Mammalian type opsin 5 preferentially activates G14 in Gq-type G proteins triggering intracellular calcium response

Mammalian type opsin 5 (Opn5m), a UV-sensitive G protein-coupled receptor opsin highly conserved in vertebrates, would provide a common basis for UV sensing from lamprey to humans. However, G protein coupled with Opn5m remains controversial due to variations in assay conditions and the origin of Opn5m across different reports. Here, we examined Opn5m from diverse species using an aequorin luminescence assay and G alpha-KO cell line. Beyond the commonly studied major G alpha classes, G alpha q, G alpha 11, G alpha 14, and G alpha 15 in the Gq class were individually investigated in this study, as they can drive distinct signaling pathways in addition to a canonical calcium response. UV light triggered a calcium response via all the tested Opn5m proteins in 293T cells, which was abolished by Gq-type G alpha deletion and rescued by cotransfection with mouse and medaka Gq-type G alpha proteins. Opn5m preferentially activated G alpha 14 and close relatives. Mutational analysis implicated specific regions, including alpha 3-beta 5 and alpha G-alpha 4 loops, alpha G and alpha 4 helices, and the extreme C terminus, in the preferential activation of G alpha 14 by Opn5m. FISH revealed co-expression of genes encoding Opn5m and G alpha 14 in the scleral cartilage of medaka and chicken eyes, supporting their physiological coupling. This suggests that the preferential activation of G alpha 14 by Opn5m is relevant for UV sensing in specific cell types

The Opsin 3/Teleost multiple tissue opsin system: mRNA localization in the retina and brain of medaka (Oryzias latipes)

The photoreceptor protein, opsin, is one of the major components for vision and photoreceptive function in animals. Although many opsins have been discovered from animal genomes, only a few nonimage‐forming functions mediated by opsins have been identified. Understanding the mRNA distribution of photoreceptor proteins is one crucial step in uncovering their photoreceptive function in animals. Here, we focus on the medaka fish (Oryzias latipes) Opsin 3 (Opn3)/Teleost multiple opsin (Tmt) system, which constitutes a separate phylogenetic group, having putative blue light photoreceptors for nonimage‐forming functions. In medaka, there is one opn3 and five tmt‐opsin orthologs. The expression pattern of the opn3/tmt‐opsins in the retina and brain was investigated by in situ hybridization. mRNAs for opn3/tmt‐opsins were distributed in the retinal ganglion cells as well as interneurons and specific brain nuclei. Specifically, hybridization signals were observed in the glutamate decarboxylase 1 (gad1)‐expressing amacrine cells for opn3, tmt1a, tmt1b, and tmt2, in the caudal lobe of the cerebellum for tmt1b and tmt2, in the cranial nerve nuclei for opn3, tmt1a, tmt1b, tmt2, and in the rostral pars distalis (adenohypophysis) for opn3. These expression patterns suggest that blue light sensing in the fish retina and brain may be involved in the integration of visual inputs, vestibular function, somatosensation, motor outputs, and pituitary endocrine regulation

The medaka mutant deficient in eyes shut homolog exhibits opsin transport defects and enhanced autophagy in retinal photoreceptors

Eyes shut homolog (EYS) encodes a proteoglycan and the human mutation causes retinitis pigmentosa type 25 (RP25) with progressive retinal degeneration. RP25 most frequently affects autosomal recessive RP patients with many ethnic backgrounds. Although studies using RP models have facilitated the development of therapeutic medications, Eys has been lost in rodent model animals. Here we examined the roles for Eys in the maintenance of photoreceptor structure and function by generating eys-null medaka fish using the CRISPR-Cas9 system. Medaka EYS protein was present near the connecting cilium of wild-type photoreceptors, while it was absent from the eys−/− retina. The mutant larvae exhibited a reduced visual motor response compared with wild-type. In contrast to reported eys-deficient zebrafish at the similar stage, no retinal cell death was detected in the 8-month post-hatching (8-mph) medaka eys mutant. Immunohistochemistry showed a significant reduction in the length of cone outer segments (OSs), retention of OS proteins in the inner segments of photoreceptors, and abnormal filamentous actin network at the base of cone OSs in the mutant retina by 8 mph. Electron microscopy revealed aberrant structure of calyceal processes, numerous vesiculation and lamellar interruptions, and autophagosomes in the eys-mutant cone photoreceptors. In situ hybridization showed an autophagy component gene, gabarap, was ectopically expressed in the eys-null retina. These results suggest eys is required for regeneration of OS, especially of cone photoreceptors, and transport of OS proteins by regulating actin filaments. Enhanced autophagy may delay the progression of retinal degeneration when lacking EYS in the medaka retina

A Sealed-bid Auction with Fund Binding: Preventing Maximum Bidding Price Leakage

In an open-bid auction, a bidder can know the budgets of other bidders. Thus, a sealed-bid auction that hides bidding prices is desirable. However, in previous sealed-bid auction protocols, it has been difficult to provide a ``fund binding'' property, which would guarantee that a bidder has funds more than or equal to the bidding price and that the funds are forcibly withdrawn when the bidder wins. Thus, such protocols are vulnerable to false bidding. As a solution, many protocols employ a simple deposit method in which each bidder sends a deposit to a smart contract, which is greater than or equal to the bidding price, before the bidding phase. However, this deposit reveals the maximum bidding price, and it is preferable to hide this information. In this paper, we propose a sealed-bid auction protocol that provides a fund binding property. Our protocol not only hides the bidding price and a maximum bidding price, but also provides fund binding, simultaneously. For hiding the maximum bidding price, we pay attention to the fact that usual Ethereum transactions and transactions for sending funds to a one-time address have the same transaction structure, and it seems that they are indistinguishable. We discuss how much bidding transactions are hidden. We also employ DECO (Zhang et al,. CCS 2020) that proves the validity of the data to a verifier in which the data are taken from a source without showing the data itself. Finally, we give our implementation which shows transaction fees required and compare it to a sealed-bid auction protocol employing the simple deposit method

Diversification processes of teleost intron-less opsin genes

Opsins are universal photosensitive proteins in animals. Vertebrates have a variety of opsin genes for visual and non-visual photoreceptions. Analysis of the gene structures shows that most opsin genes have introns in their coding regions. However, teleosts exceptionally have several intron-less opsin genes which are presumed to have been duplicated by an RNA-based gene duplication mechanism, retroduplication. Among these retrogenes, we focused on the Opn4 (melanopsin) gene responsible for non-image-forming photoreception. Many teleosts have five Opn4 genes including one intron-less gene, which is speculated to have been formed from a parental intron-containing gene in the Actinopterygii. In this study, to reveal the evolutionary history of Opn4 genes, we analyzed them in teleost (zebrafish and medaka) and non-teleost (bichir, sturgeon and gar) fishes. Our synteny analysis suggests that the intron-less Opn4 gene emerged by retroduplication after branching of the bichir lineage. In addition, our biochemical and histochemical analyses showed that, in the teleost lineage, the newly acquired intron-less Opn4 gene became abundantly used without substantial changes of the molecular properties of the Opn4 protein. This stepwise evolutionary model of Opn4 genes is quite similar to that of rhodopsin genes in the Actinopterygii. The unique acquisition of rhodopsin and Opn4 retrogenes would have contributed to the diversification of the opsin gene repertoires in the Actinopterygii and the adaptation of teleosts to various aquatic environments

Examining the Youth Multi-Sport Event Environment: Implications towards athlete development and transitioning.

Many factors are associated with a person’s attitude formation and intention towards a behavior. In this study, we examined organizational factors that helped form young athletes attitudes regarding their future plans in high performance sport. Through a mixed method survey design, data was collected from 207 young pre-elite athletes who competed during the 2017 Commonwealth Youth Games in Bahamas. Several organizationally controlled aspects of the games environment were found to contribute to young athlete’s satisfaction with the event including their accommodations, available information regarding their sport and finally, the social and cultural activities during games-time. However, satisfaction with the games environment was not predictive of young athletes future intentions to remain in sport. Qualitative thematic coding denoted two key themes related to athlete plans to continue in high performance sport: level of satisfaction and learning. Further, qualitative results revealed five main impediments to continuing in high performance sport specifically, physiological, psychological, performance, environment and life concerns. The paper contributes to our understanding of the controllable and uncontrollable social and environmental factors in a multi-sport international event. Nationally controlled factors that influence young athletes attitude formation, specifically their satisfaction and intentions to remain in sport

The effect of Rugby World Cup 2019 on residents’ psychological well-being: a mediating role of psychological capital

© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. The current study aimed to examine the relationships among residents’ impact perception of 2019 Rugby World Cup (i.e. economic impact, cultural interests and new opportunity, community pride, and environmental impact), psychological capital (i.e. hope, self-efficacy, optimism, and resilience), and psychological well-being. The two-wave time-lagged data were collected from residents in the host cities (a following week of the event and eight weeks after the event; N = 206). The results indicated that residents’ perceptions of event impacts were positively associated with psychological capital, which influenced psychological well-being over time. Cultural interests and community pride were considered particularly influential in enhancing residents’ psychological well-being. The findings are useful for policy makers, planning to utilize mega events to boost residents’ psychological well-being

Effect of difference in interaction strength on two-dimensional lattice structure in a binary system with DNA nanoparticles

Keeping two-dimensional lattice structures formed by nanoparticles covered with DNA in mind, we carry out Brownian dynamics simulations to study the effect of interaction strength on a two-dimensional lattice structure formed in a binary system. In our previous study [H. Katsuno, Y. Maegawa, and M. Sato, J. Phys. Soc. Jpn. 85, 074605 (2016)], we carried out simulations using the Lennard-Jones potential, in which the difference in interaction length was taken into account. When the interaction length between different species, σ$, is smaller than that between the same species, σ, various lattice structures were formed with changing the ratio σ$/σ. In this paper, taking the difference in the interaction strength into account, we study the effect of the difference in interaction strength on the two-dimensional lattice structure. © 2017 The Japan Society of Applied PhysicsEmbargo Period 12 month