QTL experiments in pigs are often analysed separately, although similar or same founder breeds are frequently used to establish the experimental design. The aim of the present study was to jointly analyse three porcine F 2-crosses for six growth and four muscling traits. The crosses were a Meishan × Pietrain cross, a Wild Boar × Pietrain cross, and a Wild Boar × Meishan cross. In some cases, same founder animals were used to establish the crosses. 966 F 2-individuals were genotyped for 242 genetic markers (mostly microsatellites) and phenotyped for birth weight, 21 and 35 day weight, slaughter weight, carcass length, food conversion ratio, ham meat weight, shoulder meat weight, loin and neck meat weight, and meat area. A multi-allele multi-QTL model was applied that estimated an additive QTL effect for each founder breed and parental origin (either paternally or maternally derived), and a dominant QTL effect for each cross. This model was previously introduced in plant breeding. Numerous QTL were mapped on the autosomes. Most QTL were localised on SSC1, 2, 3, 4, 6 and 8, and no QTL were on SSC9, 11, 13, 15, 17 and 18. The confidence intervals were short in many cases. QTL with an exceptionally high test statistic were found for carcass length on SSC1, 4, 7 and 17. The coefficient of variation was remarkably small for this trait, which suggests that carcass length is affected by only a few genes with large effects. Positional and functional candidates underlying promising QTL are suggested for further study.

SUMMARY Cerebral organoids, three-dimensional cultures that model organogenesis, provide a new platform to investigate human brain development. High cost, variability and tissue heterogeneity limit their broad applications. Here we developed a miniaturized spinning bioreactor (SpinΩ) to generate forebrain-specific organoids from human iPSCs. These organoids recapitulate key features of human cortical development, including progenitor zone organization, neurogenesis, gene expression, and notably, a distinct human-specific outer radial glia cell layer. We also developed protocols for midbrain and hypothalamic organoids. Finally, we employed the forebrain organoid platform to model Zika virus (ZIKV) exposure. Quantitative analyses revealed preferential, productive infection of neural progenitors with either African or Asian ZIKV strains. ZIKV infection leads to increased cell death and reduced proliferation, resulting in decreased neuronal cell layer volume resembling microcephaly. Together, our brain region-specific organoids and SpinΩ provide an accessible and versatile platform for modeling human brain development and disease, and for compound testing including potential ZIKV antiviral drugs.

Summary Tumor heterogeneity is a major barrier to effective cancer diagnosis and treatment. We recently identified cancer-specific differentially DNA-methylated regions (cDMRs) in colon cancer, which also distinguish normal tissue types from each other, suggesting that these cDMRs might be generalized across cancer types. Here we show stochastic methylation variation of the same cDMRs, distinguishing cancer from normal, in colon, lung, breast, thyroid, and Wilms tumors, with intermediate variation in adenomas. Whole genome bisulfite sequencing shows these variable cDMRs are related to loss of sharply delimited methylation boundaries at CpG islands. Furthermore, we find hypomethylation of discrete blocks encompassing half the genome, with extreme gene expression variability. Genes associated with the cDMRs and large blocks are involved in mitosis and matrix remodeling, respectively. These data suggest a model for cancer involving loss of epigenetic stability of well-defined genomic domains that underlies increased methylation variability in cancer and could contribute to tumor heterogeneity.

Methylation of cytosines in the mammalian genome represents a key epigenetic modification and is dynamically regulated during development. Compelling evidence now suggests that dynamic regulation of DNA methylation is mainly achieved through a cyclic enzymatic cascade comprised of cytosine methylation, iterative oxidation of methyl group by TET dioxygenases, and restoration of unmodified cytosines by either replication-dependent dilution or DNA glycosylase-initiated base excision repair. In this review, we discuss the mechanism and function of DNA demethylation in mammalian genomes, focusing particularly on how developmental modulation of the cytosine-modifying pathway is coupled to active reversal of DNA methylation in diverse biological processes.