Isthmin 1 (ism1) is required for normal hematopoiesis in developing zebrafish

PloS One
Arturo BerrunDavid L Stachura

Abstract

Hematopoiesis is an essential and highly regulated biological process that begins with hematopoietic stem cells (HSCs). In healthy organisms, HSCs are responsible for generating a multitude of mature blood cells every day, yet the molecular pathways that instruct HSCs to self-renew and differentiate into post-mitotic blood cells are not fully known. To understand these molecular pathways, we investigated novel genes expressed in hematopoietic-supportive cell lines from the zebrafish (Danio rerio), a model system increasingly utilized to uncover molecular pathways important in the development of other vertebrate species. We performed RNA sequencing of the transcriptome of three stromal cell lines derived from different stages of embryonic and adult zebrafish and identified hundreds of highly expressed transcripts. For our studies, we focused on isthmin 1 (ism1) due to its shared synteny with its human gene ortholog and because it is a secreted protein. To characterize ism1, we performed loss-of-function experiments to identify if mature blood cell production was disrupted. Myeloid and erythroid lineages were visualized and scored with transgenic zebrafish expressing lineage-specific markers. ism1 knockdown led to reduced numbers...Continue Reading

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Citations

Mar 28, 2019·Frontiers in Cell and Developmental Biology·Christopher B Mahony, Julien Y Bertrand
Dec 29, 2020·Frontiers in Oncology·Chuanyou XiaKlas Strååt
Feb 26, 2021·PloS One·Rebecca L BelmonteDavid L Stachura
Jul 25, 2021·Developmental and Comparative Immunology·Congjun LiGuangdong Ji
Aug 21, 2021·Bioinformatics and Biology Insights·Samrat Thapa, David L Stachura

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Methods Mentioned

BETA
transgenic
florescent microscopy
flow cytometry
fluorescence-activated cell sorting
PCR
FACS
RNA-seq

Software Mentioned

FloJo

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