Reorganization of Vegetal Cortex Microtubules and Its Role in Axis Induction in the Early Vertebrate Embryo

In vertebrate species, induction of the embryonic axis is initiated by the transport of maternally supplied determinants, initially localized to the vegetal pole of the egg, toward the prospective organizer in the animal region. This transport process remains incompletely understood. Here, we review studies involving embryonic manipulations, visualization, and functional analysis of the cytoskeleton and loss- and gain-of-function conditions, which provide insights in this process. Transport of dorsal determinants requires cytoskeletal reorganization of a vegetal array of microtubules, microtubule motors, and an off-center movement of the vegetal cortex with respect to the inner egg core, a so-called cortical rotation. Additional mechanisms may be used in specific systems, such as a more general animally directed movement found in the teleost embryo. Initial polarity of the microtubule movement depends on early asymmetries, which are amplified by the movement of the outermost cortex. An interplay between microtubule organization and axis specification has also been reported in other animal species. Altogether, these studies show the importance of cytoskeletal dynamic changes, such as bundling, force-inducing motor activity, and regulated cytoskeletal growth, for the intracellular transport of maternally inherited factors to their site of action in the zygote

Germ Cell Determinant Transmission, Segregation, and Function in the Zebrafish Embryo

Animals specify primordial germ cells (PGCs) in two alternate modes: preformation and epigenesis. Epigenesis relies on signal transduction from the surrounding tissues to instruct a group of cells to acquire PGC identity. Preformation, thought to be the more derived PGC specification mode, is instead based on the maternal inheritance of germ cell-determining factors. We use the zebrafish as a model system, in which PGCs are specified through maternal inheritance of germ plasm, to study this process in vertebrates. In zebrafish, maternally inherited germ plasm ribonucleoparticles (RNPs) have co-opted the cytoskeletal machinery to reach progressive levels of multimerization, resulting in the formation of four large masses of aggregated germ plasm RNPs. At later stages, germ plasm masses continue to use components of the cell division machinery, such as the spindles, centrosomes, and/or subcellular organelles to segregate asymmetrically during cell division and subsequently induce germ cell fate. This chapter discusses the current knowledge of germ cell specification focusing on the zebrafish as a model system. We also provide a comparative analysis of the mechanism for germ plasm RNP segregation in zebrafish versus other known vertebrate systems of germ cell preformation, such as in amphibian and avian models

Localized Products of futile cycle/ lrmp Promote Centrosome-Nucleus Attachment in the Zebrafish Zygote

SummaryBackgroundThe centrosome has a well-established role as a microtubule organizer during mitosis and cytokinesis. In addition, it facilitates the union of parental haploid genomes following fertilization by nucleating a microtubule aster along which the female pronucleus migrates toward the male pronucleus. Stable associations between the sperm aster and the pronuclei are essential during this directed movement.ResultsOur studies reveal that the zebrafish gene futile cycle (fue) is required in the zygote for male pronucleus-centrosome attachment and female pronuclear migration. We show that fue encodes a novel, maternally-provided long form of lymphoid-restricted membrane protein (lrmp), a vertebrate-specific gene of unknown function. Both maternal lrmp messenger RNA (mRNA) and protein are highly localized in the zygote, in a largely overlapping pattern at nuclear membranes, centrosomes, and spindles. Truncated Lrmp-EGFP fusion proteins identified subcellular targeting signals in the C terminus of Lrmp; however, endogenous mRNA localization is likely important to ensure strict spatial expression of the protein. Localization of both Lrmp protein and lrmp RNA is defective in fue mutant embryos, indicating that correct targeting of lrmp gene products is dependent on Lrmp function.ConclusionsLrmp is a conserved vertebrate gene whose maternally inherited products are essential for nucleus-centrosome attachment and pronuclear congression during fertilization. Precise subcellular localization of lrmp products also suggests a requirement for strict spatiotemporal regulation of their function in the early embryo

Chromatin regulators and the determination of embryonic polarity in Drosphila

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Biology, 1994.Includes bibliographical references.by Francisco Pelegri.Ph.D

A mutation in the zebrafish maternal-effect gene nebel affects furrow formation and vasa RNA localization

AbstractBackground: In many animals, embryonic patterning depends on a careful interplay between cell division and the segregation of localized cellular components. Both of these processes in turn rely on cytoskeletal elements and motor proteins. A type of localized cellular component found in most animals is the germ plasm, a specialized region of cytoplasm that specifies the germ-cell fate. The gene vasa has been shown in Drosophila to encode an essential component of the germ plasm and is thought to have a similar function in other organisms. In the zebrafish embryo, the vasa RNA is localized to the furrows of the early cellular divisions.Results: We identified the gene nebel in a pilot screen for zebrafish maternal-effect mutations. Embryos from females homozygous for a mutation in nebel exhibit defects in cell adhesion. Our analysis provides genetic evidence for a function of the microtubule array that normally develops at the furrow in the deposition of adhesive membrane at the cleavage plane. In addition, nebel mutant embryos show defects in the early localization of vasa RNA. The vasa RNA localization phenotype could be mimicked with microtubule-inhibiting drugs, and confocal microscopy suggests an interaction between microtubules and vasa-RNA-containing aggregates.Conclusions:Our data support two functions for the microtubule reorganization at the furrow, one for the exocytosis of adhesive membrane, and another for the translocation of vasa RNA along the forming furrow

Sovereign genes: wildlife conservation, genetic preservation, and Indigenous data sovereignty

The application of conservation genetics to wildlife preservation efforts are ongoing and promising. These involve the mobilization of a toolkit that ranges from monitoring the genetic diversity of rare species to more ambitious experiments in repopulating species experiencing genetic bottlenecks. All such efforts are predicated upon the deliberate and thoughtful preservation of existing genetic diversity. The history of genetic collection and conservation, however, for medical and health applications, is one that has repeatedly fallen into colonial habits, violated Indigenous sovereignty, and sown distrust with Indigenous communities. Given the importance of Indigenous communities in the future of wildlife conservation, the future of conservation genetics is best assured through the honoring of Indigenous Data Sovereignty. This paper reviews the stakes of this question, reflects on the status of recent conservation genetics efforts relative to the question of Indigenous sovereignty, and lays out a preliminary set of principles for collaborative work on wildlife conservation employing genetic tools

The Maternal-Effect Gene cellular island Encodes Aurora B Kinase and Is Essential for Furrow Formation in the Early Zebrafish Embryo

Females homozygous for a mutation in cellular island (cei) produce embryos with defects in cytokinesis during early development. Analysis of the cytoskeletal events associated with furrow formation reveal that these defects include a general delay in furrow initiation as well as a complete failure to form furrow-associated structures in distal regions of the blastodisc. A linkage mapping-based candidate gene approach, including transgenic rescue, shows that cei encodes the zebrafish Aurora B kinase homologue. Genetic complementation analysis between the cei mutation and aurB zygotic lethal mutations corroborate gene assignment and reveal a complex nature of the maternal-effect cei allele, which appears to preferentially affect a function important for cytokinesis in the early blastomeres. Surprisingly, in cei mutant embryos a short yet otherwise normal furrow forms in the center of the blastodisc. Furrow formation is absent throughout the width of the blastodisc in cei mutant embryos additionally mutant for futile cycle, which lack a spindle apparatus, showing that the residual furrow signal present in cei mutants is derived from the mitotic spindle. Our analysis suggests that partially redundant signals derived from the spindle and astral apparatus mediate furrow formation in medial and distal regions of the early embryonic blastomeres, respectively, possibly as a spatial specialization to achieve furrow formation in these large cells. In addition, our data also suggest a role for Cei/AurB function in the reorganization of the furrow-associated microtubules in both early cleavage- and somite-stage embryos. In accordance with the requirement for cei/aurB in furrow induction in the early cleavage embryo, germ plasm recruitment to the forming furrow is also affected in embryos lacking normal cei/aurB function

Intended Consequences Statement in Conservation Science and Practice

As the biodiversity crisis accelerates, the stakes are higher for threatened plants and animals. Rebuilding the health of our planet will require addressing underlying threats at many scales, including habitat loss and climate change. Conservation interventions such as habitat protection, management, restoration, predator control, trans location, genetic rescue, and biological control have the potential to help threatened or endangered species avert extinction. These existing, well-tested methods can be complemented and augmented by more frequent and faster adoption of new technologies, such as powerful new genetic tools. In addition, synthetic biology might offer solutions to currently intractable conservation problems. We believe that conservation needs to be bold and clear-eyed in this moment of great urgency

Calcium signaling in vertebrate embryonic patterning and morphogenesis

AbstractSignaling pathways that rely on the controlled release and/or accumulation of calcium ions are important in a variety of developmental events in the vertebrate embryo, affecting cell fate specification and morphogenesis. One such major developmentally important pathway is the Wnt/calcium signaling pathway, which, through its antagonism of Wnt/β-catenin signaling, appears to regulate the formation of the early embryonic organizer. In addition, the Wnt/calcium pathway shares components with another non-canonical Wnt pathway involved in planar cell polarity, suggesting that these two pathways form a loose network involved in polarized cell migratory movements that fashion the vertebrate body plan. Furthermore, left–right axis determination, neural induction and somite formation also display dynamic calcium release, which may be critical in these patterning events. Finally, we summarize recent evidence that propose a role for calcium signaling in stem cell biology and human developmental disorders