Development Genes and Evolution

, Volume 228, Issue 5, pp 189–196 | Cite as

Developmental mechanisms of migratory muscle precursors in medaka pectoral fin formation

  • Saori Tani-Matsuhana
  • Rie Kusakabe
  • Kunio Inoue
Original Article


Limb muscles are formed from migratory muscle precursor cells (MMPs) that delaminate from the ventral region of dermomyotomes and migrate into the limb bud. MMPs remain undifferentiated during migration, commencing differentiation into skeletal muscle after arrival in the limb. However, it is still unclear whether the developmental mechanisms of MMPs are conserved in teleost fishes. Here, we investigate the development of pectoral fin muscles in the teleost medaka Oryzias latipes. Expression of the MMP marker lbx1 is first observed in several somites prior to the appearance of fin buds. lbx1-positive cells subsequently move anteriorly and localize in the prospective fin bud region to differentiate into skeletal muscle cells. To address the developmental mechanisms underlying fin muscle formation, we knocked down tbx5, a gene that is required for fin bud formation. tbx5 morphants showed loss of fin buds, whereas lbx1 expression initiated normally in anterior somites. Unlike in normal embryos, expression of lbx1 was not maintained in migrating fin MMPs or within the fin buds. We suggest that fin MMPs appear to undergo two phases in their development, with an initial specification of MMPs occurring independent of fin buds and a second fin bud-dependent phase of MMP migration and proliferation. Our results showed that medaka fin muscle is composed of MMPs. It is suggested that the developmental mechanism of fin muscle formation is conserved in teleost fishes including medaka. Through this study, we also propose new insights into the developmental mechanisms of MMPs in fin bud formation.


Fish fin muscle Myogenesis lbx1 Migratory muscle precursors Medaka 



We thank National Bio Resource Project Medaka (NBRP Medaka) for providing hatching enzyme. We also thank Dr. Marianne Bronner for valuable comments on the manuscript.

Funding information

This study was supported by Grant-in-Aid for JSPS Research Fellow (24-2096), JSPS KAKENHI (16K18551) to STM, and also in part by a research grant from the Takeda Science Foundation to RK.

Supplementary material

427_2018_616_MOESM1_ESM.pptx (1.6 mb)
Fig. S1 Immunostaining with MF20 to detect skeletal muscle in medaka embryos. (a) In stage 33, MF20 signal is observed in trunk muscle and fin bud muscle. (b) In stage 35, MF20 can mark another signal positive band between post fin bud and somites, which seems to be PHM, in addition to trunk and fin bud muscle. (c) In stage 35, lbx1 is expressed in pectoral fin muscle, but not in PHM which is marked with MF20. Dorsal is to the top. Arrows and arrowheads in (a, b) show MF20 signals in pectoral fin buds and PHM, respectively. Arrows in (c) show lbx1-positive cells in pectoral fin buds. (PPTX 1669 kb)
427_2018_616_MOESM2_ESM.pptx (1.6 mb)
Fig. S2 Expression patterns of tbx5a, tbx5b and dlx2 in medaka embryos. (a) tbx5a, (c) tbx5b and (e) dlx2 around stage 31. tbx5a and tbx5b are expressed in mesenchymal cells of fin buds, and dlx2 is expressed at the AER. (b), (d) and (f) are vertical sections of (a), (c) and (e) at the pectoral fin bud level, respectively. (a, c, e) Dorsal view, anterior is to the top. (b, d, f) Dorsal is to the top. Arrows show the expression signals in pectoral fin buds. Scale bars: 100 μm (a, c, e) or 50 μm (b, d, f) (PPTX 1655 kb)
427_2018_616_MOESM3_ESM.pptx (1.4 mb)
Fig. S3 tbx5 MOs affect formation of pectoral fin buds in medaka embryos. (a) tbx5a MO was designed to be complementary to the junction of exon2 and intron2. The MO caused exon2 skipping and a frame shift. tbx5b MO was designed to be complementary to the junction of intron2 and exon3. The MO caused a premature stop codon in intron2. Arrows show primers for RT-PCR to check for the transcripts, tbx5a or tbx5b. (b) RT-PCR was performed to detect tbx5a or tbx5b transcripts in tbx5 MOs-injected embryos or uninjected embryos. Black asterisks and white asterisks show normal transcripts and abnormal transcripts, respectively. (c) Embryos injected with tbx5 MOs were inhibited from forming pectoral fin buds whereas uninjected embryos formed normal pectoral fin buds at stage 35 (shown in arrows). (d) dlx2 expression was not observed in prospective fin bud area of stage 31 embryos lacking visible fin bud by tbx5 MOs. (PPTX 1422 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Saori Tani-Matsuhana
    • 1
  • Rie Kusakabe
    • 2
  • Kunio Inoue
    • 1
  1. 1.Department of Biology, Graduate School of ScienceKobe UniversityKobeJapan
  2. 2.Laboratory for Evolutionary MorphologyRIKEN Center for Biosystems Dynamics ResearchKobeJapan

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