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International Journal of Developmental Biology

Puspendra Saswat Mahapatra, Renu Singh, Kuldeep Kumar, Nihar Ranjan Sahoo, Pranjali Agarwal, Bhabesh Mili, Kinsuk Das, Mihir Sarkar, Subrat Kumar Bhanja, B-C Das, Sujoy Kumar Dhara, Sadhan Bag
Generation of pluripotent stem cells by reprogramming somatic cells of quality animals have numerous potential applications in agricultural and biomedical sciences. Unfortunately, till now, reprogramming of buffalo fetal fibroblast cells (bFFs) has been very low despite intensive effort. Here, we attempted to enhance the reprogramming efficiency by using HDAC inhibitor valproic acid (VPA) in bFFs transfected with pLentG-KOSM pseudo virus carrying mouse specific pluripotent genes. FACS analysis revealed that VPA treatment significantly increased (p < 0...
August 16, 2016: International Journal of Developmental Biology
Rakhee Lohia, Punita Jain, Mukul Jain, Pradeep Kumar Burma, Anju Shrivastava, Shweta Saran
Sirtuins belong to class III histone deacetylases and require NAD (+)for its activity. They relate to nutritional state of the cell and directly link cellular metabolic signalling to the state of protein post-translational modifications. Sirtuins play an important role in healthy aging, longevity, age-related diseases as well as the cell survival mechanism like autophagy. Here, we investigate the functions of Dictyostelium discoideum Sir2D that show similarity to human SIRT1. This gene is expressed throughout growth and development...
August 16, 2016: International Journal of Developmental Biology
Liya He, Jiao Feng, Sha Lu, Zhiwen Chen, Chunmei Chen, Ya He, Xiuwen Yi, Liyan Xi
Transferring DNA into cells is an essential method of research on molecular cloning and gene function. As the molecular biology and materials physics develop, more and more new transformation methods have been applied in mammalian cells. Some techniques have been successfully developed for several types of fungi, but their efficiencies are extremely low. To better study on functional genes of fungi, and to improve the characteristics of fungi in an easy, safe and reliable way, many research are made to effectively develop such technologies in a wide variety of species and to increase the efficiency and reproducibility of genetic transformation...
August 16, 2016: International Journal of Developmental Biology
Sophie Khazanov, Yael Paz, Amit Hefetz, Ben Jerry Gonzales, Yaara Netser, Abed AlFatah Mansour, Nissim Ben-Arie
During embryonic development of the central nervous system (CNS), the expression of the bHLH transcription factor Nato3 (Ferd3l) is unique and restricted to the floor plate of the neural tube. In mice lacking Nato3 the floor plate cells of the spinal cord do not fully maturate, whereas in the midbrain floor plate, progenitors lose some neurogenic activity, giving rise to a reduced population of dopaminergic neurons. Since the floor plate is considered to be disintegrated at the time of birth, Nato3 expression was never tested postnatally and in adult mice...
August 16, 2016: International Journal of Developmental Biology
Gary S McDowell, Anna Philpott
The small protein modifier, ubiquitin, can be covalently attached to proteins in the process of ubiquitylation, resulting in a variety of functional outcomes. In particular, the most commonly-associated and well-studied fate for proteins modified with ubiquitin is their ultimate destruction: degradation by the 26S proteasome via the ubiquitin-proteasome system, or digestion in lysosomes by proteolytic enzymes. From the earliest days of ubiquitylation research, a reliable and versatile "cell-in-a-test-tube" system has been employed in the form of cytoplasmic extracts from the eggs and embryos of the frog Xenopus laevis...
August 16, 2016: International Journal of Developmental Biology
Dong-Hyeon Shin, Joung-Woo Hong
It remains unclear how a limited amount of maternal transcription factor Dorsal (Dl) directs broad expression of short gastrulation (sog) throughout the presumptive neurogenic ectoderm in the Drosophila early embryo. Here, we present evidence that the sog shadow enhancer employs dual modes of transcriptional synergy to produce this broad pattern. Bioinformatics analyses indicated that a minimal enhancer region, systematically mapped in vivo, contains five Dl-, three Zelda (Zld)-, and three Bicoid (Bcd)-binding sites; four of these five Dl-binding sites are closed linked to two Zld- and two Bcd-binding sites...
August 16, 2016: International Journal of Developmental Biology
Alexander A Tokmakov, Tetsushi Iwasaki, Ken-Ichi Sato, Shinji Kamada
Differentiated somatic cells and nuclei can be reprogrammed to a pluripotent undifferentiated state in the cytoplasm of oocytes and eggs. The ability of the gamete cells to induce reprogramming is not species-specific, so the extracts prepared from the oocytes and eggs of the African clawed frog Xenopus laevis can reprogram somatic mammalian cells. Thus, Xenopus egg extract-mediated reprogramming may constitute an alternative or complement other experimental reprogramming approaches, such as nuclear transfer, cell fusion, and transcription factor transduction...
June 1, 2016: International Journal of Developmental Biology
Francesco De Carli, Vincent Gaggioli, Gaël A Millot, Olivier Hyrien
DNA combing is a standard technique to map DNA replication at the single molecule level. Typically, replicating DNA is metabolically labelled with nucleoside or nucleotide analogs, purified, stretched on coverslips and treated with fluorescent antibodies to reveal tracts of newly synthesized DNA. Fibres containing a locus of interest can then be identified by fluorescent in situ hybridization (FISH) with DNA probes. These steps are complex and the throughput is low. Here, we describe a simpler, antibody-free method to reveal replication tracts and identify the locus of origin of combed DNA replication intermediates...
May 20, 2016: International Journal of Developmental Biology
Amin S M Salehi, Conner C Earl, Christina Muhlestein, Bradley C Bundy
Cell-free protein synthesis has been around for decades but never been so close to becoming a robust tool for the production of biotherapeutics. In this review, we focus on how Escherichia coli-based cellfree protein synthesis can be modified in various ways to produce challenging, complex anticancer biotherapeutics. Here we report progresses in extract preparation and its relation to cell-free cancer research. The future prospects of cell-free technology and its potentials in various areas of cancer therapeutics production are also highlighted...
May 19, 2016: International Journal of Developmental Biology
Steven Cupello, Christine Richardson, Shan Yan
In response to a variety of DNA replication stress or DNA damaging agents, the DNA damage response (DDR) pathways are triggered for cells to coordinate DNA repair, cell cycle checkpoints, apoptosis, and senescence. Cell-free Xenopus egg extracts, derived from the eggs of African clawed frogs (Xenopus laevis), have been widely used for studies concerning DDR pathways. In this review, we focus on how different experimental systems have been established using Xenopus egg extracts to investigate the DDR pathways that are activated in response to DNA replication stress, double-strand breaks (DSBs), inter-strand crosslinks (ICLs), and oxidative stress...
2016: International Journal of Developmental Biology
Dominic Poccia
Typically sperm nuclei are genetically inert and contain extremely compacted chromatin. Following fertilization, the first steps in their conversion to somatic nuclei (male pronuclei) which will support further development involve chromatin decondensation and the formation of a new nuclear envelope. We have studied the reactivation of sea urchin sperm nuclei in a cell-free system derived from homogenates of activated sea urchin egg cytoplasm. The cell-free system has provided several novel insights including requirements for sperm-specific histone phosphorylation on N- and C-terminal extensions and disassembly of the sperm nuclear lamina for decondensation, the utilization of remnant regions of the sperm nuclear envelope to direct polarized binding and fusion of egg membranes to form the new nuclear envelope, and a role for phosphoinositide metabolism in initiation of membrane fusion through binding of a minor membrane fraction enriched in PtdIns (4,5)P2, PLCγ and SFK1 which locally produces a fusigenic lipid, diacylglycerol...
April 6, 2016: International Journal of Developmental Biology
Wei-Lin Wang, David Shechter
Chromatin, primarily a complex of DNA and histone proteins, is the physiological form of the genome. Chromatin is generally repressive for transcription and other information transactions that occur on DNA. A wealth of post-translational modifications on canonical histones and histone variants encode regulatory information to recruit or repel effector proteins on chromatin, promoting and further repressing transcription and thereby form the basis of epigenetic information. During metazoan oogenesis, large quantities of histone proteins are synthesized and stored in preparation for the rapid early cell cycles of development and to elicit maternal control of chromatin assembly pathways...
2016: International Journal of Developmental Biology
Mateusz Debowski, Mohammed El Dika, Jacek Malejczyk, Robert Zdanowski, Claude Prigent, Jean-Pierre Tassan, Malgorzata Kloc, Miroslaw Lachowicz, Jacek Z Kubiak
During the cell cycle, cyclin dependent kinase 1 (CDK1) and protein phosphatase 2A (PP2A) play major roles in the regulation of mitosis. CDK1 phosphorylates a series of substrates triggering M-phase entry. Most of these substrates are dephosphorylated by PP2A. To allow phosphorylation of CDK1 substrates, PP2A is progressively inactivated upon M-phase entry. We have shown previously that the interplay between these two activities determines the timing of M-phase entry. Slight diminution of CDK1 activity by the RO3306 inhibitor delays M-phase entry in a dose-dependent manner in Xenopus embryo cell-free extract, while reduction of PP2A activity by OA inhibitor accelerates this process also in a dose-dependent manner...
2016: International Journal of Developmental Biology
Predrag Jevtić, Ana Milunović-Jevtić, Matthew R Dilsaver, Jesse C Gatlin, Daniel L Levy
Striking size variations are prominent throughout biology, at the organismal, cellular, and subcellular levels. Important fundamental questions concern organelle size regulation and how organelle size is regulated relative to cell size, also known as scaling. Uncovering mechanisms of organelle size regulation will inform the functional significance of size as well as the implications of misregulated size, for instance in the case of nuclear enlargement in cancer. Xenopus egg and embryo extracts are powerful cell-free systems that have been utilized extensively for mechanistic and functional studies of various organelles and subcellular structures...
2016: International Journal of Developmental Biology
Takashi Onikubo, David Shechter
Chromatin is the complex of DNA and histone proteins that is the physiological form of the eukaryotic genome. Chromatin is generally repressive for transcription, especially so during early metazoan development when maternal factors are explicitly in control of new zygotic gene expression. In the important model organism Xenopus laevis, maturing oocytes are transcriptionally active with reduced rates of chromatin assembly, while laid eggs and fertilized embryos have robust rates of chromatin assembly and are transcriptionally repressed...
2016: International Journal of Developmental Biology
Jacek Z Kubiak, Claude Prigent
The aim of this short review is to describe the contribution of Xenopus laevis egg extracts to the discovery and understanding of the regulation and function of the serine/threonine kinase Aurora-A. The power of these extracts to recapitulate cell cycle events makes them a precious tool to decipher complex biological processes at the molecular level, including the mechanisms that affect Aurora-A (post-translational modifications) and mechanisms in which Aurora-A plays a crucial role (bipolar spindle assembly)...
2016: International Journal of Developmental Biology
Suzanne Vigneron, Perle Robert, Khaled Hached, Lena Sundermann, Sophie Charrasse, Jean-Claude Labbé, Anna Castro, Thierry Lorca
Entry into mitosis requires the coordinated activation of various protein kinases and phosphatases that together activate sequential signaling pathways allowing entry, progression and exit of mitosis. The limiting step is thought to be the activation of the mitotic Cdk1-cyclin B kinase. However, this model has recently evolved with new data showing that in addition to the Cdk1-cyclin B complex, Greatwall (Gwl) kinase is also required to enter into and maintain mitosis. This new concept proposes that entry into mitosis is now based on the combined activation of both kinases Cdk1-cyclin B and Gwl, the former promoting massive phosphorylation of mitotic substrates and the latter inhibiting PP2A-B55 phosphatase responsible for dephosphorylation of these substrates...
2016: International Journal of Developmental Biology
Vincenzo Sannino, Arun M Kolinjivadi, Giorgio Baldi, Vincenzo Costanzo
The correct duplication of genetic information is essential to maintain genome stability, which is lost in cancer cells. Replication fork integrity is ensured by a number of DNA metabolism proteins that assist replication of chromatin regions difficult to replicate due to their intrinsic DNA sequence composition, coordinate repair of DNA molecules resulting from aberrant replication events or protect replication forks in the presence of lesions impairing their progression. Some DNA metabolism genes involved in DNA repair are essential in higher eukaryotes even in unchallenged conditions, suggesting the existence of biological processes requiring these specialized functions in organisms with complex genomes...
2016: International Journal of Developmental Biology
J Julian Blow, Ronald A Laskey
Here we discuss the important contributions that cell-free extracts have made to the study of complex biological processes. We provide a brief history of how cell-free extracts of frog eggs were developed to avoid many of the problems that can arise from the dilution and mixing of cellular components that typically occur when cell-free extracts are prepared. We briefly describe how Xenopus egg extracts have been fundamental to the study of many important cellular processes including DNA replication, cell cycle progression, nuclear protein import, nuclear assembly and chromosome organisation...
2016: International Journal of Developmental Biology
Jacek Z Kubiak, Takeo Kishimoto
Professor Takeo Kishimoto's research has an enormous impact on the cell cycle field. Although his favorite model has always been a starfish oocyte, he has used many other model organisms in his research. Cell-free extracts have been wildly used in his laboratory as a very useful tool to answer cell cycle research questions. Recently, professor Kishimoto discovered the identity of the M-phase promoting factor (MPF) that was thought for years to be cyclin-dependent kinase 1 (CDK1). However, Takeo Kishimoto found that MPF consists in fact of two kinases: CDK1 and Greatwall kinase...
2016: International Journal of Developmental Biology
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