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Article ; Online: Age-related ciliopathy: Obesogenic shortening of melanocortin-4 receptor-bearing neuronal primary cilia.

Oya, Manami / Miyasaka, Yoshiki / Nakamura, Yoshiko / Tanaka, Miyako / Suganami, Takayoshi / Mashimo, Tomoji / Nakamura, Kazuhiro

Cell metabolism

2024

Abstract: Obesity is often associated with aging. However, the mechanism of age-related obesity is unknown. The melanocortin-4 receptor (MC4R) mediates leptin-melanocortin anti-obesity signaling in the hypothalamus. Here, we discovered that MC4R-bearing primary ... ...

Abstract	Obesity is often associated with aging. However, the mechanism of age-related obesity is unknown. The melanocortin-4 receptor (MC4R) mediates leptin-melanocortin anti-obesity signaling in the hypothalamus. Here, we discovered that MC4R-bearing primary cilia of hypothalamic neurons progressively shorten with age in rats, correlating with age-dependent metabolic decline and increased adiposity. This "age-related ciliopathy" is promoted by overnutrition-induced upregulation of leptin-melanocortin signaling and inhibited or reversed by dietary restriction or the knockdown of ciliogenesis-associated kinase 1 (CILK1). Forced shortening of MC4R-bearing cilia in hypothalamic neurons by genetic approaches impaired neuronal sensitivity to melanocortin and resulted in decreased brown fat thermogenesis and energy expenditure and increased appetite, finally developing obesity and leptin resistance. Therefore, despite its acute anti-obesity effect, chronic leptin-melanocortin signaling increases susceptibility to obesity by promoting the age-related shortening of MC4R-bearing cilia. This study provides a crucial mechanism for age-related obesity, which increases the risk of metabolic syndrome.
Language	English
Publishing date	2024-02-27
Publishing country	United States
Document type	Journal Article
ZDB-ID	2176834-1
ISSN	1932-7420 ; 1550-4131
ISSN (online)	1932-7420
ISSN	1550-4131
DOI	10.1016/j.cmet.2024.02.010
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Article: Behavioral Consequences of a Combination of

Fujihara, Kazuyuki / Sato, Takumi / Higeta, Kazuya / Miyasaka, Yoshiki / Mashimo, Tomoji / Yanagawa, Yuchio

Frontiers in pharmacology

2021 Volume 12, Page(s) 646088

Abstract: Glutamate decarboxylase 67-kDa isoform (GAD67), which is encoded by ... ...

Abstract	Glutamate decarboxylase 67-kDa isoform (GAD67), which is encoded by the
Language	English
Publishing date	2021-03-30
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587355-6
ISSN	1663-9812
ISSN	1663-9812
DOI	10.3389/fphar.2021.646088
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Article ; Online: Protocol for highly selective transgene expression through the flip-excision switch system by using a unilateral spacer sequence in rodents.

Matsushita, Natsuki / Kato, Shigeki / Nishizawa, Kayo / Sugawara, Masateru / Takeuchi, Kosei / Miyasaka, Yoshiki / Mashimo, Tomoji / Kobayashi, Kazuto

STAR protocols

2023 Volume 4, Issue 4, Page(s) 102667

Abstract: We present a protocol to induce Cre-dependent transgene expression in specific cell types in the rat brain, suppressing a leak expression in off-target cells, by using a flip-excision switch system with a unilateral spacer sequence. We describe steps for ...

Abstract	We present a protocol to induce Cre-dependent transgene expression in specific cell types in the rat brain, suppressing a leak expression in off-target cells, by using a flip-excision switch system with a unilateral spacer sequence. We describe steps for construction of transfer plasmids, preparation of adeno-associated viral vectors, intracranial injection, and detection of transgene expression. Our protocol provides a useful strategy for a better understanding of the structure and function of specific cell types in the complex neural circuit. For complete details on the use and execution of this protocol, please refer to Matsushita et al.
MeSH term(s)	Animals ; Rats ; Rodentia ; Transgenes
Language	English
Publishing date	2023-10-31
Publishing country	United States
Document type	Journal Article
ISSN	2666-1667
ISSN (online)	2666-1667
DOI	10.1016/j.xpro.2023.102667
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Dimensional Accuracy of Dental Casting Patterns Fabricated Using Consumer 3D Printers.

Ishida, Yoshiki / Miura, Daisuke / Miyasaka, Taira / Shinya, Akikazu

Polymers

2020 Volume 12, Issue 10

Abstract: Consumer 3D printers have improved considerably due to the evolution of additive manufacturing. This study aimed to examine the accuracy of consumer printers in dental restoration. Cylindrical patterns mimicking a full crown were created and enlarged ... ...

Abstract	Consumer 3D printers have improved considerably due to the evolution of additive manufacturing. This study aimed to examine the accuracy of consumer printers in dental restoration. Cylindrical patterns mimicking a full crown were created and enlarged from 100% to 105% of the original size. Two types of consumer 3D printers, including a fused deposition modeling (FDM) device and a stereolithography (SLA) device, and two types of dental 3D printers, including a multi-jet device and an SLA device, were used to fabricate the patterns. Then, the outer and inner diameters and depths, and surface roughness of the patterns were measured. The changing rates of the outer diameter of models created using dental printers were significantly smaller than those of the models created using consumer printers (
Language	English
Publishing date	2020-09-29
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527146-5
ISSN	2073-4360 ; 2073-4360
ISSN (online)	2073-4360
ISSN	2073-4360
DOI	10.3390/polym12102244
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Article ; Online: Highly selective transgene expression through the flip-excision switch system by using a unilateral spacer sequence.

Matsushita, Natsuki / Kato, Shigeki / Nishizawa, Kayo / Sugawara, Masateru / Takeuchi, Kosei / Miyasaka, Yoshiki / Mashimo, Tomoji / Kobayashi, Kazuto

Cell reports methods

2023 Volume 3, Issue 2, Page(s) 100393

Abstract: The flip-excision switch (FLEX) system with an adeno-associated viral (AAV) vector allows expression of transgenes in specific cell populations having Cre recombinase. A significant issue with this system is non-specific expression of transgenes in ... ...

Abstract	The flip-excision switch (FLEX) system with an adeno-associated viral (AAV) vector allows expression of transgenes in specific cell populations having Cre recombinase. A significant issue with this system is non-specific expression of transgenes in tissues after vector injection. We show here that Cre-independent recombination events in the AAV genome carrying the FLEX sequence occur mainly during the production of viral vectors in packaging cells, which results in transgene expression in off-target populations. Introduction of a relatively longer nucleotide sequence between two recognition sites at the unilateral side of the transgene cassette, termed a unilateral spacer sequence (USS), is useful to suppress the recombination in the viral genome, leading to the protection of non-specific transgene expression with enhanced gene expression selectivity. Our FLEX/USS system offers a powerful strategy for highly specific Cre-dependent transgene expression, aiming at various applications for structural and functional analyses of target cell populations.
MeSH term(s)	Recombination, Genetic ; Transgenes ; Genetic Vectors/genetics ; Genome
Language	English
Publishing date	2023-01-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2667-2375
ISSN (online)	2667-2375
DOI	10.1016/j.crmeth.2022.100393
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Article ; Online: Improvement of lateral property of unidirectional-strengthened CFRP laminates using recycled carbon fiber.

Melinda, Annisa Prita / Yamamoto, Rino / Miyasaka, Yuichi / Yoresta, Fengky Satria / Imai, Yusuke / Sugimoto, Yoshiki / Nomura, Kazuki / Matsumoto, Yukihiro

Scientific reports

2023 Volume 13, Issue 1, Page(s) 13697

Abstract: The unidirectional carbon fiber reinforced polymer (UD-CFRP) lacks the modulus of elasticity and strength in the lateral direction. This study investigates whether matrix resin with CFRP waste, recycled carbon fiber (rCF), can improve the lateral ... ...

Abstract	The unidirectional carbon fiber reinforced polymer (UD-CFRP) lacks the modulus of elasticity and strength in the lateral direction. This study investigates whether matrix resin with CFRP waste, recycled carbon fiber (rCF), can improve the lateral properties of CFRP. In total, twelve CFRP strips specimen were prefabricated of unidirectional carbon fiber (CF) sheet by hand lay-up (HLU) method and were tested by tensile test and X-ray computed tomography (X-ray CT). Factors such as fiber direction and void distribution significantly affecting its mechanical properties are assessed by X-ray CT inspection. It can be seen that rCF is mixed in a random direction at the position filled with matrix resin without rCF. However, a similar frequency of unimpregnation and voids can be observed in both specimens. Test results showed that experimental values of CFRP laminates with rCF-mixed matrix resin increased compared to the CFRP laminates without rCF. The percentage increase in the lateral tensile strength and modulus of elasticity of the rCFRP compared to the control specimen without rCF is 27.36% and 10.62%, respectively. This study proved that rCF can increase the lateral properties of unidirectional CFRP and shows the effective use of rCF for strengthening material in construction applications.
Language	English
Publishing date	2023-08-22
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-40813-2
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Article ; Online: Identification of three distinct cell populations for urate excretion in human kidneys.

Sakaguchi, Yoshihiko M / Wiriyasermkul, Pattama / Matsubayashi, Masaya / Miyasaka, Masaki / Sakaguchi, Nau / Sahara, Yoshiki / Takasato, Minoru / Kinugawa, Kaoru / Sugie, Kazuma / Eriguchi, Masahiro / Tsuruya, Kazuhiko / Kuniyasu, Hiroki / Nagamori, Shushi / Mori, Eiichiro

The journal of physiological sciences : JPS

2024 Volume 74, Issue 1, Page(s) 1

Abstract: In humans, uric acid is an end-product of purine metabolism. Urate excretion from the human kidney is tightly regulated by reabsorption and secretion. At least eleven genes have been identified as human renal urate transporters. However, it remains ... ...

Abstract	In humans, uric acid is an end-product of purine metabolism. Urate excretion from the human kidney is tightly regulated by reabsorption and secretion. At least eleven genes have been identified as human renal urate transporters. However, it remains unclear whether all renal tubular cells express the same set of urate transporters. Here, we show renal tubular cells are divided into three distinct cell populations for urate handling. Analysis of healthy human kidneys at single-cell resolution revealed that not all tubular cells expressed the same set of urate transporters. Only 32% of tubular cells were related to both reabsorption and secretion, while the remaining tubular cells were related to either reabsorption or secretion at 5% and 63%, respectively. These results provide physiological insight into the molecular function of the transporters and renal urate handling on single-cell units. Our findings suggest that three different cell populations cooperate to regulate urate excretion from the human kidney, and our proposed framework is a step forward in broadening the view from the molecular to the cellular level of transport capacity.
MeSH term(s)	Humans ; Uric Acid/metabolism ; Kidney/metabolism ; Biological Transport
Chemical Substances	Uric Acid (268B43MJ25)
Language	English
Publishing date	2024-01-02
Publishing country	Japan
Document type	Journal Article
ZDB-ID	2234472-X
ISSN	1880-6562 ; 1880-6546
ISSN (online)	1880-6562
ISSN	1880-6546
DOI	10.1186/s12576-023-00894-0
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Uc I Zs.360: Show issues

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Article ; Online: Protocol for highly selective transgene expression through the flip-excision switch system by using a unilateral spacer sequence in rodents

Natsuki Matsushita / Shigeki Kato / Kayo Nishizawa / Masateru Sugawara / Kosei Takeuchi / Yoshiki Miyasaka / Tomoji Mashimo / Kazuto Kobayashi

STAR Protocols, Vol 4, Iss 4, Pp 102667- (2023)

2023

Abstract: Summary: We present a protocol to induce Cre-dependent transgene expression in specific cell types in the rat brain, suppressing a leak expression in off-target cells, by using a flip-excision switch system with a unilateral spacer sequence. We describe ... ...

Abstract	Summary: We present a protocol to induce Cre-dependent transgene expression in specific cell types in the rat brain, suppressing a leak expression in off-target cells, by using a flip-excision switch system with a unilateral spacer sequence. We describe steps for construction of transfer plasmids, preparation of adeno-associated viral vectors, intracranial injection, and detection of transgene expression. Our protocol provides a useful strategy for a better understanding of the structure and function of specific cell types in the complex neural circuit.For complete details on the use and execution of this protocol, please refer to Matsushita et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Keywords	Molecular Biology ; Neuroscience ; Science (General) ; Q1-390
Language	English
Publishing date	2023-12-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Dimensional Accuracy of Dental Casting Patterns Fabricated Using Consumer 3D Printers

Yoshiki Ishida / Daisuke Miura / Taira Miyasaka / Akikazu Shinya

Polymers, Vol 12, Iss 2244, p

2020 Volume 2244

Abstract	Consumer 3D printers have improved considerably due to the evolution of additive manufacturing. This study aimed to examine the accuracy of consumer printers in dental restoration. Cylindrical patterns mimicking a full crown were created and enlarged from 100% to 105% of the original size. Two types of consumer 3D printers, including a fused deposition modeling (FDM) device and a stereolithography (SLA) device, and two types of dental 3D printers, including a multi-jet device and an SLA device, were used to fabricate the patterns. Then, the outer and inner diameters and depths, and surface roughness of the patterns were measured. The changing rates of the outer diameter of models created using dental printers were significantly smaller than those of the models created using consumer printers ( p < 0.05). Significantly greater surface roughness was obtained in the tooth axis of the model fabricated using the consumer FDM device ( p < 0.05). However, no significant differences were observed on the surface roughness of both axes between the consumer SLA device and the dental devices ( p > 0.05). However, FDM showed larger surface roughness than dental devices in both axes ( p < 0.05). Thus, the SLA consumer printer can be applied to fabricate resin patterns with enlargement editing of 1–3% along the horizontal axis.
Keywords	additive manufacturing ; 3D printer ; computer-aided-design/computer-aided-manufacturing ; dimensional accuracy ; Organic chemistry ; QD241-441
Subject code	670
Language	English
Publishing date	2020-09-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Genetic deletion of the 67‐kDa isoform of glutamate decarboxylase alters conditioned fear behavior in rats

Kazuyuki Fujihara / Takumi Sato / Yoshiki Miyasaka / Tomoji Mashimo / Yuchio Yanagawa

FEBS Open Bio, Vol 11, Iss 2, Pp 340-

2021 Volume 353

Abstract: The GABAergic system is thought to play an important role in the control of cognition and emotion, such as fear, and is related to the pathophysiology of psychiatric disorders. For example, the expression of the 67‐kDa isoform of glutamate decarboxylase ( ...

Abstract	The GABAergic system is thought to play an important role in the control of cognition and emotion, such as fear, and is related to the pathophysiology of psychiatric disorders. For example, the expression of the 67‐kDa isoform of glutamate decarboxylase (GAD67), a GABA‐producing enzyme, is downregulated in the postmortem brains of patients with major depressive disorder and schizophrenia. However, knocking out the Gad1 gene, which encodes GAD67, is lethal in mice, and thus, the association between Gad1 and cognitive/emotional functions is unclear. We recently developed Gad1 knockout rats and found that some of them can grow into adulthood. Here, we performed fear‐conditioning tests in adult Gad1 knockout rats to assess the impact of the loss of Gad1 on fear‐related behaviors and the formation of fear memory. In a protocol assessing both cued and contextual memory, Gad1 knockout rats showed a partial antiphase pattern of freezing during training and significantly excessive freezing during the contextual test compared with wild‐type rats. However, Gad1 knockout rats did not show any synchronous increase in freezing with auditory tones in the cued test. On the other hand, in a contextual memory specialized protocol, Gad1 knockout rats exhibited comparable freezing behavior to wild‐type rats, while their fear extinction was markedly impaired. These results suggest that GABA synthesis by GAD67 has differential roles in cued and contextual fear memory.
Keywords	amygdala ; CRISPR/Cas9 ; fear conditioning ; GABA ; genome editing ; inhibitory neurotransmitter ; Biology (General) ; QH301-705.5
Subject code	150
Language	English
Publishing date	2021-02-01T00:00:00Z
Publisher	Wiley
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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