SpringerLink
Log in

The Cyclin-Dependent Kinase Inhibitor p27Kip1 is a Positive Regulator of Schwann Cell Differentiation In Vitro

  • Published:
Journal of Molecular Neuroscience Aims and scope Submit manuscript

Abstract

Schwann cell precursors differentiating into a myelinating phenotype are critical for peripheral nerve development and regeneration. However, little is known about the underlying molecular mechanisms of Schwann cell differentiation. In the present study, we performed a cyclic adenosine monophosphate-induced Schwann cell differentiation model in vitro. Western blot analysis showed that p27Kip1 expression was upregulated during the differentiation of Schwann cell, while the inhibition of p27Kip1 expression by short hairpin RNA-mediated knockdown significantly abolished the expression of promyelinating markers and the alteration of cellular morphology. In addition, immunofluorescence revealed a decrease of p27Kip1 nuclear staining and a concomitant increase of cytoplasmic staining in differentiated Schwann cells. In summary, our data indicated that p27Kip1 was a positive regulator of Schwann cell differentiation in vitro.

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
EUR 32.99 /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

Similar content being viewed by others

Abbreviations

cAMP:

Cyclic adenosine monophosphate

KPC:

Kip1 ubiquitylation-promoting complex

SKP2:

S-phase kinase-associated protein 2

DMEM:

Dulbecco’s modified Eagle’s medium

PBS:

Phosphate-buffered saline

SCs:

Schwann cells

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

References

  • Arroyo EJ, Bermingham JR Jr, Rosenfeld MG, Scherer SS (1998) Promyelinating Schwann cells express Tst-1/SCIP/Oct-6. J Neurosci 18:7891–7902

    PubMed  CAS  Google Scholar 

  • Bermingham JR, Shearin H, Pennington J, O’Moore J, Jaegle M, Driegen S, van Zon A, Darbas A, Ozkaynak E, Ryu EJ, Milbrandt J, Meijer D (2006) The claw paw mutation reveals a role for Lgi4 in peripheral nerve development. Nat Neurosci 9:76–84

    Article  PubMed  CAS  Google Scholar 

  • Brockes JP, Fields KL, Raff MC (1979) Studies on cultured rat Schwann cells. I. Establishment of purified populations from cultures of peripheral nerve. Brain Res 165(1):105–118

    Article  PubMed  CAS  Google Scholar 

  • Carrano AC, Eytan E, Hershko A, Pagano M (1999) SKP2 is required for ubiquitin-mediated degradation of the CDK inhibitor p27. Nat Cell Biol 1:193–199

    Article  PubMed  CAS  Google Scholar 

  • Casini T, Pelicci PG (1999) A function of p21 during promyelocytic leukemia cell differentiation independent of CDK inhibition and cell cycle arrest. Oncogene 18:3235–3243

    Article  PubMed  CAS  Google Scholar 

  • Cosgaya JM, Chan JR, Shooter EM (2002) The neurotrophin receptor p75NTR as a positive modulator of myelination. Science 298:1245–1248

    Article  PubMed  CAS  Google Scholar 

  • Durand B, Gao FB, Raff M (1997) Accumulation of the cyclin-dependent kinase inhibitor p27Kip1 and the timing of oligodendrocyte differentiation. EMBO J 16:306–317

    Article  PubMed  CAS  Google Scholar 

  • Elledge SJ, Harper JW (1994) Cdk inhibitors: on the threshold of checkpoints and development. Curr Opin Cell Biol 6:847–852

    Article  PubMed  CAS  Google Scholar 

  • Fero ML, Rivkin M, Tasch M, Porter P, Carow CE, Firpo E, Polyak K, Tsai LH, Broudy V, Perlmutter RM, Kaushansky K, Roberts JM (1996) A syndrome of multiorgan hyperplasia with features of gigantism, tumorigenesis, and female sterility in p27Kip1-deficient mice. Cell 85:733–744

    Article  PubMed  CAS  Google Scholar 

  • Ganoth D, Bornstein G, Ko TK, Larsen B, Tyers M, Pagano M, Hershko A (2001) Role of the SCFSkp2 ubiquitin ligase in the degradation of p21Cip1 in S phase. Nat Cell Biol 3:321–324

    Article  PubMed  CAS  Google Scholar 

  • Hara T, Kamura T, Kotoshiba S, Takahashi H, Fujiwara K, Onoyama I, Shirakawa M, Mizushima N, Nakayama KI (2005) Role of the UBL-UBA protein KPC2 in degradation of p27 at G1 phase of the cell cycle. Mol Cell Biol 21:9292–9303

    Article  Google Scholar 

  • Ishida N, Hara T, Kamura T, Yoshida M, Nadayama K, Nakayama KI (2002) Phosphorylation of p27Kip1 on serine 10 is required for its binding to CRM1 and nuclear export. J Biol Chem 277:14355–14358

    Article  PubMed  CAS  Google Scholar 

  • Jaegle M, Mandemaders W, Broos L, Zwart R, Karis A, Visser P, Grosveld F, Meijer D (1996) The POU factor Oct-6 and Schwann cell differentiation. Science 273:507–510

    Article  PubMed  CAS  Google Scholar 

  • Jessen KR, Mirsky R (2002) Signals that determine Schwann cell identity. J Anat 200:367–376

    Article  PubMed  CAS  Google Scholar 

  • Kamura T, Hara T, Matsumoto M, Ishida N, Okumura F, Hatakeyama S, Yoshida M, Nakayama K, Nakayama KI (2004) Cytoplasmic ubiquitin ligase KPC regulates proteolysis of p27(Kip1)at G1 phase. Nat Cell Biol 6:1229–35

    Article  PubMed  CAS  Google Scholar 

  • Kiyokawa H, Kineman RD, Manova-Todorova KO, Soares VC, Hoffman ES, Ono M, Khanam D, Hayday AC, Frohman LA, Koff A (1996) Enhanced growth of mice lacking the cyclin-dependent kinase inhibitor function of p27Kip1. Cell 85:721–732

    Article  PubMed  CAS  Google Scholar 

  • Kotoshiba S, Kamura T, Hara T, Ishida N, Nakayama KI (2005) Molecular dissection of the interaction between p27 and KPC, the ubiquitin ligase that regulates proteolysis of p27 in G1 phase. J Biol Chem 280:17694–17700

    Article  PubMed  CAS  Google Scholar 

  • Lee EY, Chang CY, Hu N, Wang YC, Lai CC, Herrup K, Lee WH, Bradley A (1992) Mice deficient for Rb are nonviable and show defects in neurogenesis and haematopoiesis [see comments]. Nature 359:288–294

    Article  PubMed  CAS  Google Scholar 

  • Mathon NF, Malcolm DS, Harrisingh MC, Cheng L, Lloyd AC (2001) Lack of replicative senescence in normal rodent glia. Science 291(5505):872–875

    Article  PubMed  CAS  Google Scholar 

  • Mi S, Miller RH, Lee X, Scott ML, Shulag-Morskaya S, Shao Z, Chang J, Thill G, Levesque M, Zhang M, Zhang M, Hession C, Sah D, Trapp B, He Z, Jung V, McCoy JM, Pepinsky RB (2005) LINGO-1 negatively regulates myelination by oligodendrocytes. Nat Neurosci 8:745–751

    Article  PubMed  CAS  Google Scholar 

  • Mirsky R, Parkinson DB, Dong Z, Meier C, Calle E, Brennan A, Topilko P, Harris BS, Stewart HJ, Jessen KR (2001) Regulation of genes involved in Schwann cell development and differentiation. Prog Brain Res 132:3–11

    Article  PubMed  CAS  Google Scholar 

  • Montagnoli A, Fiore F, Eytan E, Carrano AC, Draetta GF, Hershko A, Pagano M (1999) Ubiquitination of p27 is regulated by Cdk-dependent phosphorylation and trimeric complex formation. Genes Dev 13:1181–1189

    Article  PubMed  CAS  Google Scholar 

  • Nakayama K, Ishida N, Shirane M, Inomata A, Inoue T, Shishido N, Horii I, Loh DY, Nakayama KI (1996) Mice lacking p27kip1 display increased body size, multiple organ hyperplasia, retinal dysplasia, and pituitary tumors. Cell 85:707–720

    Article  PubMed  CAS  Google Scholar 

  • Nguyen L, Besson A, Heng JI, Schuurmans C, Teboul L, Philpott A, Roberts JM, Guillemot F (2006) p27kip1 independently promotes neuronal differentiation and migration in the cerebral cortex. Genes Dev 20:1511–1524

    Article  PubMed  CAS  Google Scholar 

  • Ohnuma S, Philpott A, Wang K, Holt CE, Harris WA (1999) p27**c1, a Cdk inhibitor, promotes the determination of glial cells in Xenopus retina. Cell 99:499–510

    Article  PubMed  CAS  Google Scholar 

  • Pagano M, Tam SW, Theodoras AM, Beer-Romero P, Del Sal G, Chau V, Yew PR, Draetta GF, Rolfe M (1995) Role of the ubiquitin–proteasome pathway in regulating abundance of the cyclin-dependent kinase. Science 269:682–685

    Article  PubMed  CAS  Google Scholar 

  • Raff MC, Hornby-Smith A, Brockes JP (1978) Cyclic AMP as a mitogenic signal for cultured rat Schwann cells. Nature 273:672–673

    Article  PubMed  CAS  Google Scholar 

  • Son YJ, Thompson WJ (1995) Schwann cell processes guide regeneration of peripheral axons. Neuron 14:125–132

    Article  PubMed  CAS  Google Scholar 

  • Spruck C, Strohmaier H, Watson M, Smith AP, Ryan A, Krek TW, Reed SI (2001) A CDK-independent function of mammalian Cks1: targeting of SCF(Skp2) to the CDK inhibitor p27Kip1. Mol Cell 7:639–650

    Article  PubMed  CAS  Google Scholar 

  • Sutterluty H, Chatelain E, Marti A, Wirbelauer C, Senften M, Muller U, Krek W (1999) p45SKP2 promotes p27Kip1 degradation and induces S phase in quiescent cells. Nat Cell Biol 1:207–214

    Article  PubMed  CAS  Google Scholar 

  • Topilko P, Schneider-Maunoury S, Leve G, Baron-Van EA, Chennoufi AB, Seitanidou T, Babinet C, Charnay P (1994) Krox-20 controls myelination in the peripheral nervous system. Nature 371:796–799

    Article  PubMed  CAS  Google Scholar 

  • Warner LE, Garcia CA, Lupski JR (1999) Hereditary peripheral neuropathies: clinical forms, genetics and molecular mechanisms. Annu Rev Med 50:263–275

    Article  PubMed  CAS  Google Scholar 

  • Yoon C, Korade Z, Carter BD (2008) Protein kinase A-induced phosphorylation of the p65 subunit of nuclear factor-kappaB promotes Schwann cell differentiation into a myelinating phenotype. J Neurosci 28(14):3738–3746

    Article  PubMed  CAS  Google Scholar 

  • Zezula J, Casaccia-Bonnefil P, Ezhevsky SA, Osterhout DJ, Levine JM, Dowdy SF, Chao MV, Koff A (2001) p21cip1 is required for the differentiation of oligodendrocytes independently of cell cycle withdrawal. EMBO Rep 2:27–34

    Article  PubMed  CAS  Google Scholar 

  • Zhang P, Wong C, Liu D, Finegold M, Harper JW, Elledge SJ (1999) p21(CIP1) and p57(KIP2) control muscle differentiation at the myogenin step. Genes Dev 13:213–224

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (nos. 30300099, 30770488, and 30870320), the Natural Science Foundation of Jiangsu province (nos. BK2009161, BK2009156, and BK2009157), the Natural Science Foundation of Jiangsu Colleges and Universities Grant (09KJD310005), and The Society and Technology Grew Project of Nantong City (S2008020).

Author information

Authors and Affiliations

  1. Department of Orthopaedics, Affiliated Hospital of Nantong University, Nantong, 226001, People’s Republic of China

    Honghui Li, Huiguang Yang, Yonghua Liu, Weipeng Huan, Shuangwei Zhang, Gang Wu, Qiuhui Lu, Qiuhong Wang & Youhua Wang

  2. Department of Orthopaedics, Medical College of Nantong University, Nantong, 226001, People’s Republic of China

    Honghui Li, Huiguang Yang, Yonghua Liu, Weipeng Huan, Shuangwei Zhang, Gang Wu, Qiuhui Lu, Qiuhong Wang & Youhua Wang

Authors
  1. Honghui Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Huiguang Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yonghua Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Weipeng Huan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shuangwei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gang Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Qiuhui Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Qiuhong Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Youhua Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Youhua Wang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, H., Yang, H., Liu, Y. et al. The Cyclin-Dependent Kinase Inhibitor p27Kip1 is a Positive Regulator of Schwann Cell Differentiation In Vitro. J Mol Neurosci 45, 277–283 (2011). https://doi.org/10.1007/s12031-011-9518-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12031-011-9518-2

Keywords

Search

Navigation