Biology subjects

Kumagai, T.

Publications and source records attributed to Kumagai, T..

4 recordsLinked to original sources

bioRxiv2025

Heat-mediated manipulation of gene expression by IR-LEGO in the developing genitalia in Drosophila

Manipulating gene expression in a tissue-specific and temporally controlled manner is essential for understanding the function of the focal genes. Still, in many cases, the limited availability of specific promotors to drive ectopic manipulation remains a restricting factor in developing organs, even in Drosophila. Developing external genitalia is one such organ with a complex anatomical structure shaped by a joint regulatory network of many transcription factors. To overcome the restriction, we employed the infrared laser-evoked gene operator system (IR-LEGO), in which infrared laser (1,480 nm) irradiation induces gene expression under the control of a heat shock promoter. Pupal genital structures were irradiated at approximately 24 or 48 hours after puparium formation. We tested a range of laser power and depth to the target structure by a reporter assay using green fluorescent protein, which was induced under the control of the heat shock protein 70 promoter (hs-GAL4). In previous studies, the IR-LEGO has been used as a tool to induce ectopic transgene expression. In this study, we attempted to knock down genes such as yellow (y) and odd-paired (opa) ectopically by RNAi using the GAL4/UAS system. The results demonstrated that this technique has a high potential in manipulating transcript abundance levels in small groups of cells in specific genital structures to unravel novel functions of genes involved in the morphogenesis of species-specific and rapidly evolving anatomical structures. Article summaryManipulating gene expression is essential for understanding the function of the genes at specific regions of the body. In particular, knowing how different genes contribute to shaping complex anatomical structures, such as insect external genitalia, is an intriguing question. While manipulating gene expression precisely at a specific time and space remains challenging, we show that we can knock down a gene in a specific group of cells in the developing genitalia of Drosophila pupae, using infrared laser irradiation. This is a new approach to investigating how activity of genes shapes the rapidly evolving male genitalia in insects.

genetics↗

bioRxiv2024

Effectiveness of Exercise Intervention in Preventing Active Arthritis Exacerbation in an SKG Mouse Model of Rheumatoid Arthritis

ObjectivesTo investigate the effects of low-intensity exercise on active arthritis in an SKG mouse model of human rheumatoid arthritis (RA) pathology. MethodsTwenty-four female SKG mice were divided into three groups: sedentary (control), AR (induced arthritis), and AREx (induced arthritis plus low-intensity exercise). Arthritis was induced via intraperitoneal administration of mannan. After a 2-week inflammation period, low-intensity treadmill exercise was performed only in the AREx group. Arthritis was assessed weekly during the rearing period. After 4 weeks of exercise, histological and bone morphometric analyses of the right ankle joint were performed. A histological analysis of the gastrocnemius muscle was also performed. Bulk mRNA sequencing was conducted on the left synovial membrane-fat pad (SM-FP) complex. ResultsThe synovitis score showed no change; however, the arthritis score was significantly lower in the AREx group than in the AR group (p<0.05), indicating that low-intensity exercise suppressed arthritis exacerbation. The calcaneal and talar bone volumes decreased in the AR group, whereas the AREx group showed no significant change. In the SM-FP complex tissue, the gene expression of inflammatory cytokines decreased in the AREx group compared with the AR group, particularly the suppression of IL6/Jak/Stat3. Immunohistochemical analysis revealed significantly decreased expression of pro-inflammatory cytokines and increased expression of anti-inflammatory cytokines in the synovium of the AREx group compared with the AR group (p<0.05). ConclusionLow-intensity exercise therapy for active RA showed anti-inflammatory and suppressive effects on arthritis exacerbation in SKG mice, a mouse model of human RA pathology. Key messagesO_LIExercise had an anti-inflammatory effect on SKG mice, a mouse model of rheumatoid arthritis. C_LIO_LIExercise suppresses pro-inflammatory cytokine pathways such as IL6/Jak/Stat3 signalling in the synovial-fat complex tissue. C_LIO_LIExercise therapy is effective in improving the pathophysiology of active rheumatoid arthritis. C_LI

molecular biology↗

bioRxiv2022

Diversity and characteristics of pTet family plasmids revealed by genomic epidemiology of Campylobacter jejuni from human patients in Toyama, Japan from 2015 to 2019

This study investigated 116 clinical isolates of Campylobacter jejuni from Toyama, Japan, which were isolated from 2015 to 2019. Antimicrobial susceptibility testing and whole-genome sequencing were used for phenotypic and genotypic characterization to compare antimicrobial resistance (AMR) profiles and phylogenic linkage. The multilocus sequence typing approach identified 37 sequence types (STs) and 15 clonal complexes (CCs), including 7 novel STs, and the high frequency CCs were CC21 (27.7%), CC48 (10.9%), and CC354 (9.9%). Overall, 58.6% of the isolates were resistant to at least one of the antibiotics and 3.4% were resistant to three or more antibiotic classes. The AMR profiles and related resistant factors were as follows; fluoroquinolones (51.7%), mutation in QRDRs (GyrA T86I), tetracyclines (27.6%), acquisition of tet(O), ampicillin (5.2%), promoter mutation in blaOXA193, aminoglycosides (1.7%), acquisition of ant(6)-Ia and aph(3)-III, chloramphenicol (0.9%), acquisition of cat. The resistance factors of fosfomycin (1 strain), sulfamethoxazole-trimethoprim (2 strain), and linezolid (1 strain) resistant isolates were unknown. The acquired resistance genes, tet(O), ant(6>)-Ia, aph(3)-III, and cat, were located on pTet family plasmids. Furthermore, three pTet family plasmids formed larger plasmids that incorporated additional genes such as the Type IV secretion system. A comparison of pTet family plasmids in Japan has not been reported, and these results imply that the diversity of pTet family plasmids has increased. The prevalence of ST4526, belonging to CC21, in Japan has been reported, and it was also the major ST type (10.9%) in this study, suggesting that the ST4526 prevalence continues in Japan.

microbiology↗

bioRxiv2019

Dynamic and synergistic influences of air temperature and rainfall on general flowering in a Bornean lowland tropical forest

Supra-annually synchronized flowering events occurring in tropical forests in Southeast Asia, known as general flowering (GF), are \"spectacular and mysterious\" forest events. Recently, studies that combined novel molecular techniques and model-based theoretical approaches suggested that cool temperature and drought synergistically drove GF. Although the novel approaches advanced our understanding of GF, it is usually difficult to know whether the mathematical formulation reasonably well represents the complex and dynamic processes involved in GF. In the present study, we collected a 17-year set of community-wide phenology data during 1993 to 2011 from Lambir Hills National Park in Borneo, Malaysia, and analyzed it using a model-free approach, empirical dynamic modeling (EDM), that does not rely on specific assumptions about the underlying mechanisms, to overcome and complement the previous limitations. By using EDM, we found that GF in the forest in Lambir Hills National Park is synergistically driven by cool air temperature and drought, which is consistent with the previous studies. Also, we found that cumulative meteorological variables, rather than instantaneous values, drive GF with delayed effects, which is also consistent with the previous studies. Interestingly, the present study showed that effects of cumulative meteorological variables on GF changed through time, which implies that the relationship between GF and meteorological variables may be influenced by other factors such as plant/soil nutrient resource dynamics. Future studies integrating novel mathematical/statistical frameworks, long-term and large spatial scale ecosystem monitoring and molecular phenology data are promising for better understanding and fore-casting of GF events in tropical forests in Southeast Asia.

ecology↗