Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Subscribe

Logo for

Development 133 (9): 1657-1671

Early, H+-V-ATPase-dependent proton flux is necessary for consistent left-right patterning of non-mammalian vertebrates

Dany S. Adams1, Kenneth R. Robinson2, Takahiro Fukumoto3,*, Shipeng Yuan4, R. Craig Albertson3, Pamela Yelick3, Lindsay Kuo3, Megan McSweeney3, and Michael Levin1,{dagger}

1 The Forsyth Center for Regenerative and Developmental Biology, and Department of Developmental Biology, Harvard School of Dental Medicine, 140 The Fenway, Boston, MA 02115, USA.
2 Department of Biological Sciences, Purdue University, West Lafayette, IN 47906, USA.
3 Department of Cytokine Biology, The Forsyth Institute, 140 The Fenway, Boston, MA 02115, USA.
4 Cardiovascular Research Center, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, USA.

{dagger} Author for correspondence (e-mail: mlevin{at}forsyth.org)

Accepted for publication 27 February 2006.

Abstract: Biased left-right asymmetry is a fascinating and medically important phenomenon. We provide molecular genetic and physiological characterization of a novel, conserved, early, biophysical event that is crucial for correct asymmetry: H+ flux. A pharmacological screen implicated the H+-pump H+-V-ATPase in Xenopus asymmetry, where it acts upstream of early asymmetric markers. Immunohistochemistry revealed an actin-dependent asymmetry of H+-V-ATPase subunits during the first three cleavages. H+-flux across plasma membranes is also asymmetric at the four- and eight-cell stages, and this asymmetry requires H+-V-ATPase activity. Abolishing the asymmetry in H+ flux, using a dominant-negative subunit of the H+-V-ATPase or an ectopic H+ pump, randomized embryonic situs without causing any other defects. To understand the mechanism of action of H+-V-ATPase, we isolated its two physiological functions, cytoplasmic pH and membrane voltage (Vmem) regulation. Varying either pH or Vmem, independently of direct manipulation of H+-V-ATPase, caused disruptions of normal asymmetry, suggesting roles for both functions. V-ATPase inhibition also abolished the normal early localization of serotonin, functionally linking these two early asymmetry pathways. The involvement of H+-V-ATPase in asymmetry is conserved to chick and zebrafish. Inhibition of the H+-V-ATPase induces heterotaxia in both species; in chick, H+-V-ATPase activity is upstream of Shh; in fish, it is upstream of Kupffer's vesicle and Spaw expression. Our data implicate H+-V-ATPase activity in patterning the LR axis of vertebrates and reveal mechanisms upstream and downstream of its activity. We propose a pH- and Vmem-dependent model of the early physiology of LR patterning.

Key Words: Left-right asymmetryH+-V-ATPaseV-ATPaseXenopusChickZebrafishAxial patterningCytoplasmic pHMembrane voltage


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Atp6ap2/(Pro)renin Receptor Interacts with Par3 as a Cell Polarity Determinant Required for Laminar Formation during Retinal Development in Mice.
A. Kanda, K. Noda, K. Yuki, Y. Ozawa, T. Furukawa, A. Ichihara, and S. Ishida (2013)
J. Neurosci. 33, 19341-19351
   Abstract »    Full Text »    PDF »
Differential roles for 3-OSTs in the regulation of cilia length and motility.
J. M. Neugebauer, A. B. Cadwallader, J. D. Amack, B. W. Bisgrove, and H. J. Yost (2013)
Development 140, 3892-3902
   Abstract »    Full Text »    PDF »
Transmembrane voltage potential is an essential cellular parameter for the detection and control of tumor development in a Xenopus model.
B. T. Chernet and M. Levin (2013)
Dis. Model. Mech. 6, 595-607
   Abstract »    Full Text »    PDF »
Serotonin has early, cilia-independent roles in Xenopus left-right patterning.
L. N. Vandenberg, J. M. Lemire, and M. Levin (2013)
Dis. Model. Mech. 6, 261-268
   Abstract »    Full Text »    PDF »
Left-right patterning: conserved and divergent mechanisms.
T. Nakamura and H. Hamada (2012)
Development 139, 3257-3262
   Abstract »    Full Text »    PDF »
Identification of Domains within the V-ATPase Accessory Subunit Ac45 Involved in V-ATPase Transport and Ca2+-dependent Exocytosis.
E. J. R. Jansen, N. H. M. van Bakel, N. F. M. O. Loohuis, T. G. M. Hafmans, T. Arentsen, A. J. M. Coenen, W. J. J. M. Scheenen, and G. J. M. Martens (2012)
J. Biol. Chem. 287, 27537-27546
   Abstract »    Full Text »    PDF »
Early, nonciliary role for microtubule proteins in left-right patterning is conserved across kingdoms.
M. Lobikin, G. Wang, J. Xu, Y.-W. Hsieh, C.-F. Chuang, J. M. Lemire, and M. Levin (2012)
PNAS 109, 12586-12591
   Abstract »    Full Text »    PDF »
The reconstructed ancestral subunit a functions as both V-ATPase isoforms Vph1p and Stv1p in Saccharomyces cerevisiae.
G. C. Finnigan, V. Hanson-Smith, B. D. Houser, H. J. Park, and T. H. Stevens (2011)
Mol. Biol. Cell 22, 3176-3191
   Abstract »    Full Text »    PDF »
Micropatterned mammalian cells exhibit phenotype-specific left-right asymmetry.
L. Q. Wan, K. Ronaldson, M. Park, G. Taylor, Y. Zhang, J. M. Gimble, and G. Vunjak-Novakovic (2011)
PNAS 108, 12295-12300
   Abstract »    Full Text »    PDF »
Transmembrane potential of GlyCl-expressing instructor cells induces a neoplastic-like conversion of melanocytes via a serotonergic pathway.
D. Blackiston, D. S. Adams, J. M. Lemire, M. Lobikin, and M. Levin (2011)
Dis. Model. Mech. 4, 67-85
   Abstract »    Full Text »    PDF »
V-ATPase-Mediated Granular Acidification Is Regulated by the V-ATPase Accessory Subunit Ac45 in POMC-Producing Cells.
E. J. R. Jansen, T. G. M. Hafmans, and G. J. M. Martens (2010)
Mol. Biol. Cell 21, 3330-3339
   Abstract »    Full Text »    PDF »
Consistent left-right asymmetry cannot be established by late organizers in Xenopus unless the late organizer is a conjoined twin.
L. N. Vandenberg and M. Levin (2010)
Development 137, 1095-1105
   Abstract »    Full Text »    PDF »
Electrical dimensions in cell science.
C. D. McCaig, B. Song, and A. M. Rajnicek (2009)
J. Cell Sci. 122, 4267-4276
   Abstract »    Full Text »    PDF »
Differential expression profiling of ovarian genes in prelaying and laying geese.
B. Kang, J. R. Guo, H. M. Yang, R. J. Zhou, J. X. Liu, S. Z. Li, and C. Y. Dong (2009)
Poultry Science 88, 1975-1983
   Abstract »    Full Text »    PDF »
The Vacuolar-ATPase Complex Regulates Retinoblast Proliferation and Survival, Photoreceptor Morphogenesis, and Pigmentation in the Zebrafish Eye.
R. J. Nuckels, A. Ng, T. Darland, and J. M. Gross (2009)
Invest. Ophthalmol. Vis. Sci. 50, 893-905
   Abstract »    Full Text »    PDF »
Is left-right asymmetry a form of planar cell polarity?.
S. Aw and M. Levin (2009)
Development 136, 355-366
   Abstract »    Full Text »    PDF »
Modulation of potassium channel function confers a hyperproliferative invasive phenotype on embryonic stem cells.
J. Morokuma, D. Blackiston, D. S. Adams, G. Seebohm, B. Trimmer, and M. Levin (2008)
PNAS 105, 16608-16613
   Abstract »    Full Text »    PDF »
A ryanodine receptor-dependent Cai2+ asymmetry at Hensen's node mediates avian lateral identity.
A. Garic-Stankovic, M. Hernandez, G. R. Flentke, M. H. Zile, and S. M. Smith (2008)
Development 135, 3271-3280
   Abstract »    Full Text »    PDF »
Tail Regeneration in Xenopus laevis as a Model for Understanding Tissue Repair.
A.-S. Tseng and M. Levin (2008)
Journal of Dental Research 87, 806-816
   Abstract »    Full Text »    PDF »
Calcium dynamics integrated into signalling pathways that influence vertebrate axial patterning.
C. M Freisinger, I. Schneider, T. A Westfall, and D. C Slusarski (2008)
Phil Trans R Soc B 363, 1377-1385
   Abstract »    Full Text »    PDF »
Biochemical and functional characterization of the actin-binding activity of the B subunit of yeast vacuolar H+-ATPase.
J. Zuo, S. Vergara, S. Kohno, and L. S. Holliday (2008)
J. Exp. Biol. 211, 1102-1108
   Abstract »    Full Text »    PDF »
Exclusion of a Proton ATPase from the Apical Membrane Is Associated with Cell Polarity and Tip Growth in Nicotiana tabacum Pollen Tubes.
A. C. Certal, R. B. Almeida, L. M. Carvalho, E. Wong, N. Moreno, E. Michard, J. Carneiro, J. Rodriguez-Leon, H.-M. Wu, A. Y. Cheung, et al. (2008)
PLANT CELL 20, 614-634
   Abstract »    Full Text »    PDF »
Calcium fluxes in dorsal forerunner cells antagonize -catenin and alter left-right patterning.
I. Schneider, D. W. Houston, M. R. Rebagliati, and D. C. Slusarski (2008)
Development 135, 75-84
   Abstract »    Full Text »    PDF »
An Arabidopsis quiescin-sulfhydryl oxidase regulates cation homeostasis at the root symplast-xylem interface.
S. Alejandro, P. L. Rodriguez, J. M. Belles, L. Yenush, M. J. Garcia-Sanchez, J. A. Fernandez, and R. Serrano (2007)
EMBO J. 26, 3203-3215
   Abstract »    Full Text »    PDF »
Na,K-ATPase {alpha}2 and Ncx4a regulate zebrafish left-right patterning.
X. Shu, J. Huang, Y. Dong, J. Choi, A. Langenbacher, and J.-N. Chen (2007)
Development 134, 1921-1930
   Abstract »    Full Text »    PDF »
H+ pump-dependent changes in membrane voltage are an early mechanism necessary and sufficient to induce Xenopus tail regeneration.
D. S. Adams, A. Masi, and M. Levin (2007)
Development 134, 1323-1335
   Abstract »    Full Text »    PDF »
Intrinsic chiral properties of the Xenopus egg cortex: an early indicator of left-right asymmetry?.
M. V. Danilchik, E. E. Brown, and K. Riegert (2006)
Development 133, 4517-4526
   Abstract »    Full Text »    PDF »

To Advertise     Find Products


Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882