Defective jagged-1 signaling affects GnRH development and contributes to congenital hypogonadotropic hypogonadism

In vertebrate species, fertility is controlled by gonadotropin-releasing hormone (GnRH) neurons. GnRH cells arise outside the central nervous system, in the developing olfactory pit, and migrate along olfactory/vomeronasal/terminal nerve axons into the forebrain during embryonic development. Congenital hypogonadotropic hypogonadism (CHH) and Kallmann syndrome are rare genetic disorders characterized by infertility, and they are associated with defects in GnRH neuron migration and/or altered GnRH secretion and signaling. Here, we documented the expression of the jagged-1/Notch signaling pathway in GnRH neurons and along the GnRH neuron migratory route both in zebrafish embryos and in human fetuses. Genetic knockdown of the zebrafish ortholog of JAG1 (jag1b) resulted in altered GnRH migration and olfactory axonal projections to the olfactory bulbs. Next-generation sequencing was performed in 467 CHH unrelated probands, leading to the identification of heterozygous rare variants in JAG1. Functional in vitro validation of JAG1 mutants revealed that 7 out of the 9 studied variants exhibited reduced protein levels and altered subcellular localization. Together our data provide compelling evidence that Jag1/Notch signaling plays a prominent role in the development of GnRH neurons, and we propose that JAG1 insufficiency may contribute to the pathogenesis of CHH in humans.

[1]  S. Radovick,et al.  Development of the gonadotropin‐releasing hormone system , 2021, Journal of neuroendocrinology.

[2]  M. Xiang,et al.  Notch signaling determines cell-fate specification of the two main types of vomeronasal neurons of rodents , 2021, bioRxiv.

[3]  G. Vannelli,et al.  Benzo[a]pyrene impairs the migratory pattern of human gonadotropin-releasinghormone- secreting neuroblasts , 2021, European journal of histochemistry : EJH.

[4]  J. Shendure,et al.  CADD-Splice—improving genome-wide variant effect prediction using deep learning-derived splice scores , 2021, Genome Medicine.

[5]  P. Fontanaud,et al.  Synaptic communication mediates the assembly of a self-organizing circuit that controls reproduction , 2021, Science Advances.

[6]  R. Quinton,et al.  Genetics of congenital hypogonadotropic hypogonadism: peculiarities and phenotype of an oligogenic disease , 2020, Human Genetics.

[7]  P. Thistlethwaite,et al.  Notch Enhances Ca2+ Entry by Activating Calcium-sensing Receptors and Inhibiting Voltage-gated K+ Channels. , 2020, American journal of physiology. Cell physiology.

[8]  I. Krantz,et al.  Alagille syndrome mutation update: Comprehensive overview of JAG1 and NOTCH2 mutation frequencies and insight into missense variant classification , 2019, Human mutation.

[9]  P. Duminuco,et al.  Evidence for a Common Genetic Origin of Classic and Milder Adult-Onset Forms of Isolated Hypogonadotropic Hypogonadism , 2019, Journal of clinical medicine.

[10]  R. Mirsky,et al.  Neural crest Notch/Rbpj signaling regulates olfactory gliogenesis and neuronal migration† , 2018, Genesis.

[11]  T. Schilling,et al.  Cell-type heterogeneity in the early zebrafish olfactory epithelium is generated from progenitors within preplacodal ectoderm , 2018, eLife.

[12]  R. Quinton,et al.  KLB, encoding β‐Klotho, is mutated in patients with congenital hypogonadotropic hypogonadism , 2017, EMBO molecular medicine.

[13]  D. Larizza,et al.  A frequent oligogenic involvement in congenital hypothyroidism. , 2017, Human molecular genetics.

[14]  Alain Chédotal,et al.  Development of the neurons controlling fertility in humans: new insights from 3D imaging and transparent fetal brains , 2016, Development.

[15]  W. Pear,et al.  Delta/Notch-Like EGF-Related Receptor (DNER) Is Not a Notch Ligand , 2016, PloS one.

[16]  Surangi N Perera,et al.  Evidence for a Notch1‐mediated transition during olfactory ensheathing cell development , 2016, Journal of anatomy.

[17]  G. Merlo,et al.  The zebrafish: an emerging animal model for investigating the hypothalamic regulation of reproduction. , 2016, Minerva endocrinologica.

[18]  C. Peano,et al.  The Dlx5 and Foxg1 transcription factors, linked via miRNA-9 and -200, are required for the development of the olfactory and GnRH system , 2015, Molecular and Cellular Neuroscience.

[19]  M. Maghnie,et al.  Expert consensus document: European Consensus Statement on congenital hypogonadotropic hypogonadism—pathogenesis, diagnosis and treatment , 2015, Nature Reviews Endocrinology.

[20]  Matan Golan,et al.  Architecture of GnRH-Gonadotrope-Vasculature Reveals a Dual Mode of Gonadotropin Regulation in Fish. , 2015, Endocrinology.

[21]  C. Ruhrberg,et al.  Dysfunctional SEMA3E signaling underlies gonadotropin-releasing hormone neuron deficiency in Kallmann syndrome. , 2015, The Journal of clinical investigation.

[22]  Bale,et al.  Standards and Guidelines for the Interpretation of Sequence Variants: A Joint Consensus Recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology , 2015, Genetics in Medicine.

[23]  Jana Marie Schwarz,et al.  MutationTaster2: mutation prediction for the deep-sequencing age , 2014, Nature Methods.

[24]  R. Fagerholm,et al.  Mutation screening of SEMA3A and SEMA7A in patients with congenital hypogonadotropic hypogonadism , 2014, Pediatric Research.

[25]  N. Wayne,et al.  Early Development of the Gonadotropin-Releasing Hormone Neuronal Network in Transgenic Zebrafish , 2013, Front. Endocrinol..

[26]  M. Wegner,et al.  Olfactory ensheathing glia are required for embryonic olfactory axon targeting and the migration of gonadotropin-releasing hormone neurons , 2013, Biology Open.

[27]  A. Vettori,et al.  Generation and application of signaling pathway reporter lines in zebrafish , 2013, Molecular Genetics and Genomics.

[28]  P. Dworzynski,et al.  Mutations in FGF17, IL17RD, DUSP6, SPRY4, and FLRT3 are identified in individuals with congenital hypogonadotropic hypogonadism. , 2013, American journal of human genetics.

[29]  K. Flegal,et al.  Association of All-Cause Mortality With Overweight and Obesity Using Standard Body Mass Index Categories , 2012 .

[30]  J. Hardelin,et al.  SEMA3A, a Gene Involved in Axonal Pathfinding, Is Mutated in Patients with Kallmann Syndrome , 2012, PLoS genetics.

[31]  S. Wray,et al.  Neural Crest and Ectodermal Cells Intermix in the Nasal Placode to Give Rise to GnRH-1 Neurons, Sensory Neurons, and Olfactory Ensheathing Cells , 2011, The Journal of Neuroscience.

[32]  M. DePristo,et al.  A framework for variation discovery and genotyping using next-generation DNA sequencing data , 2011, Nature Genetics.

[33]  M. Hammerschmidt,et al.  Zebrafish in endocrine systems: recent advances and implications for human disease. , 2011, Annual review of physiology.

[34]  S. Tobet,et al.  Gonadotropin-releasing hormone (GnRH) neuron migration: Initiation, maintenance and cessation as critical steps to ensure normal reproductive function , 2011, Frontiers in Neuroendocrinology.

[35]  C. Greer,et al.  Composition of the migratory mass during development of the olfactory nerve , 2010, The Journal of comparative neurology.

[36]  J. Gusella,et al.  Oligogenic basis of isolated gonadotropin-releasing hormone deficiency , 2010, Proceedings of the National Academy of Sciences.

[37]  P. Bork,et al.  A method and server for predicting damaging missense mutations , 2010, Nature Methods.

[38]  K. Kaestner,et al.  Jagged1 is a competitive inhibitor of Notch signaling in the embryonic pancreas , 2009, Mechanisms of Development.

[39]  P. Rakic,et al.  Interaction between Reelin and Notch Signaling Regulates Neuronal Migration in the Cerebral Cortex , 2008, Neuron.

[40]  M. Westerfield,et al.  Notch signaling regulates endocrine cell specification in the zebrafish anterior pituitary. , 2008, Developmental biology.

[41]  Y. Gothilf,et al.  Early Development of Forebrain Gonadotrophin‐Releasing Hormone (GnRH) Neurones and the Role of GnRH as an Autocrine Migration Factor , 2008, Journal of neuroendocrinology.

[42]  B. Appel,et al.  Notch signaling regulates midline cell specification and proliferation in zebrafish. , 2006, Developmental biology.

[43]  N. Illing,et al.  Development of GnRH cells: Setting the stage for puberty , 2006, Molecular and Cellular Endocrinology.

[44]  H. Nagatsuka,et al.  Expression of Notch1 and Hes5 in the developing olfactory epithelium , 2006, Acta oto-laryngologica.

[45]  A. Conigliaro,et al.  Expression analysis of jagged genes in zebrafish embryos , 2005, Developmental dynamics : an official publication of the American Association of Anatomists.

[46]  Yoshihiro Yoshihara,et al.  Mutually Exclusive Glomerular Innervation by Two Distinct Types of Olfactory Sensory Neurons Revealed in Transgenic Zebrafish , 2005, The Journal of Neuroscience.

[47]  Hong Wu,et al.  Notch Signaling, Brain Development, and Human Disease , 2005, Pediatric Research.

[48]  H. Okamoto,et al.  Robo2 is required for establishment of a precise glomerular map in the zebrafish olfactory system , 2005, Development.

[49]  K. Nibu,et al.  Notch expression in developing olfactory neuroepithelium , 2004, Neuroreport.

[50]  Steven Henikoff,et al.  SIFT: predicting amino acid changes that affect protein function , 2003, Nucleic Acids Res..

[51]  C. Haass,et al.  A γ‐secretase inhibitor blocks Notch signaling in vivo and causes a severe neurogenic phenotype in zebrafish , 2002, EMBO reports.

[52]  M. Mishina,et al.  Regulation by Protein Kinase A Switching of Axonal Pathfinding of Zebrafish Olfactory Sensory Neurons through the Olfactory Placode–Olfactory Bulb Boundary , 2002, The Journal of Neuroscience.

[53]  François Guillemot,et al.  Mash1 and Ngn1 control distinct steps of determination and differentiation in the olfactory sensory neuron lineage. , 2002, Development.

[54]  S. Ekker,et al.  Effective targeted gene ‘knockdown’ in zebrafish , 2000, Nature Genetics.

[55]  G. Weinmaster,et al.  Embryonic lethality and vascular defects in mice lacking the Notch ligand Jagged1. , 1999, Human molecular genetics.

[56]  S. Artavanis-Tsakonas,et al.  Intracellular Cleavage of Notch Leads to a Heterodimeric Receptor on the Plasma Membrane , 1997, Cell.

[57]  D. Sassoon,et al.  Notch4/int-3, a mammary proto-oncogene, is an endothelial cell-specific mammalian Notch gene. , 1996, Development.

[58]  C. Kimmel,et al.  Stages of embryonic development of the zebrafish , 1995, Developmental dynamics : an official publication of the American Association of Anatomists.

[59]  A. Becchetti,et al.  Neuroblast long-term cell cultures from human fetal olfactory epithelium respond to odors , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[60]  U. Lendahl,et al.  The novel Notch homologue mouse Notch 3 lacks specific epidermal growth factor-repeats and is expressed in proliferating neuroepithelium , 1994, Mechanisms of Development.

[61]  H. Weintraub,et al.  Mouse notch: expression in hair follicles correlates with cell fate determination , 1993, The Journal of cell biology.

[62]  U. Lendahl,et al.  Motch A and motch B--two mouse Notch homologues coexpressed in a wide variety of tissues. , 1993, Experimental cell research.

[63]  J. Rossant,et al.  Expression analysis of a Notch homologue in the mouse embryo. , 1992, Developmental biology.

[64]  G. Weinmaster,et al.  Notch2: a second mammalian Notch gene. , 1992, Development.

[65]  F. Valverde,et al.  Formation of an olfactory glomerulus: Morphological aspects of development and organization , 1992, Neuroscience.

[66]  D. Gallahan,et al.  Mouse mammary tumor gene int-3: a member of the notch gene family transforms mammary epithelial cells , 1992, Journal of virology.

[67]  G. Weinmaster,et al.  A homolog of Drosophila Notch expressed during mammalian development. , 1991, Development.

[68]  J. Sklar,et al.  TAN-1, the human homolog of the Drosophila Notch gene, is broken by chromosomal translocations in T lymphoblastic neoplasms , 1991, Cell.

[69]  F. Wondisford,et al.  Migratory arrest of gonadotropin-releasing hormone neurons in transgenic mice. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[70]  W. Harris,et al.  Xotch, the Xenopus homolog of Drosophila notch. , 1990, Science.

[71]  D. Pfaff,et al.  Luteinizing hormone-releasing hormone (LHRH)-expressing cells do not migrate normally in an inherited hypogonadal (Kallmann) syndrome. , 1989, Brain research. Molecular brain research.

[72]  H. Gainer,et al.  Evidence that cells expressing luteinizing hormone-releasing hormone mRNA in the mouse are derived from progenitor cells in the olfactory placode. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[73]  S. Wray,et al.  Spatiotemporal cell expression of luteinizing hormone-releasing hormone in the prenatal mouse: evidence for an embryonic origin in the olfactory placode. , 1989, Brain research. Developmental brain research.

[74]  D. Gallahan,et al.  Mammary tumorigenesis in feral mice: identification of a new int locus in mouse mammary tumor virus (Czech II)-induced mammary tumors , 1987, Journal of virology.

[75]  M. Maghnie,et al.  Characteristics of a nationwide cohort of patients presenting with isolated hypogonadotropic hypogonadism (IHH). , 2018, European journal of endocrinology.

[76]  D. Libri,et al.  New understandings of the genetic basis of isolated idiopathic central hypogonadism. , 2012, Asian journal of andrology.

[77]  H. Bellen,et al.  Endocytosis and intracellular trafficking of Notch and its ligands. , 2010, Current topics in developmental biology.

[78]  Y. Gothilf,et al.  Targeted gonadotropin-releasing hormone-3 neuron ablation in zebrafish: effects on neurogenesis, neuronal migration, and reproduction. , 2010, Endocrinology.

[79]  Jonathan A. Epstein,et al.  The multifaceted role of Notch in cardiac development and disease , 2008, Nature Reviews Genetics.

[80]  B. Thisse,et al.  High-resolution in situ hybridization to whole-mount zebrafish embryos , 2007, Nature Protocols.

[81]  W. Driever,et al.  Distinct delta and jagged genes control sequential segregation of pancreatic cell types from precursor pools in zebrafish. , 2007, Developmental biology.

[82]  A. Bauer,et al.  Regulation of Notch signalling by non-visual beta-arrestin. , 2005, Nature cell biology.