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Marine Biotechnology

, Volume 20, Issue 2, pp 155–167 | Cite as

Novel Isoforms of N16 and N19 Families Implicated for the Nacreous Layer Formation in the Pearl Oyster Pinctada fucata

  • Fumito Ohmori
  • Shigeharu Kinoshita
  • Daisuke Funabara
  • Hiroki Koyama
  • Kiyohito Nagai
  • Kaoru Maeyama
  • Kikuhiko Okamoto
  • Shuichi Asakawa
  • Shugo Watabe
Original Article
  • 173 Downloads

Abstract

Although a wide variety of proteins and genes possibly related to the shell formation in bivalve have been identified, their functions have been only partially approved. We have recently performed deep sequencing of expressed sequence tags (ESTs) from the pearl oyster Pinctada fucata using a next-generation sequencer, identifying a dozen of novel gene candidates which are possibly associated with the nacreous layer formation. Among the ESTs, we focused on three novel isoforms (N16-6, N16-7, and N19-2) of N16 and N19 families with reference to five known genes in the families and determined the full-length cDNA sequences of these isoforms. Reverse transcription-polymerase chain reaction revealed that N16-6 was expressed in gill, gonad, adductor muscle, and mantle, whereas N16-7 exclusively in mantle. N19-2 was expressed in all tissues examined. In situ hybridization demonstrated their regional expression in mantle and pearl sac, which well corresponded to those shown by EST analysis previously reported. Shells in the pearl oyster injected with dsRNAs of N16-7 and N19-2 showed abnormal surface appearance in the nacreous layer. Taken together, novel isoforms in N16 and N19 families shown in this study are essential to form the nacreous layer.

Keywords

Pearl oyster Pinctada fucata Nacreous layer Mantle Pearl sac Tissue distribution RNAi 

Supplementary material

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Supplementary Table 1 (DOCX 15 kb)
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Supplementary Fig. 1 The DNA nucleotide sequences of N16 isoforms, N16–1, N16–2, N16–3, N16–5, N16–6 and N16–7, from the pearl oyster Pinctada fucata. (PPTX 60.4 kb)
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Supplementary Fig. 2 The DNA nucleotide sequences of N19 isoforms, N19 and N19–2, from the pearl oyster Pinctada fucata. (PPTX 61.2 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Fumito Ohmori
    • 1
  • Shigeharu Kinoshita
    • 1
  • Daisuke Funabara
    • 2
  • Hiroki Koyama
    • 3
  • Kiyohito Nagai
    • 4
  • Kaoru Maeyama
    • 5
  • Kikuhiko Okamoto
    • 5
  • Shuichi Asakawa
    • 1
  • Shugo Watabe
    • 1
    • 6
  1. 1.Graduate School of Agricultural and Life SciencesThe University of TokyoTokyoJapan
  2. 2.Graduate School of BioresourcesMie UniversityTsuJapan
  3. 3.Graduate School of Biosphere ScienceHiroshima UniversityHiroshimaJapan
  4. 4.Pearl Research Laboratory, K. Mikimoto & Co., Ltd.ShimaJapan
  5. 5.Mikimoto Pharmaceutical Co., Ltd.IseJapan
  6. 6.School of Marine BiosciencesKitasato UniversitySagamiharaJapan

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