Request pdf ooplasmic segregation in the zebrafish zygote and early embryo. We investigated the distribution of tubulin in zebrafish oocytes and embryos using. Ooplasmic reorganization is particularly pronounced in oocytes from birds, reptiles, worms, and fish, containing a mixture of yolk granules ygs. A new micromanipulation technique permitted the scrambling of the zygote cytoplasm.
After insemination, a wave of highly elevated free calcium crosses the egg with a peak velocity of 89 micronss. Sakai1 department of biology, tokyo metropolitan university, tokyo, japan ooplasmic segregation, which generally occurs following fertilization in fish. Preformation, thought to be the more derived pgc specification mode, is instead based on the maternal inheritance of germ celldetermining factors. Changes in tubulin protein distribution in zebrafish danio rerio. On the mechanism of ooplasmic segregation in singlecell.
A few hours later, gastrulation is initiated by invagination of. Biomolecules free fulltext the zebrafish perivitelline. A guide for the laboratory use of zebrafish danio rerio. Germ cell determinant transmission, segregation, and function. The myoplasm of ascidian eggs is a localized cytoplasmic region containing a unique cytoskeletal domain. Pattern of ooplasmic movements and transport pathways. Microtubulebased movements during ooplasmic segregation in the medaka fish egg oryzias latipes tamika a. This process is experimental and the keywords may be updated as the learning algorithm improves.
Using timelapse video microscopy, we found that ooplasmic inclusions in the fertilized medaka egg displayed two types of movement during ooplasmic segregation. Such interference had no effect on preimplantation development, and when zygotes with scrambled cytoplasm were transfered to the pseudopregnant females, normal and fertile mice were born. Miller curriculum vitae marine biological laboratory. A contracting actin network drags the ooplasm toward the animal pole, while comet tails push the yolk granules in the opposite direction. The first phase occurs shortly after fertilization. Reorganization of vegetal cortex microtubules and its role. Fertilization and ooplasmic movements in the ascidian egg.
Ooplasmic segregation, initiated in the stage v oocyte, led to the formation of ooplasmic domains rich in organelles, and ribonucleoproteins. As the sperm aster grows, er accumulates in its centrosomal area and along its astral rays. The role of a vegetal array of parallel microtubules in the directed transport of dorsal determinants at the zygote and cleavage. The role of a vegetal array of parallel microtubules in the directed transport of dorsal determinants at. To this end, the organization of the ooplasm and vitelloplasm. The aim of this work is to determine when and how ooplasmic segregation is initiated in the zebrafish egg. Studies on the ooplasmic segregation in the egg of the fish. Pronounced segregation of donor mitochondria introduced by.
Calcium transients accompany ooplasmic segregation in. Hardcopies of the 4th edition of the zebrafish book can be obtained for a nominal fee from the zebrafish international resource center, 5274 university of oregon, eugene, or 97403 usa. Germ plasm anchors at tight junctions in the early zebrafish. A detailed description of the early development of the egg of b. Structure of the yolk syncytial layer in teleostei and analogous. Bulk actin dynamics drive phase segregation in zebrafish. During ooplasmic segregation, the myoplasm moves first to the vegetal pole and then to the future posterior region of the fertilized egg, where it subsequently enters the. Studies on the ooplasmic segregation in the egg of the. The zygote blastodisc grows by accumulation of ooplasm.
In the teleost egg, the embryo is immersed in an extraembryonic fluid that fills the space between the embryo and the chorion and partially isolates it from the external environment, called the perivitelline fluid pvf. Actin networks in the bulk cytoplasm, rather than cortical dynamics, drive ooplasm segregation in zebrafish oocytes. On the mechanism of ooplasmic segregation in singlecell zebrafish embryos. If the migration speeds of the oil droplets starting from various levels of the. Segregation of maternal determinants within the oocyte constitutes the first step in embryo patterning. Pattern of ooplasmic movements and transport pathways by ricardo fuentes and juan fernandez abstract. May 30, 2019 segregation of maternal determinants within the oocyte constitutes the first step in embryo patterning.
Fluck franklin and marshall college, department of biology, lancaster, pennsylvania 17604 abstract. In zebrafish oocytes, extensive ooplasmic streaming leads to the segregation of ooplasm from yolk granules along the animalvegetal axis of the oocyte. The pvf allows the embryo to avoid dehydration, to maintain a safe osmotic balance and provides. Imaging of multicellular largescale rhythmic calcium. The measurements are made on a moving picture, taken by tim. The molecular biology of fertilization 1st edition. Using light microscopy techniques, we have studied the movements that follow fertilization in the denuded egg of the ascidian phallusia mammillata. Dynamic microtubules at the vegetal cortex predict the. Hardcopies of the 4th edition of the zebrafish book can be obtained for a nominal fee from the zebrafish international resource center, 5274. The observations are made chiefly on the living egg. Here, we show that this process does not rely on cortical actin reorganization, as previously thought, but instead on a cellcycledependent bulk actin polymerization wave traveling from the animal to the vegetal pole of the oocyte.
Jul, 2010 ooplasmic segregation in the zebrafish zygote and early embryo. During ooplasmic segregation, a specific cytoskeletal domain the myoplasm is translocated first to the vegetal pole vp and then to the posterior region of the zygote. Calcium signalling in extraembryonic domains during early teleost development. The spermatozoa can fertilize the eggs whether it is genetically active or not lee et al. The subject index, species index and the abundant illustrations add extra value to this long awaited book. Biology, husbandry, diseases, and research applications is a comprehensive work that fulfills a critical need for a thorough compilation of information on this species. Calcium transients accompany ooplasmic segregation in zebrafish embryos calcium transients accompany ooplasmic segregation in zebrafish embryos leung, christina f webb, sarah e miller, andrew l.
Calcium transients accompany ooplasmic segregation in zebrafish embryos. All embryos were collected by natural spawning and raised at 28. Ooplasmic segregation in fish egg droplets are assembled at the latitude of about 60. A guide for the laboratory use of zebrafish danio rerio, eugene, university of oregon press. Imaging of multicellular largescale rhythmic calcium waves. The text provides significant updates for working vivarium professionals maintaining zebrafish colonies, veterinarians responsible for.
Pattern of ooplasmic movements and transport pathways ricardo fuentes laboratory of developmental cell biology, department of biology, faculty of sciences, university of chile, santiago, chile. Zebrafish 101 for iacucs focus of monte matthews presentation institutional oversight training and education for both iacucs and zebrafish researchers when to have a protocol counting zebrafish focus of dr. The first manifested itself as the movement of many inclusions diameter 1. This is followed by a description of ectodermal, mesodermal and entodermal derivatives and the development of various organs. Ooplasmic segregation in the zebrafish zygote and early embryo. Polarity and reorganization of the endoplasmic reticulum. The zebrafish egg has been used extensively to understand the mechanisms of ooplasmic segregation. Ooplasmic segregation in the zebrafish zygote and early. We have studied egg activation and ooplasmic segregation in the ascidian phallusia mammillata using an imaging system that let us simultaneously monitor egg morphology and calciumdependent aequorin luminescence. Biomolecules free fulltext the zebrafish perivitelline fluid. Cytoplasmic transfer between human oocytes ooplasm donation and germinal vesicle transfer, which represents a complete cytoplasmic exchange, have been performed recently as a means to try to improve the outcome of assisted reproduction methods cohen et al.
A guide for the laboratory use of zebrafish brachydanio rerio. During stage 1b, pronuclei approached each other inset in fig. Ascidian ooplasmic segregation occurs in two major phases figure 1, right. In zebra ish oocytes, extensive ooplasmic streaming leads to the. Pronuclear transfers with dba2 and c57bl6 strains reveal that the ooplasm strainspecifically modifies paternal pronuclei postfertilization latham and. Converting the zebrafish oocyte to the fertilizable. Bulk actin dynamics drive phase segregation in zebra ish oocytes. Reorganization of cytoplasm in the zebrafish oocyte and egg during early steps of ooplasmic segregation by juan fernandez, macarena valladares, ricardo fuentes and andrea ubilla cite.
The zebrafish oocyte, egg and zygote are valuable material for the study of. Formation and dynamics of cytoplasmic domains and their genetic. Transcriptome dynamics in early zebrafish embryogenesis. Dachsous1b cadherin regulates actin and microtubule. A gastrulation center is described in ascidian eggs. This demonstrates that no morphogenetic factors are prelocalized in the egg cytoplasm. Pattern of ooplasmic movements and transport pathways patterns of cytoplasmic movements and organization of.
In zebrafish, maternally inherited germ plasm ribonucleoparticles rnps have. On the mechanism of ooplasmic segregation in single. Purchase the molecular biology of fertilization 1st edition. Nussleinvolhard and dahm, 2002, and obtaining adult normal specimens or haploid larvae that survived for about five days as it occurs. Jun 11, 2015 animals specify primordial germ cells pgcs in two alternate modes. Epigenesis relies on signal transduction from the surrounding tissues to instruct a group of cells to acquire pgc identity. Our data suggest that the segregation of ooplasm from ygs along the av axis of the zebrafish oocyte depends on the concerted activities of two distinct force generating mechanisms exerted by the bulk actomyosin cytoskeleton within the oocyte. It must be mentioned that the ring eventually reaches the vegetal pole after several hours, by the morula stage omitted from fig. In zebrafish embryos, the microtubule cytoskeleton has been shown to have. The process of ooplasmic segregation in all teleost species appears. We have previously demonstrated that transient localized elevations of free cytosolic calcium in the forming blastodisc bd and animal hemisphere cortex correlate with a morphological contraction that appears to play a major role in ooplasmic segregation in zygote. Bulk actin dynamics drive phase segregation in zebrafish oocytes.
In particular, our observations show that, as a result of a series of movements described below, the mitochondriarich subcortical myoplasm is split in two parts during the second phase of ooplasmic segregation. Patterns of free calcium in zebrafish embryos journal of. Germ cell determinant transmission, segregation, and function in. During ooplasmic segregation, the myoplasm moves first to the vegetal pole and then to the future posterior region of the fertilized egg, where it subsequently enters the muscle cell lineage during cleavage. The zebrafish perivitelline fluid provides maternallyinherited defensive immunity 1. The spermatozoa can fertilize the eggs whether it is genetically active. The exact composition of the pvf remains unknown in vertebrate animals. In the newly fertilized zebrafish egg, however, ooplasm is. Reorganization of vegetal cortex microtubules and its role in. Ooplasmic segregation has been studied extensively in the zebra. These results suggest that localized domains of elevated free cytosolic calcium are essential for ooplasmic segregation in zebrafish. Fertilization and egg activation in fishes springerlink.
Extensive cytoplasmic rearrangements occur in ascidian eggs between fertilization and first cleavage. Runge 1943, hisaoka and firlit 1960, and beams et al. Reorganization of cytoplasm in the zebrafish oocyte and. Mar 01, 2010 christina ramires ferreira, jorg patrick burgstaller, felipe perecin, joaquim mansano garcia, marcos roberto chiaratti, simone cristina meo, mathias muller, lawrence charles smith, flavio vieira meirelles, ralf steinborn, pronounced segregation of donor mitochondria introduced by bovine ooplasmic transfer to the female germline, biology of. The activation wave of calcium in the ascidian egg and its. During the second phase of ooplasmic segregation, which takes place once meiosis is completed, the concentrated er domain at the vegetalcontraction pole moves with the sperm aster and the bulk of the myoplasm toward the future posterior side of the embryo. The zebrafish zygote is a convenient cell model for the study of the mechanisms involved in several processes including fertilization, cytoplasmic movements driving ooplasmic segregation. A gastrulation center in the ascidian egg development. Microtubulebased movements during ooplasmic segregation. Germ cell determinant transmission, segregation, and. George sanders presentation occupational health and safety veterinary care. Organization and function of the cytoskeleton in the zygote a. We used timelapse video microscopy to monitor the effects of cytochalasin d ccd and deme.
Sep 10, 2015 the fertilization of zebrafish and medaka embryos is followed by ooplasmic segregation wherein the embryonic yolk separates from the ooplasm and the zygotic cell protrudes, forming the animal pole. Turnover of the actomyosin complex in zebrafish embryos. Role of ooplasmic segregation in mammalian development. Apr 06, 2016 the ascidian egg undergoes dramatic cytoplasmic and cortical reorganizations between fertilization and the beginning of the first cleavage, a process that has been referred to as ooplasmic segregation 8385. Reorganization of cytoplasm in the zebrafish oocyte and egg. A guide for the laboratory use of zebrafish danio brachydanio rerio.
To this end, the organization of the ooplasm and vitelloplasm were examined in oocytes and eggs shortly after activation. Oxygen enhancement ratio catabolite activator protein operon fusion secondary oocyte ooplasmic segregation these keywords were added by machine and not by the authors. These procedures have been performed essentially in the absence of any basic research to. Microtubulebased movements during ooplasmic segregation in. Ooplasmic segregation in the medaka oryzias latipes egg. Early zebrafish development, through midblastula stages, has also been studied by using fluorescent calcium reporters and confocal microscopy 15, 16. Ooplasmic donation in humans human reproduction oxford. Zebrafish embryos danio rerio from the wild type ab strain or the transgenic strain bacmpogfp were obtained from our breeding colony. Nov 04, 2004 the book is divided into 21 chapters, starting with the egg and embryonic development up to hatching.
191 1412 451 1187 1128 1024 1318 435 1317 983 410 58 63 188 1525 241 49 1640 908 483 475 1243 1477 1534 354 1686 1380 1177 1431 111 1224 398 666 1035 328 581 238