DMRT1: An Ancient Sexual Regulator Required for Human Gonadogenesis

Transcriptional regulators related to the invertebrate sexual regulators doublesex and mab-3 occur throughout metazoans and control sex in most animal groups. Seven of these DMRT genes are found in mammals, and mouse genetics has shown that one, Dmrt1, plays a crucial role in testis differentiation, both in germ cells and somatic cells. Deletions and, more recently, point mutations affecting human DMRT1 have demonstrated that its heterozygosity is associated with 46,XY complete gonadal dysgenesis. Most of our detailed knowledge of DMRT1 function in the testis, the focus of this review, derives from mouse studies, which have revealed that DMRT1 is essential for male somatic and germ cell differentiation and maintenance of male somatic cell fate after differentiation. Moreover, ectopic DMRT1 can reprogram differentiated female granulosa cells into male Sertoli-like cells. The ability of DMRT1 to control sexual cell fate likely derives from at least 3 properties. First, DMRT1 functionally collaborates with another key male sex regulator, SOX9, and possibly other proteins to maintain and reprogram sexual cell fate. Second, and related, DMRT1 appears to function as a pioneer transcription factor, binding “closed” inaccessible chromatin and promoting its opening to allow binding by other regulators including SOX9. Third, DMRT1 binds DNA by a highly unusual form of interaction and can bind with different stoichiometries.

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Towards a Novel Computer-Aided Optimization of Microreactors: Techno-Economic Evaluation of an Immobilized Enzyme System

Symmetry ◽

10.3390/sym13030524 ◽

2021 ◽

Vol 13 (3) ◽

pp. 524

Author(s):

Philip Pietrek ◽

Manfred Kraut ◽

Roland Dittmeyer

Keyword(s):

Magnetic Beads ◽

Glucose Dehydrogenase ◽

Packed Bed ◽

Detailed Knowledge ◽

Reaction Conditions ◽

Flow Reactors ◽

Continuous Flow Reactors ◽

Enzyme Cascades ◽

And Control ◽

Physical And Chemical

Immobilized multi-enzyme cascades are increasingly used in microfluidic devices. In particular, their application in continuous flow reactors shows great potential, utilizing the benefits of reusability and control of the reaction conditions. However, capitalizing on this potential is challenging and requires detailed knowledge of the investigated system. Here, we show the application of computational methods for optimization with multi-level reactor design (MLRD) methodology based on the underlying physical and chemical processes. We optimize a stereoselective reduction of a diketone catalyzed by ketoreductase (Gre2) and Nicotinamidadenindinukleotidphosphat (NADPH) cofactor regeneration with glucose dehydrogenase (GDH). Both enzymes are separately immobilized on magnetic beads forming a packed bed within the microreactor. We derive optimal reactor feed concentrations and enzyme ratios for enhanced performance and a basic economic model in order to maximize the techno-economic performance (TEP) for the first reduction of 5-nitrononane-2,8-dione.

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SEL-5, A Serine/Threonine Kinase That Facilitates lin-12 Activity in Caenorhabditis elegans

Genetics ◽

10.1093/genetics/153.4.1641 ◽

1999 ◽

Vol 153 (4) ◽

pp. 1641-1654 ◽

Cited By ~ 1

Author(s):

Hanna Fares ◽

Iva Greenwald

Keyword(s):

Cell Fate ◽

Point Mutations ◽

Kinase Domain ◽

Terminal Region ◽

Serine Threonine Kinase ◽

Intracellular Domain ◽

Cell Fate Decisions ◽

Threonine Kinase ◽

Amino Terminal ◽

Fate Decision

Abstract Ligands present on neighboring cells activate receptors of the LIN-12/Notch family by inducing a proteolytic cleavage event that releases the intracellular domain. Mutations that appear to eliminate sel-5 activity are able to suppress constitutive activity of lin-12(d) mutations that are point mutations in the extracellular domain of LIN-12, but cannot suppress lin-12(intra), the untethered intracellular domain. These results suggest that sel-5 acts prior to or during ligand-dependent release of the intracellular domain. In addition, sel-5 suppression of lin-12(d) mutations is tissue specific: loss of sel-5 activity can suppress defects in the anchor cell/ventral uterine precursor cell fate decision and a sex myoblast/coelomocyte decision, but cannot suppress defects in two different ventral hypodermal cell fate decisions in hermaphrodites and males. sel-5 encodes at least two proteins, from alternatively spliced mRNAs, that share an amino-terminal region and differ in the carboxy-terminal region. The amino-terminal region contains the hallmarks of a serine/threonine kinase domain, which is most similar to mammalian GAK1 and yeast Pak1p.

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YAP creates epiblast responsiveness to inductive signals for germ cell fate

Development ◽

10.1242/dev.199732 ◽

2021 ◽

Author(s):

Saya Kagiwada ◽

Shinya Aramaki ◽

Guangming Wu ◽

Borami Shin ◽

Eva Kutejova ◽

...

Keyword(s):

Cell Differentiation ◽

Germ Cell ◽

Signaling Pathway ◽

Cell Fate ◽

Cell Lineage ◽

Signaling Molecules ◽

Hippo Signaling ◽

Epiblast Stem Cells ◽

Germ Cell Differentiation ◽

Cell Induction

The germ cell lineage in mammals is induced by the stimulation of pluripotent epiblast cells with signaling molecules. Previous studies have suggested that the germ cell differentiation competence or responsiveness of epiblast cells to signaling molecules is established and maintained in epiblast cells of a specific differentiation state. However, the molecular mechanism underlying this process has not been well defined. Here, using the differentiation model of epiblast stem cells (EpiSCs), we have shown that two defined EpiSC lines have robust germ cell differentiation competence. However, another defined EpiSC line has no competence. By evaluating the molecular basis of EpiSCs with distinct germ cell differentiation competence, we identified YAP/YAP1/YAP65, an intracellular mediator of the Hippo signaling pathway, as a critical mediator for establishing germ cell induction. Strikingly, deletion of YAP severely affected responsiveness to inductive stimuli, leading to a defect in WNT target activation and germ cell differentiation. In conclusion, we propose that the Hippo/YAP signaling pathway creates a potential for germ cell fate induction via mesodermal WNT signaling in pluripotent epiblast cells.

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Controlling intimacy: Sexual scripts among men and women in prostitution

Current Sociology ◽

10.1177/0011392118815945 ◽

2018 ◽

Vol 68 (3) ◽

pp. 353-371 ◽

Cited By ~ 2

Author(s):

Margaretha Järvinen ◽

Theresa Dyrvig Henriksen

Keyword(s):

Qualitative Interviews ◽

Sexual Scripts ◽

The Other ◽

Men And Women ◽

Male And Female ◽

Female Sex ◽

Emotional Distance ◽

Male Sex ◽

And Control

Inspired by sexual scripting theory, this article analyses intimacy and control in prostitution. The authors identify two strategies for maintaining control among male and female sex sellers. The first strategy is to restrict prostitution to relationships with as much sexual reciprocity as possible. The other is to maintain sexual/emotional distance from customers – yet often acting the opposite. The article questions prevailing stereotypes about male sex sellers being more agentic and autonomous than female sex sellers, arguing that control in prostitution can be achieved (and lost) in different ways. The analysis shows how scripting theory – with its differentiation between the cultural, interpersonal and intrapsychic levels of scripting – may be used to understand variations and contradictions in prostitution experiences. The article is based on 36 qualitative interviews with men and women in escort services, clinic prostitution and prostitution in private apartments in Denmark.

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The lipid metabolome of kidney cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.30_suppl.16 ◽

2012 ◽

Vol 30 (30_suppl) ◽

pp. 16-16

Author(s):

Saby George ◽

Kyoung-Soo Choi ◽

Jeff C. Miecznikowski ◽

Roberto Pili ◽

A. Latif Kazim

Keyword(s):

Kidney Cancer ◽

Treatment Strategies ◽

Charge Ratio ◽

P Value ◽

Detailed Knowledge ◽

Lipid Profiling ◽

Butyl Ether ◽

Bonferroni Adjustment ◽

Lipid Species ◽

And Control

16 Background: The commonest type of kidney cancer is CCRCC. Kidney cancer is known to accumulate lipids and a detailed knowledge of the lipid species present in these tumors could lead to a better understanding of the underlying aberrant metabolic pathways and suggest possible treatment strategies. We attempt to identify the lipidomic profile of CCRCC using a liquid chromatography MS-based approach (LC-MS). Methods: We utilized 6 fresh frozen representative samples of CCRCC and matching non-tumor areas of kidney from nephrectomy samples. Lipids and other non-polar cellular constituents were extracted from both CCRCC and control tissues by methyl-t-butyl ether /methanol. LC-MS based lipid profiling was performed on a Waters Q-ToF Premier MS coupled with Ultra Performance LC. The peak detection and alignment across all chromatograms were performed using the XCMS software (v 1.14.1, Scripps Center for Metabolomics). Statistical comparisons of the intensities of aligned peaks were performed using the XCMS-built-in Welch's t-test. The XCMS data was converted to log2 ratios (normal/tumor) in order to utilize the paired aspect of this data: each patient’s disease tissue was analyzed in conjunction with corresponding normal tissue. Significance was determined by controlling the family wise error rate (FWER) at 0.05 using a Bonferroni adjustment. All computations were performed using the R statistical software and the “limma” package. Results: The outcome of XCMS was converted to a table that contains fold change, p value and mass to charge ratio (m/z) for each peak, its corresponding retention time, and the integrated peak intensities from all samples. Controlling FWER at 0.05 using a Bonferroni scheme, we found eight statistically significant lipids. Preliminary attempts to identify the analytes included use of METLIN (Scripps Center for Metabolomics) and HMDB (Human metabolome database, Genome Alberta and Genome Canada) databases. The identified metabolites included phosphatidylcholines, cholesterol esters and triglycerides, as well as other lipid species. Conclusions: The lipid metabolomic profile varied significantly between CCRCC and control. Further studies are underway to confirm the identities and significance of the lipid species in detail.

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IgM and IgD B cell receptors differentially respond to endogenous antigens and control B cell fate

eLife ◽

10.7554/elife.35074 ◽

2018 ◽

Vol 7 ◽

Cited By ~ 23

Author(s):

Mark Noviski ◽

James L Mueller ◽

Anne Satterthwaite ◽

Lee Ann Garrett-Sinha ◽

Frank Brombacher ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Cell Fate ◽

Plasma Cells ◽

Autoreactive B Cells ◽

Binding Domains ◽

Bcr Signaling ◽

And Control

Naive B cells co-express two BCR isotypes, IgM and IgD, with identical antigen-binding domains but distinct constant regions. IgM but not IgD is downregulated on autoreactive B cells. Because these isotypes are presumed to be redundant, it is unknown how this could impose tolerance. We introduced the Nur77-eGFP reporter of BCR signaling into mice that express each BCR isotype alone. Despite signaling strongly in vitro, IgD is less sensitive than IgM to endogenous antigen in vivo and developmental fate decisions are skewed accordingly. IgD-only Lyn−/− B cells cannot generate autoantibodies and short-lived plasma cells (SLPCs) in vivo, a fate thought to be driven by intense BCR signaling induced by endogenous antigens. Similarly, IgD-only B cells generate normal germinal center, but impaired IgG1+ SLPC responses to T-dependent immunization. We propose a role for IgD in maintaining the quiescence of autoreactive B cells and restricting their differentiation into autoantibody secreting cells.

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Efficient generation of targeted large insertions in mouse embryos using 2C-HR-CRISPR

10.1101/204339 ◽

2017 ◽

Cited By ~ 2

Author(s):

Bin Gu ◽

Eszter Posfai ◽

Janet Rossant

Keyword(s):

Cell Fate ◽

Protein Function ◽

Rapid Assessment ◽

Mouse Genetics ◽

Mouse Embryos ◽

Open Chromatin ◽

Cell Stage ◽

Cell Fate Decisions ◽

Efficient Generation ◽

Cell Embryo

Rapid and efficient generation of large fragment targeted knock-in mouse models is still a major hurdle in mouse genetics. Here we developed 2C-HR-CRISPR, a highly efficient gene editing method based on introducing CRISPR reagents into mouse embryos at the 2-cell stage, taking advantage of the likely increase in HR efficiency during the long G2 phase and open chromatin structure of the 2-cell embryo. With 2C-HR-CRISPR and a modified biotin-streptavidin approach to localize repair templates to target sites, we rapidly targeted 20 endogenous genes that are expressed in mouse blastocysts with fluorescent reporters and generated reporter mouse lines. We showcase the first live triple-color blastocyst with all three lineages differentially reported. Additionally, we demonstrated efficient double targeting, enabling rapid assessment of the auxin-inducible degradation system for probing protein function in mouse embryos. These methods open up exciting avenues for exploring cell fate decisions in the blastocyst and later stages of development. We also suggest that 2C-HR-CRISPR can be a better alternative to random transgenesis by ensuring transgene insertions at defined ‘safe harbor’ sites.

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Fibronectin-Enriched Biomaterials, Biofunctionalization, and Proactivity: A Review

Applied Sciences ◽

10.3390/app112412111 ◽

2021 ◽

Vol 11 (24) ◽

pp. 12111

Author(s):

Carla Palomino-Durand ◽

Emmanuel Pauthe ◽

Adeline Gand

Keyword(s):

Cell Fate ◽

Tissue Repair ◽

Cell Attachment ◽

Advanced Materials ◽

Tissue Repair And Regeneration ◽

Biological Responses ◽

Cell Material Interactions ◽

And Control ◽

Implanted Device ◽

Adhesion Process

Modern innovation in reconstructive medicine implies the proposition of material-based strategies suitable for tissue repair and regeneration. The development of such systems necessitates the design of advanced materials and the control of their interactions with their surrounding cellular and molecular microenvironments. Biomaterials must actively engage cellular matter to direct and modulate biological responses at implant sites and beyond. Indeed, it is essential that a true dialogue exists between the implanted device and the cells. Biomaterial engineering implies the knowledge and control of cell fate considering the globality of the adhesion process, from initial cell attachment to differentiation. The extracellular matrix (ECM) represents a complex microenvironment able to meet these essential needs to establish a relationship between the material and the contacting cells. The ECM exhibits specific physical, chemical, and biochemical characteristics. Considering the complexity, heterogeneity, and versatility of ECM actors, fibronectin (Fn) has emerged among the ECM protagonists as the most pertinent representative key actor. The following review focuses on and synthesizes the research supporting the potential to use Fn in biomaterial functionalization to mimic the ECM and enhance cell–material interactions.

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Regulation of neurogenesis and gliogenesis by the matricellular protein CCN2 in the mouse retina

10.1101/2021.04.01.438112 ◽

2021 ◽

Author(s):

Golam Mohiuddin ◽

Genesis Lopez ◽

Jose Sinon ◽

M. Elizabeth Hartnett ◽

Anastasiia Bulakhova ◽

...

Keyword(s):

Glial Cell ◽

Cell Fate ◽

Matrix Protein ◽

Ex Vivo ◽

Mouse Genetics ◽

Extracellular Matrix Protein ◽

Retinal Cell ◽

Mouse Retina ◽

Matricellular Protein ◽

Retinal Progenitor

AbstractCellular communication network (CCN) 2 is an extracellular matrix protein with cell type- and context-dependent functions. Using a combination of mouse genetics and omic approaches, we show that CCN2 is expressed in early embryonic retinal progenitor cells (RPCs) and becomes restricted to fully differentiated Müller glial cells (MGCs) thereafter. Germline deletion of CCN2 in mice decreases BrdU labeling, reduces RPC pool, and impairs the competency of remaining RPCs to generate early and late born retinal cell types. Retinal hypocellularity and microphthalmia ensue. The transcriptomic changes associated with CCN2 inactivation include reduced marker and transcriptional regulator genes of retinal ganglion cells, photoreceptors and MGCs. Yap (Yes-associated protein), a singular node for transcriptional regulation of growth and differentiation genes, is also a target of CCN2 signals. In an organotypic model of ex vivo cultured embryonic retinas, CCN2 and YAP immunoreactivity signals overlap. Lentivirus-mediated YAP expression in CCN2-deficient retinal explants increases the number of differentiating Sox9-positive MGCs. Taken together, our data indicate that CCN2 controls the proliferative and differentiation potentials of RPCs ultimately endowing, a subpopulation thereof, with Müller glial cell fate.Summary statementA CCN2-YAP regulatory axis controls retinal progenitor cell growth and lineage commitment to neuronal and glial cell fates.

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"Like a porn movie": Notes on boundaries and bodies that matter in male sex clubs

Vibrant Virtual Brazilian Anthropology ◽

10.1590/s1809-43412012000100005 ◽

2012 ◽

Vol 9 (1) ◽

pp. 131-153

Author(s):

Camilo Braz

Keyword(s):

Alcohol Consumption ◽

Condom Use ◽

Sao Paulo ◽

Sexual Practices ◽

Ethnographic Research ◽

São Paulo ◽

Male Sex ◽

And Control ◽

Drug And Alcohol ◽

The City

This article is based on ethnographic research carried out in male sex clubs in São Paulo between 2006 and 2008. Drawing on interviews conducted with club-goers and club owners, it discusses the recent segmentation of the sexual leisure market for men in the city, and the processes by which stereotypes and characteristics associated with virility are valued and performed. The possible effects of these processes on subjectivity constitution are also investigated. In sex clubs, sexual practices considered borderline, such as fist-fucking and other practices associated with BDSM, are material for specific and refined learning. The data gathered from interviews show that condom use and drug and alcohol consumption are subject to a sort of surveillance and control, especially when it comes to their questions of 'excess'. This control provides sex clubs with a sense of legitimacy, making them part of a viable erotic market. The intention here is to demonstrate the analytical interest of this control in the context of the construction of subjects and bodies that matter in these venues. Like practices which evoke control or loss thereof, bodies and clubs need to have their excesses checked so that they are intelligible and desirable.

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