title: ZC4H2, an XLID gene, is required for the generation of a specific subset of CNS interneurons.
abstract: Miles-Carpenter syndrome (MCS) was described in 1991 as an XLID syndrome with fingertip arches and contractures and mapped to proximal Xq. Patients had microcephaly, short stature, mild spasticity, thoracic scoliosis, hyperextendable MCP joints, rocker-bottom feet, hyperextended elbows and knees. A mutation, p.L66H, in ZC4H2, was identified in a XLID re-sequencing project. Additional screening of linked families and next generation sequencing of XLID families identified three ZC4H2 mutations: p.R18K, p.R213W and p.V75in15aa. The families shared some relevant clinical features. In silico modeling of the mutant proteins indicated all alterations would destabilize the protein. Knockout mutations in zc4h2 were created in zebrafish and homozygous mutant larvae exhibited abnormal swimming, increased twitching, defective eye movement and pectoral fin contractures. Because several of the behavioral defects were consistent with hyperactivity, we examined the underlying neuronal defects and found that sensory neurons and motoneurons appeared normal. However, we observed a striking reduction in GABAergic interneurons. Analysis of cell-type-specific markers showed a specific loss of V2 interneurons in the brain and spinal cord, likely arising from mis-specification of neural progenitors. Injected human wt ZC4H2 rescued the mutant phenotype. Mutant zebrafish injected with human p.L66H or p.R213W mRNA failed to be rescued, while the p.R18K mRNA was able to rescue the interneuron defect. Our findings clearly support ZC4H2 as a novel XLID gene with a required function in interneuron development. Loss of function of ZC4H2 thus likely results in altered connectivity of many brain and spinal circuits.
authors.affiliation: Greenwood Genetic Center, Greenwood, SC 29646, USA.
authors.forename: Melanie
authors.initials: M
authors.lastname: May
authors.affiliation: Department of Biology, Chungnam National University, Daejeon 305-764, Korea.
authors.forename: Kyu-Seok
authors.initials: KS
authors.lastname: Hwang
authors.affiliation: Department of Child Health, University of Missouri School of Medicine, Columbia, MO 65212, USA.
authors.forename: Judith
authors.initials: J
authors.lastname: Miles
authors.affiliation: Division of Genetics and Metabolism, University of Florida College of Medicine, Gainesville, FL 33612, USA.
authors.forename: Charlie
authors.initials: C
authors.lastname: Williams
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, MD 21287, USA.
authors.forename: Tejasvi
authors.initials: T
authors.lastname: Niranjan
authors.affiliation: Arkansas Children's Hospital, Little Rock, AR 72202, USA.
authors.forename: Stephen G
authors.initials: SG
authors.lastname: Kahler
authors.affiliation: Institute of Medical Genetics, Catholic University, Rome 00-168, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Chiurazzi
authors.affiliation: Institute of Medical Genetics, University of Zurich, Schwerzenbach 8006, Switzerland.
authors.forename: Katharina
authors.initials: K
authors.lastname: Steindl
authors.affiliation: Department of Bioinformatics, Erasmus University Medical Center, Rotterdam 3015, Netherlands.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Van Der Spek
authors.affiliation: Department of Bioinformatics, Erasmus University Medical Center, Rotterdam 3015, Netherlands.
authors.forename: Sigrid
authors.initials: S
authors.lastname: Swagemakers
authors.affiliation: Division of Genetics and Metabolism, University of Florida College of Medicine, Gainesville, FL 33612, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Mueller
authors.affiliation: Department of Physics and Astronomy, Clemson University, Clemson, SC 29634, USA.
authors.forename: Shannon
authors.initials: S
authors.lastname: Stefl
authors.affiliation: Department of Physics and Astronomy, Clemson University, Clemson, SC 29634, USA.
authors.forename: Emil
authors.initials: E
authors.lastname: Alexov
authors.affiliation: Department of Biology, Chungnam National University, Daejeon 305-764, Korea.
authors.forename: Jeong-Im
authors.initials: JI
authors.lastname: Ryu
authors.affiliation: Department of Biology, Chungnam National University, Daejeon 305-764, Korea.
authors.forename: Jung-Hwa
authors.initials: JH
authors.lastname: Choi
authors.affiliation: Department of Biology, Chungnam National University, Daejeon 305-764, Korea.
authors.forename: Hyun-Taek
authors.initials: HT
authors.lastname: Kim
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire CB10 1SA, UK and.
authors.forename: Patrick
authors.initials: P
authors.lastname: Tarpey
authors.affiliation: Institute of Medical Genetics, Catholic University, Rome 00-168, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Neri
authors.affiliation: Greenwood Genetic Center, Greenwood, SC 29646, USA.
authors.forename: Lynda
authors.initials: L
authors.lastname: Holloway
authors.affiliation: Greenwood Genetic Center, Greenwood, SC 29646, USA.
authors.forename: Cindy
authors.initials: C
authors.lastname: Skinner
authors.affiliation: Greenwood Genetic Center, Greenwood, SC 29646, USA.
authors.forename: Roger E
authors.initials: RE
authors.lastname: Stevenson
authors.affiliation: Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112, USA.
authors.forename: Richard I
authors.initials: RI
authors.lastname: Dorsky
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, MD 21287, USA.
authors.forename: Tao
authors.initials: T
authors.lastname: Wang
authors.affiliation: Greenwood Genetic Center, Greenwood, SC 29646, USA, ceschwartz@ggc.org zebrakim@cnu.ac.kr.
authors.forename: Charles E
authors.initials: CE
authors.lastname: Schwartz
authors.affiliation: Department of Biology, Chungnam National University, Daejeon 305-764, Korea, ceschwartz@ggc.org zebrakim@cnu.ac.kr.
authors.forename: Cheol-Hee
authors.initials: CH
authors.lastname: Kim
mesh_terms: D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000235:genetics*; D002460:Cell Line; D002490:Central Nervous System / Q000166:cytology* / Q000378:metabolism*; D002522:Chlorocebus aethiops; D019295:Computational Biology; D005260:Female; D015870:Gene Expression; D050172:Genes, X-Linked; D006801:Humans; D007395:Interneurons / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D009154:Mutation; D009687:Nuclear Proteins; D009928:Organ Specificity / Q000235:genetics; D010375:Pedigree; D015027:Zebrafish
chemical_list: D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; C582075:ZC4H2 protein, human