scholarly journals The Pluripotency-Associated Gene Dppa4 Is Dispensable for Embryonic Stem Cell Identity and Germ Cell Development but Essential for Embryogenesis

2009 ◽  
Vol 29 (11) ◽  
pp. 3186-3203 ◽  
Author(s):  
Babita Madan ◽  
Vikas Madan ◽  
Odile Weber ◽  
Philippe Tropel ◽  
Carmen Blum ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Function ◽  
Es Cells ◽  
Embryonic Stem ◽  
Cell Development ◽  
Es Cell ◽  
Pluripotent Cells ◽  
Cell Identity ◽  
Germ Cell Development ◽  
Skeletal Defects

ABSTRACT Dppa4 (developmental pluripotency-associated 4) has been identified in several high-profile screens as a gene that is expressed exclusively in pluripotent cells. It encodes a nuclear protein with an SAP-like domain and appears to be associated preferentially with transcriptionally active chromatin. Its exquisite expression pattern and results of RNA interference experiments have led to speculation that Dppa4, as well as its nearby homolog Dppa2, might play essential roles in embryonic stem (ES) cell function and/or germ cell development. To rigorously assess suggested roles, we have generated Dppa4-deficient and Dppa4/Dppa2 doubly deficient ES cells, as well as mice lacking Dppa4. Contrary to predictions, we find that Dppa4 is completely dispensable for ES cell identity and germ cell development. Instead, loss of Dppa4 in mice results in late embryonic/perinatal death and striking skeletal defects with partial penetrance. Thus, surprisingly, Dppa4-deficiency affects tissues that apparently never transcribed the gene, and at least some loss-of-function defects manifest phenotypically at an embryonic stage long after physiologic Dppa4 expression has ceased. Concomitant with targeted gene inactivation, we have introduced into the Dppa4 locus a red fluorescent marker (tandem-dimer red fluorescent protein) that is compatible with green fluorescent proteins and allows noninvasive visualization of pluripotent cells and reprogramming events.

scholarly journals Survival Motor Neuron Protein Participates in Mouse Germ Cell Development and Spermatogonium Maintenance

2020 ◽  
Vol 21 (3) ◽  
pp. 794 ◽  
Author(s):  
Wei-Fang Chang ◽  
Jie Xu ◽  
Tzu-Ying Lin ◽  
Jing Hsu ◽  
Hsiu-Mei Hsieh-Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
Motor Neuron ◽  
Cell Biology ◽  
Critical Role ◽  
Embryonic Stem ◽  
Cell Development ◽  
Survival Motor Neuron ◽  
Specific Marker ◽  
Germ Cell Development

The defective human survival motor neuron 1 (SMN1) gene leads to spinal muscular atrophy (SMA), the most common genetic cause of infant mortality. We previously reported that loss of SMN results in rapid differentiation of Drosophila germline stem cells and mouse embryonic stem cells (ESCs), indicating that SMN also plays important roles in germ cell development and stem cell biology. Here, we show that in healthy mice, SMN is highly expressed in the gonadal tissues, prepubertal spermatogonia, and adult spermatocytes, whereas low SMN expression is found in differentiated spermatid and sperm. In SMA-like mice, the growth of testis tissues is retarded, accompanied with gamete development abnormalities and loss of the spermatogonia-specific marker. Consistently, knockdown of Smn1 in spermatogonial stem cells (SSCs) leads to a compromised regeneration capacity in vitro and in vivo in transplantation experiments. In SMA-like mice, apoptosis and accumulation of the R-loop structure were significantly elevated, indicating that SMN plays a critical role in the survival of male germ cells. The present work demonstrates that SMN, in addition to its critical roles in neuronal development, participates in mouse germ cell and spermatogonium maintenance.

scholarly journals Effects of environmental Bisphenol A exposures on germ cell development and Leydig cell function in the human fetal testis

PLoS ONE ◽  
2018 ◽  
Vol 13 (1) ◽  
pp. e0191934 ◽  
Author(s):  
Soria Eladak ◽  
Delphine Moison ◽  
Marie-Justine Guerquin ◽  
Gabriele Matilionyte ◽  
Karen Kilcoyne ◽  
...  
Keyword(s):  
Bisphenol A ◽  
Germ Cell ◽  
Leydig Cell ◽  
Cell Function ◽  
Cell Development ◽  
Germ Cell Development ◽  
Fetal Testis ◽  
Leydig Cell Function

scholarly journals Nodal secures pluripotency upon embryonic stem cell progression from the ground state

2016 ◽  
Author(s):  
Carla Mulas ◽  
Tüzer Kalkan ◽  
Austin Smith
Keyword(s):  
Ground State ◽  
Es Cells ◽  
Embryonic Stem ◽  
Es Cell ◽  
Lineage Specification ◽  
Cell Identity ◽  
Neural Fate ◽  
Cell Progression ◽  
Adherent Culture ◽  
Kinetics Of

SUMMARYNaïve mouse embryonic stem (ES) cells can readily acquire specific fates, but the cellular and molecular processes that enable lineage specification are poorly characterised. Here we investigated progression from the ES cell ground state in adherent culture. We utilised down-regulation of Rex1::GFPd2 to track loss of ES cell identity. We found that cells that have newly down-regulated this reporter have acquired competence for germline induction. They can also be efficiently specified for different somatic lineages, responding more rapidly than naïve cells to inductive cues. Nodal is a candidate autocrine regulator of pluripotency. Abrogation of Nodal signalling did not substantially alter kinetics of exit from the ES cell state, but accelerated subsequent adoption of neural fate at the expense of other lineages. This effect was evident if Nodal was inhibited prior to extinction of ES cell identity. We suggest that Nodal is pivotal for non-neural competence in cells departing naïve pluripotency.

scholarly journals Embryonic stem cells commit to differentiation by symmetric divisions following a variable lag period

2020 ◽  
Author(s):  
Stanley E Strawbridge ◽  
Guy B Blanchard ◽  
Austin Smith ◽  
Hillel Kugler ◽  
Graziano Martello
Keyword(s):  
Cell Cycle ◽  
Cell Fate ◽  
Es Cells ◽  
Embryonic Stem ◽  
Es Cell ◽  
Cell Identity ◽  
Daughter Cells ◽  
Population Dynamics Model ◽  
Developmental Progression ◽  
Lag Period

ABSTRACTMouse embryonic stem (ES) cells are derived from the epiblast of the preimplantation embryo and retain the capacity to give rise to all embryo lineages. ES cells can be released into differentiation from a near-homogeneous maintenance condition. Exit from the ES cell state can be accurately monitored using the Rex1-GFPd2 transgenic reporter, providing a powerful system for examining a mammalian cell fate transition. Here, we performed live-cell imaging and tracking of ES cells during entry into differentiation for 48 hours in defined conditions. We observed a greater cell surface area and a modest shortening of the cell cycle prior to exit and subsequently a reduction in cell size and increase in motility. We did not see any instance of cells regaining ES cell identity, consistent with unidirectional developmental progression. Transition occurred asynchronously across the population but genealogical tracking revealed a high correlation in cell-cycle length and Rex1-GFPd2 expression between daughter cells. A population dynamics model was consistent with symmetric divisions during exit from naive pluripotency. Collapse of ES cell identity occurred acutely in individual cells but after a variable delay. The variation in lag period can extend up to three generations, creating marked population asynchrony.

FMR1 Deficiency Alters Self-Renewal and Proliferation of Mouse Embryonic Cells

Nano LIFE ◽  
2015 ◽  
Vol 05 (02) ◽  
pp. 1550003
Author(s):  
Shyam Sundhar Bale ◽  
Nima Saeidi ◽  
Srivatsan Kidambi ◽  
Martin L. Yarmush ◽  
Monica Casali
Keyword(s):  
Rna Binding ◽  
Cell Function ◽  
Fragile X ◽  
Es Cells ◽  
Embryonic Stem ◽  
Embryonic Cells ◽  
Es Cell ◽  
Self Renewal ◽  
Translational Repressor ◽  
Undifferentiated State

The regulation of embryonic stem (ES) cell self-renewal and pluripotency is based upon highly orchestrated transcription factor networks. RNA inhibition has been demonstrated to affect ES cell function by altering gene expression levels that are critical to the maintenance and differentiation of ES cells. Fragile X mental retardation protein (FMRP) is a selective RNA-binding protein that can act as a translational repressor for bound mRNA and regulates the expression of a variety of gene transcripts in numerous adult cells. The absence of FMRP results in the most common form of inherited intellectual disability, Fragile X syndrome. In an effort to determine the role of FMRP during development, we silenced the FMRP gene (FMR1) using short hairpin RNA (shRNA). Prior to differentiation induction, we analyzed the phenotype of FMR1 knock down (FMR1-kd) mouse ES cells in their undifferentiated state. Herein, we report that FMR1-kd ES cells proliferate at a greater rate than wild-type ES cells resulting in a 25% reduction in doubling time. FMR1-kd ES cells were found to have an increased expression of three self-renewal genes (OCT-4, Sox2, Nanog) in the undifferentiated state. Moreover, FMR1-kd ES cells failed to downregulate OCT-4 during differentiation programs resulting in abnormal fate decisions in vitro. These results demonstrate an unexpected correlation between FMR1 expression and OCT-4 regulation suggesting that FMRP is involved in the silencing of OCT-4 during the commencement of differentiation programs.

scholarly journals Disorganization of the germ cell pool leads to primary ovarian insufficiency

Reproduction ◽  
2017 ◽  
Vol 153 (6) ◽  
pp. R205-R213 ◽  
Author(s):  
Ikko Kawashima ◽  
Kazuhiro Kawamura
Keyword(s):  
Germ Cell ◽  
Cell Biology ◽  
Embryonic Stem ◽  
Primordial Follicle ◽  
Cell Development ◽  
Primary Ovarian Insufficiency ◽  
Ovarian Insufficiency ◽  
Germ Cell Development ◽  
Female Germ Cell

The mammalian ovary is an organ that controls female germ cell development, storing them and releasing mature oocytes for transporting to the oviduct. During the fetal stage, female germ cells change from a proliferative state to meiosis before forming follicles with the potential for the growth of surrounding somatic cells. Understanding of molecular and physiological bases of germ cell development in the fetal ovary contributed not only to the elucidation of genetic disorders in primary ovarian insufficiency (POI), but also to the advancement of novel treatments for patients with POI. Accumulating evidence indicates that mutations inNOBOX,DAZLandFIGLAgenes are associated with POI. In addition, cell biology studies revealed the important roles of these genes as essential translational factors for germ cell development. Recent insights into the role of the PI3K (phosphatidylinositol 3-kinase)-Akt signaling pathway in primordial follicle activation allowed the development of a new infertility treatment, IVA (in vitroactivation), leading to successful pregnancy/delivery in POI patients. Furthermore, elucidation of genetic dynamics underlying female germ cell development could allow regeneration of oocytes from ES (embryonic stem)/iPS (induced pluripotent stem) cells in mammals. The purpose of this review is to summarize basic findings related to female germ cell development and potential clinical implications, especially focusing on POI etiologies. We also summarize evolving new POI therapies based on IVA as well as oocyte regeneration.

Murine embryonic stem cell differentiation is promoted by SOCS-3 and inhibited by the zinc finger transcription factor Klf4

Blood ◽  
2005 ◽  
Vol 105 (2) ◽  
pp. 635-637 ◽  
Author(s):  
Yanjun Li ◽  
Jeanette McClintick ◽  
Li Zhong ◽  
Howard J. Edenberg ◽  
Mervin C. Yoder ◽  
...  
Keyword(s):  
Cell Function ◽  
Embryoid Body ◽  
Es Cells ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Northern Blotting ◽  
Cytokine Signaling ◽  
Embryonic Stem Cell Differentiation ◽  
Es Cell ◽  
Self Renewal

Abstract Embryonic stem (ES) cells homozygous for a Shp-2 mutation (Shp-2Δ46-110) demonstrate leukemia inhibitory factor (LIF) hypersensitivity and increased LIF-stimulated phosphorylation of signal transducer and activator of transcription (STAT3). We hypothesized that LIF-responsive genes in Shp-2Δ46-110 cells would represent potential candidates for molecules vital for ES cell self-renewal. Using microarray analysis, we detected 41 genes whose expression was modified by LIF in Shp-2Δ46-110 ES cells. Induction of 2 significantly up-regulated genes, suppressor of cytokine signaling–3 (SOCS-3) and Krüppel-like factor 4 (Klf4), was verified using Northern blotting. ES cells overexpressing SOCS-3 had an increased capacity to differentiate to hematopoietic progenitors, rather than to self-renew. In contrast, ES cells overexpressing Klf4 had a greater capacity to self-renew based on secondary embryoid body (EB) formation. Klf4-transduced d6 EBs expressed higher levels of Oct-4, consistent with the notion that Klf4 promotes ES cell self-renewal. These findings verify the negative role of SOCS-3 on LIF signaling and provide a novel role for Klf4 in ES cell function.

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