Biology subjects

Nagasaka, Y.

Publications and source records attributed to Nagasaka, Y..

3 recordsLinked to original sources

bioRxiv2025

Sex differences in human IgG1-mediated angiogenesis inhibition depend on Y chromosome-encoded DDX3Y

Prevalent diseases of angiogenesis such as age-related macular degeneration (AMD) exhibit sex-specific prevalence, with female sex being an independent risk factor for the advanced neovascular form of AMD. The basis for these sex differences is poorly understood. Here, we quantify the impact of sex on suppression of aberrant choroidal angiogenesis by human immunoglobulin 1 (hIgG1), which possesses class-wide, antigen-independent anti-angiogenic activity. Males exhibited significantly more robust hIgG1-induced chemotaxis inhibition in primary human and mouse macrophages and anti-angiogenic activity in mouse laser-induced choroidal neovascularization (CNV). Four core genotypes and XY* Turner Syndrome mouse models revealed the Y chromosome as a sex-biasing factor contributing to hIgG1 responses. Transcriptomic and RNAi screening identified DEAD-Box Helicase 3 Y-Linked (DDX3Y) as necessary for the robust inhibitory effects of hIgG1 in males. Male mice as well as human and mouse macrophages lacking DDX3Y exhibited blunted hIgG1 responses in CNV and macrophage chemotaxis, resembling those of females. These results unveil a novel mechanism through which sex chromosome complement differences can impact key processes involved in AMD, carrying implications for various sex-related disorders and conditions involving angiogenesis.

pathology↗

bioRxiv2024

Flow zoometry of Drosophila

Drosophila serves as a highly valuable model organism across numerous fields including genetics, immunology, neuroscience, cancer biology, and developmental biology. Central to Drosophila-based biological research is the ability to perform comprehensive genetic or chemical screens. However, this research is often limited by its dependence on laborious manual handling and analysis, making it prone to human error and difficult to discern statistically significant or rare events amid the noise of individual variations resulting from genetic and environmental factors. In this article we present flow zoometry, a whole-animal equivalent of flow cytometry for large-scale, individual-level, high-content screening of Drosophila. Our flow zoometer automatically clears the tissues of Drosophila melanogaster, captures three-dimensional (3D) multi-color fluorescence tomograms of single flies with single-cell volumetric resolution at an unprecedented throughput of over 1,000 animals within 48 hours (24 hr for clearing; 24 hr for imaging), and performs AI-enhanced data-driven analysis - a task that would traditionally take months or years with manual techniques. To demonstrate its broad applications, we employed the flow zoometer in various laborious screening assays, including those in toxicology, genotyping, and tumor screening. Flow zoometry represents a pivotal evolution in high-throughput screening technology: previously from molecules to cells, now from cells to whole animals. This advancement serves as a foundational platform for "statistical spatial biology", to improve empirical precision and enable serendipitous discoveries across various fields of biology.

bioengineering↗

bioRxiv2021

The learning curve of murine subretinal injection among clinically trained ophthalmic surgeons

PO_SCPLOWURPOSEC_SCPLOWSubretinal injection (SRI) in mice is widely used in retinal research, yet the learning curve (LC) of this surgically challenging technique is unknown. MO_SCPLOWETHODSC_SCPLOWTo evaluate the LC for SRI in a murine model, we analyzed training data from 3 clinically trained ophthalmic surgeons from 2018 to 2020. Successful SRI was defined as either the absence of retinal pigment epithelium (RPE) degeneration after phosphate buffer saline injection and the presence of RPE degeneration after Alu RNA injection. Multivariable survival-time regression models were used to evaluate the association between surgeon experience and success rate, with adjustment for injection agents, and to calculate an approximate case number to achieve a 95% success rate. A Cumulative Sum (CUSUM) analysis was performed and plotted individually to monitor each surgeons simultaneous performance. RO_SCPLOWESULTSC_SCPLOWDespite prior microsurgery experience, the combined average success rate of the first 50 cases in mice was only 27%. The predicted SRI success rate did not reach a plateau above 95% until approximately 364 prior cases. Using the 364-training case as a "cutoff" point, the predicted probability of success before and after the 364th case was 65.38% and 99.32%, respectively (P < 0.0001). CUSUM analysis showed an initial upward slope and then remained within the decision intervals with an acceptable success rate set at 95% in the late stage. CO_SCPLOWONCLUSIONSC_SCPLOWThis study demonstrates the complexity and substantial LC for successful SRI in mice with high confidence. A systematic training system could improve the reliability and reproducibility of SRI-related experiments and improve the interpretation of experimental results using this technique. Translational RelevanceOur prediction model and monitor system allow objective quantification of technical proficiency in the field of subretinal drug delivery and gene therapy for the first time.

neuroscience↗