:HL-60 Cells; D006801:Humans; D034904:Lamin Type A / Q000096:biosynthesis / Q000235:genetics; D046210:Microfluidic Analytical Techniques; D020556:Neutrophil Infiltration; D009504:Neutrophils / Q000378:metabolism / Q000502:physiology; D009685:Nuclear Envelope / Q000378:metabolism* / Q000502:physiology; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology / Q000502:physiology; D000097682:Lamin B Receptor
D034904:Lamin Type A; D018160:Receptors, Cytoplasmic and Nuclear; D014212:Tretinoin; D000097682:Lamin B Receptor