Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
???displayArticle.abstract???
An early step in establishing left-right (LR) symmetry in zebrafish is the generation of asymmetric fluid flow by Kupffer's vesicle (KV). As a result of fluid flow, a signal is generated and propagated from the KV to the leftlateral plate mesoderm, activating a transcriptional response of Nodal expression in the leftlateral plate mesoderm (LPM). The mechanisms and molecules that aid in this transfer of information from the KV to the leftLPM are still not clear. Here we provide several lines of evidence demonstrating a role for a member of the TGFβ family member, Dvr1, a zebrafish Vg1 ortholog. Dvr1 is expressed bilaterally between the KV and the LPM. Knockdown of Dvr1 by morpholino causes dramatically reduced or absent expression of southpaw (spaw, a Nodal homolog), in LPM, and corresponding loss of downstream Lefty (lft1 and lft) expression, and aberrant brain and heart LR patterning. Dvr1 morphant embryos have normal KV morphology and function, normal expression of southpaw (spaw) and charon (cha) in the peri-KV region and normal expression of a variety of LPM markers in LPM. Additionally, Dvr1 knockdown does not alter the capability of LPM to respond to signals that initiate and propagate spaw expression. Co-injection experiments in Xenopus and zebrafish indicate that Dvr1 and Spaw can enhance each other's ability to activate the Nodal response pathway and co-immunoprecipitation experiments reveal differential relationships among activators and inhibitors in this pathway. These results indicate that Dvr1 is responsible for enabling the transfer of a left-right signal from KV to the LPM.
Fig. 5. Dvr1 and Spaw mutually enhance the activation of the Nodal response pathway in Xenopus. (A) Method of injection of mRNAs for Xenopus animal cap assay. D is dorsal and V is ventral. Embryos were evaluated for ectopic xbra expression at stage 10.5. The black arrow on the diagram indicates ectopic xbra expression and the red arrowhead indicates endogenous xbra expression. (B)E) Evaluation of ectopic xbra expression in embryos injected with (B) control GFP mRNA, (C) dvr1 mRNA, (D) spaw mRNA and (E) dvr1 and spaw mRNA. (F) Quantification of the area of ectopic xbra expression in dvr1 (n=53; 17/53 had no ectopic expression), spaw (n=51; 19/51 had no ectopic expression) and dvr1+spaw (n=76) mRNA injected embryos. Results are averages from 4 different batches of embryos, and include embryos that exhibited no ectopic xbra expressionfor. Error bars: s.e.m.
Fig. 6. Spaw and Dvr1 do not Co-Immunoprecipitate. (A) Scheme of protein co-precipitation assay. Xenopus embryos were injected with mRNAs at 2 cell stage and collected at stage 10. Cleared lysates were loaded directly onto gels (nput or in parallel were used for immunoprecipitation by anti-myc or anti-Flag epitopes, which were then loaded on gels and processed for Western Blot analysis. (B) Western result with anti-HA antibody shows that there is a strong interaction between Spaw and Charon, but not Spaw and Dvr1. Anti-FLAG IP was used as a control for IP.
Amack,
Two T-box genes play independent and cooperative roles to regulate morphogenesis of ciliated Kupffer's vesicle in zebrafish.
2007, Pubmed,
Xenbase
Amack,
Two T-box genes play independent and cooperative roles to regulate morphogenesis of ciliated Kupffer's vesicle in zebrafish.
2007,
Pubmed
,
Xenbase
Bisgrove,
Multiple pathways in the midline regulate concordant brain, heart and gut left-right asymmetry.
2000,
Pubmed
Bisgrove,
Regulation of midline development by antagonism of lefty and nodal signaling.
1999,
Pubmed
Branford,
Lefty-dependent inhibition of Nodal- and Wnt-responsive organizer gene expression is essential for normal gastrulation.
2002,
Pubmed
,
Xenbase
Dohrmann,
Induction of axial mesoderm by zDVR-1, the zebrafish orthologue of Xenopus Vg1.
1996,
Pubmed
,
Xenbase
Essner,
Kupffer's vesicle is a ciliated organ of asymmetry in the zebrafish embryo that initiates left-right development of the brain, heart and gut.
2005,
Pubmed
Essner,
Conserved function for embryonic nodal cilia.
2002,
Pubmed
,
Xenbase
Hamada,
Establishment of vertebrate left-right asymmetry.
2002,
Pubmed
Hashimoto,
The Cerberus/Dan-family protein Charon is a negative regulator of Nodal signaling during left-right patterning in zebrafish.
2004,
Pubmed
,
Xenbase
Helde,
The DVR-1 (Vg1) transcript of zebrafish is maternally supplied and distributed throughout the embryo.
1993,
Pubmed
,
Xenbase
Hyatt,
The left-right coordinator: the role of Vg1 in organizing left-right axis formation.
1998,
Pubmed
,
Xenbase
Hyatt,
Initiation of vertebrate left-right axis formation by maternal Vg1.
1996,
Pubmed
,
Xenbase
Kramer,
PKCgamma regulates syndecan-2 inside-out signaling during xenopus left-right development.
2002,
Pubmed
,
Xenbase
Levin,
Left-right asymmetry in embryonic development: a comprehensive review.
2005,
Pubmed
Link,
Proteomics of early zebrafish embryos.
2006,
Pubmed
Long,
The zebrafish nodal-related gene southpaw is required for visceral and diencephalic left-right asymmetry.
2003,
Pubmed
Marjoram,
Rapid differential transport of Nodal and Lefty on sulfated proteoglycan-rich extracellular matrix regulates left-right asymmetry in Xenopus.
2011,
Pubmed
,
Xenbase
Nakamura,
Fluid flow and interlinked feedback loops establish left-right asymmetric decay of Cerl2 mRNA.
2012,
Pubmed
Neugebauer,
FGF signalling during embryo development regulates cilia length in diverse epithelia.
2009,
Pubmed
,
Xenbase
Nonaka,
Randomization of left-right asymmetry due to loss of nodal cilia generating leftward flow of extraembryonic fluid in mice lacking KIF3B motor protein.
1998,
Pubmed
Okada,
Mechanism of nodal flow: a conserved symmetry breaking event in left-right axis determination.
2005,
Pubmed
Ramsdell,
Molecular mechanisms of vertebrate left-right development.
1998,
Pubmed
,
Xenbase
Rankin,
Regulation of left-right patterning in mice by growth/differentiation factor-1.
2000,
Pubmed
Robu,
p53 activation by knockdown technologies.
2007,
Pubmed
Schier,
The one-eyed pinhead gene functions in mesoderm and endoderm formation in zebrafish and interacts with no tail.
1997,
Pubmed
Schneider,
Zebrafish Nkd1 promotes Dvl degradation and is required for left-right patterning.
2010,
Pubmed
Schulte-Merker,
Expression of zebrafish goosecoid and no tail gene products in wild-type and mutant no tail embryos.
1994,
Pubmed
,
Xenbase
Schweickert,
Cilia-driven leftward flow determines laterality in Xenopus.
2007,
Pubmed
,
Xenbase
Schweickert,
The nodal inhibitor Coco is a critical target of leftward flow in Xenopus.
2010,
Pubmed
,
Xenbase
Srivastava,
A subclass of bHLH proteins required for cardiac morphogenesis.
1995,
Pubmed
Tanaka,
Long-range action of Nodal requires interaction with GDF1.
2007,
Pubmed
,
Xenbase
Tannahill,
Localized synthesis of the Vg1 protein during early Xenopus development.
1989,
Pubmed
,
Xenbase
Thomsen,
Processed Vg1 protein is an axial mesoderm inducer in Xenopus.
1993,
Pubmed
,
Xenbase
Vonica,
The left-right axis is regulated by the interplay of Coco, Xnr1 and derrière in Xenopus embryos.
2007,
Pubmed
,
Xenbase
Wall,
Mesendoderm induction and reversal of left-right pattern by mouse Gdf1, a Vg1-related gene.
2000,
Pubmed
,
Xenbase
Wang,
Initiation and propagation of posterior to anterior (PA) waves in zebrafish left-right development.
2008,
Pubmed
Weinberg,
Developmental regulation of zebrafish MyoD in wild-type, no tail and spadetail embryos.
1996,
Pubmed
Yamamoto,
Nodal signaling induces the midline barrier by activating Nodal expression in the lateral plate.
2003,
Pubmed
Ye,
Mutation of the bone morphogenetic protein GDF3 causes ocular and skeletal anomalies.
2010,
Pubmed
