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Critical role of connexin43 in zebrafish late primitive and definitive hematopoiesis

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Abstract

In vitro studies have suggested that connexin43 (cx43) expression is of particular importance during establishment and regeneration of the mammalian hematopoietic system. However, little is known about its in vivo functions during hematopoiesis due to the embryonic lethality of mammalian knockout models. In this study, we observed that zebrafish cx43 is not only expressed in the eyes, cerebellum, heart, and vasculature, but also expressed, albeit at low levels, in intermediate cell mass (ICM, the primitive hematopoietic site). Knockdown of cx43 leads to vacuolization in the wedge of the ICM and an apparent reduction in the number of circulating blood cells, but does not affect their cellular morphology. Whole-mount in situ hybridization analysis revealed that the hemangioblastic marker flk-1 and the primitive hematopoietic markers lmo2 and scl are basically maintained at normal levels in cx43 morphant embryos at 12–13 h postfertilization (hpf) compared with the con-MO injected embryos. However, subsequent expression of the definitive hematopoietic stem cell (HSC) marker c-myb was severely downregulated in the ventral wall of the dorsal aorta of cx43-depleted embryos at 36 hpf. Furthermore, we confirmed this phenotype by injection of cx43-MO into Tg(gata1:EGFP) embryos. Together, our results show that cx43 contributes to late primitive and definitive hematopoiesis in zebrafish embryos.

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References

  • Alves LA, Campos de Carvalho AC, Cirne Lima EO, Rocha e Souza CM, Dardenne M, Spray DC et al (1995) Functional gap junctions in thymic epithelial cells are formed by connexin 43. Eur J Immunol 25(2):431–437

    Article  CAS  PubMed  Google Scholar 

  • Amatruda JF, Zon LI (1999) Dissecting hematopoiesis and disease using the zebrafish. Dev Biol 216(1):1–15

    Article  CAS  PubMed  Google Scholar 

  • Bahary N, Zon LI (1998) Use of the zebrafish (Danio rerio) to define hematopoiesis. Stem Cells 16(Suppl 2):67–78

    PubMed  Google Scholar 

  • Bergoffen J, Scherer SS, Wang S, Scott MO, Bone LJ, Paul DL et al (1993) Connexin mutations in X-linked Charcot-Marie-Tooth disease. Science 262(5142):2039–2042

    Article  CAS  PubMed  Google Scholar 

  • Bruzzone R, White TW, Paul DL (1996) Connections with connexins: the molecular basis of direct intercellular signaling. Eur J Biochem 238(1):1–27

    Article  CAS  PubMed  Google Scholar 

  • Burns CE, DeBlasio T, Zhou Y, Zhang J, Zon L, Nimer SD (2002) Isolation and characterization of runxa and runxb, zebrafish members of the runt family of transcriptional regulators. Exp Hematol 30(12):1381–1389

    Article  CAS  PubMed  Google Scholar 

  • Cancelas JA, Koevoet WL, de Koning AE, Mayen AE, Rombouts EJ, Ploemacher RE (2000) Connexin-43 gap junctions are involved in multiconnexin-expressing stromal support of hemopoietic progenitors and stem cells. Blood 96(2):498–505

    CAS  PubMed  Google Scholar 

  • Chatterjee B, Chin AJ, Valdimarsson G, Finis C, Sonntag JM, Choi BY et al (2005) Developmental regulation and expression of the zebrafish connexin43 gene. Dev Dyn 233(3):890–906

    Article  CAS  PubMed  Google Scholar 

  • Detrich HW 3rd, Kieran MW, Chan FY, Barone LM, Yee K, Rundstadler JA et al (1995) Intraembryonic hematopoietic cell migration during vertebrate development. Proc Natl Acad Sci USA 92(23):10713–10717

    Article  CAS  PubMed  Google Scholar 

  • Dooley K, Zon LI (2000) Zebrafish: a model system for the study of human disease. Curr Opin Genet Dev 10(3):252–256

    Article  CAS  PubMed  Google Scholar 

  • Dorshkind K, Green L, Godwin A, Fletcher WH (1993) Connexin-43-type gap junctions mediate communication between bone marrow stromal cells. Blood 82(1):38–45

    CAS  PubMed  Google Scholar 

  • Galloway JL, Zon LI (2003) Ontogeny of hematopoiesis: examining the emergence of hematopoietic cells in the vertebrate embryo. Curr Top Dev Biol 53:139–158

    Article  CAS  PubMed  Google Scholar 

  • Hsia N, Zon LI (2005) Transcriptional regulation of hematopoietic stem cell development in zebrafish. Exp Hematol 33(9):1007–1014

    Article  CAS  PubMed  Google Scholar 

  • Iovine MK, Higgins EP, Hindes A, Coblitz B, Johnson SL (2005) Mutations in connexin43 (GJA1) perturb bone growth in zebrafish fins. Dev Biol 278(1):208–219

    Article  CAS  PubMed  Google Scholar 

  • Kelsell DP, Dunlop J, Stevens HP, Lench NJ, Liang JN, Parry G et al (1997) Connexin 26 mutations in hereditary non-syndromic sensorineural deafness. Nature 387(6628):80–83

    Article  CAS  PubMed  Google Scholar 

  • Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF (1995) Stages of embryonic development of the zebrafish. Dev Dyn 203(3):253–310

    CAS  PubMed  Google Scholar 

  • Krenacs T, Rosendaal M (1998) Connexin43 gap junctions in normal, regenerating, and cultured mouse bone marrow and in human leukemias: their possible involvement in blood formation. Am J Pathol 152(4):993–1004

    CAS  PubMed  Google Scholar 

  • Liu D, Wang YX, Hu JY, Sun SN, Song HY (2008) The effects of connexin43 down regulation on the development of the embryonic heart and vasculature in zebrafish. Progr Biochem Biophys 35(7):766–771

    CAS  Google Scholar 

  • Nasevicius A, Ekker SC (2000) Effective targeted gene ‘knockdown’ in zebrafish. Nat Genet 26(2):216–220

    Article  CAS  PubMed  Google Scholar 

  • North TE, Zon LI (2003) Modeling human hematopoietic and cardiovascular diseases in zebrafish. Dev Dyn 228(3):568–583

    Article  CAS  PubMed  Google Scholar 

  • Rosendaal M (1995) Gap junctions in blood forming tissues. Microsc Res Tech 31(5):396–407

    Article  CAS  PubMed  Google Scholar 

  • Rosendaal M, Gregan A, Green CR (1991) Direct cell-cell communication in the blood-forming system. Tissue Cell 23(4):457–470

    Article  CAS  PubMed  Google Scholar 

  • Rosendaal M, Green CR, Rahman A, Morgan D (1994) Up-regulation of the connexin43+ gap junction network in haemopoietic tissue before the growth of stem cells. J Cell Sci 107(Pt 1):29–37

    CAS  PubMed  Google Scholar 

  • Saez JC, Branes MC, Corvalan LA, Eugenin EA, Gonzalez H, Martinez AD et al (2000) Gap junctions in cells of the immune system: structure, regulation and possible functional roles. Braz J Med Biol Res 33(4):447–455

    Article  CAS  PubMed  Google Scholar 

  • Shiels A, Mackay D, Ionides A, Berry V, Moore A, Bhattacharya S (1998) A missense mutation in the human connexin50 gene (GJA8) underlies autosomal dominant “zonular pulverulent” cataract, on chromosome 1q. Am J Hum Genet 62(3):526–532

    Article  CAS  PubMed  Google Scholar 

  • Sohl G, Willecke K (2004) Gap junctions and the connexin protein family. Cardiovasc Res 62(2):228–232

    Article  PubMed  Google Scholar 

  • Thompson MA, Ransom DG, Pratt SJ, MacLennan H, Kieran MW, Detrich HW 3rd et al (1998) The cloche and spadetail genes differentially affect hematopoiesis and vasculogenesis. Dev Biol 197(2):248–269

    Article  CAS  PubMed  Google Scholar 

  • Westerfield M (2000) The zebrafish book: a guide for the laboratory use of zebrafish (Danio rerio). University of Oregon Press, Oregon

    Google Scholar 

Download references

Acknowledgments

We thank Prof. Tingxi Liu for his generous gift of Tg(gata1:EGFP) fish, Dr. Linxi Qian for his technical assistance. This work was supported by grants from National Natural Science Foundation of China (30600489) and Shanghai Municipal Science and Technology Commission (074319111).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Shanghai Medical College and Key Laboratory of Molecular Medicine, Ministry of Education, Fudan University, 200032, Shanghai, People’s Republic of China

    Qiu Jiang, Dong Liu, Jingying Hu, Li Tan, Yuexiang Wang, Min Yu & Houyan Song

  2. Children’s Hospital, Fudan University, 200032, Shanghai, People’s Republic of China

    Shuna Sun & Yonghao Gui

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  1. Qiu Jiang
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  2. Dong Liu
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  3. Shuna Sun
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  4. Jingying Hu
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  5. Li Tan
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  6. Yuexiang Wang
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  7. Yonghao Gui
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  8. Min Yu
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  9. Houyan Song
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Correspondence to Houyan Song.

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Qiu Jiang and Dong Liu contributed equally to this work.

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Jiang, Q., Liu, D., Sun, S. et al. Critical role of connexin43 in zebrafish late primitive and definitive hematopoiesis. Fish Physiol Biochem 36, 945–951 (2010). https://doi.org/10.1007/s10695-009-9371-1

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  • DOI: https://doi.org/10.1007/s10695-009-9371-1

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