Critical function of AP-2gamma/TCFAP2C in mouse embryonic germ cell maintenance

Formation of the germ cell lineage involves multiple processes, including repression of somatic differentiation and reacquisition of pluripotency as well as a unique epigenetic constitution. The transcriptional regulator Prdm1 has been identified as a main coordinator of this process, controlling epigenetic modification and gene expression. Here we report on the expression pattern of the transcription factor Tcfap2c, a putative downstream target of Prdm1, during normal mouse embryogenesis and the consequences of its specific loss in primordial germ cells (PGCs) and their derivatives. Tcfap2c is expressed in PGCs from Embryonic Day 7.25 (E 7.25) up to E 12.5, and targeted disruption resulted in sterile animals, both male and female. In the mutant animals, PGCs were specified but were lost around E 8.0. PGCs generated in vitro from embryonic stem cells lacking TCFAP2C displayed induction of Prdm1 and Dppa3. Upregulation of Hoxa1, Hoxb1, and T together with lack of expression of germ cell markers such Nanos3, Dazl, and Mutyh suggested that the somatic gene program is induced in TCFAP2C-deficient PGCs. Repression of TCFAP2C in TCam-2, a human PGC-resembling seminoma cell line, resulted in specific upregulation of HOXA1, HOXB1, MYOD1, and HAND1, indicative of mesodermal differentiation. Expression of genes indicative of ectodermal, endodermal, or extraembryonic differentiation, as well as the finding of no change to epigenetic modifications, suggested control by other factors. Our results implicate Tcfap2c as an important effector of Prdml activity that is required for PGC maintenance, most likely mediating Prdm1-induced suppression of mesodermal differentiation

Principles of early human development and germ cell program from conserved model systems

Human primordial germ cells (hPGCs), the precursors of sperm and eggs, originate during week 2-3 of early postimplantation development(1). Using in vitro models of hPGC induction(2-4), recent studies suggest striking mechanistic differences in specification of human and mouse PGCs(5). This may partly be due to the divergence in their pluripotency networks, and early postimplantation development(6-8). Since early human embryos are inaccessible for direct studies, we considered alternatives, including porcine embryos that, as in humans, develop as bilaminar embryonic discs. Here we show that porcine PGCs (pPGCs) originate from the posterior pre-primitive streak competent epiblast by sequential upregulation of SOX17 and BLIMP1 in response to WNT and BMP signalling. Together with human and monkey in vitro models simulating peri-gastrulation development, we show conserved principles for epiblast development for competency for PGC fate, followed by initiation of the epigenetic program(9-11), regulated by a balanced SOX17–BLIMP1 gene dosage. Our combinatorial approach using human, porcine and monkey in vivo and in vitro models, provides synthetic insights on early human development

Lineage Conversion of Murine Extraembryonic Trophoblast Stem Cells to Pluripotent Stem Cells▿†

In mammals, the first cell fate decision is initialized by cell polarization at the 8- to 16-cell stage of the preimplantation embryo. At this stage, outside cells adopt a trophectoderm (TE) fate, whereas the inside cell population gives rise to the inner cell mass (ICM). Prior to implantation, transcriptional interaction networks and epigenetic modifications divide the extraembryonic and embryonic fate irrevocably. Here, we report that extraembryonic trophoblast stem cell (TSC) lines are converted to induced pluripotent stem cells (TSC-iPSCs) by overexpressing Oct4, Sox2, Klf4, and cMyc. Methylation studies and gene array analyses indicated that TSC-iPSCs had adopted a pluripotent potential. The rate of conversion was lower than those of somatic reprogramming experiments, probably due to the unique genetic network controlling extraembryonic lineage fixation. Both in vitro and in vivo, TSC-iPSCs differentiated into tissues representing all three embryonic germ layers, indicating that somatic cell fate could be induced. Finally, TSC-iPSCs chimerized the embryo proper and contributed to the germ line of mice, indicating that these cells had acquired full somatic differentiation potential. These results lead to a better understanding of the molecular processes that govern the first lineage decision in mammals

Diagnose nutricional com o uso de sensor óptico ativo em algodoeiro Nutritional diagnosis with the use of active optical sensor in cotton

Este trabalho objetivou avaliar a resposta espectral do dossel de plantas à variação de doses de nitrogênio (N) e sua relação com os teores foliares de N, índice de clorofila e produtividade na cultura do algodoeiro. O experimento foi conduzido em Chapadão do Sul, MS, no delineamento em blocos casualizados, com quatro repetições. Os tratamentos consistiram de cinco doses de N: 0, 30, 70, 110 e 150 kg ha-1, divididas em duas aplicações aos 28 e 41 dias após a emergência utilizando-se, como fonte, o fertilizante ureia. As maiores correlações dos índices de clorofila e do índice de vegetação por diferença normalizada com a produtividade foram observadas na quarta leitura, aos 56 dias após a emergência, indicando que neste período a produtividade já pode estar comprometida caso haja falhas no suprimento de N à cultura. Os resultados obtidos indicaram que o sensor se torna mais sensível à variação do teor foliar de N conforme a planta se desenvolve mas não quando ela atinge um alto índice de área foliar e começa a entrar em senescência. Concluiu-se, então, que o sensoriamento remoto ao nível terrestre permite estimar indiretamente a quantidade de N absorvido, o índice de clorofila e a produtividade do algodoeiro.<br>This research aimed to evaluate the spectral response to variation of nitrogen levels and its relationship with leaf nitrogen, chlorophyll and yield in cotton crop. The experiment was conducted in Chapadão do Céu, MS, in a randomized block design with four replications. The treatments consisted of five N rates of 0, 30, 70, 110 and 150 kg ha-1, divided in two applications at 28 and 41 days after emergence, using urea fertilizer as a source. The highest correlations of the chlorophyll index and normalized difference vegetation index with yield were observed in the fourth observation, at 56 days after emergence, indicating that in this period, yield may already be compromised if there is shortage in the supply of N to the crop. The results obtained indicated that the sensor becomes more sensitive to variations of nitrogen levels as the crop grows, but not when it reaches a high leaf area index and in the begining of the senescence. It is concluded that ground level remote sensing permits to estimate indirectly the amount of N absorbed, chlorophyll index and cotton yield