Skip to main content
Springer Nature Link
Log in

DDX6 localizes to nuage structures and the annulus of mammalian spermatogenic cells

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

The localization of DEAD (Asp-Glu-Ala-Asp) box helicase 6 (DDX6) in spermatogenic cells from the mouse, rat, and guinea pig was studied by immunofluorescence (IF) and immunoelectron microscopy (IEM). Spermatogenic cells from these species yielded similar DDX6 localization pattern. IF microscopy results showed that DDX6 localizes to both the nucleus and cytoplasm. In the cytoplasm of spermatogenic cells, diffuse cytosolic and discrete granular staining was observed, with the staining pattern changing during cell differentiation. IEM revealed that DDX6 localized to the five different types of nuage structures and non-nuage structures, including small granule aggregate and late spermatid annuli. Nuclear labeling was strongest in leptotene and zygotene spermatocytes and moderately strong in the nuclear pocket of late spermatids. DDX6 also localized to the surface of outer dense fibers, which comprise of flagella. The results show that DDX6 is present in nuage and non-nuage structures as well as nuclei, suggesting that DDX6 has diverse functions in spermatogenic cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Canada)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Akao Y, Nakagawa Y (2006) Expression of the DEAD-box/RNA helicase rck/p54 in mouse tissues: implications for heterogeneous protein expression. J Histochem Cytochem 54:955–960

    Article  CAS  PubMed  Google Scholar 

  • Andrei MA, Ingelfinger D, Heitzmann R, Achel T, Rivera-Pomar R, Lührmann R (2005) A role for elF4E and elF4E-transporter in targeting mRNPs to mammalian processing bodies. RNA 11:717–727

    Article  CAS  PubMed  Google Scholar 

  • Aravin AA, Van der Heijden GW, Castaneda J, Vagin VV, Hannon GJ, Bortvin A (2009) Cytoplasmic compartmentalization of the fetal piRNA pathway in mice. PLoS Genet 5:1–12

    Article  Google Scholar 

  • Clermont Y, Oko R, Hermo L (1993) Cell biology of mammalian spermiogenesis. In: Desjardins C, Ewing LL (eds) Cell and molecular biology of the testis. Oxford University Press, New York, pp 332–376

    Google Scholar 

  • Coller JM, Tucker M, Sheth U, Valecia-Sanchez MA, Parker R (2001) The DEAD box helicase, Dhh1p, functions in mRNA decapping and interacts with both the decapping and deadenylase complexes. RNA 7:1717–1727

    Google Scholar 

  • Eulalio A, Behm-Ansmant L, Schweizer D, Izaurralde E (2007) P-body formation is a consequence, not the cause, of RNA-mediated gene silencing. Mol Cell Biol 27:3970–3981

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Fawcett DW, Eddy EM, Phillips DM (1970) Observations on the fine structure and relationships of the chromatoid body in mammalian spermatogenesis. Biol Reprod 2:129–153

    Article  CAS  PubMed  Google Scholar 

  • Fenger-Groen M, Fillman C, Norrild B, Lykke-Andersen J (2005) Multiple processing body factors and the ARE binding protein TTP activate mRNA decapping. Mol Cell 20:905–915

    Article  Google Scholar 

  • Ferraiuolo M, Basak S, Dostie J, Murray EL, Schoenberg DR, Sonnenberg N (2005) A role for the elF4E-binding protein 4E-T in P-body formation and mRNA decay. J Cell Biol 170:913–924

    Article  CAS  PubMed  Google Scholar 

  • Franks T, Lukke-Andersen J (2008) The control of mRNA decapping and P-body formation. Mol Cell 32:605–615

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Greer AE, Hearing P, Ketner G (2011) The adenovirus E4 11 k protein binds and localizes the cytoplasmic P-body component Ddx6 to aggresomes. Virology 417:161–168

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Haraguchi CM, Mabuchi T, Yokota S (2003) Localization of a mitochondrial type of NADP-dependent isocitrate dehydrogenase in kidney and heart of rat: an immunocytochemical and biochemical study. J Histochem Cytochem 51:215–226

    Article  CAS  PubMed  Google Scholar 

  • Huys A, Thibault PA, Wilson JA (2013) Modulation of hepatitis C virus RNA accumulation and translation by DDX6 and miR-122 are mediated by separate mechanism. PLoS One 8(6):e67437

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • James V, Zhang Y, Foxler DE, de Moor CH, Kong YW, Webb TM, Self TJ, Feng Y, Lagos D, Chu CY, Rana TM, Morley SJ, Longmore GD, Bushell M, Sharp TV (2010) LIM-domain proteins, LIMD1, Ajuba, and WTIP are required for microRNA-mediated gene silencing. Proc Natl Acad Sci USA 107:12499–12504

    Article  CAS  PubMed  Google Scholar 

  • Kotaja N, Sassone-Corsi P (2007) The chromatoid body: a germ-cell-specific RNA-processing centre. Nat Rev Mol Cell Biol 8:85–90

    Article  CAS  PubMed  Google Scholar 

  • Kotaja N, Bhattacharyya SN, Jaskiewicz L, Kimmins S, Parvinen M, Filipowicz W, Sassone-Corsi P (2006) The chromatoid body of male germ cells: similarity with processing bodies and presence of dicer and microRNA pathway components. Proc Natl Acad Sci USA 103:2647–2652

    Article  CAS  PubMed  Google Scholar 

  • Ladomery M, Wade E, Sommerville J (1997) Xp54, the Xenopus homologue of human RNA helicase p54, is an integral component of stored mRNP particles in oocytes. Nucleic Acids Res 25:965–973

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Lalli MF, Clermont Y (1981) Structural changes of the head components of rat spermatid during spermatogenesis. Am J Anat 160:419–434

    Article  CAS  PubMed  Google Scholar 

  • Matsumoto K, Kwon O-Y, Kim H, Akao Y (2005) Epression of rck/p54, a DEAD-box RNA helicase, in gametogenesis and early embryogenesis of mice. Dev Dyn 233:1149–1156

    Article  CAS  PubMed  Google Scholar 

  • Meikar O, DaRos M, Korhonen H, Kotaja N (2011) Chromatoid body and small RNAs in male germ cells. Reproduction 142:195–209

    Article  CAS  PubMed  Google Scholar 

  • Minshall N, Kress M, Weil D, Standart N (2009) Role of p54 RNA helicase activity and its C-terminal domain in translational repression, P-body localization and assembly. Mol Biol Cell 20:2464–2472

    Article  PubMed Central  PubMed  Google Scholar 

  • Naarmann IS, Narnisch C, Müller-Newen G, Urlaub H, Ostareck-Lederer A, Ostareck DH (2010) DDX6 recruits translational silenced human reticulocyte 15-lipoxygenase mRNA to RNP granules. RNA 16:2189–2204

    Article  CAS  PubMed  Google Scholar 

  • Onohara Y, Yokota S (2012) Expression of DDX25 in nuage components of mammalian spermatogenic cells: immunofluorescence and immunoelectron microscopic study. Histochem Cell Biol 137:37–51

    Article  CAS  PubMed  Google Scholar 

  • Onohara Y, Fujiwara T, Yasukochi T, Himeno M, Yokota S (2010) Localization of mouse vasa homolog protein in chromatoid body and related nuage structures of mammalian spermatogenic cells during spermatogenesis. Histochem Cell Biol 133:627–639

    Article  CAS  PubMed  Google Scholar 

  • Rickwood D, Ford TC (1983) Preparation and fractionation of nuclei, nucleoli and deoxyribonucleoproteins. In: Rickwood D (ed) Iodinated density gradient media. A practical approach. IRL PRESS, Oxford, pp 69–89

    Google Scholar 

  • Russell L, Frank B (1978) Ultrastructural characterization of nuage in spermatocytes of the rat testis. Anat Rec 190:79–98

    Article  CAS  PubMed  Google Scholar 

  • Russell LD, Ettlin RA, Hikim APS, Clegg ED (1990) Histological and histopathological evaluation of the testis. Cache River Press, Florida

    Google Scholar 

  • Smillie DA, Sommerville J (2002) RNA helicase p54 (DDX6) is a shuttling protein involved in nuclear assembly of stores mRNP particles. J Cell Sci 115:395–407

    CAS  PubMed  Google Scholar 

  • Takebe M, Onohara Y, Yokota S (2013) Miki Takebe, Expression of MAEL in Nuage and non-Nuage compartments of rat spermatogenic cells and colocalization with DDX4, DDX25 and MIWI. Histochem Cell Biol 140:169–181

    Article  CAS  PubMed  Google Scholar 

  • Tritschler F, Braun JE, Eulalio A, Truffault V, Izaurralde E, Weichenrieder O (2009) Structural basis for the mutually exclusive anchoring of P body components EDC3 and Tral to the DEAD box protein DDX6/Me31B. Mol Cell 33:661–668

    Article  CAS  PubMed  Google Scholar 

  • Voronina E, Seydoux G, Nagamori N, Sassone-Corsi P (2013) RNA granules in germ cells. Cold Spring Harb Perspect Biol 2011(3):1–27

    Google Scholar 

  • Weston A, Sommerville J (2006) Xp54 and related (DDX6-like) RNA helicases: roles in messenger RNP assembly, translation regulation and RNA degradation. Nucleic Acids Res 34:3082–3094

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yokota S (2012a) Preparation of colloidal gold particles and conjugation of protein A, IgG, F(ab’)2, and streptavidin. In: Schwarzbach SD, Osafune T (eds) Immunoelectron microscopy. Methods and protocols. Springer, New York, pp 109–119

    Google Scholar 

  • Yokota S (2012b) Nuage proteins: their localization in subcellular structures of spermatogenic cells as revealed by immunoelectron microscopy. Histochem Cell Biol 138:1–11

    Article  CAS  PubMed  Google Scholar 

  • Yokota S, Onohara Y (2013) Expression sites of NANOS1 in rat spermatogenic cells as revealed by immunoelectron microscopy. Cell Bio 2:1–10

    Google Scholar 

  • Yonetamari H, Onohara Y, Yokota S (2012) Localization of BRUNNOL2 in rat spermatogenic cells as revealed by immunofluorescence and immunoelectron microscopic techniques. Open J Cell Biol 2:11–20

    Article  Google Scholar 

  • Zhang W-J, Wu JY (1996) Functional properties of p54, a novel SR protein active in constitutive and alternative splicing. Mol Cell Biol 16:5400–5408

    CAS  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

The work was supported in part by research funds from Nagasaki International University, a grant-in-aid from the Ministry of Education, Science, Culture and Sport, and Sport (17570158), by science research promotion fund from the Promotion and Mutual Aid Corporation for Private Schools of Japan.

Author information

Authors and Affiliations

  1. Section of Functional Morphology, Faculty of Pharmaceutical Sciences, Nagasaki International University, Huis Ten Bosch 2825-7, Sasebo, Nagasaki, 859-329, Japan

    Chika Kawahara, Sadaki Yokota & Hideaki Fujita

Authors
  1. Chika Kawahara
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sadaki Yokota
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hideaki Fujita
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sadaki Yokota.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kawahara, C., Yokota, S. & Fujita, H. DDX6 localizes to nuage structures and the annulus of mammalian spermatogenic cells. Histochem Cell Biol 141, 111–121 (2014). https://doi.org/10.1007/s00418-013-1153-2

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00418-013-1153-2

Keywords

Search

Navigation