Complementary Volume Electron Microscopy-based approaches reveal ultrastructural changes in germline intercellular bridges of D. melanogaster DOI Creative Commons
Irina Kolotuev, Antony Williams,

Caroline Kizilyaprak

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 23, 2025

ABSTRACT Intercellular bridges are essential to connect developing germline cells. The Drosophila melanogaster egg chamber is a powerful model system study intercellular bridges, or ring canals (RCs). RCs the oocyte supporting nurse cells, and defects in their stability growth lead infertility. Despite importance, it has been technically difficult use electron microscopy-based approaches monitor changes RC structure during oogenesis. Here, we describe application of complementary set volume EM-based visualize ultrastructural RCs. combination array tomography (AT) focused ion beam (FIB) scanning microscopy (SEM) allowed us gain insight into previously unappreciated aspects structure. We were able quantify differences size thickness within between germ cell clusters at different developmental stages. Within cluster, correlates with lineage; largest formed first division, smallest fourth mitotic division. observed formation membrane interdigitations vicinity much earlier than reported, reconstruction from mid-stage EC provided 3D orientation these extensive cell-cell contacts. Our imaging also revealed novel that appeared line interior lumen. Although focus was on RCs, our dataset contains valuable details additional types structures, including fusome, stem cells niche, migrating border This framework could be applied other tissues samples face similar technical challenges, where small interest located large sample volume.

Language: Английский

Complementary Volume Electron Microscopy-based approaches reveal ultrastructural changes in germline intercellular bridges of D. melanogaster DOI Creative Commons
Irina Kolotuev, Antony Williams,

Caroline Kizilyaprak

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 23, 2025

ABSTRACT Intercellular bridges are essential to connect developing germline cells. The Drosophila melanogaster egg chamber is a powerful model system study intercellular bridges, or ring canals (RCs). RCs the oocyte supporting nurse cells, and defects in their stability growth lead infertility. Despite importance, it has been technically difficult use electron microscopy-based approaches monitor changes RC structure during oogenesis. Here, we describe application of complementary set volume EM-based visualize ultrastructural RCs. combination array tomography (AT) focused ion beam (FIB) scanning microscopy (SEM) allowed us gain insight into previously unappreciated aspects structure. We were able quantify differences size thickness within between germ cell clusters at different developmental stages. Within cluster, correlates with lineage; largest formed first division, smallest fourth mitotic division. observed formation membrane interdigitations vicinity much earlier than reported, reconstruction from mid-stage EC provided 3D orientation these extensive cell-cell contacts. Our imaging also revealed novel that appeared line interior lumen. Although focus was on RCs, our dataset contains valuable details additional types structures, including fusome, stem cells niche, migrating border This framework could be applied other tissues samples face similar technical challenges, where small interest located large sample volume.

Language: Английский

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