Pax3 lineage-specific deletion of Gpr161 is associated with spinal neural tube and craniofacial malformations during embryonic development

Shh signaling is the morphogen signaling that regulates embryonic craniofacial and neural tube development. G protein-coupled receptor 161 (Gpr161) is a negative regulator of Shh signaling, and its inactivation in mice results in embryo lethality with craniofacial and neural tube defects (NTDs). However, the structural defects of later embryonic stages in Gpr161 null mice and cell lineages underlying abnormalities were not well characterized due to their limited lifespan. We found the Pax3 lineage-specific deletion of Gpr161 in mice presented with tectal hypertrophy (anterior dorsal neuroepithelium), cranial vault and facial bone hypoplasia (cranial neural crest (CNC)), vertebral abnormalities (somite), and the closed form of spina bifida (posterior dorsal neuroepithelium). In particular, the closed form of spina bifida is partly due to the reduced Pax3 and Cdx4 gene expression of the posterior dorsal neural tubes of Gpr161 mutant embryos involving decreased Wnt signaling whereas Shh signaling was increased. This study provides the novel role of Gpr161 in the posterior neural tube development and confirms its role on CNC- and somite-derived skeletogenesis and midbrain morphogenesis in mice.

[1]  R. Pasi,et al.  Neural tube defects: Different types and brief review of neurulation process and its clinical implication , 2021, Journal of family medicine and primary care.

[2]  B. Wlodarczyk,et al.  Wnt1 Lineage Specific Deletion of Gpr161 Results in Embryonic Midbrain Malformation and Failure of Craniofacial Skeletal Development , 2021, Frontiers in Genetics.

[3]  G. Konopka,et al.  Derepression of sonic hedgehog signaling upon Gpr161 deletion unravels forebrain and ventricular abnormalities. , 2019, Developmental biology.

[4]  M. Ross,et al.  Dominant negative GPR161 rare variants are risk factors of human spina bifida , 2018, Human molecular genetics.

[5]  B J Buchan,et al.  Spina bifida. , 2018, The Journal of the Arkansas Medical Society.

[6]  Tommaso Mazza,et al.  Hypomorphic Recessive Variants in SUFU Impair the Sonic Hedgehog Pathway and Cause Joubert Syndrome with Cranio-facial and Skeletal Defects. , 2017, American journal of human genetics.

[7]  M. Qiu,et al.  Suppressor of Fused restraint of Hedgehog activity level is critical for osteogenic proliferation and differentiation during calvarial bone development , 2017, The Journal of Biological Chemistry.

[8]  S. Wakana,et al.  A spontaneous and novel Pax3 mutant mouse that models Waardenburg syndrome and neural tube defects. , 2017, Gene.

[9]  James Briscoe,et al.  Morphogen rules: design principles of gradient-mediated embryo patterning , 2015, Development.

[10]  T. Matise,et al.  The orphan GPCR, Gpr161, regulates the retinoic acid and canonical Wnt pathways during neurulation. , 2015, Developmental biology.

[11]  Jonathan J. Wilde,et al.  Genetic, epigenetic, and environmental contributions to neural tube closure. , 2014, Annual review of genetics.

[12]  J. Epstein,et al.  β-catenin regulates Pax3 and Cdx2 for caudal neural tube closure and elongation , 2014, Development.

[13]  V. Palma,et al.  Proliferation of Murine Midbrain Neural Stem Cells Depends upon an Endogenous Sonic Hedgehog (Shh) Source , 2013, PloS one.

[14]  L. Rangell,et al.  The Ciliary G-Protein-Coupled Receptor Gpr161 Negatively Regulates the Sonic Hedgehog Pathway via cAMP Signaling , 2013, Cell.

[15]  E. Martí,et al.  Dorsal–ventral patterning of the neural tube: A tale of three signals , 2012, Developmental neurobiology.

[16]  C. Carter,et al.  The role of primary cilia in the pathophysiology of neural tube defects. , 2012, Neurosurgical focus.

[17]  N. Manley,et al.  Whole mount in situ hybridization of E8.5 to E11.5 mouse embryos. , 2011, Journal of visualized experiments : JoVE.

[18]  A. Lassar,et al.  A gradient of Shh establishes mutually repressing somitic cell fates induced by Nkx3.2 and Pax3. , 2008, Developmental biology.

[19]  E. Willems,et al.  An optimized procedure for whole-mount in situ hybridization on mouse embryos and embryoid bodies , 2008, Nature Protocols.

[20]  H. Williams,et al.  A unifying hypothesis for hydrocephalus, Chiari malformation, syringomyelia, anencephaly and spina bifida , 2008, Cerebrospinal Fluid Research.

[21]  M. Lauth,et al.  Genetic elimination of Suppressor of fused reveals an essential repressor function in the mammalian Hedgehog signaling pathway. , 2006, Developmental cell.

[22]  G. Shaw,et al.  Genes encoding catalytic subunits of protein kinase A and risk of spina bifida. , 2005, Birth defects research. Part A, Clinical and molecular teratology.

[23]  J. Epstein,et al.  Insertion of Cre into the Pax3 locus creates a new allele of Splotch and identifies unexpected Pax3 derivatives. , 2005, Developmental biology.

[24]  Toyoaki Tenzen,et al.  Hedgehog signaling in the neural crest cells regulates the patterning and growth of facial primordia. , 2004, Genes & development.

[25]  P. Mehlen,et al.  Inhibition of Neuroepithelial Patched-Induced Apoptosis by Sonic Hedgehog , 2003, Science.

[26]  G. McKnight,et al.  Protein Kinase A Deficiency Causes Axially Localized Neural Tube Defects in Mice* , 2002, The Journal of Biological Chemistry.

[27]  M. Loeken,et al.  Rescue of neural tube defects in Pax-3-deficient embryos by p53 loss of function: implications for Pax-3- dependent development and tumorigenesis. , 2002, Genes & development.

[28]  M. Bronner‐Fraser,et al.  Inhibition of Sonic hedgehog signaling in vivo results in craniofacial neural crest cell death , 1999, Current Biology.

[29]  J. Charrow,et al.  Myelomeningocele and Waardenburg syndrome (type 3) in patients with interstitial deletions of 2q35 and the PAX3 gene: possible digenic inheritance of a neural tube defect. , 1998, American journal of medical genetics.

[30]  M. Loeken,et al.  Neural Tube Defects in Embryos of Diabetic Mice: Role of the Pax-3 Gene and Apoptosis , 1997, Diabetes.

[31]  E. Mariman,et al.  A frameshift mutation in the gene for PAX3 in a girl with spina bifida and mild signs of Waardenburg syndrome. , 1995, Journal of medical genetics.

[32]  K. Vogan,et al.  The splotch-delayed (Spd) mouse mutant carries a point mutation within the paired box of the Pax-3 gene. , 1993, Genomics.

[33]  T. Franz.  The Splotch (Sp1H) and Splotch-delayed (Spd) alleles: differential phenotypic effects on neural crest and limb musculature , 1993, Anatomy and Embryology.

[34]  R. Balling,et al.  Variations of cervical vertebrate after expression of a Hox-1.1 transgene in mice , 1990, Cell.

[35]  Madeline G. Andrews,et al.  New perspectives on the mechanisms establishing the dorsal-ventral axis of the spinal cord. , 2019, Current topics in developmental biology.

[36]  J. Lupski,et al.  Whole-exome sequencing identifies homozygous GPR161 mutation in a family with pituitary stalk interruption syndrome. , 2015, The Journal of clinical endocrinology and metabolism.