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Results for “Developmental Biology”

Search indexed bioRxiv preprints in genomics, neuroscience, cell biology and bioinformatics. Read source abstracts and check manuscript versions; preprints are not peer reviewed.

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At least 253 records · Page 14Linked to original sources

bioRxiv2016

PRDX6 inhibits neurogenesis of neural precursor cells through downregulation of wdfy1 mediated TLR4 signal

Impaired neurogenesis has been associated with brain disorders. The role of peroxiredoxin 6 (PRDX6) in the neurodegenerative diseases is very controversial. To demonstrate the role of PRDX6 in neurogenesis, we compared neurogenesis ability and studied the molecular mechanisms. It was found that the neurogenesis of neural stem cells and expression of the marker protein were lowered in PRDX6 Tg-mice compared with non-tg mice. Moreover, the expression of wdfy1 was dramatically decreased in PRDX6-Tg mice, also, we observed that wdfy1 siRNA decreases the differentiation ability of primary neural stem cells to astrocyte and neuronal cells as well as PC12 cells. However, knockdown of PRDX6 recovered neurogenesis in the brain of PRDX6-Tg mice as well as PC-12 cells. We also showed that TLR4 was dramatically reduced in PRDX6 Tg mice as well as PC-12 cells and PRDX6 overexpression reduced neurogenesis was rescued after treatment of TLR4 siRNA. We further found that reduced TLR4 expression and neurogenesis was reversed in the neuron from PRDX6-Tg mice as well as PC12 cells by introduction of wdfy1 plasmid. Moreover, TLR4 siRNA reduced neurogenesis and wdfy1 expression. This study indicated that PRDX6 inhibits neurogenesis of neural precursor cells through TLR4 dependent downregulation of wdfy1.

developmental biology↗

bioRxiv2016

DNA-polymerase guided elimination of paternal mitochondrial genomes:An escape-proof obstacle to their transmission

Mitochondrial DNA is predominantly inherited from only one parent. In animals this is usually the mother. This program is not in the interest of the paternal mitochondrial genome whose potential to contribute to future generations is restricted. However, in a dramatic example of genetic conflict, nuclear programs ensure the outcome. Two large mitochondria extend the length of Drosophila sperm tails. The hundreds of nucleoids in these mitochondria vanish during spermatogenesis eliminating their potential for transmission. Our previous work showed that mutational inactivation of EndoG, a nuclear encoded mitochondrial endonuclease, slows elimination of mitochondrial genomes. Here, we show that knockdown of the nuclearly encoded mitochondrial DNA polymerase, Tamas, produces a much more complete block of mtDNA loss. Recruitment of Tamas to the nucleoid at the time of its disappearance suggests a direct contribution to the elimination, but the 3'-exonuclease function of the polymerase is not needed. While DNA elimination is a surprising function for DNA polymerase, its use to restrict paternal genomes provides a strategy that cannot easily be evaded by the mitochondrial genome without compromising its replication.

developmental biology↗

bioRxiv2016

Pulse of α-2-macroglobulin and lipocalin-1 in the pregnant uterus of European polecats (Mustela putorius) at the time of implantation

Uterine secretory proteins protect the uterus and conceptuses against infection, facilitate implantation, control cellular damage resulting from implantation, and supply embryos with nutrients. The early conceptus of the European polecat (Mustela putorius) grows and develops free in the uterus until implanting at about 12 days after mating. Using a proteomics approach we found that the proteins appearing in the uterus leading up to and including the time of implantation changed dramatically with time. Several of the proteins identified have been found in pregnant uteri of other placental mammals, such as 1-antitrypsin, serum albumin, lactoferrin, cathepsin L1, uteroferrin, and ectonucleotide pyrophosphatase. The broad-spectrum proteinase inhibitor 2-macroglobulin rose from relatively low abundance initially to dominate the protein profile by the time of implantation. Its functions may be to limit damage caused by the release of proteinases during implantation, and to control other processes around the site of implantation. Lipocalin-1 (also known as tear lipocalin) has not previously been recorded as a uterine secretion in pregnancy, and also increased substantially in concentration. If polecat lipocalin-1 has similar biochemical properties to the human form, then it may have a combined function in transporting or scavenging lipids, and antimicrobial activities. The changes in the uterine secretory proteome of Euroepan polecats may be similar in those species of mustelid that engage in embryonic diapause, but possibly only following reactivation of the embryo.

developmental biology↗

bioRxiv2016

DIV-1/PolA2 Promotes GLP-1/Notch-Mediated Cellular Events in Caenorhabditis elegans

Notch signaling is a highly conserved cell signaling system in most multicellular organisms and plays a critical role in animal development. In various tumor cells, Notch signaling is elevated and has been considered as an important target in cancer treatments. In C. elegans, GLP-1 (one of two C. elegans Notch receptors) activity is required for cell fate specification in germline and somatic tissues. In this study, we have identified div-1 gene as a positive regulator for GLP-1/Notch-mediated cellular events. C. elegans div-1 encodes the B subunit of the DNA polymerase alpha-primase complex and is highly expressed in proliferative germ cells. Functional analyses demonstrated that i) DIV-1 is required for the robust proliferation typical of the germline, ii) loss of DIV-1 enhances and suppresses specific phenotypes that are associated with reduced and elevated GLP-1/Notch activity in germline and somatic tissues, and iii) DIV-1 works together with FBF/PUF proteins, downstream regulators of GLP-1/Notch signaling, to promote germline stem cell (GSC) maintenance and germline proliferation. To maintain GSCs and proliferative cell fate, GLP-1/Notch activity must remain above a threshold for proliferation/differentiation decision. Our results propose that DIV-1 may control the level of threshold for GLP-1/Notch-mediated germline proliferation. PolA2, a mammalian homolog of the C. elegans DIV-1, has been emerged as a therapeutic target for non-small cell lung cancer (NSCLC). Notably, Notch signaling is altered in approximately one third of NSCLCs. Therefore, the discovery of the DIV-1 effect on GLP-1/Notch-mediated cellular events has implications for our understanding of vertebrate PolA2 protein and its influence on stem cell maintenance and tumorigenesis.

developmental biology↗

bioRxiv2016

A new coactivator complex required for retinoic acid-dependent regulation of embryonic symmetry

Bilateral symmetry is a striking feature of the vertebrate body plan organization. Vertebral precursors, called somites, provide one of the best illustrations of embryonic symmetry. Maintenance of somitogenesis symmetry requires Retinoic acid (RA) and its coactivator Rere/Atrophin2. Here, using a proteomic approach we identify a protein complex, containing Wdr5, Hdac1, Hdac2 and Rere (named WHHERE), which regulates RA signalling and controls embryonic symmetry. We demonstrate that Wdr5, Hdac1 and Hdac2 are required for RA signalling in vitro and in vivo. Mouse mutants for Wdr5 and Hdac1 exhibit asymmetrical somite formation characteristic of RA-deficiency. We also identify the Rere-binding histone methyltransferase Ehmt2/G9a, as a RA coactivator controlling somite symmetry. Upon RA treatment, WHHERE and Ehmt2 become enriched at RA target genes to promote RNA Polymerase II recruitment. Our work identifies a novel protein complex linking key epigenetic regulators acting in the molecular control of embryonic bilateral symmetry.

developmental biology↗

bioRxiv2016

Germ granules prevent accumulation of somatic transcripts in the adult C. elegans germline

The germ cells of multicellular organisms protect their developmental potential through specialized mechanisms. A shared feature of germ cells from worms to humans is the presence of non-membrane-bound ribonucleoprotein organelles called germ granules. Depletion of germ granules in Caenorhabditis elegans (i.e., P granules) leads to sterility and in some germlines expression of the neuronal transgene unc-119::gfp and the muscle myosin MYO-3. Thus, P granules are hypothesized to maintain germ cell totipotency by preventing somatic development, although the mechanism by which P granules carry out this function is unknown. In this study, we performed transcriptome and single molecule RNA-FISH analyses of dissected P-granule-depleted gonads at different developmental stages. Our results demonstrate that P granules are necessary for adult germ cells to down-regulate spermatogenesis RNAs and to prevent the accumulation of numerous soma-specific RNAs. P-granule-depleted gonads that express the unc-119::gfp transgene also express many other genes involved in neuronal development and concomitantly lose expression of germ cell fate markers. Finally, we show that removal of either of two critical P-granule components, PGL-1 or GLH-1, is sufficient to cause germ cells to express UNC-119::GFP and MYO-3 and to display RNA accumulation defects similar to those observed after depletion of P granules. Our data identify P granules as critical modulators of the germline transcriptome and guardians of germ cell fate.

developmental biology↗

bioRxiv2016

foxQ2 evolved a key role in anterior head and central brain patterning in protostomes

Anterior patterning of animals is based on a set of highly conserved transcription factors but the interactions within the protostome anterior gene regulatory network (aGRN) remain enigmatic. Here, we identify the foxQ2 ortholog of the red flour beetle Tribolium castaneum as novel upstream component of the insect aGRN. It is required for the development of the labrum and higher order brain structures, namely the central complex and the mushroom bodies. We reveal Tc-foxQ2 interactions by RNAi and heat shock-mediated misexpression. Surprisingly, Tc-foxQ2 and Tc-six3 mutually activate each other forming a novel regulatory module at the top of the insect aGRN. Comparisons of our results with those of sea urchins and cnidarians suggest that foxQ2 has acquired functions in head and brain patterning during protostome evolution. Our findings expand the knowledge on foxQ2 gene function to include essential roles in epidermal development and central brain patterning.\n\nAuthor summaryThe development of the anterior most part of any animal embryo - for instance the brain of vertebrates and the head of insects - depends on a very similar set of genes present in all animals. This is true for the two major lineages of bilaterian animals, the deuterostomes (including sea urchin and humans) and protostomes (including annelids and insects) and the cnidarians (e.g. the sea anemone), which are representatives of more ancient animals. However, the interaction of these genes has been studied in deuterostomes and cnidarians but not in protostomes. Here, we present the first study the function of the gene foxQ2 in protostomes. We found that the gene acts at the top level of the genetic network and when its function is knocked down, the labrum (a part of the head) and higher order brain centers do not develop. This is in contrast to the other animal groups where foxQ2 appears to play a less central role. We conclude that foxQ2 has acquired additional functions in the course of evolution of protostomes.

developmental biology↗

bioRxiv2016

Effective Cell Immunoablation in Undisrupted Developing Avian Embryos

In birds the construction of germline chimeras by grafting exogenous primordial germ cells (PGCs) during embryonic development is feasible since they migrate to the gonads through the blood. Up to date, the efficiencies are highly variable, in part dependent on the destruction of endogenous PGCs in the recipient embryo. We show an almost complete ablation of the endogenous PGCs in stage X embryos using a baby rabbit serum (BRS), with previous cellular signaling by specific antibodies (SSEA1). The application of the treatments, either on epiblast or subgerminaly, produced the reduction of the PGCs in the embryos in a dose dependent manner. No malformations or damages were detected in the treated embryos. However, subgerminal injection of this cocktail produced a massive cellular destruction in all embryos. Therefore, sequential application is a selective and effective method to produce receptor embryos. Nevertheless, it can also be highly destructive if the mixture is applied locally, this could be useful in the treatment of malignancies.\n\nSUMMARY STATEMENTAn immunosurgery procedure is described that yields an almost complete ablation of primordial germ cells in early developing chick embryos, thus increasing the expected rates of chimerism when foreign PGCs are grafted onto these embryos

developmental biology↗

bioRxiv2016

Senescence gives insights into the morphogenetic evolution of anamniotes.

Summary statementWe report the presence of senescent cells in several transient structures in developing amphibian and teleost fish, suggesting novel mechanisms of morphogenesis that appeared early in vertebrate evolution.\n\nAbstractSenescence represents a mechanism to avoid undesired cell proliferation that plays a role in tumor suppression, wound healing and embryonic development. In order to gain insight on the evolution of senescence, we looked at its presence in developing axolotls (urodele amphibians) and in zebrafish (teleost fish), which are both anamniotes. Our data indicate that cellular senescence is present in various developing structures in axolotls (pronephros, olfactory nerve fascicles, lateral organs, gums) and in zebrafish (epithelium of the yolk sac and in the lower part of the gut). Senescence was particularly associated with transient structures (pronephros in axolotls & yolk sac in zebrafish) suggesting that it plays a role in the elimination of these tissues. Our data supports the notion that cellular senescence evolved early in vertebrate evolution to influence embryonic development.

developmental biology↗

bioRxiv2016

Zebrafish Nanog is not required in embryonic cells

SUMMARY STATEMENTThe study of nanog mutants reveals that Nanog is required only for extraembryonic tissue development, not in embryonic cells.\n\nABSTRACTThe role of the zebrafish transcription factor Nanog has been controversial. It has been suggested that Nanog is primarily required for the formation of the extraembryonic yolk syncytial layer (YSL) and only indirectly regulates gene expression in embryonic cells. By contrast, a more recent study has proposed that Nanog directly regulates transcription in embryonic cells during zygotic genome activation. To clarify the roles of Nanog, we performed a detailed analysis of zebrafish nanog mutants. While zygotic nanog mutants survive to adulthood, maternal-zygotic and maternal mutants exhibit developmental arrest at the blastula stage. In the absence of Nanog, the YSL fails to form and embryonic tissue detaches from the yolk. Zygotic transcription of a subset of embryonic genes is affected in nanog mutants but both the YSL and embryonic phenotype can be rescued by providing nanog mRNA in YSL precursors. Notably, nanog mutant cells transplanted into wild-type hosts proliferate and contribute to embryonic tissues from all germ layers. These results indicate that zebrafish Nanog is necessary for YSL formation but is not directly required for embryonic cell differentiation.

developmental biology↗

bioRxiv2016

Specific expression and function of the Six3 optix in serially homologous organs

Organ size and pattern results from the integration of two positional information systems. One global, encoded by the Hox genes, links organ type with position along the main body axis. Within specific organs, local information is conveyed by signaling molecules that regulate organ growth and pattern. The mesothoracic (T2) wing and the metathoracic (T3) haltere of Drosophila represent a paradigmatic example of this coordination. The Hox gene Ultrabithorax (Ubx), expressed in the developing T3, selects haltere identity by, among other processes, modulating the production and signaling efficiency of Dpp, a BMP2-like molecule that acts as a major regulator of size and pattern. Still, the mechanisms of the Hox-signal integration even in this well-studied system, are incomplete. Here we have investigated this issue by studying the expression and function of the Six3 transcription factor optix during the development of the Drosophila wing and haltere development. We find that in both organs Dpp defines the expression domain of optix through repression, and that the specific position of this domain in wing and haltere seems to reflect the differential signaling profile among these organs. We show that optix expression in wing and haltere primordia is conserved beyond Drosophila in other higher diptera. Despite the similar expression pattern, optix plays different roles in wing and haltere. In the wing, optix is required for the growth of the most anterior/proximal region (the \"marginal cell\") and for the correct formation of sensory structures along the proximal anterior wing margin. In contrast, in the haltere optix is necessary for the suppression of sensory bristles without any noticeable effect on organ growth. Therefore, optix shows an organ-specific function. Beyond dipterans, optix expression in the anterior wing has been shown also in butterflies. We propose that the ancestral role of optix might have been structural in the anterior wing. Once expressed in the wing, optix expression had been re-deployed for wing spot formation in other parts of the wing of Heliconius butterflies.

developmental biology↗

bioRxiv2016

Ras/ERK-signalling promotes tRNA synthesis and growth via the RNA polymerase III repressor Maf1 in Drosophila

The small G-protein Ras is a conserved regulator of cell and tissue growth. These effects of Ras are mediated largely through activation of a canonical RAF-MEK-ERK kinase cascade. An important challenge is to identify how this Ras/ERK pathway alters cellular metabolism to drive growth. Here we report on stimulation of RNA polymerase III (Pol III)-mediated tRNA synthesis as a growth effector of Ras/ERK signalling in Drosophila. We find that activation of Ras/ERK signalling promotes tRNA synthesis both in vivo and in cultured Drosophila S2 cells. We also show that Pol III function is required for Ras/ERK signalling to drive proliferation in both epithelial and stem cells in Drosophila tissues. We find that the transcription factor Myc is required but not sufficient for Ras-mediated stimulation of tRNA synthesis. Instead we show that the main way that Ras promotes Pol III function and tRNA synthesis is by inhibiting the nuclear localization and function of the Pol III repressor Maf1. We propose that inhibition of Maf1 and stimulation of tRNA synthesis is one way by which Ras signalling enhances protein synthesis to promote cell and tissue growth.

developmental biology↗

bioRxiv2016

Hoxa1 and TALE proteins display cross-regulatory interactions and form a combinatorial binding code on Hoxa1 targets

Hoxa1 has diverse functional roles in differentiation and development. We have identified and characterized properties of regions bound by Hoxa1 on a genome-wide basis in differentiating mouse ES cells. Hoxa1 bound regions are enriched for clusters of consensus binding motifs for Hox, Pbx and Meis and many display co-occupancy of Pbx and Meis. Pbx and Meis are members of the TALE family and genome-wide analysis of multiple TALE members (Pbx, Meis, TGIF, Prep1 and Prep2) show that nearly all Hoxa1 targets display occupancy of one or more TALE members. The combinatorial binding patterns of TALE proteins defines distinct classes of Hoxa1 targets and indicates a role as cofactors in modulating the specificity of Hox proteins. We also discovered extensive auto- and cross-regulatory interactions among the Hoxa1 and TALE genes. This study provides new insight into a regulatory network involving combinatorial interactions between Hoxa1 and TALE proteins.

developmental biology↗

bioRxiv2016

The architectural balance of the Ventral Nerve Cord depends on the level of JNK signaling activity

Central Nervous System (CNS) organogenesis is a complex process that obeys precise architectural rules. The impact that nervous system architecture may have on its functionality remains, however, relatively unexplored. To clarify this problem, we analyzed the development of the Drosophila embryonic Ventral Nerve Cord (VNC). VNC morphogenesis requires the tight control of Jun kinase (JNK) signaling, exerted in part via a negative feedback loop mediated by the dual specificity phosphatase Puckered, in a subset of pioneer neurons. Here we show that the JNK pathway autonomously regulates neuronal electrophysiological properties without affecting synaptic vesicle transport. Manipulating, during early embryogenesis, JNK signaling activity in pioneer neurons, directly influences their function as organizers of VNC architecture and, moreover, uncovers a role in the coordination of the embryonic motor circuitry that is required for hatching. Together, our data reveal critical links, mediated by the control of the JNK signaling cascade by Puckered, between the structural organization of the VNC and its functional optimization.

developmental biology↗

bioRxiv2016

Tracking the embryonic stem cell transition from ground state pluripotency

Mouse embryonic stem (ES) cells are locked into self-renewal by shielding from inductive cues. Release from this ground state in minimal conditions offers a system for delineating developmental progression from naive pluripotency. Here we examined the initial transition of ES cells. The population behaves asynchronously. We therefore exploited a short-half-life Rex1::GFP reporter to isolate cells either side of exit from naive status. Extinction of ES cell identity in single cells is acute. It occurs only after near-complete elimination of naive pluripotency factors, but precedes appearance of lineage specification markers. Cells newly departed from the ES cell state exhibit global transcriptome features consistent with features of early post-implantation epiblast and distinct from primed epiblast. They also exhibit a genome-wide increase in DNA methylation, intermediate between early and late epiblast. These findings are consistent with the proposition that naive cells transition to a discrete formative phase of pluripotency preparatory to lineage priming.\n\nHighlightsO_LIThe Rex1 destabilized GFP reporter demarcates naive pluripotency.\nC_LIO_LIExit from the naive state is asynchronous in the population.\nC_LIO_LITransition is relatively acute in individual cells and precedes lineage priming.\nC_LIO_LITranscriptome and DNA methylome reflect events in the pre-gastrulation embryo.\nC_LI

developmental biology↗

bioRxiv2016

The Dachsous/Fat/Four-jointed pathway implements axial long-range cell orientation

Despite a cumulative body of knowledge describing short-range cell interactions in morphogenetic processes, relatively little is known on the mechanism involved in the long-range spatial and temporal coordination of cells to build functional and structurally organized tissues. In particular, the attainment of a functionally optimized epithelia must require directional cues to instruct cell movements and cell orientations throughout the tissue field. In Drosophila, the adult epidermis of the abdominal segments is created de novo by the replacement of obsolete larval epidermal cells (LECs) by histoblasts (imaginal founder cells). As these proliferate, expand and fuse, they uniformly organize orienting on the surface along the antero-posterior axis. We found that the coordinated, axially oriented changes in shape of histoblasts respond to a dynamic, yet stereotyped redesign of the epithelial field mediated by the Dachsous/Fat/Four-jointed (Ds/Ft/Fj) pathway. The establishment and refinement of the expression gradients of the atypical cadherins Ds and Ft result in their axial polarization across cell interfaces and differential adhesiveness. We suggest that the role of Ds/Ft/Fj in long-range axially oriented planar cell alignment is a general function and that the regulation of the expression of its components would be crucial in the achievement of tissue uniformity in many other morphogenetic models or during tissue repair.

developmental biology↗

bioRxiv2016

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

Naive 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 naive 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 naive pluripotency.

developmental biology↗

bioRxiv2016

Blood flow directs arterial-venous remodeling through Notch activation and endothelial cell migration

Arteries and veins are formed independently by different types of endothelial cells (ECs). In vascular remodeling, arteries and veins become connected and some arteries become veins. It is unclear how ECs in transforming vessels change their type and how fates of individual vessels are determined. In embryonic trunk, vascular remodeling transforms arterial intersegmental vessels (ISVs) into a functional network of arteries and veins. We found that, once an ISV is connected to venous circulation, venous blood flow promotes upstream migration of ECs that results in displacement of arterial ECs by venous ECs, completing the transformation of this ISV into a vein without trans-differentiation of ECs. Arterial blood flow initiated in two neighboring ISVs prevents their transformation into veins by activating Notch signaling in ECs. Together, different responses of ECs to arterial and venous blood flow lead to the formation of a balanced network with equal numbers of arteries and veins.

developmental biology↗