Skip to main content

Advertisement

SpringerLink
Log in

Profilin-1 negatively controls osteoclast migration by suppressing the protrusive structures based on branched actin filaments

  • Original Article
  • Published:
Journal of Bone and Mineral Metabolism Aims and scope Submit manuscript

A Correction to this article was published on 19 August 2023

This article has been updated

Abstract

Background

Profilin-1 (Pfn1), an evolutionarily conserved actin-binding protein, is an important regulator of the cytoskeleton. We previously reported the osteoclast-specific Pfn1-conditional knockout (cKO) mice had postnatal osteolytic phenotype with craniofacial and long-bone deformities associated with increased migration of cultured osteoclasts. We hypothesized the increased cellular processes structured with branched actin filaments may underlies the mechanism of increased bone resorption in these mutant mice.

Materials and methods

The morphological structure and cell migration of the cultured osteoclasts were analyzed using fluorescent microscopy and time-lapse image capturing. Fractional migration distances, as well as the index of protrusive structures (%-PB) that evaluates relative border length of the protrusion were compared between the cells from control and Pfn1-cKO mice.

Results

Time-lapse image analysis showed that %-PB was significantly larger in Pfn1-cKO osteoclasts. In addition, the fractional migration distance was positively correlated with the index. When the branched actin filament organization was suppressed by chemical inhibitors, the osteoclast migration was declined. Importantly, the suppression was more extensive in Pfn1-cKO than in control osteoclasts.

Conclusion

Our results indicated the causative involvement of the increased branched actin filament formation at least in part for their excessive migration. Our findings provide a mechanistic rationale for testing novel therapeutic approaches targeting branched actin filaments in osteolytic disorders.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Change history

References

  1. Ralston SH, Layfield R (2012) Pathogenesis of paget disease of bone. Calcif Tissue Int 91:97–113

    Article  CAS  PubMed  Google Scholar 

  2. Ralston SH, Albagha OM (2014) Genetics of paget’s disease of bone. Curr Osteoporos Rep 12:263–271

    Article  PubMed  Google Scholar 

  3. Gennari L, Rendina D, Falchetti A, Merlotti D (2019) Paget’s disease of bone. Calcif Tissue Int 104:483–500

    Article  CAS  PubMed  Google Scholar 

  4. Merlotti D, Materozzi M, Bianciardi S, Guarnieri V, Rendina D, Volterrani L, Bellan C, Mingiano C, Picchioni T, Frosali A, Orfanelli U, Cenci S, Gennari L (2020) Mutation of PFN1 gene in an early onset, polyostotic paget-like disease. J Clin Endocrinol Metab 105:2553–2565

    Article  Google Scholar 

  5. Scotto di Carlo F, Pazzaglia L, Esposito T, Gianfrancesco F (2020) The loss of profilin 1 causes early onset paget’s disease of bone. J Bone Miner Res 35:1387–1398

    Article  CAS  PubMed  Google Scholar 

  6. Böttcher RT, Wiesner S, Braun A, Wimmer R, Berna A, Elad N, Medalia O, Pfeifer A, Aszodi A, Costell M, Fassler R (2009) Profilin 1 is required for abscission during late cytokinesis of chondrocytes. EMBO J 28:1157–1169

    Article  PubMed  PubMed Central  Google Scholar 

  7. Miyajima D, Hayata T, Suzuki T, Hemmi H, Nakamoto T, Notomi T, Amagasa T, Bottcher RT, Costell M, Fassler R, Ezura Y, Noda M (2012) Profilin1 regulates sternum development and endochondral bone formation. J Biol Chem 287:33545–33553

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Lin W, Izu Y, Smriti A, Kawasaki M, Pawaputanon C, Bottcher RT, Costell M, Moriyama K, Noda M, Ezura Y (2018) Profilin1 is expressed in osteocytes and regulates cell shape and migration. J Cell Physiol 233:259–268

    Article  CAS  PubMed  Google Scholar 

  9. Shirakawa J, Kajikawa S, Bottcher RT, Costell M, Izu Y, Hayata T, Noda M, Ezura Y (2019) Profilin 1 negatively regulates osteoclast migration in postnatal skeletal growth, remodeling, and homeostasis in mice. JBMR Plus 3:e10130

    Article  PubMed  PubMed Central  Google Scholar 

  10. Schachtner H, Calaminus SD, Thomas SG, Machesky LM (2013) Podosomes in adhesion, migration, mechanosensing and matrix remodeling. Cytoskeleton 70:572–589

    Article  CAS  PubMed  Google Scholar 

  11. Henty-Ridilla JC, Goode BL (2015) Global resource distribution: allocation of actin building blocks by profilin. Dev Cell 32:5–6

    Article  CAS  PubMed  Google Scholar 

  12. Takeshita S, Kaji K, Kudo A (2000) Identification and characterization of the new osteoclast progenitor with macrophage phenotypes being able to differentiate into mature osteoclasts. J Bone Miner Res 15:1477–1488

    Article  CAS  PubMed  Google Scholar 

  13. Kajikawa S, Taguchi Y, Hayata T, Ezura Y, Ueta R, Arimura S, Inoue JI, Noda M, Yamanashi Y (2018) Dok-3 and Dok-1/-2 adaptors play distinctive roles in cell fusion and proliferation during osteoclastogenesis and cooperatively protect mice from osteopenia. Biochem Biophys Res Commun 498:967–974

    Article  CAS  PubMed  Google Scholar 

  14. Hetrick B, Han MS, Helgeson LA, Nolen BJ (2013) Small molecules CK-666 and CK-869 inhibit actin-related protein 2/3 complex by blocking an activating conformational change. Chem Biol 20:701–712

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Beckham Y, Vasquez RJ, Stricker J, Sayegh K, Campillo C, Gardel ML (2014) Arp2/3 inhibition induces amoeboid-like protrusions in MCF10A epithelial cells by reduced cytoskeletal-membrane coupling and focal adhesion assembly. PLoS ONE 9:e100943

    Article  PubMed  PubMed Central  Google Scholar 

  16. Skruber K, Warp PV, Shklyarov R, Thomas JD, Swanson MS, Henty-Ridilla JL, Read TA, Vitriol EA (2020) Arp2/3 and Mena/VASP require profilin 1 for actin network assembly at the leading edge. Curr Biol 30:2651–2664 (e2655)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Destaing O, Saltel F, Géminard JC, Jurdic P, Bard F (2003) Podosomes display actin turnover and dynamic self-organization in osteoclasts expressing actin-green fluorescent protein. Mol Biol Cell 14:407–416

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. van den Dries K, Linder S, Maridonneau-Parini I, Poincloux R (2019) Probing the mechanical landscape - new insights into podosome architecture and mechanics. J Cell Sci 132:jcs236828

    Article  PubMed  Google Scholar 

  19. Bae YH, Ding Z, Das T, Wells A, Gertler F, Roy P (2010) Profilin1 regulates PI(3,4)P2 and lamellipodin accumulation at the leading edge thus influencing motility of MDA-MB-231 cells. Proc Natl Acad Sci USA 107:21547–21552

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Bender M, Stritt S, Nurden P, van Eeuwijk JM, Zieger B, Kentouche K, Schulze H, Morbach H, Stegner D, Heinze KG, Dütting S, Gupta S, Witke W, Falet H, Fischer A, Hartwig JH, Nieswandt B (2014) Megakaryocyte-specific profilin1-deficiency alters microtubule stability and causes a Wiskott-Aldrich syndrome-like platelet defect. Nat Commun 5:4746

    Article  CAS  PubMed  Google Scholar 

  21. Schoppmeyer R, Zhao R, Cheng H, Hamed M, Liu C, Zhou X, Schwarz EC, Zhou Y, Knorck A, Schwar G, Ji S, Liu L, Long J, Helms V, Hoth M, Yu X, Qu B (2017) Human profilin 1 is a negative regulator of CTL mediated cell-killing and migration. Eur J Immunol 47:1562–1572

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Dr. Reinhard Fässler and Dr. Ralph Böttcher for their support providing the mutant Pfn1-flox mouse line for this research, and Dr. Sunao Takeshita for kindly providing CMG14-12 cell lines. Dr. Koichiro Komatsu and Dr. Hisashi Ideno at Tsurumi University for kind support and discussion.

Funding

This study was funded by grants-in-aid (KAKENHI: 16K10892, 19K09617 and 19K24056), and grant for Joint Usage/Research Program of Medical Research Institute, Tokyo Medical and Dental University, from the Ministry of Education, Culture, Sports, Science and Technology. This study was also supported by the research and education funds from Ehime prefecture and Imabari City, Japan.

Author information

Authors and Affiliations

  1. Department of Veterinary Medicine, The Faculty of Veterinary Medicine, Okayama University of Science, Imabari, Ehime, 794-8555, Japan

    Shuhei Kajikawa & Yayoi Izu

  2. Department of Joint Surgery and Sports Medicine, Tokyo Medical and Dental University (TMDU), Tokyo, 113-8510, Japan

    Yoichi Ezura

  3. Department of Occupational Therapy, Faculty of Health and Medical Sciences, Teikyo Heisei University, Tokyo, 170-0013, Japan

    Yoichi Ezura

  4. Department of Pharmacology, Tsurumi University School of Dental Medicine, Tsurumi, Kanagawa, 230-8501, Japan

    Kazuhisa Nakashima & Akira Nifuji

  5. Department of Orthopedics, TMDU, Tokyo, 113-8510, Japan

    Masaki Noda

  6. Center for Stem Cell and Regenerative Medicine, TMDU, Tokyo, 113-8510, Japan

    Masaki Noda

Authors
  1. Shuhei Kajikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yoichi Ezura
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yayoi Izu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kazuhisa Nakashima
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Masaki Noda
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Akira Nifuji
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

SK was responsible for data collection, data analysis, and manuscript writing. YE was responsible for the conception, designing, data analysis, manuscript writing, and editing. YI and KN was responsible for critical reviewing and discussion. MN supervised this project. AN was responsible for the conception, designing, and reviewing. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Yoichi Ezura.

Ethics declarations

Conflict of interest

All authors declare no competing financial interests. The funders had no role in the design of the study, in the collection, analyses, or interpretation of data, in the writing of the manuscript, or in the decision to publish the results.

Ethical approval

All experimental protocols were approved by the animal welfare committee of the Tokyo Medical and Dental University (A2017-082A, A2018-180A, and A2019-222A).

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 4212 KB)

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kajikawa, S., Ezura, Y., Izu, Y. et al. Profilin-1 negatively controls osteoclast migration by suppressing the protrusive structures based on branched actin filaments. J Bone Miner Metab 40, 561–570 (2022). https://doi.org/10.1007/s00774-022-01320-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00774-022-01320-y

Keywords

Search

Navigation