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Cited by 91melanin and skin pigmentationDevelopmental Biology and Gene RegulationConnexins and lens biology

Gap junctions composed of connexins 41.8 and 39.4 are essential for colour pattern formation in zebrafish

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Irion, Uwe, Frohnhöfer, Hans Georg, Krauss, Jana, Çolak Champollion, Tuǧba, Maischein, Hans-Martin, Geiger-Rudolph, Silke, Weiler, Christian, Nüsslein-Volhard, Christiane·2014·eLife
DOI 10.7554/elife.05125PMC PMC4296512
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Abstract

Interactions between all three pigment cell types are required to form the stripe pattern of adult zebrafish (Danio rerio), but their molecular nature is poorly understood. Mutations in leopard (leo), encoding Connexin 41.8 (Cx41.8), a gap junction subunit, cause a phenotypic series of spotted patterns. A new dominant allele, leotK3, leads to a complete loss of the pattern, suggesting a dominant negative impact on another component of gap junctions. In a genetic screen, we identified this component as Cx39.4 (luchs). Loss-of-function alleles demonstrate that luchs is required for stripe formation in zebrafish; however, the fins are almost not affected. Double mutants and chimeras, which show that leo and luchs are only required in xanthophores and melanophores, but not in iridophores, suggest that both connexins form heteromeric gap junctions. The phenotypes indicate that these promote homotypic interactions between melanophores and xanthophores, respectively, and those cells instruct the patterning of the iridophores.

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  • Gja4 and Gja5b are gap junction proteins that are required autonomously by melanophores and xanthophores for homotypic and heterotypic interactions.

    “Gja4 and Gja5b are gap junction proteins that are required autonomously by melanophores and xanthophores for homotypic and heterotypic interactions, i.e., interactions between the same or different pigment cell types, respectively15,16.”

    From Genomic and genetic insights into speciation and pigment pattern diversification in <i>Danio</i> fishes (2025) · cites this paper as [15]

    The Results section on Cx41.8 and Cx39.4 reports that leo and luc function is required in xanthophores and melanophores; transplantation experiments specifically support a cell-autonomous requirement in xanthophores. The Discussion supports homotypic melanophore interactions but says melanophore–xanthophore interaction via gap junctions might occur and that direct interaction in vivo is not known. The claim presents that heterotypic interaction as established, and our copy does not identify the connexins by the names Gja4 and Gja5b. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

  • Gap junctions containing Gja5b are critical for stripe patterning in D. rerio, with mutants developing spots similar to the wild-type pattern of D. tinwini.

    “Gap junctions containing Gja5b are critical for stripe patterning in D. rerio with mutants developing spots similar to the wild-type pattern of D. tinwini15,16,54.”

    From Genomic and genetic insights into speciation and pigment pattern diversification in <i>Danio</i> fishes (2025) · cites this paper as [15]

    The Introduction (page unavailable) identifies connexin 41.8/GJA5 as a gap-junction subunit and says leopard mutants in Danio rerio have stripes broken into dark spots. The Abstract (page unavailable) likewise reports spotted patterns from leo mutations. Our copy does not identify the spots as similar to the wild-type pattern of D. tinwini, so that comparison is unsupported. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • The common ancestor of Danio likely had horizontal stripes dependent on gap junctions between and amongst melanophores and xanthophores involving Gja4 and Gja5b as in D. rerio.

    “Given that the common ancestor of Danio likely had horizontal stripes presumably dependent on gap junctions between and amongst melanophores and xanthophores involving Gja4 and Gja5b as in D. rerio15,16”

    From Genomic and genetic insights into speciation and pigment pattern diversification in <i>Danio</i> fishes (2025) · cites this paper as [15]

    The Introduction describes horizontal stripes in D. rerio, and the Discussion (‘Gap junction communication between chromatophores’; page not provided) links connexins Cx41.8 and Cx39.4 to interactions among melanophores and xanthophores. The paper does not establish that the common ancestor of Danio had horizontal stripes or depended on these gap junctions; that ancestral inference goes beyond its findings. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

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Cites

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Research works printed but not linked: 44

  1. Brand M Granato M Nüsslein-Volhard C 2002 Keeping and raising zebrafish Nüsslein-Volhard C Dahm R Zebrafish: a practical approach Oxford, New York, NY Oxford University Press
  2. Budi EH Patterson LB Parichy DM 2011 Post-embryonic nerve-associated precursors to adult pigment cells: genetic requirements and dynamics of morphogenesis and differentiation PLOS Genetics 7 e1002044 10.1371/journal.pgen.1002044 21625562 PMC3098192 · DOI · Paper page
  3. Cruciani V Mikalsen SO 2006 The vertebrate connexin family Cellular and Molecular Life Sciences 63 1125 1140 10.1007/s00018-005-5571-8 16568237 PMC11136263 · DOI
  4. Dooley CM Mongera A Walderich B Nüsslein-Volhard C 2013 On the embryonic origin of adult melanophores: the role of ErbB and Kit signalling in establishing melanophore stem cells in zebrafish Development 140 1003 1013 10.1242/dev.087007 23364329 · DOI
  5. Eastman SD Chen TH Falk MM Mendelson TC Iovine MK 2006 Phylogenetic analysis of three complete gap junction gene families reveals lineage-specific duplications and highly supported gene classes Genomics 87 265 274 10.1016/j.ygeno.2005.10.005 16337772 · DOI
  6. Eom DS Inoue S Patterson LB Gordon TN Slingwine R Kondo S Watanabe M Parichy DM 2012 Melanophore migration and survival during zebrafish adult pigment stripe development require the immunoglobulin superfamily adhesion molecule Igsf11 PLOS Genetics 8 e1002899 10.1371/journal.pgen.1002899 22916035 PMC3420941 · DOI · Paper page
  7. Frankel JS 1979 Inheritance of spotting in the leopard danio The Journal of Heredity 70 287 288 489959
  8. Frohnhofer HG Krauss J Maischein HM Nüsslein-Volhard C 2013 Iridophores and their interactions with other chromatophores are required for stripe formation in zebrafish Development 140 2997 3007 10.1242/dev.096719 23821036 PMC3912879 · DOI · Paper page
  9. Haffter P Odenthal J Mullins MC Lin S Farrell MJ Vogelsang E Haas F Brand M van Eeden FJ Furutani-Seiki M Granato M Hammerschmidt M Heisenberg CP Jiang YJ Kane DA Kelsh RN Hopkins N Nüsslein-Volhard C 1996 Mutations affecting pigmentation and shape of the adult zebrafish Development Genes and Evolution 206 260 276 10.1007/s004270050051 24173565 · DOI · Paper page
  10. Hamada H Watanabe M Lau HE Nishida T Hasegawa T Parichy DM Kondo S 2014 Involvement of Delta/Notch signaling in zebrafish adult pigment stripe patterning Development 141 318 324 10.1242/dev.099804 24306107 PMC3879813 · DOI
  11. Hirata M Nakamura K Kanemaru T Shibata Y Kondo S 2003 Pigment cell organization in the hypodermis of zebrafish Developmental Dynamics 227 497 503 10.1002/dvdy.10334 12889058 · DOI
  12. Hirata M Nakamura K Kondo S 2005 Pigment cell distributions in different tissues of the zebrafish, with special reference to the striped pigment pattern Developmental Dynamics 234 293 300 10.1002/dvdy.20513 16110504 · DOI
  13. Inaba M Yamanaka H Kondo S 2012 Pigment pattern formation by contact-dependent depolarization Science 335 677 10.1126/science.1212821 22323812 · DOI · Paper page
  14. Inoue S Kondo S Parichy DM Watanabe M 2014 Tetraspanin 3c requirement for pigment cell interactions and boundary formation in zebrafish adult pigment stripes Pigment Cell & Melanoma Research 27 190 200 10.1111/pcmr.12192 24734316 PMC3988474 · DOI
  15. Iwashita M Watanabe M Ishii M Chen T Johnson SL Kurachi Y Okada N Kondo S 2006 Pigment pattern in jaguar/obelix zebrafish is caused by a Kir7.1 mutation: implications for the regulation of melanosome movement PLOS Genetics 2 e197 10.1371/journal.pgen.0020197 17121467 PMC1657052 · DOI · Paper page
  16. Kane DA Kishimoto T 2002 Cell labeling and transplantation techniques Nüsslein-Volhard C Dahm R Zebrafish: A Practical Approach New York Oxford University Press 95 119
  17. Kar R Batra N Riquelme MA Jiang JX 2012 Biological role of connexin intercellular channels and hemichannels Archives of Biochemistry and Biophysics 524 2 15 10.1016/j.abb.2012.03.008 22430362 PMC3376239 · DOI
  18. Kirschbaum F 1975 Untersuchungen über das farbmuster der zebrabarbe 'brachydanio rerio' (Cyprinidae, Teleostei) Wilhelm Roux's Archives of Developmental Biology 177 129 152 10.1007/BF00848526 28304951 · DOI
  19. Koval M Molina SA Burt JM 2014 Mix and match: investigating heteromeric and heterotypic gap junction channels in model systems and native tissues FEBS Letters 588 1193 1204 10.1016/j.febslet.2014.02.025 24561196 PMC3992227 · DOI
  20. Krauss J Astrinidis P Frohnhöfer HG Walderich B Nüsslein-Volhard C 2013 transparent, a gene affecting stripe formation in Zebrafish, encodes the mitochondrial protein Mpv17 that is required for iridophore survival Biology Open 2 703 710 10.1242/bio.20135132 23862018 PMC3711038 · DOI · Paper page
  21. Krauss J Frohnhöfer HG Walderich B Maischein HM Weiler C Irion U Nüsslein-Volhard C 2014 Endothelin signalling in iridophore development and stripe pattern formation of zebrafish Biology Open 3 503 509 10.1242/bio.20148441 24857848 PMC4058085 · DOI · Paper page
  22. Lister JA Robertson CP Lepage T Johnson SL Raible DW 1999 nacre encodes a zebrafish microphthalmia-related protein that regulates neural-crest-derived pigment cell fate Development 126 3757 3767 10433906
  23. Lopes SS Yang X Müller J Carney TJ McAdow AR Rauch GJ Jacoby AS Hurst LD Delfino-Machín M Haffter P Geisler R Johnson SL Ward A Kelsh RN 2008 Leukocyte tyrosine kinase functions in pigment cell development PLOS Genetics 4 e1000026 10.1371/journal.pgen.1000026 18369445 PMC2265441 · DOI · Paper page
  24. Maderspacher F Nüsslein-Volhard C 2003 Formation of the adult pigment pattern in zebrafish requires leopard and obelix dependent cell interactions Development 130 3447 3457 10.1242/dev.00519 12810592 · DOI · Paper page
  25. Mahalwar P Walderich B Singh AP Nüsslein-Volhard C 2014 Local reorganization of xanthophores fine-tunes and colors the striped pattern of zebrafish Science 345 1362 1364 10.1126/science 25214630 · DOI · Paper page
  26. McMenamin SK Bain EJ McCann AE Patterson LB Eom DS Waller ZP Hamill JC Kuhlman JA Eisen JS Parichy DM 2014 Thyroid hormone-dependent adult pigment cell lineage and pattern in zebrafish Science 345 1358 1361 10.1126/science.1256251 25170046 PMC4211621 · DOI · Paper page
  27. Milks LC Kumar NM Houghten R Unwin N Gilula NB 1988 Topology of the 32-kd liver gap junction protein determined by site-directed antibody localizations The EMBO Journal 7 2967 2975 2460334 PMC454678
  28. Musa H Fenn E Crye M Gemel J Beyer EC Veenstra RD 2004 Amino terminal glutamate residues confer spermine sensitivity and affect voltage gating and channel conductance of rat connexin40 gap junctions The Journal of Physiology 557 863 878 10.1113/jphysiol.2003.059386 15107469 PMC1665163 · DOI
  29. Nakamasu A Takahashi G Kanbe A Kondo S 2009 Interactions between zebrafish pigment cells responsible for the generation of Turing patterns Proceedings of the National Academy of Sciences of USA 106 8429 8434 10.1073/pnas.0808622106 PMC2689028 19433782 · DOI · Paper page
  30. Odenthal J Rossnagel K Haffter P Kelsh RN Vogelsang E Brand M van Eeden FJ Furutani-Seiki M Granato M Hammerschmidt M Heisenberg CP Jiang YJ Kane DA Mullins MC Nüsslein-Volhard C 1996 Mutations affecting xanthophore pigmentation in the zebrafish, Danio rerio Development 123 391 398 9007257
  31. Parichy DM Mellgren EM Rawls JF Lopes SS Kelsh RN Johnson SL 2000a Mutational analysis of endothelin receptor b1 (rose) during neural crest and pigment pattern development in the zebrafish Danio rerio Developmental Biology 227 294 306 10.1006/dbio.2000.9899 11071756 · DOI · Paper page
  32. Parichy DM Ransom DG Paw B Zon LI Johnson SL 2000b An orthologue of the kit-related gene fms is required for development of neural crest-derived xanthophores and a subpopulation of adult melanocytes in the zebrafish, Danio rerio Development 127 3031 3044 10862741
  33. Parichy DM Rawls JF Pratt SJ Whitfield TT Johnson SL 1999 Zebrafish sparse corresponds to an orthologue of c-kit and is required for the morphogenesis of a subpopulation of melanocytes, but is not essential for hematopoiesis or primordial germ cell development Development 126 3425 3436 10393121
  34. Parichy DM Turner JM 2003 Temporal and cellular requirements for Fms signaling during zebrafish adult pigment pattern development Development 130 817 833 10.1242/dev.00307 12538511 · DOI · Paper page
  35. Patterson LB Parichy DM 2013 Interactions with iridophores and the tissue environment required for patterning melanophores and xanthophores during zebrafish adult pigment stripe formation PLOS Genetics 9 e1003561 10.1371/journal.pgen.1003561 23737760 PMC3667786 · DOI · Paper page
  36. Rohner N Perathoner S Frohnhöfer HG Harris MP 2011 Enhancing the efficiency of N-ethyl-N-nitrosourea-induced mutagenesis in the zebrafish Zebrafish 8 119 123 10.1089/zeb.2011.0703 21861612 · DOI
  37. Singh AP Schach U Nüsslein-Volhard C 2014 Proliferation, dispersal and patterned aggregation of iridophores in the skin prefigure striped colouration of zebrafish Nature Cell Biology 16 607 614 10.1038/ncb2955 24776884 · DOI · Paper page
  38. Thévenin AF Kowal TJ Fong JT Kells RM Fisher CG Falk MM 2013 Proteins and mechanisms regulating gap-junction assembly, internalization, and degradation Physiology 28 93 116 10.1152/physiol.00038.2012 23455769 PMC3768091 · DOI
  39. Unwin PN Zampighi G 1980 Structure of the junction between communicating cells Nature 283 545 549 10.1038/283545a0 7354837 · DOI
  40. Watanabe M Kondo S 2012 Changing clothes easily: connexin41.8 regulates skin pattern variation Pigment Cell & Melanoma Research 25 326 330 10.1111/j.1755-148X.2012.00984.x 22313791 · DOI
  41. Watanabe M Iwashita M Ishii M Kurachi Y Kawakami A Kondo S Okada N 2006 Spot pattern of leopard Danio is caused by mutation in the zebrafish connexin41.8 gene EMBO Reports 7 893 897 10.1038/sj.embor.7400757 16845369 PMC1559663 · DOI · Paper page
  42. Watanabe M Watanabe D Kondo S 2012 Polyamine sensitivity of gap junctions is required for skin pattern formation in zebrafish Scientific Reports 2 473 10.1038/srep00473 22737406 PMC3382735 · DOI
  43. Yamaguchi M Yoshimoto E Kondo S 2007 Pattern regulation in the stripe of zebrafish suggests an underlying dynamic and autonomous mechanism Proceedings of the National Academy of Sciences of USA 104 4790 4793 10.1073/pnas.0607790104 PMC1820822 17360399 · DOI
  44. Yamanaka H Kondo S 2014 In vitro analysis suggests that difference in cell movement during direct interaction can generate various pigment patterns in vivo Proceedings of the National Academy of Sciences of USA 111 1867 1872 10.1073/pnas.1315416111 PMC3918784 24449859 · DOI

Cited by

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Authors
Irion, U., Frohnhöfer, H. G., Krauss, J., Çolak Champollion, T., Maischein, H.-M., Geiger-Rudolph, S., Weiler, C., & Nüsslein-Volhard, C.
Affiliations
  • Max Planck Institute for Developmental Biology (DE)
  • Max Planck Institute for Developmental Biology, Tübingen, Germany
Corresponding author
Irion, Uwe
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2014-12-23
Volume
3
Pages
e05125-e05125
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.05125
PMC
PMC4296512
OpenAlex
W2159198630
Cited by
91
References
1
Citations by year
2026: 4 · 2025: 6 · 2024: 4 · 2023: 5 · 2021: 17 · 2020: 11 · 2019: 10 · 2018: 8 · 2017: 9 · 2016: 8 · 2015: 9
FWCI
4.00
Type
journal article
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Max Planck Institute for Plasma Physics (submittedVersion)
Funders
European Commission (ZF-HEALTH grant, 242048)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › melanin and skin pigmentation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Connexins and lens biology
Keywords
gap junctions, xanthophores, melanophores, iridophores, dominant negative effect, chimeras, double mutants
MeSH terms
Alleles, Amino Acid Sequence, Animals, Chimera, Color, Embryo, Nonmammalian, Female, Genotype, Male, Molecular Sequence Data, Mutation, Phenotype, Pigmentation, Pigments, Biological, Zebrafish, Sequence Alignment, Gap Junctions, Connexins, Gene Expression Regulation, Developmental, Protein Isoforms, Zebrafish Proteins

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