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Single-cell RNA sequencing reveals heterogeneous response of interlamellar cell mass to hypoxic stress in crucian carp (Carassius auratus) gills.

Chengbin Wu,Zhi-Guang Hou,Xin Zhao,Yi-Huan Xu,Chunguang Gong

Published 2025 in Fish and Shellfish Immunology

ABSTRACT

Gills are the main respiratory organs of fish, and they are highly sensitive to dissolved oxygen levels. Here, we investigated the heterogenous response of interlamellar cell mass (ILCM) to hypoxic stress in crucian carp (Carassius auratus) gill using single-cell RNA sequencing (scRNA-seq). The scRNA-seq and histological analyses demonstrated that the cell proportion of ILCM in all cell types decreased from 49.4% to 26.7% under hypoxia, and its immune and proliferative functions were suppressed, indicating that ILCM might be the major cell type responding to hypoxic stress in crucian carp gill. Cell-cell communication analysis suggested that ligand-receptor tnfb-cd40 in ILCM might play an important role in immune response. The enrichment analysis of differentially expressed genes (DEGs) revealed that misfolded proteins were accumulated in the endoplasmic reticulum (ER) of ILCM during hypoxia, eventually leading to ER stress. Further exploration of ILCM apoptosis mechanism during hypoxia revealed that ER stress might be the main pathway responsible for ILCM apoptosis. In addition, gsdma3/caspase-3-mediated pyroptosis might contribute to gill remodeling. In summary, our scRNA-seq provides an insight into heterogeneous response of gill ILCM to hypoxic stress, revealing the hypoxic adaptation strategy and gill remodeling molecular mechanisms in crucian carp.

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