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| Citation: |
LI J, ZHANG Y L, YANG F, et al. Comparative transcriptomic analysis of different hemocyte subpopulations in crayfish Cherax quadricarinatus [J]. Fujian Journal of Agricultural Sciences，2025，X（X）：1−11.
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Semigranular cell (SGC) and granular cell (GC) are the two primary subpopulations of circulating hemocytes in crayfish, representing distinct stages of immune cell differentiation. This study aims to further elucidate the functions of SGC and GC, providing essential data to facilitate future research on hemocyte subpopulations in crustaceans.
Using Cherax quadricarinatus as the experimental model, SGC and GC were isolated and purified via Percoll discontinuous density gradient centrifugation. Transcriptomic sequencing was then conducted on SGC and GC, followed by differential expression analysis, Gene Ontology (GO) enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis to investigate their functional divergence. Finally, real-time quantitative RT-PCR was employed to validate the expression of selected differentially expressed genes.
Sequencing identified 116,199 non-redundant unique genes (unigenes) in Cherax quadricarinatus hemocytes. These unigenes had an average length of 763 bp and an N50 of 1,313 bp. A total of 4,488 differentially expressed genes (DEGs) were identified, with 3,951 genes significantly upregulated in SGC and 537 genes showing higher expression in GC. The GO enrichment analysis revealed the following results: in Cellular Component, 7 terms enriched in SGC vs. 10 terms in GC; in Molecular Function, 31 terms enriched in SGC vs. 60 terms in GC; in Biological Process, 154 terms enriched in SGC vs. 102 terms in GC. KEGG analysis identified 44 pathways enriched in SGC and 10 pathways in GC.
Ultimately, the analysis revealed that: the DEGs enriched in SGC are primarily involved in cellular processes such as proliferation, differentiation, transcriptional regulation, enzyme synthesis, endocytosis, and cell adhesion. In contrast, DEGs enriched in GC are mainly associated with transmembrane transport, metabolic pathways, the prophenoloxidase system, phagocytosis, and antimicrobial peptide synthesis.
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