# Transcriptome analysis of atad3-null zebrafish embryos elucidates possible disease mechanisms

## TL;DR

Researchers used zebrafish to study a gene linked to human neurological diseases and found mitochondrial dysfunction is central to the disorder.

## Contribution

A zebrafish model of ATAD3A deficiency was created, revealing disease mechanisms and potential therapeutic readouts.

## Key findings

- Atad3-null zebrafish showed microcephaly, eye and heart defects, and reduced mitochondrial activity.
- Transcriptome analysis showed reduced mitochondrial pathways and increased cytosolic tRNA synthetases.
- Findings in zebrafish were corroborated in human patient fibroblasts, confirming relevance to human disease.

## Abstract

ATAD3A, a nuclear gene encoding the ATAD3A protein, has diverse roles in mitochondrial processes, encompassing mitochondrial dynamics, mitochondrial DNA maintenance, metabolic pathways and inter-organellar interactions. Pathogenic variants in this gene cause neurological diseases in humans with recognizable genotype-phenotype correlations. Yet, gaps in knowledge remain regarding the underlying pathogenesis.

To further investigate the gene function and its implication in health and disease, we utilized CRISPR/Cas9 genome editing to generate a knockout model of the zebrafish ortholog gene, atad3. We characterized the phenotype of the null model, performed mitochondrial and functional tests, and compared the transcriptome of null embryos to their healthy siblings.

Analysis of atad3-null zebrafish embryos revealed microcephaly, small eyes, pericardial edema and musculature thinning, closely mirroring the human rare disease phenotype. Larvae exhibited delayed hatching and embryonic lethality by 13 days post-fertilization (dpf). Locomotor activity, ATP content, mitochondrial content, and mitochondrial activity were all reduced in the mutant embryos. Transcriptome analysis at 3 dpf via RNA-sequencing indicated decline in most mitochondrial pathways, accompanied by a global upregulation of cytosolic tRNA synthetases, presumably secondary to mitochondrial stress and possibly endoplasmic reticulum (ER)-stress. Differential expression of select genes was corroborated in fibroblasts from an affected individual.

The atad3-null zebrafish model emerges as a reliable representation of human ATAD3A-associated disorders, with similarities in differentially expressed pathways and processes. Furthermore, our study underscores mitochondrial dysfunction as the primary underlying pathogenic mechanism in ATAD3A-associated disorders and identifies potential readouts for therapeutic studies.

The online version contains supplementary material available at 10.1186/s13023-025-03709-0.