Biology subjects

Inoue, E.

Publications and source records attributed to Inoue, E..

9 recordsLinked to original sources

bioRxiv2026

Protective effects of heat shock protein 70 induction against global warming by using oriental bezoar and ginseng

The interest in compounds that protect against heat stress-induced damage has been heightened due to world global warming. We found the protective effects of a Japanese natural drug named BG, containing oriental bezoar and ginseng, against heat stress in Drosophila. BG suppressed the heat-induced shortened lifespan and reduced fertility in Drosophila. Interestingly, the protective effects of BG against heat stress were abolished in heat shock protein 70 (HSP70) mutant flies. To see the protective effects in humans, we applied BG on the cytotoxicity in heat-stressed human hepatic cell line, HepG2. BG suppressed heat stress-induced cytotoxicity at 43{degrees}C, and increased HSP70 and heat shock factor 1 (HSF1) mRNA expression in HepG2 cells. These findings indicate that BG protects against heat stress-induced damage via the HSF1/HSP70 pathway and has potential as a therapeutic agent for heat stress-induced disorders, including heatstroke even in human.

pharmacology and toxicology↗

bioRxiv2026

Genome compartments guide protamine replacement and genome stability during spermiogenesis

Compartment-scale genome organisation persists in mammalian sperm, yet how histone-protamine replacement is orchestrated in space and time during spermiogenesis remains unclear. Here we combine stage-resolved purification of mouse spermatids with spike-in-normalised ATAC-seq and PRM1 CUT&Tag to map chromatin accessibility and protamine incorporation across spermiogenesis. We uncover a transient, genome-wide hyper-accessible phase coincident with replacement that is uncoupled from transcription and suppressed in catalytic PHF7-mutant spermatids. PRM1 loading initiates within accessible A-compartment chromatin and later spreads across both A- and B-compartments, whereas protamine-null mice demonstrated that PRM1/ PRM2 deficiency selectively destabilises A-compartment closure. Sequencing of short DNA fragments from Prm1/Prm2 dosage-reduced epididymal sperm reveals early enrichment of breakage at protamine-targeted A-compartment regions, which dissipates as fragmentation becomes genome-wide during epididymal transit. Together, our data place histone-protamine replacement within a compartment-centred framework that links 3D genome domains to the timing, targeting and integrity of the sperm genome.

genomics↗

bioRxiv2024

Electric-field induced sleep promotion and lifespan extension in Gaucher's disease model flies: Association with improving ER stress and autophagy

Gauchers disease (GD), one kind of genetic disease, is characterized by a mutation in metabolic enzyme, glucocerebrosidase (GBA), leading to accumulation of its substrate, glucosylceramide in tissues. We previously discovered that the minos-inserted mutation of the GBA gene in fruit flies, Drosophila melanogaster, reproduced human GD characteristics and can be used as a promising model for molecular mechanisms study. Recently, we reported that extremely low-frequency electric fields (ELF-EFs) promoted sleep and extended the lifespan of normal flies. In this study, GD model flies were exposed to ELF-EFs, and sleep parameters, gene expressions, and longevity were evaluated. The amount of sleep and lifespan increased after EF exposure. Simultaneously, the expression of the endoplasmic reticulum (ER) stress-related gene, PERK and the autophagy-related gene, p62 was elevated. The data suggest that the healthy effects of EFs on GD flies are associated with improved adaptation to cellular stress.

physiology↗

bioRxiv2023

Isolation of stage-specific spermatogenic cells by dynamic histone incorporation and removal in spermatogenesis.

Due to the lack of an in vitro spermatogenesis system, studies on mammalian spermatogenesis require the isolation of specific germ cell populations for further analyses. BSA gradient and elutriation have been used for several decades to purify testicular germ cells; more recently, fluorescence-activated cell sorting (FACS) has become popular. Although each method has its advantages and disadvantages and is used depending on the purpose of the experiment, reliance on FACS is expected to be more prevalent because fewer cells can be managed. However, the currently used FACS method for testicular germ cells relies on karyotypic differences via DNA staining. Thus, it remains challenging to separate post-meiotic haploid cells (spermatids) according to their differentiation stage despite significant variations in morphology and chromatin state. In this study, we developed a method for finely separating testicular germ cells using VC mice carrying fluorescently tagged histones. This method enables the separation of spermatogonia, spermatocytes, and spermatids based on the intensity of histone fluorescence and cell size. Combined with a DNA staining dye, this method separates spermatids after elongation according to each spermiogenic stage. Although the necessity for a specific transgenic mouse line is less versatile, this method is expected to be helpful for the isolation of testicular germ cell populations because it is highly reproducible and independent of complex cell sorter settings and staining conditions.

developmental biology↗

bioRxiv2023

A novel mouse allele of the DNA/RNA helicase senataxin (Setxspcar3) causing meiotic arrest of spermatocytes and male infertility

An unbiased screen for discovering novel genes for fertility identified the spcar3, spermatocyte arrest 3, mutant phenotype. The spcar3 mutation identified a new allele of the Setx gene, encoding senataxin, a DNA/RNA helicase that regulates transcription termination by resolving DNA/RNA hybrid R-loop structures. Although mutations in the human SETX gene cause neural disorders, Setxspcar3 mutant mice do not show any apparent neural phenotype, but instead exhibit male infertility and female subfertility. Histology of the Setxspcar3mutant testes revealed absence of spermatids and mature spermatozoa in the seminiferous tubules. Cytological analysis of chromosome spread preparations of the Setxspcar3 mutant spermatocytes revealed normal synapsis, but aberrant DNA damage in the autosomes, and defective formation of the sex body. Furthermore, Setxspcar3 testicular cells exhibited abnormal accumulation of R-loops compared to wild type testicular cells. Transient expression assays identified regions of the senataxin protein required for sub-nuclear localization. Together, these results not only confirm that senataxin is required for normal meiosis and spermatogenesis but also provide a new resource for determination of its role in maintaining R-loop formation and genome integrity.

cell biology↗

bioRxiv2023

Comparisons of paternity success for resident and non-resident males and their influences on paternal sibling cohorts in Japanese macaques on Shodoshima Island

In group-living mammals, reproductive success can be attributed to both resident and non-resident males. However, the reproductive success of non-resident males has rarely been investigated at an individual level. As male reproductive success is highly skewed towards specific males, often the most dominant males, the percentage of similar-aged paternal siblings within groups is expected to increase. However, the extent to which each male contributes to the production of cohorts of paternal siblings remains unclear. Here we examined the paternity of 46 offspring born over five consecutive years in a group of Japanese macaques Macaca fuscata on Shodoshima Island. We quantitatively assessed paternity success for males, including both resident and non-resident males, and the percentages of paternal sibling dyads in the same age cohorts produced by those males. Non-resident males sired neither higher nor lower percentage of offspring compared to resident males, indicating that various males, including non-resident males, usually partake in the within-group breeding of macaque groups. These are possibly because female preference of mating partners can change over time. Subadult males had a lower percentage of paternity success, which may be because females may not prefer the physically immature subadult males. Various males, including non-resident males, contributed to the creation of paternal sibling in the same age cohort. The overall results suggest that not only resident but also non-resident males play an important role in shaping within-group kin structures. Future studies are required to examine how paternal siblings interact with each other.

ecology↗

bioRxiv2021

Induction of antioxidant and detoxifying enzymes by oriental bezoar

ObjectivesOriental bezoar, a gallstone formed in the gall sac of Bos taurus Linne var. domesticus Gmelin (Bovidae), has been used as a crude drug in oriental medicine for stroke, convulsion, epilepsy, swelling and pain in the throat, and high fever. Despite reports on the antioxidative effect of oriental bezoar-containing drugs, in vitro-cultured calculus bovis (a substitute for oriental bezoar), and its components (namely, bilirubin and biliverdin), the intracellular mechanism of the antioxidative action of oriental bezoar remains unclear. Therefore, we studied the effect of oriental bezoar on the nuclear factor erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway, which transcriptionally regulates genes encoding antioxidant and detoxifying enzymes, such as heme oxygenase-1 (HO-1), glutathione-S-transferase alpha 1 (GSTA1), and quinone oxidoreductase 1 (NQO1), to elucidate its antioxidative effect. MethodsARE promoter activity and mRNA expression (HO-1, GSTA1, and NQO1) in HepG2 cells supplemented with test drugs were examined in vitro by luciferase reporter assay and real-time PCR, respectively. ResultsOriental bezoar induced ARE promoter activity and increased the mRNA expression level of HO-1, GSTA1, and NQO1. Its components, namely, bilirubin and biliverdin, also increased the ARE promoter activity. ConclusionThese results suggest that oriental bezoar can induce antioxidative and detoxification effects via the Nrf2-ARE pathway.

pharmacology and toxicology↗

bioRxiv2020

Protective effects of saffron and its constituent crocetin to motor symptoms, short life span and rough-eyed phenotypes in the fly models of Parkinson's disease in vivo

Parkinsons disease (PD) is a common neurodegenerative disorder with motor symptoms linked to the loss of dopaminergic neurons in the brain. -Synuclein is an aggregation-prone neural protein that plays a role in the pathogenesis of PD. In our previous paper, we found that saffron; the stigma of Crocus sativus Linne (Iridaceae), and its constituents (crocin and crocetin) suppressed aggregation of -synuclein and promoted the dissociation of -synuclein fibrils in vitro. In this study, we investigated the effect of dietary saffron and its constituent, crocetin, in vivo on a fly PD model overexpressing several mutant -synuclein in a tissue-specific manner. Saffron and crocetin significantly suppressed the decrease of climbing ability in the Drosophila overexpressing A30P (A30P fly PD model) or G51D (G51D fly PD model) mutated -synuclein in neurons. Saffron and crocetin extended the life span in the G51D fly PD model. Saffron suppressed the rough-eyed phenotype and the dispersion of the size histogram of the ocular long axis in A30P fly PD model in eye. Saffron had a cytoprotective effect on a human neuronal cell line with -synuclein fibrils. These data showed that saffron and its constituent crocetin have protective effects on the progression of PD disease in animals in vivo and suggest that saffron and crocetin can be used to treat PD.

pharmacology and toxicology↗

bioRxiv2018

Metabolomic analyses of plasma and liver of mice fed with immature Citrus tumida peel

Supplementing food with functional small molecules has been shown to prevent diseases and improve the quality of life, especially in elderly people. Citrus fruits and citrus fruit-products are popular food supplements across the world. In this study, we focused on a Japanese citrus fruit, Citrus tumida hort. ex Tanaka (C. tumida), and elucidated the effects of supplementation of the peels of immature C. tumida (PIC) on food intake, body and fat tissue weights, and metabolic profiles of plasma and liver in mice. Supplementation with 5% (w/w) PIC for 4 weeks significantly suppressed body weight gain and decreased adipose tissue weight, including that of the epididymal, perirenal, and subcutaneous fats. Metabolome analyses using capillary electrophoresis time-of-flight mass spectrometry showed that the level of 2-hydroxyvaleric acid was reduced in the blood plasma of mice fed with PIC. Supplementation with PIC significantly elevated the levels of dipeptides (Thr-Asp, Ser-Glu, and Ala-Ala), glucuronic acid (and/or galacturonic acid-2), and S-methylglutathione, and significantly reduced the levels of betaine aldehyde in the liver. In conclusion, PIC supplementation affects the metabolism of fatty acids, pectin, glutathione, and choline. Our study demonstrates the potential beneficial effects of PIC, especially in metabolic syndrome and obesity. PIC may be developed as a functional food and used in the treatment of these diseases. Nutritional and metabolome studies are effective in studying the effects of specific dietary supplements and will contribute to the development of functional foods.

biochemistry↗