Tools to analyze the organization and formation of the germline cyst in zebrafish oogenesis

Vineet Kumar; Yaniv M. Elkouby

doi:10.1242/dev.201349

Tools to analyze the organization and formation of the germline cyst in zebrafish oogenesis

Vineet Kumar, Yaniv M. Elkouby^*

^*Corresponding author for this work

Department of Developmental Biology and Cancer Research

Research output: Contribution to journal › Article › peer-review

Abstract

Oocytes develop in the germline cyst, a cellular organization in which germ cells are tightly interconnected and surrounded by somatic cells. The cyst produces oocytes for follicle formation and is a hub for essential processes in meiosis and oocyte differentiation. However, the formation and organization of the cyst, and their contribution to oocyte production in vertebrates remain unclear. Here, we provide tools for three-dimensional and functional in vivo analyses of the germline cyst in the zebrafish ovary. We describe the use of serial block-face scanning electron microscopy (SBF-SEM) to resolve the three-dimensional architecture of cells and organelles in the cyst at ultrastructural resolution. We present a deep learning-based pipeline for high-throughput quantitative analysis of three-dimensional confocal datasets of cysts in vivo. We provide a method for laser ablation of cellular components for manipulating cyst cells in ovaries. These methods will facilitate the investigation of the cyst cellular organization, expand the toolkit for the study of the zebrafish ovary, and advance our understanding of female developmental reproduction. They could also be further applied to the investigation of other developmental systems.

Original language	American English
Article number	dev201349
Journal	Development (Cambridge)
Volume	150
Issue number	13
DOIs	https://doi.org/10.1242/dev.201349
State	Published - Jul 2023

Bibliographical note

Funding Information:
We thank A. Beckett, from the Biomedical EM unit at the University of Liverpool, for her help in processing our samples for SBF-SEM imaging. We also thank Z. Manevich, from the Faculty of Medicine Microscopy Core at the Hebrew University, and Y. Addadi, from the core facility at the Weizmann Institute, for their support of our multi-photon microscopy operation in our laser ablation experiments. This work was supported by the Israel Science Foundation (3291/19 and 558/19 to Y.E.) and a Hebrew University of Jerusalem (HUJI) International PhD Talent Scholarship (V.K.). Open Access funding provided by Hebrew University of Jerusalem. Deposited in PMC for immediate release.

Funding Information:
This work was supported by the Israel Science Foundation (3291/19 and 558/19 to Y.E.) and a Hebrew University of Jerusalem (HUJI) International PhD Talent Scholarship (V.K.). Open Access funding provided by Hebrew University of Jerusalem. Deposited in PMC for immediate release.

Publisher Copyright:
© 2023. Published by The Company of Biologists Ltd.

Keywords

3D deep-learning data
Germ cell development and morphogenesis
Germline cyst
Live manipulation by laser-ablation
Microscopy image analysis
Oogenesis
Zebrafish ovary

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10.1242/dev.201349

Cite this

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title = "Tools to analyze the organization and formation of the germline cyst in zebrafish oogenesis",

abstract = "Oocytes develop in the germline cyst, a cellular organization in which germ cells are tightly interconnected and surrounded by somatic cells. The cyst produces oocytes for follicle formation and is a hub for essential processes in meiosis and oocyte differentiation. However, the formation and organization of the cyst, and their contribution to oocyte production in vertebrates remain unclear. Here, we provide tools for three-dimensional and functional in vivo analyses of the germline cyst in the zebrafish ovary. We describe the use of serial block-face scanning electron microscopy (SBF-SEM) to resolve the three-dimensional architecture of cells and organelles in the cyst at ultrastructural resolution. We present a deep learning-based pipeline for high-throughput quantitative analysis of three-dimensional confocal datasets of cysts in vivo. We provide a method for laser ablation of cellular components for manipulating cyst cells in ovaries. These methods will facilitate the investigation of the cyst cellular organization, expand the toolkit for the study of the zebrafish ovary, and advance our understanding of female developmental reproduction. They could also be further applied to the investigation of other developmental systems.",

keywords = "3D deep-learning data, Germ cell development and morphogenesis, Germline cyst, Live manipulation by laser-ablation, Microscopy image analysis, Oogenesis, Zebrafish ovary",

author = "Vineet Kumar and Elkouby, {Yaniv M.}",

note = "Funding Information: We thank A. Beckett, from the Biomedical EM unit at the University of Liverpool, for her help in processing our samples for SBF-SEM imaging. We also thank Z. Manevich, from the Faculty of Medicine Microscopy Core at the Hebrew University, and Y. Addadi, from the core facility at the Weizmann Institute, for their support of our multi-photon microscopy operation in our laser ablation experiments. This work was supported by the Israel Science Foundation (3291/19 and 558/19 to Y.E.) and a Hebrew University of Jerusalem (HUJI) International PhD Talent Scholarship (V.K.). Open Access funding provided by Hebrew University of Jerusalem. Deposited in PMC for immediate release. Funding Information: This work was supported by the Israel Science Foundation (3291/19 and 558/19 to Y.E.) and a Hebrew University of Jerusalem (HUJI) International PhD Talent Scholarship (V.K.). Open Access funding provided by Hebrew University of Jerusalem. Deposited in PMC for immediate release. Publisher Copyright: {\textcopyright} 2023. Published by The Company of Biologists Ltd.",

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N2 - Oocytes develop in the germline cyst, a cellular organization in which germ cells are tightly interconnected and surrounded by somatic cells. The cyst produces oocytes for follicle formation and is a hub for essential processes in meiosis and oocyte differentiation. However, the formation and organization of the cyst, and their contribution to oocyte production in vertebrates remain unclear. Here, we provide tools for three-dimensional and functional in vivo analyses of the germline cyst in the zebrafish ovary. We describe the use of serial block-face scanning electron microscopy (SBF-SEM) to resolve the three-dimensional architecture of cells and organelles in the cyst at ultrastructural resolution. We present a deep learning-based pipeline for high-throughput quantitative analysis of three-dimensional confocal datasets of cysts in vivo. We provide a method for laser ablation of cellular components for manipulating cyst cells in ovaries. These methods will facilitate the investigation of the cyst cellular organization, expand the toolkit for the study of the zebrafish ovary, and advance our understanding of female developmental reproduction. They could also be further applied to the investigation of other developmental systems.

AB - Oocytes develop in the germline cyst, a cellular organization in which germ cells are tightly interconnected and surrounded by somatic cells. The cyst produces oocytes for follicle formation and is a hub for essential processes in meiosis and oocyte differentiation. However, the formation and organization of the cyst, and their contribution to oocyte production in vertebrates remain unclear. Here, we provide tools for three-dimensional and functional in vivo analyses of the germline cyst in the zebrafish ovary. We describe the use of serial block-face scanning electron microscopy (SBF-SEM) to resolve the three-dimensional architecture of cells and organelles in the cyst at ultrastructural resolution. We present a deep learning-based pipeline for high-throughput quantitative analysis of three-dimensional confocal datasets of cysts in vivo. We provide a method for laser ablation of cellular components for manipulating cyst cells in ovaries. These methods will facilitate the investigation of the cyst cellular organization, expand the toolkit for the study of the zebrafish ovary, and advance our understanding of female developmental reproduction. They could also be further applied to the investigation of other developmental systems.

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DO - 10.1242/dev.201349

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SN - 0950-1991

VL - 150

JO - Development (Cambridge)

JF - Development (Cambridge)

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M1 - dev201349

ER -

Tools to analyze the organization and formation of the germline cyst in zebrafish oogenesis

Abstract

Bibliographical note

Keywords

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