Alternative titles; symbols
HGNC Approved Gene Symbol: NODAL
Cytogenetic location: 10q22.1 Genomic coordinates (GRCh38): 10:70,431,936-70,447,951 (from NCBI)
| Location | Phenotype |
Phenotype MIM number |
Inheritance |
Phenotype mapping key |
|---|---|---|---|---|
| 10q22.1 | Heterotaxy, visceral, 5 | 270100 | Autosomal dominant | 3 |
NODAL is a member of the TGF-beta gene family and is expressed during mouse gastrulation (Zhou et al., 1993). Nodal has a left-sided expression pattern that is disrupted in mouse models of LR axis development (see Lowe et al. (1996)).
Gebbia et al. (1997) stated that they had characterized the human homolog of mouse Nodal to search for mutations in individuals with LR axis malformations.
Collignon et al. (1996) showed that some mice manifest LR axis malformations when doubly heterozygous for null mutations in Nodal and in the transcription factor Hnf3b (600288).
Olson and Srivastava (1996) reviewed the role of the morphogen Nodal in the control of the direction of cardiac looping and in the development of left to right asymmetry in chick and mouse.
Brennan et al. (2001) demonstrated that signals from the epiblast are responsible for the initiation of the proximal-distal polarity in the mouse embryo. Nodal acts to promote posterior cell fates in the epiblast and to maintain molecular pattern in the adjacent extra-embryonic ectoderm. Both of these functions are independent of SMAD2 (601366). Moreover, Nodal signals from the epiblast also pattern the visceral endoderm by activating the SMAD2-dependent pathway required for specification of anterior identity in overlying epiblast cells. Brennan et al. (2001) concluded that proximal-distal and subsequent anterior-posterior polarity of the pregastrulation embryo result from reciprocal cell-cell interactions between the epiblast and the 2 extraembryonic tissues.
By analyzing mouse mutants lacking left-sided expression of Lefty2 (601877), Meno et al. (2001) observed that Nodal is able to diffuse over a large distance in the absence of Lefty2. They concluded that Nodal is a long-range signaling molecule but that its range of action is normally limited by the feedback inhibitor Lefty2.
Iratni et al. (2002) demonstrated that the transcriptional corepressor DRAP1 (602289) has a very specific role in regulation of Nodal activity during mouse embryogenesis. Iratni et al. (2002) found that loss of DRAP1 leads to severe gastrulation defects that are consistent with increased expression of Nodal and can be partially suppressed by Nodal heterozygosity. Biochemical studies indicated that DRAP1 interacts with and inhibits DNA binding by the winged-helix transcription factor FOXH1 (FAST1; 603621), a critical component of a positive feedback loop for Nodal activity. Iratni et al. (2002) proposed that DRAP1 limits the spread of a morphogenetic signal by downmodulating the response to the Nodal autoregulatory loop.
Mohapatra et al. (2009) noted that the NODAL gene contains 3 exons.
Gross (2022) mapped the NODAL gene to chromosome 10q22.1 based on an alignment of the NODAL sequence (GenBank BC104976) with the genomic sequence (GRCh38).
The mouse Nodal gene maps to chromosome 10 (Zhou et al., 1993).
In a family in which several members had situs ambiguus shown to be due to mutation in the ZIC3 gene (300265), Gebbia et al. (1997) found a normal male with a daughter with situs ambiguus; neither the father nor the daughter carried any mutation in the coding region of ZIC3 and both parents were anatomically normal. Paternity was confirmed, the mother was unrelated to the rest of the family, and the daughter had a 46,XX karyotype. Because observations in mice had suggested that heterozygous mutations in human NODAL may be associated with human situs abnormalities, Gebbia et al. (1997) searched for mutations in the NODAL gene. In the affected daughter and her unaffected mother, they found an arg183-to-gln substitution (R183Q; 601265.0001) in the prodomain of NODAL. None of more than 200 control chromosomes carried this substitution, and no other NODAL mutations were identified in other members of that family or in any other individual harboring a mutant ZIC3 allele.
In 14 of 269 patients with either classic heterotaxy or looping cardiovascular malformations (CVM), Mohapatra et al. (2009) identified 4 different missense variants (see, e.g., 601265.0002), 1 in-frame insertion/deletion (601265.0003), and 2 conserved splice site variants (see, e.g., 601265.0004) in the NODAL gene. Although similar with regard to other associated defects, individuals with the NODAL mutations had a significantly higher occurrence of pulmonary valve atresia (p = 0.001) compared with individuals without a detectable NODAL mutation. Functional analysis demonstrated that the missense variant forms of NODAL exhibited significant impairment of signaling as measured by decreased Cripto (TDGF1; 187395) coreceptor-mediated activation of artificial reporters. Expression of these NODAL proteins also led to reduced induction of SMAD2 phosphorylation and impaired SMAD2 nuclear import. Mohapatra et al. (2009) proposed a role for mutations and rare deleterious variants in NODAL as a cause for sporadic human left-right patterning defects.
Krebs et al. (2003) showed that mouse embryos mutant for the Notch ligand Dll1 (606582) or doubly mutant for Notch1 (190198) and Notch2 (600275) exhibited multiple defects in left-right asymmetry. Dll1 -/- embryos did not express Nodal in the region around the node. Analysis of the enhancer regulating node-specific Nodal expression revealed binding sites for Rbpj (RBPSUH; 147183). Mutation of these sites destroyed the ability of the enhancer to direct node-specific gene expression in transgenic mice. Krebs et al. (2003) concluded that Dll1-mediated Notch signaling is essential for generation of left-right asymmetry, and that perinodal expression of Nodal is an essential component of left-right asymmetry determination in mice.
Using gain- and loss-of-function experiments in zebrafish and mouse, Raya et al. (2003) showed that activity of the Notch pathway was necessary and sufficient for Nodal expression around the node and for proper left-right determination. They also identified critical Rbpj-binding sequences in the Nodal promoter.
Ware et al. (2006) found that mice compound heterozygous for a Zic3 and a Nodal mutation had significantly increased lethality compared to Zic3-null mice. No males and a reduced number of females survived. Accordingly, the laterality defects observed in compound heterozygous mice were more severe than in mice with either mutant alone. The findings indicated that the 2 genes interact in the same pathway. Further studies indicated that Zic3 activates an upstream enhancer of Nodal.
Yang et al. (2010) reported that holoprosencephaly (see HPE, 236100) in mice can result from simultaneous reduction in both Nodal (601265) signaling and expression levels of the Bmp antagonists chordin (CHRD; 603475) or Noggin (NOG; 602991). HPE defects are the result of reduced production of tissues that promote forebrain and craniofacial development. Nodal promotes the expression of genes in the anterior primitive streak important for the development of these tissues, whereas Bmp inhibits their expression. Pharmacologic and transgenic manipulation of these signaling pathways suggested that the Bmp and Nodal pathways antagonize each other prior to intracellular signal transduction. In vitro experiments indicated that secreted Bmp2 (112261) and Nodal can form extracellular complexes, potentially interfering with receptor activation. Yang et al. (2010) concluded that the patterning of forebrain and medial craniofacial elements requires a fine balance between BMP and NODAL signaling during primitive streak development.
Reaction-diffusion models postulated that differences in signaling range are caused by differential diffusivity of inhibitor and activator. Other models suggested that differential clearance underlies different signaling ranges. To test these models, Muller et al. (2012) measured the biophysical properties of the Nodal/Lefty (see 603037) activator/inhibitor system during zebrafish embryogenesis. Analysis of Nodal and Lefty gradients revealed that Nodals have a shorter range than Lefty proteins. Pulse-labeling analysis indicated that Nodals and Leftys have similar clearance kinetics, whereas fluorescence recovery assays revealed that Leftys have a higher effective diffusion coefficient than Nodals. Muller et al. (2012) concluded that their results indicated that differential diffusivity is the major determinant of the differences in Nodal/Lefty range and provided biophysical support for reaction-diffusion models of activator/inhibitor-mediated patterning.
Montague et al. (2018) found that zebrafish homozygous for single and double frameshift null mutations in spaw (the zebrafish ortholog of Nodal), dand5 (609068), and lefty1 (603037) were viable and lacked gross phenotypes. However, all mutant combinations displayed randomized or symmetric heart looping and jogging, similar to their respective mouse mutants. Further analysis revealed that spaw induced its expression in lateral plate mesoderm and was required for proper heart asymmetry in zebrafish, whereas dand5 and lefty1 were inhibitors of spaw and regulated the timing and speed of spaw propagation.
In a kindred in which several members had X-linked abnormalities of left-right body axis formation (306955) due to mutation in the ZIC3 gene (300265), Gebbia et al. (1997) found a normal male who had fathered a daughter with situs ambiguus (HTX5; 270100). Neither she nor the father carried any mutation in ZIC3, and both parents were anatomically normal. The daughter and her unaffected mother who was unrelated to the father were found to be heterozygous for an arg182-to-gln (R183Q) mutation in the prodomain of the NODAL gene.
In 8 of 82 unrelated Hispanic patients with heterotaxy (HTX5; 270100), Mohapatra et al. (2009) identified a heterozygous 778G-A transition in exon 2 of the NODAL gene, resulting in a gly260-to-arg (G260R) substitution in a highly conserved residue. Most patients had d-transposition of the great arteries in addition to other cardiac anomalies, 3 had abdominal situs inversus, and 2 had asplenia. The detection of this variant in the unaffected father of 1 patient and 1 of 108 Hispanic controls, suggested incomplete penetrance. None of the 190 Caucasian or African American controls examined carried the variant. Functional analysis showed reduced NODAL signaling through both FOXH1-dependent and -independent pathways, as well as reduced SMAD2 phosphorylation and impaired nuclear import.
In a male Hispanic patient with heterotaxy (HTX5; 270100), Mohapatra et al. (2009) identified an in-frame 9-bp insertion (700insTTGACTTCC) and 24-bp deletion (nucleotide 700-723) in exon 2 of the NODAL gene. The patient had d-transposition of the great arteries, pulmonary atresia, and double-inlet left ventricle. Parental DNA was not available for testing, and the mutation was not detected in 298 controls. Functional analysis showed reduced NODAL signaling through both FOXH1-dependent and -independent pathways, as well as reduced SMAD2 phosphorylation and impaired nuclear import.
In a male Hispanic patient with heterotaxy (HTX5; 270100), Mohapatra et al. (2009) identified a G-to-A transition (891+1G-A) in the donor splice site of intron 2 of the NODAL gene, predicted to result in altered splicing activity. The patient had dextrocardia, levo-transposition of the great arteries, double-outlet right ventricle, pulmonary valve stenosis, and a ventricular septal defect. The mutation was not detected in 298 controls.
Brennan, J., Lu, C. C., Norris, D. P., Rodriguez, T. A., Beddington, R. S. P., Robertson, E. J. Nodal signalling in the epiblast patterns the early mouse embryo. Nature 411: 965-969, 2001. [PubMed: 11418863] [Full Text: https://doi.org/10.1038/35082103]
Collignon, J., Varlet, I., Robertson, E. J. Relationship between asymmetric nodal expression and the direction of embryonic turning. Nature 381: 155-158, 1996. [PubMed: 8610012] [Full Text: https://doi.org/10.1038/381155a0]
Gebbia, M., Ferrero, G. B., Pilia, G., Bassi, M. T., Aylsworth, A. S., Penman-Splitt, M., Bird, L. M., Bamforth, J. S., Burn, J., Schlessinger, D., Nelson, D. L., Casey, B. X-linked situs abnormalities result from mutations in ZIC3. Nature Genet. 17: 305-308, 1997. [PubMed: 9354794] [Full Text: https://doi.org/10.1038/ng1197-305]
Gross, M. B. Personal Communication. Baltimore, Md. 9/9/2022.
Iratni, R., Yan, Y.-T., Chen, C., Ding, J., Zhang, Y., Price, S. M., Reinberg, D., Shen, M. M. Inhibition of excess Nodal signaling during mouse gastrulation by the transcriptional corepressor DRAP1. Science 298: 1996-1999, 2002. [PubMed: 12471260] [Full Text: https://doi.org/10.1126/science.1073405]
Krebs, L. T., Iwai, N., Nonaka, S., Welsh, I. C., Lan, Y., Jiang, R., Saijoh, Y., O'Brien, T. P., Hamada, H., Gridley, T. Notch signaling regulates left-right asymmetry determination by inducing Nodal expression. Genes Dev. 17: 1207-1212, 2003. [PubMed: 12730124] [Full Text: https://doi.org/10.1101/gad.1084703]
Lowe, L. A., Supp, D. M., Sampath, K., Yokoyama, T., Wright, C. V., Potter, S. S., Overbeek, P., Kuehn, M. R. Conserved left-right asymmetry of nodal expression and alterations in murine situs inversus. Nature 381: 158-161, 1996. [PubMed: 8610013] [Full Text: https://doi.org/10.1038/381158a0]
Meno, C., Takeuchi, J., Sakuma, R., Koshiba-Takeuchi, K., Ohishi, S., Saijoh, Y., Miyazaki, J., ten Dijke, P., Ogura, T., Hamada, H. Diffusion of nodal signaling activity in the absence of the feedback inhibitor lefty2. Dev. Cell 1: 127-138, 2001. [PubMed: 11703930] [Full Text: https://doi.org/10.1016/s1534-5807(01)00006-5]
Mohapatra, B., Casey, B., Li, H., Ho-Dawson, T., Smith, L., Fernbach, S. D., Molinari, L., Niesh, S. R., Jefferies, J. L., Craigen, W. J., Towbin, J. A., Belmont, J. W., Ware, S. M. Identification and functional characterization of NODAL rare variants in heterotaxy and isolated cardiovascular malformations. Hum. Molec. Genet. 18: 861-871, 2009. [PubMed: 19064609] [Full Text: https://doi.org/10.1093/hmg/ddn411]
Montague, T. G., Gagnon, J. A., Schier, A. F. Conserved regulation of Nodal-mediated left-right patterning in zebrafish and mouse. Development 145: dev171090, 2018. [PubMed: 30446628] [Full Text: https://doi.org/10.1242/dev.171090]
Muller, P., Rogers, K. W., Jordan, B. M., Lee, J. S., Robson, D., Ramanathan, S., Schier, A. F. Differential diffusivity of Nodal and Lefty underlies a reaction-diffusion patterning system. Science 336: 721-724, 2012. [PubMed: 22499809] [Full Text: https://doi.org/10.1126/science.1221920]
Olson, E., Srivastava, D. Molecular pathways controlling heart development. Science 272: 671-676, 1996. [PubMed: 8614825] [Full Text: https://doi.org/10.1126/science.272.5262.671]
Raya, A., Kawakami, Y., Rodriguez-Esteban, C., Buscher, D., Koth, C. M., Itoh, T., Morita, M., Raya, R. M., Dubova, I., Bessa, J. G., de la Pompa, J. L., Belmonte, J. C. I. Notch activity induces Nodal expression and mediates the establishment of left-right asymmetry in vertebrate embryos. Genes Dev. 17: 1213-1218, 2003. [PubMed: 12730123] [Full Text: https://doi.org/10.1101/gad.1084403]
Ware, S. M., Harutyunyan, K. G., Belmont, J. W. Heart defects in X-linked heterotaxy: evidence for a genetic interaction of Zic3 with the Nodal signaling pathway. Dev. Dyn. 235: 1631-1637, 2006. [PubMed: 16496285] [Full Text: https://doi.org/10.1002/dvdy.20719]
Yang, Y.-P., Anderson, R. M., Klingensmith, J. BMP antagonism protects Nodal signaling in the gastrula to promote the tissue interactions underlying mammalian forebrain and craniofacial patterning. Hum. Molec. Genet. 19: 3030-3042, 2010. [PubMed: 20508035] [Full Text: https://doi.org/10.1093/hmg/ddq208]
Zhou, X., Sasaki, H., Lowe, L., Hogan, B. L. M., Kuehn, M. R. Nodal is a novel TGF-beta-like gene expressed in the mouse node during gastrulation. Nature 361: 543-547, 1993. [PubMed: 8429908] [Full Text: https://doi.org/10.1038/361543a0]