# The functional organization of chromosome territories in single nuclei during zygotic genome activation

## TL;DR

This study explores how chromosomes are organized in single nuclei during early development, revealing how chromosome structure and pairing influence gene activity.

## Contribution

The study introduces a high-resolution imaging method to observe chromosome organization and pairing during zygotic genome activation.

## Key findings

- Chromosome territories show large-scale packaging differences and high homolog pairing during development.
- Spatial variability in chromosome conformations reduces homolog pairing locally.
- Transcription inhibition decreases CT opening but does not affect pairing levels.

## Abstract

Chromosome territories (CTs) are intricately organized and regulated within the nucleus. Despite remarkable advances in our understanding of genome packaging and gene expression, the interplay among CTs, pairing of parental homologous chromosomes, and genome function during development remains elusive. Here, we employ an Oligopaints-based high-resolution imaging approach to examine variable CT organization in single nuclei during the developmental process of zygotic genome activation. We reveal large-scale chromosome packaging differences and high levels of homolog pairing at the whole-chromosome scale that decreases locally due to spatial variability in chromosome conformations. In the absence of one homolog copy, the dynamics of CT compaction and RNA polymerase II recruitment are associated with the transcriptional changes in haploid embryos. Finally, global inhibition of transcription results in decreased CT opening and no significant impact on CT pairing levels. These findings enhance our understanding of variable parental genome folding and regulation during development, which may inform strategies for chromosome-based diseases.

The online version contains supplementary material available at 10.1038/s41598-026-35953-0.