Claudin-10 is required for relay of left-right patterning cues from Hensen's node to the lateral plate mesoderm.
Species-specific symmetry-breaking events at the left-right organizer (LRO) drive an evolutionarily-conserved cascade of gene expression in the lateral plate mesoderm that is required for the asymmetric positioning of organs within the body cavity. The mechanisms underlying the transfer of the left and right laterality information from the LRO to the lateral plate mesoderm are poorly understood. Here, we investigate the role of Claudin-10, a tight junction protein, in facilitating the transfer of left-right identity from the LRO to the lateral plate mesoderm. Claudin-10 is asymmetrically expressed on the right side of the chick LRO, Hensen's node. Gain- and loss-of-function studies demonstrated that right-sided expression of Claudin-10 is essential for normal rightward heart tube looping, the first morphological asymmetry during organogenesis. Manipulation of Claudin-10 expression did not perturb asymmetric gene expression at Hensen's node, but did disrupt asymmetric gene expression in the lateral plate mesoderm. Bilateral expression of Claudin-10 at Hensen's node prevented expression of Nodal, Lefty-2 and Pitx2c in the left lateral plate mesoderm, while morpholino knockdown of Claudin-10 inhibited expression of Snail1 in the right lateral plate mesoderm. We also determined that amino acids that are predicted to affect ion selectivity and protein interactions that bridge Claudin-10 to the actin cytoskeleton were essential for its left-right patterning function. Collectively, our data demonstrate a novel role for Claudin-10 during the transmission of laterality information from Hensen's node to both the left and right sides of the embryo and demonstrate that tight junctions have a critical role during the relay of left-right patterning cues from Hensen's node to the lateral plate mesoderm.
Departments of Human Genetics, McGill University, Canada; The Research Institute of the McGill University Health Centre, Montréal, Québec, Canada.
Michelle M
MM
Collins
Departments of Human Genetics, McGill University, Canada; The Research Institute of the McGill University Health Centre, Montréal, Québec, Canada.
Amanda I
AI
Baumholtz
The Research Institute of the McGill University Health Centre, Montréal, Québec, Canada.
Annie
A
Simard
INRS-Institut Armand Frappier, Laval, Québec, Canada.
Mary
M
Gregory
Departments of Human Genetics, McGill University, Canada.
Daniel G
DG
Cyr
Departments of Human Genetics, McGill University, Canada; Departments of Paediatrics, McGill University, Canada; The Research Institute of the McGill University Health Centre, Montréal, Québec, Canada. Electronic address: aimee.ryan@mcgill.ca.
Aimee K
AK
Ryan
D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D019521:Body Patterning / Q000235:genetics*; D002642:Chick Embryo; D057167:Claudins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D006321:Heart / Q000196:embryology; D055458:Left-Right Determination Factors / Q000096:biosynthesis; D008648:Mesoderm / Q000378:metabolism*; D060172:Morpholinos / Q000235:genetics; D055457:Nodal Protein / Q000096:biosynthesis; D020897:Organizers, Embryonic / Q000378:metabolism*; D038081:Organogenesis / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics; D000071250:Snail Family Transcription Factors; D019108:Tight Junctions / Q000378:metabolism*; D014157:Transcription Factors / Q000096:biosynthesis; D029961:Zebrafish Proteins / Q000096:biosynthesis
D057167:Claudins; D055458:Left-Right Determination Factors; D060172:Morpholinos; D055457:Nodal Protein; C572614:Pitx2 protein, zebrafish; C000604407:SNAI1 protein, human; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors; D029961:Zebrafish Proteins; C502009:claudin 10
Asymmetry; Chick development; Claudin-10; Claudins; Heart tube looping; Hensen’s node; Lateral plate mesoderm; Laterality cues; Left–right patterning; Nodal; Pitx2c; Tight junctions