PUBLICATION

Zebrafish B Cell Development without a Pre-B Cell Stage, Revealed by CD79 Fluorescence Reporter Transgenes

Authors
Liu, X., Li, Y.S., Shinton, S.A., Rhodes, J., Tang, L., Feng, H., Jette, C.A., Look, A.T., Hayakawa, K., Hardy, R.R.
ID
ZDB-PUB-170726-12
Date
2017
Source
Journal of immunology (Baltimore, Md. : 1950)   199(5): 1706-1715 (Journal)
Registered Authors
Feng, Hui, Jette, Cicely A., Liu, Xingjun, Look, A. Thomas, Rhodes, Jennifer
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • B-Lymphocytes/physiology*
  • CD79 Antigens/genetics
  • CD79 Antigens/metabolism*
  • Cell Differentiation
  • Cloning, Molecular
  • DNA-Binding Proteins/genetics
  • Fish Proteins/genetics
  • Fish Proteins/metabolism*
  • Genes, Reporter/genetics
  • Green Fluorescent Proteins/genetics
  • Immunoglobulin Heavy Chains/genetics
  • Immunoglobulin Heavy Chains/metabolism
  • Immunoglobulin Light Chains/genetics
  • Immunoglobulin Light Chains/metabolism
  • Kidney/physiology*
  • Lymphocyte Activation
  • Precursor Cells, B-Lymphoid/physiology*
  • Transgenes/genetics
  • Zebrafish/immunology*
  • Zebrafish Proteins/genetics
(all 22)
PubMed
28739882 Full text @ J. Immunol.
Abstract
CD79a and CD79b proteins associate with Ig receptors as integral signaling components of the B cell Ag receptor complex. To study B cell development in zebrafish, we isolated orthologs of these genes and performed in situ hybridization, finding that their expression colocalized with IgH-μ in the kidney, which is the site of B cell development. CD79 transgenic lines were made by linking the promoter and upstream regulatory segments of CD79a and CD79b to enhanced GFP to identify B cells, as demonstrated by PCR analysis of IgH-μ expression in sorted cells. We crossed these CD79-GFP lines to a recombination activating gene (Rag)2:mCherry transgenic line to identify B cell development stages in kidney marrow. Initiation of CD79:GFP expression in Rag2:mCherry+ cells and the timing of Ig H and L chain expression revealed simultaneous expression of both IgH-μ- and IgL-κ-chains, without progressing through the stage of IgH-μ-chain alone. Rag2:mCherry+ cells without CD79:GFP showed the highest Rag1 and Rag2 mRNAs compared with CD79a and CD79b:GFP+ B cells, which showed strongly reduced Rag mRNAs. Thus, B cell development in zebrafish does not go through a Raghi CD79+IgH-μ+ pre-B cell stage, different from mammals. After the generation of CD79:GFP+ B cells, decreased CD79 expression occurred upon differentiation to Ig secretion, as detected by alteration from membrane to secreted IgH-μ exon usage, similar to in mammals. This confirmed a conserved role for CD79 in B cell development and differentiation, without the requirement of a pre-B cell stage in zebrafish.
Genes / Markers
Marker Marker Type Name
cd79aGENECD79a molecule, immunoglobulin-associated alpha
cd79bGENECD79b molecule, immunoglobulin-associated beta
ighmGENEimmunoglobulin heavy constant mu
ightGENEimmunoglobulin heavy constant tau
ighv1-4GENEimmunoglobulin heavy variable 1-4
igic1s1GENEimmunoglobulin light iota constant 1, s1
lckGENELCK proto-oncogene, Src family tyrosine kinase
mitfaGENEmelanocyte inducing transcription factor a
rag1GENErecombination activating 1
rag2GENErecombination activating gene 2
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Figures
Figure Gallery (4 images)
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
fcc88Tgbacterial treatment by injection: Pseudomonas aeruginosa PA14Adult
fcc89Tgbacterial treatment by injection: Pseudomonas aeruginosa PA14Adult
fcc89Tgbacterial treatment by injection: Pseudomonas aeruginosa PA14Adult
fcc89Tgbacterial treatment by injection: Pseudomonas aeruginosa PA14Adult
1 - 4 of 4
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
bu3TgTransgenic Insertion
    cz2TgTransgenic Insertion
      fcc88TgTransgenic Insertion
        fcc89TgTransgenic Insertion
          fcc90TgTransgenic Insertion
            fcc97TgTransgenic Insertion
              1 - 6 of 6
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              Human Disease / Model
              No data available
              Sequence Targeting Reagents
              No data available
              Fish
              Fish
              bu3Tg; fcc89Tg
              cz2Tg
              fcc88Tg
              fcc89Tg
              fcc90Tg
              fcc97Tg
              WT
              1 - 7 of 7
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              Antibodies
              No data available
              Orthology
              No data available
              Engineered Foreign Genes
              Marker Marker Type Name
              EGFPEFGEGFP
              mCherryEFGmCherry
              1 - 2 of 2
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              Mapping
              No data available
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              Citation
              Liu, X., Li, Y.S., Shinton, S.A., Rhodes, J., Tang, L., Feng, H., Jette, C.A., Look, A.T., Hayakawa, K., Hardy, R.R. (2017) Zebrafish B Cell Development without a Pre-B Cell Stage, Revealed by CD79 Fluorescence Reporter Transgenes. Journal of immunology (Baltimore, Md. : 1950). 199(5):1706-1715.