Search bioRxiv⌕ Search

Biology subjects

Kamata, S.

Publications and source records attributed to Kamata, S..

4 recordsLinked to original sources

Integrating spatial and single-cell multi-omics analysis of induced pluripotent stem cell-derived cervical adenocarcinoma model

Human papillomavirus 18 (HPV18) preferentially infects cervical stem cell-like cells and is strongly associated with adenocarcinoma. However, the mechanisms underlying differentiation into cervical adenocarcinoma remain unclear due to the lack of appropriate experimental models. We aimed to establish a model of HPV18-associated cervical adenocarcinoma and elucidate its molecular and cellular differentiation mechanisms. HPV18 E6/E7 were introduced into induced pluripotent stem cell-derived reserve cell-like cells (iRCs) to generate tumor models. Spatial transcriptomics and single-cell multi-omics analyses were performed to integrate histological and molecular data. A distinct component (Gland_A) exhibited morphological and immunohistochemical features of cervical adenocarcinoma and was efficiently induced in iRC-18 tumors. Gland_A showed increased chromatin accessibility and elevated expression of FOXA1, FOXA2, and ALDH1A1. Analysis of clinical samples confirmed enrichment of ALDH1A1 in HPV-associated adenocarcinomas. This model recapitulates key features of HPV18-associated cervical adenocarcinoma and provides insights into its differentiation mechanisms.

cancer biology↗

Analysis of molecular and cellular bases of honey bee mushroom body development

Honey bee mushroom bodies (MBs) comprise three class I Kenyon cell (KC) subtypes (lKC, mKC, and sKC) with distinct somata sizes and locations and gene expression profiles. While these KC subtypes have been suggested to function in different behavioral regulations, the molecular and cellular basis of their development remains obscure. Here, we showed that lKCs, mKCs, and sKCs are produced in that order at different pupal stages by labeling proliferating MB cells with 5-ethynil-2-deoxyuridine at various pupal stages. RNA-sequencing analysis of FACS-sorted pupal MB cells identified genes that were upregulated in proliferating and non-proliferating MB cells, respectively. Furthermore, in situ hybridization of some of these genes labeled the proliferating cells or young KCs in the pupal MBs. Our findings reveal the basic scheme of the molecular and cellular processes of honey bee MB development and suggest that they are at least partially different from those of Drosophila MB development.

developmental biology↗

Analysis of antennal responses to motion stimuli in the honey bee by automated tracking using DeepLabCut

Considering recent developments in gene manipulation methods for honey bees, establishing simple, robust, and indoor assay systems which can analyze behavioral components in detail is important for the rise of honey bee behavioral genetics. We focused on the movements of antennae of the honey bee, which are used for not only multimodal sensory perception but also interactions between individuals. We developed an experimental system for analyzing the antennal responses (ARs) of the honey bee using DeepLabCut, a markerless posture-tracking tool using deep learning. The tracking of antennal movements during the presentation of vertical (downward and upward) motion stimuli using DeepLabCut successfully detected the ARs reported in the previous studies, where bees tilted their antennae in the direction opposite to the motion stimuli. In addition, we successfully detected ARs in response to horizontal (forward and backward) motion stimuli. An investigation of the developmental maturation of honey bee ARs showed that ARs to motion stimuli were not detected in bees immediately after emergence but became detectable through post-emergence development in an experience-independent manner. Furthermore, unsupervised clustering analysis using multidimensional data created by processing tracking data using DeepLabCut classified antennal movements into different clusters, suggesting that data-driven behavioral classification can apply to AR paradigms. These results reveal novel AR to visual stimuli and developmental maturation of ARs and suggest the efficacy of data-driven analysis for behavioral classification in behavioral studies of the honey bee. Summary statementAutomated tracking using DeepLabCut was successfully applied to measure the antennal response to motion stimuli and unsupervised classification of antennal movements in honey bees.

animal behavior and cognition↗

Stress-dependent dynamic and reversible formation of cytoskeleton-like filaments and gel-transition by tardigrade tolerance proteins.

Tardigrades are able to tolerate almost complete dehydration by entering a reversible ametabolic state called anhydrobiosis and resume their animation upon rehydration. Dehydrated tardigrades are exceptionally stable and withstand various physical extremes. Although trehalose and late embryogenesis abundant (LEA) proteins have been extensively studied as potent protectants against dehydration in other anhydrobiotic organisms, tardigrades produce high amounts of tardigrade-unique protective proteins. Cytoplasmic-abundant heat-soluble (CAHS) proteins are uniquely invented in the lineage of eutardigrades, a major class of the phylum Tardigrada and are essential for their anhydrobiotic survival. However, the precise mechanisms of their action in this protective role are not fully understood. In the present study, we first postulated the presence of tolerance proteins that form protective condensates via phase separation in a stress-dependent manner and searched for tardigrade proteins that reversibly form condensates upon dehydration-like stress. Through comprehensive analysis, we identified 336 such proteins, collectively dubbed "Desolvataion-induced ReversiblY condensing Proteins (DRYPs)". Unexpectedly, we rediscovered CAHS proteins as highly enriched in DRYPs, 3 of which were major components of DRYPs. We revealed that these CAHS proteins reversibly polymerize into many cytoskeleton-like filaments depending on hyperosmotic stress in cultured cells and undergo reversible gel-transition in vitro. CAHS filamentation increases cell stiffness to resist deformation and improves resistance to dehydration-like stress. The conserved putative helical C-terminal region is necessary and sufficient for filament formation by CAHS proteins, and mutations disrupting the secondary structure of this region impaired both the filament formation and the gel transition. On the basis of these results, we propose that CAHS proteins are novel cytoskeletal proteins that form filamentous networks and undergo gel-transition in a stress-dependent manner to provide on-demand physical stabilization of cell integrity against deformative forces during dehydration and also contribute to the exceptional physical stability in a dehydrated state.

biochemistry↗