Quantitative volumetric analysis of the Golgi apparatus following X-ray irradiation by super-resolution 3D-SIM microscopy

Takahiro Oike; Yuki Uchihara; Tiara Bunga Mayang Permata; Soehartati Gondhowiardjo; Tatsuya Ohno; Atsushi Shibata

doi:10.1007/s00795-020-00277-z

Quantitative volumetric analysis of the Golgi apparatus following X-ray irradiation by super-resolution 3D-SIM microscopy

Takahiro Oike, Yuki Uchihara, Tiara Bunga Mayang Permata, Soehartati Gondhowiardjo, Tatsuya Ohno, Atsushi Shibata

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

Abstract

To obtain quantitative volumetric data for the Golgi apparatus after ionizing radiation (IR) using super-resolution three-dimensional structured illumination (3D-SIM) microscopy. Normal human retinal pigment epithelial (RPE) cells were irradiated with X-rays (10 Gy), followed by immunofluorescence staining of the Golgi marker RCAS1. 3D-SIM imaging was performed using DeltaVision OMX version 4 and SoftWoRx 6.1. Polygon rendering and spot signal identification were performed using Imaris 8.1.2. Differences between groups were assessed by Welch’s t test. RCAS1 signals in untreated cells were located adjacent to nuclei and showed a reticular morphology. Upon IR, the area of RCAS1 signals expanded while retaining the reticular morphology. Polygon rendering imaging revealed that the volume of RCAS1 at 48 h post-IR was greater than that for unirradiated cells (93.7 ± 19.0 μm³ vs. 33.0 ± 4.2 μm³, respectively; P < 0.001): a 2.8-fold increase. Spot signal imaging showed that the number of RCAS1 spot signals post-IR was greater than that for unirradiated cells [3.4 ± 0.8 (× 10³) versus 1.3 ± 0.2 (× 10³), respectively; P < 0.001]: a 2.7-fold increase. This is the first study to report quantitative volumetric data of the Golgi apparatus in response to IR using super-resolution 3D-SIM microscopy.

Original language	English
Pages (from-to)	166-172
Number of pages	7
Journal	Medical Molecular Morphology
Volume	54
Issue number	2
DOIs	https://doi.org/10.1007/s00795-020-00277-z
Publication status	Published - Jun 2021

Keywords

3D-SIM
Golgi
Ionizing radiation
RCAS1
Super-resolution microscopy

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10.1007/s00795-020-00277-z

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@article{476c45180a154441bc4090ed7f3b0a73,

title = "Quantitative volumetric analysis of the Golgi apparatus following X-ray irradiation by super-resolution 3D-SIM microscopy",

abstract = "To obtain quantitative volumetric data for the Golgi apparatus after ionizing radiation (IR) using super-resolution three-dimensional structured illumination (3D-SIM) microscopy. Normal human retinal pigment epithelial (RPE) cells were irradiated with X-rays (10 Gy), followed by immunofluorescence staining of the Golgi marker RCAS1. 3D-SIM imaging was performed using DeltaVision OMX version 4 and SoftWoRx 6.1. Polygon rendering and spot signal identification were performed using Imaris 8.1.2. Differences between groups were assessed by Welch{\textquoteright}s t test. RCAS1 signals in untreated cells were located adjacent to nuclei and showed a reticular morphology. Upon IR, the area of RCAS1 signals expanded while retaining the reticular morphology. Polygon rendering imaging revealed that the volume of RCAS1 at 48 h post-IR was greater than that for unirradiated cells (93.7 ± 19.0 μm3 vs. 33.0 ± 4.2 μm3, respectively; P < 0.001): a 2.8-fold increase. Spot signal imaging showed that the number of RCAS1 spot signals post-IR was greater than that for unirradiated cells [3.4 ± 0.8 (× 103) versus 1.3 ± 0.2 (× 103), respectively; P < 0.001]: a 2.7-fold increase. This is the first study to report quantitative volumetric data of the Golgi apparatus in response to IR using super-resolution 3D-SIM microscopy.",

keywords = "3D-SIM, Golgi, Ionizing radiation, RCAS1, Super-resolution microscopy",

author = "Takahiro Oike and Yuki Uchihara and Permata, {Tiara Bunga Mayang} and Soehartati Gondhowiardjo and Tatsuya Ohno and Atsushi Shibata",

note = "Funding Information: This work was supported by JSPS KAKENHI Grant numbers JP17H04713 and JP20H04879 (to AS), the Takeda Science Foundation, the Sumitomo Foundation, and by the Suntory foundation for Life Sciences Bioorganic Research Institute. The work was also supported by the Program of the network-type Joint Usage/Research Center for Radiation Disaster Medical Science of Hiroshima University, Nagasaki University, and Fukushima Medical University, and by Gunma University Heavy Ion Medical Center, Hibah Kolaborasi Riset Internasional UI, and by the non-profit organization Radiotherapy Moonshot. Part of the study was conducted through the Joint Usage/Research Center Program of the Radiation Biology Center, Kyoto University. All the authors declare that (i) no support, financial or otherwise, has been received from any organization that may have an interest in the submitted work, and (ii) there are no other relationships or activities that influenced the submitted work. Publisher Copyright: {\textcopyright} 2021, The Author(s). Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",

year = "2021",

month = jun,

doi = "10.1007/s00795-020-00277-z",

language = "English",

volume = "54",

pages = "166--172",

journal = "Medical Molecular Morphology",

issn = "1860-1480",

publisher = "Springer Verlag",

number = "2",

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T1 - Quantitative volumetric analysis of the Golgi apparatus following X-ray irradiation by super-resolution 3D-SIM microscopy

AU - Oike, Takahiro

AU - Uchihara, Yuki

AU - Permata, Tiara Bunga Mayang

AU - Gondhowiardjo, Soehartati

AU - Ohno, Tatsuya

AU - Shibata, Atsushi

N1 - Funding Information: This work was supported by JSPS KAKENHI Grant numbers JP17H04713 and JP20H04879 (to AS), the Takeda Science Foundation, the Sumitomo Foundation, and by the Suntory foundation for Life Sciences Bioorganic Research Institute. The work was also supported by the Program of the network-type Joint Usage/Research Center for Radiation Disaster Medical Science of Hiroshima University, Nagasaki University, and Fukushima Medical University, and by Gunma University Heavy Ion Medical Center, Hibah Kolaborasi Riset Internasional UI, and by the non-profit organization Radiotherapy Moonshot. Part of the study was conducted through the Joint Usage/Research Center Program of the Radiation Biology Center, Kyoto University. All the authors declare that (i) no support, financial or otherwise, has been received from any organization that may have an interest in the submitted work, and (ii) there are no other relationships or activities that influenced the submitted work. Publisher Copyright: © 2021, The Author(s). Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/6

Y1 - 2021/6

N2 - To obtain quantitative volumetric data for the Golgi apparatus after ionizing radiation (IR) using super-resolution three-dimensional structured illumination (3D-SIM) microscopy. Normal human retinal pigment epithelial (RPE) cells were irradiated with X-rays (10 Gy), followed by immunofluorescence staining of the Golgi marker RCAS1. 3D-SIM imaging was performed using DeltaVision OMX version 4 and SoftWoRx 6.1. Polygon rendering and spot signal identification were performed using Imaris 8.1.2. Differences between groups were assessed by Welch’s t test. RCAS1 signals in untreated cells were located adjacent to nuclei and showed a reticular morphology. Upon IR, the area of RCAS1 signals expanded while retaining the reticular morphology. Polygon rendering imaging revealed that the volume of RCAS1 at 48 h post-IR was greater than that for unirradiated cells (93.7 ± 19.0 μm3 vs. 33.0 ± 4.2 μm3, respectively; P < 0.001): a 2.8-fold increase. Spot signal imaging showed that the number of RCAS1 spot signals post-IR was greater than that for unirradiated cells [3.4 ± 0.8 (× 103) versus 1.3 ± 0.2 (× 103), respectively; P < 0.001]: a 2.7-fold increase. This is the first study to report quantitative volumetric data of the Golgi apparatus in response to IR using super-resolution 3D-SIM microscopy.

AB - To obtain quantitative volumetric data for the Golgi apparatus after ionizing radiation (IR) using super-resolution three-dimensional structured illumination (3D-SIM) microscopy. Normal human retinal pigment epithelial (RPE) cells were irradiated with X-rays (10 Gy), followed by immunofluorescence staining of the Golgi marker RCAS1. 3D-SIM imaging was performed using DeltaVision OMX version 4 and SoftWoRx 6.1. Polygon rendering and spot signal identification were performed using Imaris 8.1.2. Differences between groups were assessed by Welch’s t test. RCAS1 signals in untreated cells were located adjacent to nuclei and showed a reticular morphology. Upon IR, the area of RCAS1 signals expanded while retaining the reticular morphology. Polygon rendering imaging revealed that the volume of RCAS1 at 48 h post-IR was greater than that for unirradiated cells (93.7 ± 19.0 μm3 vs. 33.0 ± 4.2 μm3, respectively; P < 0.001): a 2.8-fold increase. Spot signal imaging showed that the number of RCAS1 spot signals post-IR was greater than that for unirradiated cells [3.4 ± 0.8 (× 103) versus 1.3 ± 0.2 (× 103), respectively; P < 0.001]: a 2.7-fold increase. This is the first study to report quantitative volumetric data of the Golgi apparatus in response to IR using super-resolution 3D-SIM microscopy.

KW - 3D-SIM

KW - Golgi

KW - Ionizing radiation

KW - RCAS1

KW - Super-resolution microscopy

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U2 - 10.1007/s00795-020-00277-z

DO - 10.1007/s00795-020-00277-z

M3 - Article

AN - SCOPUS:85099941107

SN - 1860-1480

VL - 54

SP - 166

EP - 172

JO - Medical Molecular Morphology

JF - Medical Molecular Morphology

IS - 2

ER -

Quantitative volumetric analysis of the Golgi apparatus following X-ray irradiation by super-resolution 3D-SIM microscopy

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Keywords

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