title: Ovarian stimulation perturbs methylation status of placental imprinting genes and reduces blood pressure in the second generation offspring.
abstract: OBJECTIVE
  Assisted reproductive technology (ART) is associated with DNA methylation dysfunction of offspring. However, it is unclear whether ovarian stimulation (OS) is responsible for DNA methylation dysfunction of offspring STUDY DESIGN: We built the first-generation (F1) and second-generation (F2) offspring mice model of ovarian stimulation. Bodyweight of F1 and F2 were measured. Expression levels of several imprinted genes (Impact, H19, Igf2, Plagl1, Mest, and Snrpn) in F1 placenta were tested. Methylation status of Plagl1 and H19 promoters was examined with bisulfite sequencing. Glucose tolerance, blood pressure, and heart rate were evaluated in F2 mice.
  
  
  RESULTS
  The OS F1 showed elevated bodyweights in the 2nd, 3rd and 4th weeks, but the difference disappeared in the 5th week. Plagl1 was down-regulated in OS F1. Promoters of Plagl1 and H19 were also hypermethylated in OS F1. F2 of OS mice had the similar bodyweight and glucose tolerance compared with the control F2. However, F2 of OS ♂F1+OS♀ F1 showed the decreased systolic pressure, diastolic pressure, and heart rate.
  
  
  CONCLUSIONS
  Ovarian stimulation perturbs expression levels and methylation status of imprinted genes in offspring. The effect of ovarian stimulation may be passed to F2.
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China; Key Laboratory of Reproductive Genetics, Ministry of Education (Zhejiang University), Hangzhou, China.
authors.forename: Gu-Feng
authors.initials: GF
authors.lastname: Xu
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Liao
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China; Key Laboratory of Reproductive Genetics, Ministry of Education (Zhejiang University), Hangzhou, China.
authors.forename: Jing-Yi
authors.initials: JY
authors.lastname: Li
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China; Key Laboratory of Reproductive Genetics, Ministry of Education (Zhejiang University), Hangzhou, China.
authors.forename: Yi-Feng
authors.initials: YF
authors.lastname: Liu
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China; Key Laboratory of Reproductive Genetics, Ministry of Education (Zhejiang University), Hangzhou, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Yi-Qing
authors.initials: YQ
authors.lastname: Wu
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China; Key Laboratory of Reproductive Genetics, Ministry of Education (Zhejiang University), Hangzhou, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China; Key Laboratory of Reproductive Genetics, Ministry of Education (Zhejiang University), Hangzhou, China.
authors.forename: Ping-Ping
authors.initials: PP
authors.lastname: Lv
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China; Key Laboratory of Reproductive Genetics, Ministry of Education (Zhejiang University), Hangzhou, China.
authors.forename: Run-Jv
authors.initials: RJ
authors.lastname: Zhang
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China; Key Laboratory of Reproductive Genetics, Ministry of Education (Zhejiang University), Hangzhou, China. Electronic address: zhangdan@zju.edu.cn.
authors.forename: Dan
authors.initials: D
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001794:Blood Pressure / Q000502:physiology*; D001835:Body Weight / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000235:genetics; D019175:DNA Methylation; D005260:Female; D016147:Genes, Tumor Suppressor; D018392:Genomic Imprinting; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D010062:Ovulation Induction; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000235:genetics; D062085:RNA, Long Noncoding / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D055517:snRNP Core Proteins / Q000235:genetics
chemical_list: D018797:Cell Cycle Proteins; C094335:H19 long non-coding RNA; C513121:IGF2 protein, mouse; C107747:Impact protein, mouse; D047908:Intracellular Signaling Peptides and Proteins; C498423:Plagl1 protein, mouse; D011506:Proteins; D062085:RNA, Long Noncoding; D014157:Transcription Factors; C103637:mesoderm specific transcript protein; D055517:snRNP Core Proteins; D007335:Insulin-Like Growth Factor II
keywords: Imprinted gene; Methylation; Offspring; Ovarian stimulation