Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Mutations in Rab3a alter circadian period and homeostatic response to sleep loss in the mouse

Nature Genetics volume 32, pages 290–295 (2002)Cite this article

Abstract

Rab3a is the most abundant Rab (ras-associated binding) protein in the brain and has a regulatory role in synaptic vesicle trafficking1. Mice with a targeted loss-of-function mutation in Rab3a have defects in Ca2+-dependent synaptic transmission: the number of vesicles released in response to an action potential is greater than in wildtype mice, resulting in greater synaptic depression2,3 and the abolishment of CA3 mossy-fiber long term potentiation4. The effect of these changes on behavior is unknown. In a screen for mouse mutants with abnormal rest–activity and sleep patterns, we identified a semidominant mutation, called earlybird, that shortens the circadian period of locomotor activity. Sequence analysis of Rab3a identified a point mutation in the conserved amino acid (Asp77Gly) within the GTP-binding domain of this protein in earlybird mutants, resulting in significantly reduced levels of Rab3a protein. Phenotypic assessment of earlybird mice and a null allele of Rab3a revealed anomalies in circadian period and sleep homeostasis, providing evidence that Rab3a-mediated synaptic transmission is involved in these behaviors.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Identification and genetic localization of the earlybird (Ebd) mutation.
Figure 2: Sequence analysis and identification of a mutation in Rab3a in Ebd/Ebd mice.
Figure 3: Evidence for allelism between earlybird and Rab3a.
Figure 4: Analysis of rest parameters in earlybird and Rab3a−/− mice at different zeitgeber times (ZT).
Figure 5: The effect of Rab3a on the circadian system.

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

Protein Data Bank

References

  1. Sudhof, T.C. The synaptic vesicle cycle: a cascade of protein–protein interactions. Nature 375, 645–653 (1995).

    Article  CAS  Google Scholar 

  2. Geppert, M. et al. The role of Rab3A in neurotransmitter release. Nature 369, 493–497 (1994).

    Article  CAS  Google Scholar 

  3. Geppert, M., Goda, Y., Stevens, C.F. & Sudhof, T.C. The small GTP-binding protein Rab3A regulates a late step in synaptic vesicle fusion. Nature 387, 810–814 (1997).

    Article  CAS  Google Scholar 

  4. Castillo, P.E. et al. Rab3A is essential for mossy fibre long-term potentiation in the hippocampus. Nature 388, 590–593 (1997).

    Article  CAS  Google Scholar 

  5. King, D.P. & Takahashi, J.S. Molecular genetics of circadian rhythms in mammals. Annu. Rev. Neurosci. 23, 713–742 (2000).

    Article  CAS  Google Scholar 

  6. Borbely, A.A. A two process model of sleep regulation. Hum. Neurobiol. 1, 195–204 (1982).

    CAS  PubMed  Google Scholar 

  7. Klein, D.C., Moore, R.Y. & Reppert, S.M. Suprachiasmatic Nucleus: The Mind's Clock (Oxford University Press, 1991).

    Google Scholar 

  8. van der Horst, G.T. et al. Mammalian Cry1 and Cry2 are essential for maintenance of circadian rhythms. Nature 398, 627–630 (1999).

    Article  CAS  Google Scholar 

  9. Zheng, B. et al. The mPer2 gene encodes a functional component of the mammalian circadian clock. Nature 400, 169–173 (1999).

    Article  CAS  Google Scholar 

  10. Bunger, M.K. et al. Mop3 is an essential component of the master circadian pacemaker in mammals. Cell 103, 1009–1017 (2000).

    Article  CAS  Google Scholar 

  11. Vitaterna, M.H. et al. Mutagenesis and mapping of a mouse gene, Clock, essential for circadian behavior. Science 264, 719–725 (1994).

    Article  CAS  Google Scholar 

  12. King, D.P. et al. Positional cloning of the mouse circadian clock gene. Cell 89, 641–653 (1997).

    Article  CAS  Google Scholar 

  13. Nolan, P.M., Kapfhamer, D. & Bucan, M. Random mutagenesis screen for dominant behavioral mutations in mice. Methods 13, 379–395 (1997).

    Article  CAS  Google Scholar 

  14. Dumas, J.J., Zhu, Z., Connolly, J.L. & Lambright, D.G. Structural basis of activation and GTP hydrolysis in Rab proteins. Structure Fold Des. 7, 413–423 (1999).

    Article  CAS  Google Scholar 

  15. Ostermeier, C. & Brunger, A.T. Structural basis of Rab effector specificity: crystal structure of the small G protein Rab3A complexed with the effector domain of rabphilin-3A. Cell 96, 363–374 (1999).

    Article  CAS  Google Scholar 

  16. Bourne, H.R., Sanders, D.A. & McCormick, F. The GTPase superfamily: conserved structure and molecular mechanism. Nature 349, 117–127 (1991).

    Article  CAS  Google Scholar 

  17. John, J. et al. Kinetic and structural analysis of the Mg2+-binding site of the guanine nucleotide-binding protein p21H-ras. J. Biol. Chem. 268, 923–929 (1993).

    CAS  PubMed  Google Scholar 

  18. Jung, V. et al. Two types of RAS mutants that dominantly interfere with activators of RAS. Mol. Cell Biol. 14, 3707–3718 (1994).

    Article  CAS  Google Scholar 

  19. Panda, S. et al. Coordinated transcription of key pathways in the mouse by the circadian clock. Cell 109, 307–320 (2002).

    Article  CAS  Google Scholar 

  20. Pennartz, C., de Jeu, M., Bos, N., Schaap, J. & Geurtsen, A. Diurnal modulation of pacemaker potentials and calcium current in the mammalian circadian clock. Nature 416, 286–290 (2002).

    Article  CAS  Google Scholar 

  21. Tobler, I. et al. Altered circadian activity rhythms and sleep in mice devoid of prion protein. Nature 380, 639–642 (1996).

    Article  CAS  Google Scholar 

  22. Naylor, E. et al. The circadian clock mutation alters sleep homeostasis in the mouse. J. Neurosci. 20, 8138–8143 (2000).

    Article  CAS  Google Scholar 

  23. Franken, P., Lopez-Molina, L., Marcacci, L., Schibler, U. & Tafti, M. The transcription factor DBP affects circadian sleep consolidation and rhythmic EEG activity. J. Neurosci. 20, 617–625 (2000).

    Article  CAS  Google Scholar 

  24. D'Adamo, P. et al. Mutations in GDI1 are responsible for X-linked non-specific mental retardation. Nature Genet. 19, 134–139 (1998).

    Article  CAS  Google Scholar 

  25. Nolan, P.M., Houpt, T.A. & Bucan, M. Chemical mutagenesis and screening for mouse mutations with an altered rest–activity pattern. in Molecular Regulation of Arousal States (ed. Lydic, R.) 143–156 (CRC, 1998).

    Google Scholar 

  26. Veasey, S.C. et al. An automated system for recording and analysis of sleep in mice. Sleep 23, 1025–1040 (2000).

    CAS  PubMed  Google Scholar 

  27. Bennington, J.H., Kodali, S.K. & Heller, H.C. Scoring transitions to REM-sleep in rats based on the EEG phenomena of pre-REM-sleep: an improved analysis of sleep structure. Sleep 17, 28–36 (1994).

    Article  Google Scholar 

  28. Thompson, J.D., Higgins, D.G. & Gibson, T.J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22, 4673–4680 (1994).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank G. Pickard for his contribution in the early stages of this project; A. Dixon-White for technical assistance; R. Buono, A. deBrunier, J. Eberwine and K. Miyashiro for assistance with immunohistochemistry, western analysis and in situ analysis; L. Maltais for assistance with gene nomenclature; and S. Poethig, M. Chou, T. Abel and G. Leach for comments on the manuscript. This work was supported by grants from the US National Institutes of Health.

Author information

Authors and Affiliations

  1. Center for Neurobiology of Behavior of the Department of Psychiatry, University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA

    David Kapfhamer, Otto Valladares, Yi Sun, Patrick M. Nolan, Steven E. Arnold & Maja Bućan

  2. The Wistar Institute, 3601 Spruce Street, Philadelphia, Pennsylvania, USA

    John J. Rux

  3. Center for Sleep and Respiratory Neurobiology, School of Medicine, University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA

    Sigrid C. Veasey & Maja Bućan

  4. Department of Medicine, School of Medicine, University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA

    Sigrid C. Veasey

  5. Department of Genetics, School of Medicine, University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA

    Maja Bućan

Authors
  1. David Kapfhamer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Otto Valladares
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yi Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Patrick M. Nolan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. John J. Rux
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Steven E. Arnold
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Sigrid C. Veasey
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Maja Bućan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Maja Bućan.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kapfhamer, D., Valladares, O., Sun, Y. et al. Mutations in Rab3a alter circadian period and homeostatic response to sleep loss in the mouse. Nat Genet 32, 290–295 (2002). https://doi.org/10.1038/ng991

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ng991

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing