Cell Lineage Analysis of the Mammalian Female Germline

Fundamental aspects of embryonic and post-natal development, including maintenance of the mammalian female germline, are largely unknown. Here we employ a retrospective, phylogenetic-based method for reconstructing cell lineage trees utilizing somatic mutations accumulated in microsatellites, to study female germline dynamics in mice. Reconstructed cell lineage trees can be used to estimate lineage relationships between different cell types, as well as cell depth (number of cell divisions since the zygote). We show that, in the reconstructed mouse cell lineage trees, oocytes form clusters that are separate from hematopoietic and mesenchymal stem cells, both in young and old mice, indicating that these populations belong to distinct lineages. Furthermore, while cumulus cells sampled from different ovarian follicles are distinctly clustered on the reconstructed trees, oocytes from the left and right ovaries are not, suggesting a mixing of their progenitor pools. We also observed an increase in oocyte depth with mouse age, which can be explained either by depth-guided selection of oocytes for ovulation or by post-natal renewal. Overall, our study sheds light on substantial novel aspects of female germline preservation and development

Muscle-Bound Primordial Stem Cells Give Rise to Myofiber-Associated Myogenic and Non-Myogenic Progenitors

Myofiber cultures give rise to myogenic as well as to non-myogenic cells. Whether these myofiber-associated non-myogenic cells develop from resident stem cells that possess mesenchymal plasticity or from other stem cells such as mesenchymal stem cells (MSCs) remain unsolved. To address this question, we applied a method for reconstructing cell lineage trees from somatic mutations to MSCs and myogenic and non-myogenic cells from individual myofibers that were cultured at clonal density

N-Doped TiO2-Coated Ceramic Membrane for Carbamazepine Degradation in Different Water Qualities

The photocatalytic degradation of the model pollutant carbamazepine (CBZ) was investigated under simulated solar irradiation with an N-doped TiO2-coated Al2O3 photocatalytic membrane, using different water types. The photocatalytic membrane combines photocatalysis and membrane filtration in a single step. The impact of each individual constituent such as acidity, alkalinity, dissolved organic matter (DOM), divalent cations (Mg2+ and Ca2+), and Cl− on the degradation of CBZ was examined. CBZ in water was efficiently degraded by an N-doped TiO2-coated Al2O3 membrane. However, elements added to the water, which simulate the constituents of natural water, had an impact on the CBZ degradation. Water alkalinity inhibited CBZ degradation mostly due to increase in pH while radical scavenging by carbonate was more dominant at higher values (>200 mg/L as CaCO3). A negative effect of Ca2+ addition on photocatalytic degradation was found only in combination with phosphate buffer, probably caused by deposition of CaHPO4 or CaHPO4·2H2O on the catalyst surface. The presence of Cl− and Mg2+ ions had no effect on CBZ degradation. DOM significantly inhibited CBZ degradation for all tested background organic compounds. The photocatalytic activity of N-doped TiO2-coated Al2O3 membranes gradually decreased after continuous use; however, it was successfully regenerated by 0.1% HCl chemical cleaning. Nevertheless, dissolution of metals like Al and Ti should be monitored following acid cleaning

One‐step deposition of nano‐Ag‐TiO 2 coatings by atmospheric pressure plasma jet for water treatment: Application to trace pharmaceutical removal using solar photocatalysis

International audienceIn this study, micrometer thick Ag‐TiO2 coatings were deposited in a single and facile step by spraying the precursor in an atmospheric pressure plasma jet with different concentrations of Ag nanoparticles. The homogenous distribution of Ag decreased the TiO 2 crystal size and increased the surface area. The coatings were characterized to be porous with an anatase phase with improved charge separation and visible light absorption. The photocatalytic activity of the materials was investigated for degrading rhodamine B using a white lamp as a screening method to optimize Ag‐TiO 2 coatings. Then the photodegradation of trace pharmaceutical compounds (TrPCs) was investigated by using a solar light simulator at the optimal condition of a TiO 2 coating with 0.4wt% A

Microsatellite mutations are replication dependent.

<p>a) Shown are two scenarios for microsatellite mutations – replication dependent mutations that occur during mitotic divisions at a cell stage with two chromosomal copies (left), and spontaneous, replication independent mutations occurring at the quiescent oocyte stage with four chromosomal copies (right). While replication dependent mutations would result in two alleles, spontaneous mutations would mostly result in more than two alleles. The bottom plots show representative capillary signals. b) The increase with age in fraction of alleles with spontaneous mutations is significantly smaller than the increase in fraction of alleles with replication dependent mutations. <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002477#s2" target="_blank">Results</a> are for the three longitudinal experiments.</p

Oocytes form a cluster distinct from bone marrow cells.

<p>Reconstructed cell lineage tree of mice in several ages, each showing that GV oocytes from large antral follicles (red) form a cluster that is distinct from cells of bone marrow origin (mesenchymal stem cells- yellow and lymphocytes-blue). Ovarian cumulus cells are in green. Shaded boxes denote subtrees that are statistically enriched for cells of a certain cell population, using a hyper-geometric enrichment test. Mouse names represent their age in days. Y axis is depth (number of divisions since the zygote). One cell in M278 was cropped for visual clarity.</p

Accelerated increase in GV oocyte from large antral follicles depth following unilateral ovariectomy.

<p>a–c) Reconstructed lineage trees of oocytes from both ovaries of unilaterally ovariectomized mice. Blue circles are oocytes extracted from the ovary removed at a young age – 23 days (a) 29 days (b) and 26 days (c). Green circles are oocytes extracted from the contralateral ovary removed at an older age – 139 days (a) 161 days (b) and 150 days in which the ovulated oocytes were marked with blue dots (c). Horizontal lines are the average depths of young oocytes (blue) and old oocytes (green). d) Increase in depth with age following ovariectomy is accelerated relative to the increase observed for non-ovariectomized mice. Last two boxes include pooled data from the indicated mice. * p<0.05, ** p<0.01.</p

Oocyte depth increases with age.

<p>a–c) Reconstructed lineage trees of GV oocytes from large antral follilces in three mice demonstrate an increase in oocyte depth (Y axis) with age. The putative zygote is at depth 0. Median depth of oocytes is 13 divisions in M27, a 27 day old mouse (Mouse names represent their age in days) (a), 16 divisions in mouse M159 (b) and 19 in mouse M268 which contains 6 ovulated oocytes which were marked on the tree with blue dots (c). Horizontal red lines denote the median depth. All lineage trees were reconstructed using the maximum-likelihood neighbor joining method and rooted with the median identifier of all cells. d) Median depth of GV oocytes from large antral follicels increases with age. Each blue point is the median of the depths of all oocytes sampled from a single mouse. The Pearson correlation of median depth and age is 0.81 (p = 0.007). High inter-mouse variability can be seen. Errorbars are standard errors of the means. (e) Depth of pancreatic islet cells does not increase with age. Shown are the depths of pancreatic islet cells from M36, a 1-month old mouse and from M280, a 9-month old mouse.</p