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Article ; Online: Moderate‐intensity exercise increases renalase levels in the blood and skeletal muscle of rats

Katsuyuki Tokinoya / Yasuko Yoshida / Takehito Sugasawa / Kazuhiro Takekoshi

FEBS Open Bio, Vol 10, Iss 6, Pp 1005-

2020 Volume 1012

Abstract: Renalase is predominantly expressed in the kidney, where it plays a role in catecholamine metabolism and blood pressure regulation. Moderate‐intensity exercise (MEX) has been shown to increase the concentration of renalase in the blood and to reduce ... ...

Abstract	Renalase is predominantly expressed in the kidney, where it plays a role in catecholamine metabolism and blood pressure regulation. Moderate‐intensity exercise (MEX) has been shown to increase the concentration of renalase in the blood and to reduce renal function in humans. Moreover, such exercise was also reported to increase catecholamine levels. Here, we examined renalase concentration in the blood and renalase expression levels in different organs after MEX in rats. Twelve male Wistar rats were made to run on a treadmill (MEX group) for 60 min at 20 m·min−1, after resting for 15 min. The control group rats were euthanized after resting on the treadmill. Tissue and blood samples were analyzed using western blotting, real‐time RT‐PCR and ELISA. Overall, the concentrations of renalase in the blood were significantly higher in the MEX group than that in the control group. Renalase expression was decreased in the kidney after 60 min of exercise, whereas the expression of renalase mRNA and protein in the extensor digitorum longus and plantaris muscles, respectively, increased after exercise. However, the expression of renalase in the other tissues examined did not change after acute exercise. In conclusion, we report that MEX for 60 min increases both renalase concentration in the blood and its expression in skeletal muscle.
Keywords	catecholamine ; exercise ; kidney ; renalase ; Sp1 ; STAT3 ; Biology (General) ; QH301-705.5
Subject code	796
Language	English
Publishing date	2020-06-01T00:00:00Z
Publisher	Wiley
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Generation of a MyoD knock-in reporter mouse line to study muscle stem cell dynamics and heterogeneity

Ryo Fujita / Seiya Mizuno / Taketaro Sadahiro / Takuto Hayashi / Takehito Sugasawa / Fumihiro Sugiyama / Yusuke Ono / Satoru Takahashi / Masaki Ieda

iScience, Vol 26, Iss 5, Pp 106592- (2023)

2023

Abstract: Summary: Myoblast determination protein 1 (MyoD) dynamics define the activation status of muscle stem cells (MuSCs), aiding in muscle tissue regeneration after injury. However, the lack of experimental platforms to monitor MyoD dynamics in vitro and in ... ...

Abstract	Summary: Myoblast determination protein 1 (MyoD) dynamics define the activation status of muscle stem cells (MuSCs), aiding in muscle tissue regeneration after injury. However, the lack of experimental platforms to monitor MyoD dynamics in vitro and in vivo has hampered the investigation of fate determination and heterogeneity of MuSCs. Herein, we report a MyoD knock-in (MyoD-KI) reporter mouse expressing tdTomato at the endogenous MyoD locus. Expression of tdTomato in MyoD-KI mice recapitulated the endogenous MyoD expression dynamics in vitro and during the early phase of regeneration in vivo. Additionally, we showed that tdTomato fluorescence intensity defines MuSC activation status without immunostaining. Based on these features, we developed a high-throughput screening system to assess the effects of drugs on the behavior of MuSCs in vitro. Thus, MyoD-KI mice are an invaluable resource for studying the dynamics of MuSCs, including their fate decisions and heterogeneity, and for drug screening in stem cell therapy.
Keywords	Cell biology ; Molecular biology ; Stem cells research ; Science ; Q
Subject code	570
Language	English
Publishing date	2023-05-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Development of a gene doping detection method to detect overexpressed human follistatin using an adenovirus vector in mice

Koki Yanazawa / Takehito Sugasawa / Kai Aoki / Takuro Nakano / Yasushi Kawakami / Kazuhiro Takekoshi

PeerJ, Vol 9, p e

2021 Volume 12285

Abstract: Background Gene doping is the misuse of genome editing and gene therapy technologies for the purpose of manipulating specific genes or gene functions in order to improve athletic performance. However, a non-invasive detection method for gene doping using ...

Abstract	Background Gene doping is the misuse of genome editing and gene therapy technologies for the purpose of manipulating specific genes or gene functions in order to improve athletic performance. However, a non-invasive detection method for gene doping using recombinant adenoviral (rAdV) vectors containing human follistatin (hFST) genes (rAdV<hFST>) has not yet been developed. Therefore, the aim of this study was to develop a method to detect gene doping using rAdV<hFST>. Methods First, we generated rAdV<hFST> and evaluated the overexpression of the hFST gene, FST protein, and muscle protein synthesis signaling using cell lines. Next, rAdV<hFST> was injected intravenously or intramuscularly into mice, and whole blood was collected, and hFST and cytomegalovirus promoter (CMVp) gene fragments were detected using TaqMan-quantitative polymerase chain reaction (qPCR). Finally, to confirm the specificity of the primers and the TaqMan probes, samples from each experiment were pooled, amplified using TaqMan-qPCR, and sequenced using the Sanger sequencing. Results The expression of hFST and FST proteins and muscle protein synthesis signaling significantly increased in C2C12 cells. In long-term, transgene fragments could be detected until 4 days after intravenous injection and 3 days after intramuscular injection. Finally, the Sanger sequencing confirmed that the primers and TaqMan probe specifically amplified the gene sequence of interest. Conclusions These results indicate the possibility of detecting gene doping using rAdV<hFST> using TaqMan-qPCR in blood samples. This study may contribute to the development of detection methods for gene doping using rAdV<hFST>.
Keywords	Gene doping ; Follistatin ; Adenovirus vector ; Medicine ; R ; Biology (General) ; QH301-705.5
Subject code	572
Language	English
Publishing date	2021-10-01T00:00:00Z
Publisher	PeerJ Inc.
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Variabilities of salivary human herpesvirus 6/7 and cortisol levels during a three-day training camp in judo athletes

Shinsuke Tamai / Hiroaki Hiraoka / Kazuhiro Shimizu / Keisuke Miyake / Daisuke Hoshi / Kai Aoki / Koki Yanazawa / Takehito Sugasawa / Kazuhiro Takekoshi / Koichi Watanabe

Journal of Physical Fitness and Sports Medicine, Vol 11, Iss 1, Pp 43-

2022 Volume 49

Abstract: Physical fatigue accompanying athletic training has been a problem for ages. While salivary cortisol has traditionally been used to assess physical stressors, salivary human herpesvirus 6 and/or 7 (HHV-6/7) have recently been presented as novel ... ...

Abstract	Physical fatigue accompanying athletic training has been a problem for ages. While salivary cortisol has traditionally been used to assess physical stressors, salivary human herpesvirus 6 and/or 7 (HHV-6/7) have recently been presented as novel microbiological markers. Thus, we examined differences in the short-term variabilities of salivary HHV-6/7 and cortisol levels in athletes. We collected saliva samples from 14 healthy male university judo athletes who participated in a three-day training camp pre- and post-training each day to measure salivary HHV-6/7 and cortisol levels. Simultaneously, the plasma superoxide dismutase (SOD) inhibition rates and total mood disturbance (TMD) scores in the Profile of Mood States were measured as indicators of physical and psychological stressors, respectively. The plasma SOD inhibition rates significantly increased post-training, but the TMD scores did not change; thus, the physical stressors were relatively higher than the psychological stressors during the training camp. Salivary HHV-6/7 levels increased post-training with a significant main effect of training, but no changes were observed in the daily levels. Only salivary HHV-7 levels showed a significant training × elapsed day interaction. Salivary cortisol levels showed a significant main effect of training, but its levels decreased post-training. These findings suggest that salivary HHV-6/7 are sensitive markers of physical fatigue more than salivary cortisol.
Keywords	fatigue ; assessment ; saliva ; biomarker ; virus ; hormone ; Sports medicine ; RC1200-1245 ; Physiology ; QP1-981
Subject code	796
Language	English
Publishing date	2022-01-01T00:00:00Z
Publisher	Japanese Society of Physical Fitness and Sports Medicine
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: One Week of CDAHFD Induces Steatohepatitis and Mitochondrial Dysfunction with Oxidative Stress in Liver

Takehito Sugasawa / Seiko Ono / Masato Yonamine / Shin-ichiro Fujita / Yuki Matsumoto / Kai Aoki / Takuro Nakano / Shinsuke Tamai / Yasuko Yoshida / Yasushi Kawakami / Kazuhiro Takekoshi

International Journal of Molecular Sciences, Vol 22, Iss 5851, p

2021 Volume 5851

Abstract: The prevalence of nonalcoholic fatty liver disease (NAFLD) has been rapidly increasing worldwide. A choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) has been used to create a mouse model of nonalcoholic steatohepatitis (NASH). There are ... ...

Abstract	The prevalence of nonalcoholic fatty liver disease (NAFLD) has been rapidly increasing worldwide. A choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) has been used to create a mouse model of nonalcoholic steatohepatitis (NASH). There are some reports on the effects on mice of being fed a CDAHFD for long periods of 1 to 3 months. However, the effect of this diet over a short period is unknown. Therefore, we examined the effect of 1-week CDAHFD feeding on the mouse liver. Feeding a CDAHFD diet for only 1-week induced lipid droplet deposition in the liver with increasing activity of liver-derived enzymes in the plasma. On the other hand, it did not induce fibrosis or cirrhosis. Additionally, it was demonstrated that CDAHFD significantly impaired mitochondrial respiration with severe oxidative stress to the liver, which is associated with a decreasing mitochondrial DNA copy number and complex proteins. In the gene expression analysis of the liver, inflammatory and oxidative stress markers were significantly increased by CDAHFD. These results demonstrated that 1 week of feeding CDAHFD to mice induces steatohepatitis with mitochondrial dysfunction and severe oxidative stress, without fibrosis, which can partially mimic the early stage of NASH in humans.
Keywords	CDAHFD ; NASH ; mitochondrial dysfunction ; liver ; oxidative stress ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Subject code	610
Language	English
Publishing date	2021-05-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: The transcriptional corepressor CtBP2 serves as a metabolite sensor orchestrating hepatic glucose and lipid homeostasis

Motohiro Sekiya / Kenta Kainoh / Takehito Sugasawa / Ryunosuke Yoshino / Takatsugu Hirokawa / Hiroaki Tokiwa / Shogo Nakano / Satoru Nagatoishi / Kouhei Tsumoto / Yoshinori Takeuchi / Takafumi Miyamoto / Takashi Matsuzaka / Hitoshi Shimano

Nature Communications, Vol 12, Iss 1, Pp 1-

2021 Volume 19

Abstract: Sensing of nutrient status coordinates the regulation of liver glucose and lipid metabolism, and is important for metabolic homeostasis. Here the authors report that transcriptional the corepressor CtBP2 can sense nutrient status and coordinate ... ...

Abstract	Sensing of nutrient status coordinates the regulation of liver glucose and lipid metabolism, and is important for metabolic homeostasis. Here the authors report that transcriptional the corepressor CtBP2 can sense nutrient status and coordinate repression of liver glucose and lipid metabolism via Fox01 and SREBP1, respectively.
Keywords	Science ; Q
Language	English
Publishing date	2021-11-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Transient changes in serum renalase concentration during long-distance running

Yasuko Yoshida / Takehito Sugasawa / Masaya Hoshino / Katsuyuki Tokinoya / Keisuke Ishikura / Hajime Ohmori / Kazuhiro Takekoshi

Journal of Physical Fitness and Sports Medicine, Vol 6, Iss 3, Pp 159-

The case of an amateur runner under continuous training

2017 Volume 166

Abstract: Renalase is a recently discovered flavin adenine dinucleotide (FAD)-dependent soluble monoamine oxidase. It is expressed principally in the kidney, but its expression in the heart, brain, and skeletal muscle has also been confirmed. It is thought that ... ...

Abstract	Renalase is a recently discovered flavin adenine dinucleotide (FAD)-dependent soluble monoamine oxidase. It is expressed principally in the kidney, but its expression in the heart, brain, and skeletal muscle has also been confirmed. It is thought that renalase is involved in cardiac function and control of systemic blood pressure primarily by metabolizing circulating catecholamines. Animal experiments have suggested the possibility that exercise is involved in renalase secretion, but this has not been confirmed in human studies. The principal aim of the present study was to investigate the relationship between exercise and renalase concentration in humans. Subjects were young men who were continuously training to run distances of 10-20 km; the total distance that was run in the present study was 30 km. Measurements were performed at four points (before running and after running 10 km, 20 km, and 30 km). The results of one-way analysis of variance examining repeated measurements of serum renalase concentration revealed significant differences (P = 0.007). A significant negative correlation between serum renalase concentration and estimated glomerular filtration rate, which indicates renal function, calculated using serum cystatin C concentration was noted. A significant positive correlation between serum renalase concentration and oxidative stress was observed in the period before exercise and after running 20 km. The present study verified the exercise-induced increase in serum renalase concentration in amateur runners who were continuously training.
Keywords	renalase ; exercise ; cystatin c ; tbars ; egfr ; Sports medicine ; RC1200-1245 ; Physiology ; QP1-981
Subject code	796
Language	English
Publishing date	2017-05-01T00:00:00Z
Publisher	Japanese Society of Physical Fitness and Sports Medicine
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Influence of acute exercise on renalase and its regulatory mechanism

Tokinoya, Katsuyuki / Jun Shiromoto / Takehito Sugasawa / Yasuko Yoshida / Kai Aoki / Yoshimi Nakagawa / Hajime Ohmori / Kazuhiro Takekoshi

Life sciences. 2018 Oct. 01, v. 210

2018

Abstract: Renalase expression in the kidneys and liver is regulated by nuclear factor (NF)-κB, Sp1, and hypoxia-inducible factor (HIF)-1α. The dynamics of renalase expression in acute exercise, and its mechanism and physiological effects are unclear. We evaluated ... ...

Abstract	Renalase expression in the kidneys and liver is regulated by nuclear factor (NF)-κB, Sp1, and hypoxia-inducible factor (HIF)-1α. The dynamics of renalase expression in acute exercise, and its mechanism and physiological effects are unclear. We evaluated the effect of different exercise intensities on renalase expression and examined its mechanism and physiological effects.21 male Wistar rats ran for 30 min on a treadmill after resting for 15 min. The sedentary group rested on the treadmill while the exercise group ran for 30 min at 10 or 30 m/min. Skeletal muscles, the kidney, heart, liver, and blood samples were collected after exercise. The expression of renalase and phosphate IkB-α and Akt was measured by western blotting, while HIF-1α, Sp1, MuRF-1, and MAFbx were measured in the skeletal muscle by real-time RT-PCR.Renalase expression in skeletal muscles increased after acute exercise, while its expression in the kidneys, heart, and liver decreased. NF-κB regulated renalase expression in the plantaris muscle and that of HIF-1α in the soleus muscle. Phosphate Akt in the plantaris muscle significantly increased in the 30 m/min group compared with that in the sedentary group. MuRF-1 in the plantaris did not change between these groups.Renalase expression in skeletal muscles increased after acute exercise but decreased in other tissues. This increase may be a response to exercise-induced oxidative stress. Furthermore, NF-κB in the plantaris muscle may mainly regulate renalase expression, and support a relationship with the cell protective effects of renalase.
Keywords	Western blotting ; blood sampling ; exercise ; heart ; hypoxia-inducible factor 1 ; kidneys ; laboratory animals ; liver ; males ; muscles ; oxidative stress ; phosphates ; protective effect ; rats ; renalase ; skeletal muscle ; transcription factor NF-kappa B
Language	English
Dates of publication	2018-1001
Size	p. 235-242.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	3378-9
ISSN	1879-0631 ; 0024-3205
ISSN (online)	1879-0631
ISSN	0024-3205
DOI	10.1016/j.lfs.2018.07.042
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Characterization of Osteoarthritis in a Medial Meniscectomy-Induced Animal Model Using Contrast-Enhanced X-ray Microtomography

Takehito Sugasawa / Tomoaki Kuji / Kai Aoki / Koki Yanazawa / Akiko Takenouchi / Makoto Watanabe / Yoshiya Tome / Yoshinori Takeuchi / Yuichi Aita / Naoya Yahagi / Yasuhiro Shishikura / Seiko Ono / Yasuko Yoshida / Yasushi Kawakami / Kazuhiro Takekoshi

Biomedicines, Vol 8, Iss 3, p

2020 Volume 56

Abstract: The aim of this study was to clarify degradation characteristics in each tissue of the knee complex of a medial meniscectomy (MMx)-induced knee osteoarthritis (KOA) animal model using classical methods and an alternative comprehensive evaluation method ... ...

Abstract	The aim of this study was to clarify degradation characteristics in each tissue of the knee complex of a medial meniscectomy (MMx)-induced knee osteoarthritis (KOA) animal model using classical methods and an alternative comprehensive evaluation method called contrast-enhanced X-ray micro-computed tomography (CEX-μCT), which was developed in the study. Surgical MMx was performed in the right knee joints of five male Wistar rats to induce KOA. At four weeks post-surgery, the synovitis was evaluated using quantitative polymerase chain reaction (qPCR). Degradations of the articular cartilage of the tibial plateau were evaluated using classical methods and CEX-μCT. Evaluation of the synovitis demonstrated significantly increased expression levels of inflammation-associated marker genes in MMx-treated knees compared with those in sham-treated knees. Evaluation of the articular cartilage using classical methods showed that MMx fully induced degradation of the cartilage. Evaluation using CEX-μCT showed that local areas of the medial cartilage of the tibial plateau were significantly reduced in MMx-treated knees compared with those in sham-treated knees. On the other hand, total cartilage volumes were significantly increased in MMx-treated knees. On the basis of the findings of this study, the method could be relevant to study new treatments in KOA research.
Keywords	osteoarthritis ; synovitis ; articular cartilage ; microfocus x-ray ct ; 3d analysis ; Biology (General) ; QH301-705.5
Subject code	616
Language	English
Publishing date	2020-03-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Influence of Intermittent Cold Stimulations on CREB and Its Targeting Genes in Muscle

Takehito Sugasawa / Yoshiya Tome / Yoshinori Takeuchi / Yasuko Yoshida / Naoya Yahagi / Rahul Sharma / Yuichi Aita / Haruna Ueda / Reina Maruyama / Kaoru Takeuchi / Shohei Morita / Yasushi Kawamai / Kazuhiro Takekoshi

International Journal of Molecular Sciences, Vol 21, Iss 4588, p

Investigations into Molecular Mechanisms of Local Cryotherapy

2020 Volume 4588

Abstract: Local cryotherapy is widely used as a treatment for sports-related skeletal muscle injuries. The molecular mechanisms are unknown. To clarify these mechanisms, we applied one to three 15-min cold stimulations at 4 °C to various cell lines (in vitro), the ...

Abstract	Local cryotherapy is widely used as a treatment for sports-related skeletal muscle injuries. The molecular mechanisms are unknown. To clarify these mechanisms, we applied one to three 15-min cold stimulations at 4 °C to various cell lines (in vitro), the tibialis anterior (TA) muscle (ex vivo), and mouse limbs (in vivo). In the in vitro assay, cyclic AMP (cAMP) response element binding protein 1 (CREB1) was markedly phosphorylated (p-CREB1), and the CREB-binding protein (CBP) was recruited to p-CREB-1 in response to two or three cold stimulations. In a reporter assay with the cAMP-responsive element, the signals significantly increased after two to three cold stimulations at 4 °C. In the ex vivo study, CREB-targeting genes were significantly upregulated following two or three cold stimulations. The in vivo experiment disclosed that cold stimulation of a mouse limb for 9 days significantly increased mitochondrial DNA copy number and upregulated genes involved in mitochondrial biogenesis. The results suggest that local cryotherapy increases CREB transcription and upregulates CREB-targeting genes, in a manner dependent on cold stimulation frequency and duration. This information will inform further investigations into local cryotherapy as a treatment for sports-related skeletal muscle trauma.
Keywords	CREB ; cryotherapy ; gene expression ; icing ; mitochondria ; Pgc-1α ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Subject code	570
Language	English
Publishing date	2020-06-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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