著者
Satoru Takahashi Chieko Saito Ikuko Koyama-Honda Noboru Mizushima
出版者
Japan Society for Cell Biology
雑誌
Cell Structure and Function (ISSN:03867196)
巻号頁・発行日
pp.22071, (Released:2022-11-22)
被引用文献数
17

In macroautophagy, disk-shaped double-membrane structures called phagophores elongate to form cup-shaped structures, becoming autophagosomes upon closure. These autophagosomes then fuse with lysosomes to become autolysosomes and degrade engulfed material. Autophagosome formation is reported to involve other organelles, including the endoplasmic reticulum (ER) and mitochondria. Organelles are also taken up by autophagosomes as autophagy cargos. However, few studies have performed systematic spatiotemporal analysis of inter-organelle relationships during macroautophagy. Here, we investigated the organelles in contact with phagophores, autophagosomes, and autolysosomes by using three-dimensional correlative light and electron microscopy with array tomography in cells starved 30 min. As previously reported, all phagophores associate with the ER. The surface area of phagophores in contact with the ER decreases gradually as they mature into autophagosomes and autolysosomes. However, the ER still associates with 92% of autophagosomes and 79% of autolysosomes, suggesting that most autophagosomes remain on the ER after closure and even when they fuse with lysosomes. In addition, we found that phagophores form frequently near other autophagic structures, suggesting the presence of potential hot spots for autophagosome formation. We also analyzed the contents of phagophores and autophagosomes and found that the ER is the most frequently engulfed organelle (detected in 65% of total phagophores and autophagosomes). These quantitative three-dimensional ultrastructural data provide insights into autophagosome–organelle relationships during macroautophagy. Key words: 3D-CLEM, autophagosome, electron microscopy, endoplasmic reticulum, lysosome
著者
Shin Hirota Satoru Takahashi Masataka Yoshimura Sakyo Hirai Takamaro Takei Asumi Orihara Hirotaka Sagawa Hikaru Wakabayashi Shoko Fuji Shinji Yamamoto Kazutaka Sumita
出版者
The Japanese Society for Neuroendovascular Therapy
雑誌
Journal of Neuroendovascular Therapy (ISSN:18824072)
巻号頁・発行日
pp.cr.2023-0041, (Released:2023-09-01)
参考文献数
20

Objective: Basilar artery occlusion (BAO) secondary to traumatic vertebral artery (VA) dissection caused by vertebral fracture is a rare cause of acute ischemic stroke, and optimal management, such as antithrombotic agents, surgical fixation, and parent artery occlusion (PAO), has been controversial. We report a case in which mechanical thrombectomy and PAO were performed for a BAO due to right VA dissection caused by a transverse foramen fracture of the axis vertebra.Case Presentation: A patient in her 80s suffered from a backward fall, and a neck CT revealed a fracture and dislocation of the right lateral mass of the axis and a compressed transverse foramen. The patient was instructed to admit and to remain in bed rest; however, she suddenly lost consciousness the following day. The CTA revealed right VA occlusion and BAO; therefore, the patient underwent mechanical thrombectomy and the BAO was successfully reperfused but the VA stenotic dissection remained. PAO of the right VA was performed on the fifth day after the accident to prevent BAO recurrence.Conclusion: Mechanical thrombectomy is an effective treatment for BAO caused by VA dissection, and PAO may contribute to the prevention of stroke recurrence.
著者
Masaharu YOSHIHARA Teppei NISHINO Naoto SAMBE Takahiro NAYAKAMA Freddy RADTKE Seiya MIZUNO Satoru TAKAHASHI
出版者
Japanese Association for Laboratory Animal Science
雑誌
Experimental Animals (ISSN:13411357)
巻号頁・発行日
pp.21-0202, (Released:2022-04-21)
被引用文献数
4

Cell labeling technologies, including the Cre/loxP system, are powerful tools in developmental biology. Although the conventional Cre/loxP system has been extensively used to label the expression of specific genes, it is less frequently used for labeling protein-protein interactions owing to technical difficulties. In the present study, we generated a new Gal4-dependent transgenic reporter mouse line that expressed Cre recombinase and a near-infrared fluorescent protein, miRFP670. To examine whether this newly generated transgenic mouse line is applicable in labeling of protein-protein interaction, we used a previously reported transgenic mouse lines that express Notch1 receptor with its intracellular domain replaced with a yeast transcription factor, Gal4. Upon the binding of this artificial Notch1 receptor and endogenous Notch1 ligands, Gal4 would be cleaved from the cell membrane to induce expression of Cre recombinase and miRFP670. Indeed, we observed miRFP670 signal in the mouse embryos (embryonic day 14.5). In addition, we examined whether our Cre recombinase was functional by using another transgenic mouse line that express dsRed after Cre-mediated recombination. We observed dsRed signal in small intestine epithelial cells where Notch1 signal was suggested to be involved in the crypt stem cell maintenance, suggesting that our Cre recombinase was functional. As our newly generated mouse line required only the functioning of Gal4, it could be useful for labeling several types of molecular activities in vivo.
著者
Satoru Takahashi Chieko Saito Ikuko Koyama-Honda Noboru Mizushima
出版者
Japan Society for Cell Biology
雑誌
Cell Structure and Function (ISSN:03867196)
巻号頁・発行日
vol.47, no.2, pp.89-99, 2022 (Released:2022-12-22)
参考文献数
48
被引用文献数
17

In macroautophagy, disk-shaped double-membrane structures called phagophores elongate to form cup-shaped structures, becoming autophagosomes upon closure. These autophagosomes then fuse with lysosomes to become autolysosomes and degrade engulfed material. Autophagosome formation is reported to involve other organelles, including the endoplasmic reticulum (ER) and mitochondria. Organelles are also taken up by autophagosomes as autophagy cargos. However, few studies have performed systematic spatiotemporal analysis of inter-organelle relationships during macroautophagy. Here, we investigated the organelles in contact with phagophores, autophagosomes, and autolysosomes by using three-dimensional correlative light and electron microscopy with array tomography in cells starved 30 min. As previously reported, all phagophores associate with the ER. The surface area of phagophores in contact with the ER decreases gradually as they mature into autophagosomes and autolysosomes. However, the ER still associates with 92% of autophagosomes and 79% of autolysosomes, suggesting that most autophagosomes remain on the ER after closure and even when they fuse with lysosomes. In addition, we found that phagophores form frequently near other autophagic structures, suggesting the presence of potential hot spots for autophagosome formation. We also analyzed the contents of phagophores and autophagosomes and found that the ER is the most frequently engulfed organelle (detected in 65% of total phagophores and autophagosomes). These quantitative three-dimensional ultrastructural data provide insights into autophagosome–organelle relationships during macroautophagy.Key words: 3D-CLEM, autophagosome, electron microscopy, endoplasmic reticulum, lysosome
著者
Kumiko KIKUKAWA Kazuhiro KITAJIMA Tetsuo MAEDA Yoshiko UENO Satoru TAKAHASHI Yoshiharu OHNO Shigeki YOSHIDA Hideto YAMADA Fumi KAWAKAMI Kazuro SUGIMURA
出版者
Japanese Society for Magnetic Resonance in Medicine
雑誌
Magnetic Resonance in Medical Sciences (ISSN:13473182)
巻号頁・発行日
vol.11, no.1, pp.71-74, 2012 (Released:2012-03-26)
参考文献数
12
被引用文献数
2 2

Microscopically, ovarian malignant Brenner tumor shows components of malignant transitional cell tumor intermixed with benign and borderline elements. We report a case of this tumor with components that transition from benign to malignant on diffusion-weighted images (DWI) and in histologic findings. The benign component showed low signal intensity on T2-weighted images, whereas the malignant component showed high signal intensity on T2-weighted images and high signal intensity on DWI with low apparent diffusion coefficient (ADC) value.
著者
Natalia GOGOLEVA Zeynab Javanfekr SHAHRI Atsushi NODA Ching-Wei LIAO Arata WAKIMOTO Yuri INOUE Hyojung JEON Satoru TAKAHASHI Michito HAMADA
出版者
Japanese Association for Laboratory Animal Science
雑誌
Experimental Animals (ISSN:13411357)
巻号頁・発行日
pp.23-0044, (Released:2023-05-12)
被引用文献数
1

Adeno-associated virus serotype 9 (AAV9) has become a popular tool for gene transfer because of its ability to cross the blood-brain barrier and efficiently transduce genetic material into a variety of cell types. The study utilized GRR (Green-to-Red Reporter) mouse embryos, in which the expression of iCre results in the disappearance of Green Fluorescent Protein (GFP) expression and the detection of Discosoma sp. Red Fluorescent Protein (DsRed) expression by intraplacental injection. Our results demonstrate that AAV9-CMV-iCre can transduce multiple organs in embryos at developmental stages E9.5–E11.5, including the liver, heart, brain, thymus, and intestine. These findings suggest that intraplacental injection of AAV9-CMV-iCre is a viable method for the widespread transduction of GRR mouse embryos.
著者
Miyuki KOMURA Mayuko SUZUKI Natthanan SANGSRIRATANAKUL Mariko ITO Satoru TAKAHASHI Md. Shahin ALAM Mizuki ONO Chisato DAIO Dany SHOHAM Kazuaki TAKEHARA
出版者
JAPANESE SOCIETY OF VETERINARY SCIENCE
雑誌
Journal of Veterinary Medical Science (ISSN:09167250)
巻号頁・発行日
vol.81, no.3, pp.466-472, 2019 (Released:2019-03-30)
参考文献数
39
被引用文献数
1 16

The inhibitory activities of grapefruit seed extract (GSE) on avian influenza virus (AIV), Newcastle disease virus (NDV), infectious bursal disease virus (IBDV), Salmonella Infantis (SI) and Escherichia coli (EC) were evaluated. Original GSE contained 0.24% benzalkonium chloride (BZC), however, 0.0025% BZC solution could not inactivate bacteria. The activity of diluted GSE (×100, ×500 and ×1,000 with redistilled water) against selected viruses and bacteria was evaluated in this study. The GSE solutions were incubated with the pathogens over a period of time after which the remaining viruses were titrated and the bacterial colonies were counted. In the presence of organic material—5% fetal bovine serum (FBS), the test solutions were sprayed at 1 cm and 30 cm distances to test the efficacy of GSE in a spray form. Furthermore, the efficacy of GSE against bacteria on clothes was tested using non-woven cloth. GSE×100 reduced the viral titer of both AIV and NDV even in 5% FBS condition. IBDV showed high resistance to GSE. GSE×1,000 inactivated both SI and EC within 5 sec, even in the presence of 5% FBS. The disinfectant was able to maintain its efficacy in the spray form at 30 cm distance. GSE was also effective against SI and EC inoculated on fabric. GSE is a potential novel disinfectant against viruses and bacteria, effective even within a short contact time.
著者
高橋 智 Xiao Mao Stephanie Byrum Nina Nishiyama Michael Pecaut Vijayalakshmi Sridharan Marjan Boerma Alan Tackett Dai Shiba Masaki Shirakawa Satoru TAKAHASHI Michael Delp
出版者
MDPI
雑誌
International journal of molecular sciences (ISSN:14220067)
巻号頁・発行日
vol.19, no.9, 2018-08

Astronauts are reported to have experienced some impairment in visual acuity during their mission on the International Space Station (ISS) and after they returned to Earth. There is emerging evidence that changes in vision may involve alterations in ocular structure and function. To investigate possible mechanisms, changes in protein expression profiles and oxidative stress-associated apoptosis were examined in mouse ocular tissue after spaceflight. Nine-week-old male C57BL/6 mice (n = 12) were launched from the Kennedy Space Center on a SpaceX rocket to the ISS for a 35-day mission. The animals were housed in the mouse Habitat Cage Unit (HCU) in the Japan Aerospace Exploration Agency (JAXA) "Kibo" facility on the ISS. The flight mice lived either under an ambient microgravity condition (µg) or in a centrifugal habitat unit that produced 1 g artificial gravity (µg + 1 g). Habitat control (HC) and vivarium control mice lived on Earth in HCUs or normal vivarium cages, respectively. Quantitative assessment of ocular tissue demonstrated that the µg group induced significant apoptosis in the retina vascular endothelial cells compared to all other groups (p < 0.05) that was 64% greater than that in the HC group. Proteomic analysis showed that many key pathways responsible for cell death, cell repair, inflammation, and metabolic stress were significantly altered in µg mice compared to HC animals. Additionally, there were more significant changes in regulated protein expression in the µg group relative to that in the µg + 1 g group. These data provide evidence that spaceflight induces retinal apoptosis of vascular endothelial cells and changes in retinal protein expression related to cellular structure, immune response and metabolic function, and that artificial gravity (AG) provides some protection against these changes. These retinal cellular responses may affect blood–retinal barrier (BRB) integrity, visual acuity, and impact the potential risk of developing late retinal degeneration.
著者
杉山 文博 高橋 智 水野 聖哉 Channabasavaiah B. Gurumurthy Aidan R. O'Brien Rolen M. Quadros John Adams Pilar Alcaide Shinya Ayabe Johnathan Ballard Surinder K. Batra Marie-Claude Beauchamp Kathleen A. Becker Guillaume Bernas David Brough Francisco Carrillo-Salinas Wesley Chan Hanying Chen Ruby Dawson Victoria DeMambro Jinke D'Hont Katharine M. Dibb James D. Eudy Lin Gan Jing Gao Amy Gonzales Anyonya R. Guntur Huiping Guo Donald W. Harms Anne Harrington Kathryn E. Hentges Neil Humphreys Shiho Imai Hideshi Ishii Mizuho Iwama Eric Jonasch Michelle Karolak Bernard Keavney Nay-Chi Khin Masamitsu Konno Yuko Kotani Yayoi Kunihiro Imayavaramban Lakshmanan Catherine Larochelle Catherine B. Lawrence Lin Li Volkhard Lindner Xian-De Liu Gloria Lopez-Castejon Andrew Loudon Jenna Lowe Loydie A. Jerome-Majewska Taiji Matsusaka Hiromi Miura Yoshiki Miyasaka Benjamin Morpurgo Katherine Motyl Yo-ichi Nabeshima Koji Nakade Toshiaki Nakashiba Kenichi Nakashima Yuichi Obata Sanae Ogiwara Mariette Ouellet Leif Oxburgh Sandra Piltz Ilka Pinz Moorthy P. Ponnusamy David Ray Ronald J. Redder Clifford J. Rosen Nikki Ross Mark T. Ruhe Larisa Ryzhova Ane M. Salvador Sabrina Shameen Alam Radislav Sedlacek Karan Sharma Chad Smith Katrien Staes Lora Starrs Fumihiro SUGIYAMA Satoru TAKAHASHI Tomohiro Tanaka Andrew W. Trafford Yoshihiro Uno Leen Vanhoutte Frederique Vanrockeghem Brandon J. Willis Christian S. Wright Yuko Yamauchi Xin Yi Kazuto Yoshimi Xuesong Zhang Yu Zhang Masato Ohtsuka Satyabrata Das Daniel J. Garry Tino Hochepied Paul Thomas Jan Parker-Thornburg Antony D. Adamson Atsushi Yoshiki Jean-Francois Schmouth Andrei Golovko William R. Thompson K. C. Kent Lloyd Joshua A. Wood Mitra Cowan Tomoji Mashimo Seiya MIZUNO Hao Zhu Petr Kasparek Lucy Liaw Joseph M. Miano Gaetan Burgio
出版者
BMC
雑誌
Genome Biology (ISSN:1474760X)
巻号頁・発行日
vol.20, no.1, 2019-08

BackgroundCRISPR-Cas9 gene-editing technology has facilitated the generation of knockout mice, providing an alternative to cumbersome and time-consuming traditional embryonic stem cell-based methods. An earlier study reported up to 16% efficiency in generating conditional knockout (cKO or floxed) alleles by microinjection of 2 single guide RNAs (sgRNA) and 2 single-stranded oligonucleotides as donors (referred herein as "two-donor floxing" method).ResultsWe re-evaluate the two-donor method from a consortium of 20 laboratories across the world. The dataset constitutes 56 genetic loci, 17,887 zygotes, and 1718 live-born mice, of which only 15 (0.87%) mice contain cKO alleles. We subject the dataset to statistical analyses and a machine learning algorithm, which reveals that none of the factors analyzed was predictive for the success of this method. We test some of the newer methods that use one-donor DNA on 18 loci for which the two-donor approach failed to produce cKO alleles. We find that the one-donor methods are 10- to 20-fold more efficient than the two-donor approach.ConclusionWe propose that the two-donor method lacks efficiency because it relies on two simultaneous recombination events in cis, an outcome that is dwarfed by pervasive accompanying undesired editing events. The methods that use one-donor DNA are fairly efficient as they rely on only one recombination event, and the probability of correct insertion of the donor cassette without unanticipated mutational events is much higher. Therefore, one-donor methods offer higher efficiencies for the routine generation of cKO animal models.
著者
Yu-Hsin CHANG Megumi C. KATOH Ahmed M. ABDELLATIF Guli XIAFUKAITI Abdelaziz ELZEFTAWY Masami OJIMA Seiya MIZUNO Akihiro KUNO Satoru TAKAHASHI
出版者
Japanese Association for Laboratory Animal Science
雑誌
Experimental Animals (ISSN:13411357)
巻号頁・発行日
pp.19-0105, (Released:2019-12-02)
被引用文献数
1

Cre/loxP is a site-specific recombination system extensively used to enable the conditional deletion or activation of target genes in a spatial- and/or temporal-specific manner. A number of pancreatic-specific Cre driver mouse lines have been broadly established for studying the development, function and pathology of pancreatic cells. However, only a few models are currently available for glucagon-producing α-cells. Disagreement exists over the role of the MAFB transcription factor in glucagon expression during postnatal life, which might be due to the lack of α-cell-specific Cre driver mice. In the present study, we established a novel Gcg-Cre knock-in mouse line with the Cre transgene expressed under the control of the preproglucagon (Gcg) promoter without disrupting the endogenous Gcg gene expression. Then, we applied this newly developed Gcg-Cre mouse line to generate a new α-cell-specific Mafb conditional knockout mouse model (MafbΔGcg). Not only α-cell number but also glucagon production were significantly decreased in MafbΔGcg mice compared to control littermates, suggesting an indispensable role of MAFB in both α-cell development and function. Taken together, our newly developed Gcg-Cre mouse line, which was successfully utilized to uncover the role of MAFB in α-cells, is a useful tool for genetic manipulation in pancreatic α-cells, providing a new platform for future studies in this field.
著者
Saki TOME Tetsuya SASAKI Satoru TAKAHASHI Yosuke TAKEI
出版者
Japanese Association for Laboratory Animal Science
雑誌
Experimental Animals (ISSN:13411357)
巻号頁・発行日
pp.19-0038, (Released:2019-06-10)
被引用文献数
3

T helper 17 (Th17) cells have been suggested to play a crucial role in various complications during pregnancy by participating in maternal immune activation (MIA). To test a possible role for Th17 cells in MIA-mediated abortion, we analyzed transgenic mice overexpressing retinoic acid-related orphan receptor gamma-t (RORγt), a master regulator of IL-17 producing cell development. These mutant mice (RORγt Tg mice) exhibited a constitutive upregulation of serum interleukin-17A (IL-17A) and decreased E-cadherin expression in cell–cell junctions of placental tissues. Abortion after the administration of a viral-mimicking synthetic double-stranded RNA polyinosinic–polycytidylic acid was more frequent in RORγt Tg mice than wild-type mice. These results suggest that excessive Th17 cell activity alters immune responsiveness and increases the rate of abortion during gestation.
著者
Shayida MAIMAITI Ryusuke KOSHIDA Masami OJIMA Kaushalya KULATHUNGA Hisashi OISHI Satoru TAKAHASHI
出版者
Japanese Association for Laboratory Animal Science
雑誌
Experimental Animals (ISSN:13411357)
巻号頁・発行日
pp.18-0182, (Released:2019-05-16)
被引用文献数
3

Mammalian postnatal growth is regulated primarily by the growth hormone (GH)/insulin-like growth factor I (IGF-I) axis. MafB is a basic leucine zipper (bZip) transcription factor that has pleiotropic functions. Although MafB plays a critical role in fetal brain development, such as in guidance for hindbrain segmentation, its postnatal role in neurons remains to be elucidated. To investigate this, we used neuron-specific Mafb conditional knockout (cKO) mice. In addition to an approximately 50% neonatal viability, the Mafb cKO mice exhibited growth retardation without apparent signs of low energy intake. Notably, serum IGF-I levels of these mice in the postnatal stage were lower than those of control mice. They seemed to have a neuroendocrine dysregulation, as shown by the upregulation of serum GH levels in the resting state and an inconsistent secretory response of GH upon administration of growth hormone-releasing hormone. These findings reveal that neuronal MafB plays an important role in postnatal development regulated by the GH/IGF-I axis.