# Single-cell analysis of a salmonid immune system (river brown trout Salmo trutta fario) reveals evolutionary divergence and hatchery-induced transcriptional reprogramming

**Authors:** James Ord, Helena Saura Martinez, Monica Hongroe Solbakken, Anastasiia Berezenko, Simone Oberhaensli, Stephanie Talker, Heike Schmidt-Posthaus, Irene Adrian-Kalchhauser

PMC · DOI: 10.1186/s12915-026-02554-2 · BMC Biology · 2026-03-18

## TL;DR

This study uses single-cell analysis to explore the immune system of river brown trout, revealing evolutionary changes and how hatchery rearing affects immune gene expression.

## Contribution

The study provides the first single-cell transcriptome of immune cells in river brown trout and identifies hatchery-induced transcriptional changes.

## Key findings

- The study identified 34 distinct immune cell populations and novel markers in neutrophils, macrophages, T-cells, and B-cells.
- Transcriptional divergence was observed between WGD-derived ohnologue pairs, suggesting sub- and neofunctionalization.
- Hatchery-reared fish showed altered immune gene expression, indicating rearing history shapes immune cell identity.

## Abstract

Vertebrate immune systems exhibit striking evolutionary diversity, yet our understanding remains biased toward mammalian models. Here, we generate a single-cell transcriptome of immune cells from the ecologically and economically important salmonid Salmo trutta fario (river brown trout), a lineage characterized by an ancestral whole-genome duplication (WGD).

Profiling over 83,000 kidney-derived immune cells, we resolved 34 transcriptionally distinct populations, identified core immune lineages, and uncovered novel markers in neutrophils, macrophages, T-cells, and B-cells. We detected pervasive transcriptional divergence between WGD-derived ohnologue pairs, indicating putative sub- and neofunctionalization in immune gene regulation. We further show that the transcriptional identity of immune cells is shaped by rearing history: fish raised in hatcheries—whether for one or multiple generations—showed shifts in immune gene expression across cell types.

Our findings provide baseline data for the healthy brown trout cellular immune system, insight into the evolution of vertebrate immunity, and avenues for understanding the molecular basis of reduced immunological fitness of hatchery-reared fish.

The online version contains supplementary material available at 10.1186/s12915-026-02554-2.

## Linked entities

- **Species:** Salmo trutta fario (taxon 99804)

## Full-text entities

- **Genes:** CPA1 (carboxypeptidase A1) [NCBI Gene 1357] {aka CPA}, SSR4 (signal sequence receptor subunit 4) [NCBI Gene 6748] {aka CDG1Y, TRAPD}, cd74b (CD74 molecule, major histocompatibility complex, class II invariant chain b) [NCBI Gene 30645] {aka Iclp-2, cd74, iclp2, zgc:123161}, lyz (lysozyme) [NCBI Gene 677744] {aka lys-C, zgc:136734}, STK10 (serine/threonine kinase 10) [NCBI Gene 6793] {aka LOK, PRO2729}, FABP7 (fatty acid binding protein 7) [NCBI Gene 2173] {aka B-FABP, BLBP, FABPB, MRG}, ITGA5 (integrin subunit alpha 5) [NCBI Gene 3678] {aka CD49e, FNRA, VLA-5, VLA5A}, MARCO (macrophage receptor with collagenous structure) [NCBI Gene 8685] {aka SCARA2, SR-A6}, TALDO1 (transaldolase 1) [NCBI Gene 6888] {aka TAL, TAL-H, TALDOR, TALH}, RAG1 (recombination activating 1) [NCBI Gene 5896] {aka RAG-1, RNF74}, DKK3 (dickkopf Wnt signaling pathway inhibitor 3) [NCBI Gene 27122] {aka CRRL, REIC, RIG}, mpeg1.1 (macrophage expressed 1, tandem duplicate 1) [NCBI Gene 335407] {aka fj09e08, mpeg1, wu:fb14d06, wu:fj09e08, zgc:66409}, PRF1 (perforin 1) [NCBI Gene 5551] {aka HPLH2, P1, PFP}, IL2RB (interleukin 2 receptor subunit beta) [NCBI Gene 3560] {aka CD122, IL15RB, IMD63, P70-75}, ALAS2 (5'-aminolevulinate synthase 2) [NCBI Gene 212] {aka ALAS-E, ALASE, ANH1, ASB, SIDBA1, XLDPP}, KRTAP1-3 (keratin associated protein 1-3) [NCBI Gene 81850] {aka KAP1.2, KAP1.3, KAP1.6, KAP1.8A, KAP1.8B, KAP1.9}, TEK (TEK receptor tyrosine kinase) [NCBI Gene 7010] {aka CD202B, GLC3E, TIE-2, TIE2, VMCM, VMCM1}, FBP1 (fructose-bisphosphatase 1) [NCBI Gene 2203] {aka FBP}, GP1BA (glycoprotein Ib platelet subunit alpha) [NCBI Gene 2811] {aka BDPLT1, BDPLT3, BSS, CD42B, CD42b-alpha, DBPLT3}, PLK3 (polo like kinase 3) [NCBI Gene 1263] {aka CNK, FNK, PLK-3, PRK}, CD8A (CD8 subunit alpha) [NCBI Gene 925] {aka CD8, CD8alpha, IMD116, Leu2, p32}, JUND (JunD proto-oncogene, AP-1 transcription factor subunit) [NCBI Gene 3727] {aka AP-1}, H2AZ1 (H2A.Z variant histone 1) [NCBI Gene 3015] {aka H2A.Z-1, H2A.z, H2A/z, H2AFZ, H2AZ}, MRC1 (mannose receptor C-type 1) [NCBI Gene 4360] {aka CD206, CLEC13D, CLEC13DL, MMR, MRC1L1, bA541I19.1}, TRBC1 (T cell receptor beta constant 1) [NCBI Gene 28639] {aka BV05S1J2.2, TCRB, TCRBC1}, IRF8 (interferon regulatory factor 8) [NCBI Gene 3394] {aka H-ICSBP, ICSBP, ICSBP1, IMD32A, IMD32B, IRF-8}, SCT (secretin) [NCBI Gene 6343], PCNA (proliferating cell nuclear antigen) [NCBI Gene 5111] {aka ATLD2}, irf8 (interferon regulatory factor 8) [NCBI Gene 436895] {aka zgc:92252}, IL7R (interleukin 7 receptor) [NCBI Gene 3575] {aka CD127, CDW127, IL-7R-alpha, IL-7Ralpha, IL7RA, IL7Ralpha}, CD59 (CD59 molecule (CD59 blood group)) [NCBI Gene 966] {aka 16.3A5, 1F5, EJ16, EJ30, EL32, G344}, CD37 (CD37 molecule) [NCBI Gene 951] {aka GP52-40, TSPAN26}, CD28 (CD28 molecule) [NCBI Gene 940] {aka IMD123, Tp44}, Act79B (Actin 79B) [NCBI Gene 40444] {aka 143060_f_at, ACT4, Actin, ArpF, CG7478, D}, CXCR6 (C-X-C motif chemokine receptor 6) [NCBI Gene 10663] {aka BONZO, CD186, CDw186, STRL33, TYMSTR}, MPL (MPL proto-oncogene, thrombopoietin receptor) [NCBI Gene 4352] {aka C-MPL, CD110, MPLV, THCYT2, THPOR, TPOR}, CD4 (CD4 molecule) [NCBI Gene 920] {aka CD4mut, IMD79, Leu-3, OKT4D, T4}, NHSL2 (NHS like 2) [NCBI Gene 340527], dac (dachshund) [NCBI Gene 34982] {aka BG:DS02780.3, CG4952, Dac2-3, Dach, Dachshund, Dm-DAC}, SH2D1A (SH2 domain containing 1A) [NCBI Gene 4068] {aka DSHP, EBVS, IMD5, LYP, MTCP1, SAP}, SLAMF1 (signaling lymphocytic activation molecule family member 1) [NCBI Gene 6504] {aka CD150, CDw150, IPO3, SLAM}, DEF6 (DEF6 guanine nucleotide exchange factor) [NCBI Gene 50619] {aka IBP, IMD87, SLAT, SWAP70L}, RNGTT (RNA guanylyltransferase and 5'-phosphatase) [NCBI Gene 8732] {aka CAP1A, HCE, HCE1, hCAP}, plaat1l (phospholipase A and acyltransferase 1-like) [NCBI Gene 794087] {aka zgc:194264, zgc:194275}, TRGC1 (T cell receptor gamma constant 1) [NCBI Gene 6966] {aka C1, TCRG, TCRGC1}, CD79B (CD79b molecule) [NCBI Gene 974] {aka AGM6, B29, IGB, Igbeta}, OLFM4 (olfactomedin 4) [NCBI Gene 10562] {aka GC1, GW112, OLM4, OlfD, UNQ362, bA209J19.1}, XPA (XPA, DNA damage recognition and repair factor) [NCBI Gene 7507] {aka XP1, XPAC}, APC (APC regulator of Wnt signaling pathway) [NCBI Gene 324] {aka BTPS2, DESMD, DP2, DP2.5, DP3, GS}, adam28 (ADAM metallopeptidase domain 28) [NCBI Gene 767749] {aka si:ch211-133n22.1, zgc:153755}, MKI67 (marker of proliferation Ki-67) [NCBI Gene 4288] {aka KIA, MIB-, MIB-1, PPP1R105}, EGR1 (early growth response 1) [NCBI Gene 1958] {aka AT225, G0S30, KROX-24, NGFI-A, TIS8, ZIF-268}, CXCR5 (C-X-C motif chemokine receptor 5) [NCBI Gene 643] {aka BLR1, CD185, MDR15}, CD8B (CD8 subunit beta) [NCBI Gene 926] {aka CD8B1, CD8beta, LEU2, LY3, LYT3, Ly-3}, CD79A (CD79a molecule) [NCBI Gene 973] {aka IGA, IGAlpha, MB-1, MB1}, Cd74 (CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated)) [NCBI Gene 16149] {aka CLIP, DHLAG, HLADG, Ia-GAMMA, Ii}, CD3E (CD3 epsilon subunit of T-cell receptor complex) [NCBI Gene 916] {aka CD3epsilon, IMD18, T3E, TCRE}, CD2 (CD2 molecule) [NCBI Gene 914] {aka LFA-2, SRBC, T11}, FOS (Fos proto-oncogene, AP-1 transcription factor subunit) [NCBI Gene 2353] {aka AP-1, C-FOS, p55}, HBE1 (hemoglobin subunit epsilon 1) [NCBI Gene 3046] {aka HBE}
- **Diseases:** infection (MESH:D007239), kidney disease (MESH:D007674), inflammatory (MESH:D007249)
- **Chemicals:** iron (MESH:D007501), MS 222 (MESH:C003636), calcium (MESH:D002118), L-15 (-), Fatty acid (MESH:D005227), carbohydrate (MESH:D002241), LPS (MESH:D008070), PBS (MESH:D007854)
- **Species:** Salmo trutta fario (river trout, subspecies) [taxon 99804], Homo sapiens (human, species) [taxon 9606], Esox lucius (northern pike, species) [taxon 8010], Salmonella phage IKe (no rank) [taxon 10867], Drosophila melanogaster (fruit fly, species) [taxon 7227], Tetracapsuloides bryosalmonae (species) [taxon 271932], Mus musculus (house mouse, species) [taxon 10090], Oncorhynchus mykiss (rainbow trout, species) [taxon 8022], Danio rerio (leopard danio, species) [taxon 7955], Coregoninae (whitefishes, subfamily) [taxon 504567], Salmonidae (salmonids, family) [taxon 8015], Artemia (brine shrimps, genus) [taxon 6660], Salmo trutta (river trout, species) [taxon 8032], Rubroshorea almon (species) [taxon 292004]
- **Cell lines:** S2 — Drosophila melanogaster (Fruit fly), Spontaneously immortalized cell line (CVCL_Z232), M3 — Homo sapiens (Human), Amelanotic melanoma, Cancer cell line (CVCL_8336), N6 — Mus musculus (Mouse), Transformed cell line (CVCL_D461), B6 — Homo sapiens (Human), Finite cell line (CVCL_L814)

## Full text

_Full body text omitted from this summary view._ Fetch the complete paper as Markdown: https://tomesphere.com/paper/PMC12997680/full.md

## Figures

10 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12997680/full.md

## References

3 references — full list in the complete paper: https://tomesphere.com/paper/PMC12997680/full.md

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Source: https://tomesphere.com/paper/PMC12997680