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Title: Mutations in TUBB8 and Human Oocyte Meiotic Arrest

BACKGROUND: We present that human reproduction depends on the fusion of a mature oocyte with a sperm cell to form a fertilized egg. The genetic events that lead to the arrest of human oocyte maturation are unknown. METHODS: We sequenced the exomes of five members of a four-generation family, three of whom had infertility due to oocyte meiosis I arrest. We performed Sanger sequencing of a candidate gene, TUBB8, in DNA samples from these members, additional family members, and members of 23 other affected families. The expression of TUBB8 and all other β-tubulin isotypes was assessed in human oocytes, early embryos, sperm cells, and several somatic tissues by means of a quantitative reverse- transcriptase-polymerase-chain-reaction assay. We evaluated the effect of the TUBB8 mutations on the assembly of the heterodimer consisting of one α-tubulin polypeptide and one β-tubulin polypeptide (α/β-tubulin heterodimer) in vitro, on microtubule architecture in HeLa cells, on microtubule dynamics in yeast cells, and on spindle assembly in mouse and human oocytes. RESULTSL: We identified seven mutations in the primate-specific gene TUBB8 that were responsible for oocyte meiosis I arrest in 7 of the 24 families. TUBB8 expression is unique to oocytes and the early embryo, in which thismore » gene accounts for almost all the expressed β-tubulin. The mutations affect chaperone-dependent folding and assembly of the α/β-tubulin heterodimer, disrupt microtubule behavior on expression in cultured cells, alter microtubule dynamics in vivo, and cause catastrophic spindle-assembly defects and maturation arrest on expression in mouse and human oocytes. CONCLUSIONS: Lastly, TUBB8 mutations have dominant-negative effects that disrupt microtubule behavior and oocyte meiotic spindle assembly and maturation, causing female infertility.« less
Authors:
 [1] ;  [1] ;  [2] ;  [3] ;  [2] ;  [2] ;  [4] ;  [5] ;  [6] ;  [7] ;  [2] ;  [3] ;  [3] ;  [1] ;  [8] ;  [1] ;  [1] ;  [9] ;  [9] ;  [9] more »;  [1] ;  [10] ;  [11] ;  [12] ;  [1] ;  [1] ;  [13] ;  [14] ;  [1] ;  [15] ;  [16] ;  [5] ;  [1] « less
  1. Fudan Univ., Shanghai (China). State Key Laboratory of Genetic Engineering, Institutes of Biomedical Sciences, MOE Key Laboratory of Contemporary Anthropology, and Collaborative Innovation Center of Genetics and Development, School of Life Sciences
  2. Shanghai Jiao Tong Univ. (China). Reproductive Medicine Center, Shanghai Ninth Hospital
  3. Fudan Univ., Shanghai (China). Shanghai Ji Ai Genetics and IVF Institute, Obstetrics and Gynecology Hospital
  4. Shanghai, Reproductive Medicine Center, Shaanxi Maternal and Child Care Service Center, Shaanxi (China)
  5. New York University Langone Medical Center, NY (United States). Department of Biochemistry and Molecular Pharmacology
  6. Univ. of Chicago, IL (United States). Department of Molecular Genetics and Cell Biology
  7. Univ. of Chicago, IL (United States). Department of Molecular Genetics and Cell Biology
  8. Fudan Univ., Shanghai (China). Department of Otolaryngology, Eye and ENT Hospital
  9. Shanghai Institute of Planned Parenthood Research (China)
  10. Fudan Univ., Shanghai (China). Department of Integrative Medicine and Neurobiology, State Key Laboratory of Medical Neurobiology, and Shanghai Medical College
  11. University of Gothenburg (Sweden). Department of Physiology–Endocrinology, Institute of Neuroscience and Physiology
  12. Southeast University, Nanjing (China). Key Laboratory for Developmental Genes and Human Disease, Ministry of Education, Institute of Life Sciences
  13. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Sciences Division
  14. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Sciences Division; Univ. of California, Berkeley, CA (United States). Molecular and Cell Biology Department and the Howard Hughes Medical Institute
  15. Fudan Univ., Shanghai (China). State Key Laboratory of Genetic Engineering, Institutes of Biomedical Sciences, MOE Key Laboratory of Contemporary Anthropology, and Collaborative Innovation Center of Genetics and Development, School of Life Sciences; Shanghai Jiao Tong Univ. (China). Bio-X Center, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education
  16. Univ. of Chicago, IL (United States). Department of Molecular Genetics and Cell Biology; Iowa State Univ., Ames, IA (United States). Department of Genetics, Development and Cell Biology
Publication Date:
Grant/Contract Number:
AC02-05CH11231
Type:
Accepted Manuscript
Journal Name:
New England Journal of Medicine
Additional Journal Information:
Journal Volume: 374; Journal Issue: 3; Journal ID: ISSN 0028-4793
Research Org:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
USDOE; National Institutes of Health (NIH)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES
OSTI Identifier:
1379043