TY - JOUR
T1 - Actin nano-architecture of phagocytic podosomes
AU - Herron, J. Cody
AU - Hu, Shiqiong
AU - Watanabe, Takashi
AU - Nogueira, Ana T.
AU - Liu, Bei
AU - Kern, Megan E.
AU - Aaron, Jesse
AU - Taylor, Aaron
AU - Pablo, Michael
AU - Chew, Teng Leong
AU - Elston, Timothy C.
AU - Hahn, Klaus M.
N1 - Funding Information:
We thank Tony Perdue from the Department of Biology Microscopy Core at UNC for assistance with Nikon N-SIM microscopy. The Imaris workstation was provided by the Neuroscience Center Microscopy Core Facility, supported in part by funding from the NIH-NINDS Neuroscience Center Support Grant P30 NS045892 and the NIH-NICHD Intellectual and Developmental Disabilities Research Center Support Grant U54 HD079124. We thank Wolfgang Bergmeier and Juan Song of the University of North Carolina for providing mouse tissue. iPALM and TIRF-SIM imaging were conducted in collaboration with the Advanced Imaging Center at Janelia Research Campus, a facility jointly supported by the Gordon and Betty Moore Foundation and the Howard Hughes Medical Institute. We are grateful to Satya Khuon for cell culture help, Richard Superfine, Michael Falvo, and Timothy O’Brien for help with micropatterning, and Ellen C. O’Shaughnessy for macrophage culture. This work was supported by grants from the National Institute of General Medical Sciences (NIGMS) to KMH (R35 GM122596) and to TCE (R35 GM127145), as well as from the National Institute of Biomedical Imaging and Bioengineering to TCE (U01 EB018816). JCH received support from the NIGMS (5T32 GM067553), MP from the National Institutes of Health (T32 GM008570-21A1), and MEK from the National Science Foundation (NSF-MCB 2005341).
Funding Information:
We thank Tony Perdue from the Department of Biology Microscopy Core at UNC for assistance with Nikon N-SIM microscopy. The Imaris workstation was provided by the Neuroscience Center Microscopy Core Facility, supported in part by funding from the NIH-NINDS Neuroscience Center Support Grant P30 NS045892 and the NIH-NICHD Intellectual and Developmental Disabilities Research Center Support Grant U54 HD079124. We thank Wolfgang Bergmeier and Juan Song of the University of North Carolina for providing mouse tissue. iPALM and TIRF-SIM imaging were conducted in collaboration with the Advanced Imaging Center at Janelia Research Campus, a facility jointly supported by the Gordon and Betty Moore Foundation and the Howard Hughes Medical Institute. We are grateful to Satya Khuon for cell culture help, Richard Superfine, Michael Falvo, and Timothy O’Brien for help with micropatterning, and Ellen C. O’Shaughnessy for macrophage culture. This work was supported by grants from the National Institute of General Medical Sciences (NIGMS) to KMH (R35 GM122596) and to TCE (R35 GM127145), as well as from the National Institute of Biomedical Imaging and Bioengineering to TCE (U01 EB018816). JCH received support from the NIGMS (5T32 GM067553), MP from the National Institutes of Health (T32 GM008570-21A1), and MEK from the National Science Foundation (NSF-MCB 2005341).
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - Podosomes are actin-enriched adhesion structures important for multiple cellular processes, including migration, bone remodeling, and phagocytosis. Here, we characterize the structure and organization of phagocytic podosomes using interferometric photoactivated localization microscopy, a super-resolution microscopy technique capable of 15–20 nm resolution, together with structured illumination microscopy and localization-based super-resolution microscopy. Phagocytic podosomes are observed during frustrated phagocytosis, a model in which cells attempt to engulf micropatterned IgG antibodies. For circular patterns, this results in regular arrays of podosomes with well-defined geometry. Using persistent homology, we develop a pipeline for semi-automatic identification and measurement of podosome features. These studies reveal an hourglass shape of the podosome actin core, a protruding knob at the bottom of the core, and two actin networks extending from the core. Additionally, the distributions of paxillin, talin, myosin II, α-actinin, cortactin, and microtubules relative to actin are characterized.
AB - Podosomes are actin-enriched adhesion structures important for multiple cellular processes, including migration, bone remodeling, and phagocytosis. Here, we characterize the structure and organization of phagocytic podosomes using interferometric photoactivated localization microscopy, a super-resolution microscopy technique capable of 15–20 nm resolution, together with structured illumination microscopy and localization-based super-resolution microscopy. Phagocytic podosomes are observed during frustrated phagocytosis, a model in which cells attempt to engulf micropatterned IgG antibodies. For circular patterns, this results in regular arrays of podosomes with well-defined geometry. Using persistent homology, we develop a pipeline for semi-automatic identification and measurement of podosome features. These studies reveal an hourglass shape of the podosome actin core, a protruding knob at the bottom of the core, and two actin networks extending from the core. Additionally, the distributions of paxillin, talin, myosin II, α-actinin, cortactin, and microtubules relative to actin are characterized.
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U2 - 10.1038/s41467-022-32038-0
DO - 10.1038/s41467-022-32038-0
M3 - Article
C2 - 35896550
AN - SCOPUS:85135009256
SN - 2041-1723
VL - 13
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 4363
ER -