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Visualizing Multiciliated Cells in the Zebrafish Through a Combined Protocol of Whole Mount Fluorescent In Situ Hybridization and Immunofluorescence

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IMPDH forms the cytoophidium in zebrafish.

Gerson Dierley Keppeke¹, Chia-Chun Chang², Christopher L Antos²

¹School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, China; Rheumatology Division, Escola Paulista de Medicina, Universidade Federal de Sao Paulo, Sao Paulo, SP, 04023-062, Brazil.

Developmental Biology

|May 28, 2021

Summary

Inosine monophosphate dehydrogenase (IMPDH) forms cellular structures called cytoophidia, which may help regulate guanine nucleotide synthesis. This polymerization is conserved across vertebrates and occurs in various fish tissues.

Keywords:

Adult zebrafish Cytoophidium IMPDH Larval zebrafish

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Last Updated: Nov 4, 2025

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Published on: November 18, 2017

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Published on: March 25, 2009

Area of Science:

Biochemistry
Cell Biology
Molecular Biology

Background:

Inosine monophosphate dehydrogenase (IMPDH) is crucial for de novo guanine nucleotide biosynthesis.
IMPDH activity is inhibited by GTP and can form cellular structures called cytoophidia.
Cytoophidium formation is linked to increased GTP biosynthesis due to GTP-bound IMPDH resistance to inhibition.

Purpose of the Study:

To investigate the cytoophidium-forming capabilities of zebrafish IMPDH isoforms.
To explore the conservation and physiological relevance of IMPDH polymerization and cytoophidium assembly.

Main Methods:

Expression and analysis of zebrafish IMPDH1a, IMPDH1b, and IMPDH2 isoforms in cultured cells.
Introduction of point mutations disrupting cytoophidium structure.
Histological examination of IMPDH cytoophidia in various zebrafish tissues.

Main Results:

Zebrafish IMPDH1b and IMPDH2 efficiently form cytoophidia in most cell types.
Zebrafish IMPDH1a exhibits cell-type-specific cytoophidium formation.
Mutations disrupting mammalian IMPDH cytoophidium assembly also affect zebrafish IMPDH aggregation.
IMPDH cytoophidia are present in multiple tissues of larval and adult zebrafish under normal conditions.

Conclusions:

IMPDH polymerization and cytoophidium assembly represent a conserved regulatory mechanism in vertebrates.
This process likely serves specific physiological functions related to nucleotide metabolism.
Zebrafish IMPDH isoforms display distinct behaviors in cytoophidium formation, highlighting isoform-specific roles.