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Mechanical tensions regulate gene expression in the Xenopus laevis neuroectoderm

ABSTRACT: During gastrulation and neurulation, chordamesoderm and overlying neuroectoderm of vertebrate embryo are converged under the control of specific genetic program to the dorsal midline, simultaneously extending along it. To reveal genes regulated by mechanical tensions developed as a result of these morphogenetic movements we compared transcriptomes of artificially stretched neuroectodermal explants of Xenopus laevis midgastrula embryos with the non-stretched one. Total RNA was extracted from cell lysed with Trisol reagent and PureLink RNA Micro Kit according to manufacture instruction. Quality was checked with BioAnalyser and RNA 6000 Nano Kit. PolyA RNA was purified with Dynabeads mRNA Purification Kit. Illumina library was made from polyA RNA with NEBNextNEBNext RNA Library Prep Kit for Illumina according to manual. Sequencing was performed on HiSeq1500 with 50 bp read length. At least 10 millions of reads were generated for each sample. Reads mapping was done using tophat2 software v.2.0.9. As a reference genome we used Xenopus Laevis build 6 (http://www.xenbase.org/genomes/static/laevis.jsp). For quantification, we used cufflinks v2.1.1.

ORGANISM(S): Xenopus laevis

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PROVIDER: S-BSST981 | biostudies-other |

REPOSITORIES: biostudies-other

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Project description:BackgroundReproductive isolation is a defining characteristic of populations that represent unique biological species, yet we know very little about the gene expression basis for reproductive isolation. The advent of powerful molecular biology tools provides the ability to identify genes involved in reproductive isolation and focuses attention on the molecular mechanisms that separate biological species. Herein we quantify the sterility pattern of hybrid males in African Clawed Frogs (Xenopus) and apply microarray analysis of the expression pattern found in testes to identify genes that are misexpressed in hybrid males relative to their two parental species (Xenopus laevis and X. muelleri).Methodology/principal findingsPhenotypic characteristics of spermatogenesis in sterile male hybrids (X. laevis x X. muelleri) were examined using a novel sperm assay that allowed quantification of live, dead, and undifferentiated sperm cells, the number of motile vs. immotile sperm, and sperm morphology. Hybrids exhibited a dramatically lower abundance of mature sperm relative to the parental species. Hybrid spermatozoa were larger in size and accompanied by numerous undifferentiated sperm cells. Microarray analysis of gene expression in testes was combined with a correction for sequence divergence derived from genomic hybridizations to identify candidate genes involved in the sterility phenotype. Analysis of the transcriptome revealed a striking asymmetric pattern of misexpression. There were only about 140 genes misexpressed in hybrids compared to X. laevis but nearly 4,000 genes misexpressed in hybrids compared to X. muelleri.Conclusions/significanceOur results provide an important correlation between phenotypic characteristics of sperm and gene expression in sterile hybrid males. The broad pattern of gene misexpression suggests intriguing mechanisms creating the dominance pattern of the X. laevis genome in hybrids. These findings significantly contribute to growing evidence for allelic dominance in hybrids and have implications for the mechanism of species differentiation at the transcriptome level.