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Critical roles of lysine demethylase Kdm3a in craniofacial and neural development during Xenopus embryogenesis

ABSTRACT: The epigenetic modifier lysine-specific histone demethylase 3a (kdm3a) is specific for demethylation of mono- and di-methylated 9th lysine of histone H3 (H3K9me1/2) and belongs to the Jumonji domain-containing group of demethylases. Kdm3a is known to play significant roles during various biological and pathophysiological processes such as spermatogenesis and metabolism, determination of sex, androgen receptor-mediated transcription, and embryonic carcinoma cell differentiation. In the current study, we evaluated the physiological functions of kdm3a during Xenopus laevis embryogenesis. Spatiotemporal expression pattern indicated maternal nature of kdm3a, exhibiting its expression from early embryonic stages until tadpole stage, however considerable high level of expression were observed during the neurula stage of Xenopus development. Depleting kdm3a using kdm3a antisense morpholino oligonucleotides induced anomalies, such as head deformities and small-sized eyes. Our whole-mount in situ hybridization results demonstrated that kdm3a knockdown is associated with defects in neural crest formation and migration. Further, reverse transcription polymerase chain reaction revealed abnormal expression of neural markers in kdm3a morphants. RNA sequencing of kdm3a morphants indicated that kdm3a is implicated in mesoderm formation, cell adhesion, and metabolic processes of embryonic development. In conclusion, we suggest that kdm3a is critical for neural development during Xenopus embryogenesis and can be targeted for treatment of developmental disorders. The further detailed investigation is required to understand the molecular mechanism involved in neural development regulation by kdm3a.

ORGANISM(S): Xenopus laevis

PROVIDER: GSE117754 | GEO | 2018/11/23

REPOSITORIES: GEO

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Project description:Gene expression profiling has provided critical insights into the molecular pathways underlying development of model organisms, however little information is available on regulated gene expression during human embryogenesis. We have now filled this important gap of knowledge by performing genome-wide microarray analysis of the Homo sapiens gene expression during the 4-9th week, a period when most organs develop. We analyzed individually 3 embryos for each of the 4th, 5th, 6th, 7th, 8th, and 9th week of human embryonic development by using the Affymetrix U133 plus 2.0 human GeneChip array.About half of all human genes are expressed and 18.6% of the expressed genes were significantly regulated during this important period. We further identified over 5000 regulated genes, most of which were previously not known to be associated with animal development. Our study also revealed that the genes involved here are distinct from those during early embryogenesis, which include three groups of maternal genes. Furthermore, we discovered that genes in a given developmental process are coordinately regulated. This led us to develop an easily searchable database of this entire collection of gene expression profiles, allowing for identification new genes important for a particular developmental process/pathway and deducing the potential function of a novel gene. The validity of the predictions from the database was demonstrated with two examples through spatiotemporal analyses of the two novel genes. Such a database should serve as a highly valuable resource for the molecular analysis of human development and pathogenesis. Human embryos were obtained from therapeutic termination of pregnancy induced by Mifepristone. Morphologically normal embryos were selected and staged according to the Carnegie classification and the Chinese embryonic developmental characteristics. Each whole embryo was micro-dissected from an intact trophoblast and stored directly in liquid nitrogen or homogenized using Trizol solution and then stored at -80oC. Total RNA was isolated from each whole embryo individually and each of the 3 RNA samples for each developmental time point was independently analyzed on a Human U133 plus 2.0 Genome Oligonucleotide Array.

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Critical roles of lysine demethylase Kdm3a in craniofacial and neural development during Xenopus embryogenesis

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