Characterization of sea urchin unconventional myosins and analysis of their patterns of expression during early embryogenesis

Sirotkin, Vladimir; Seipel, Susan A.; Krendel, Mira; Bonder, Edward M.

doi:10.1002/1098-2795(200010)57:2<111::aid-mrd2>3.0.co;2-t

Cited by 11 publications

(7 citation statements)

References 70 publications

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“…Sea urchin eggs and staged embryos were cultured as previously described (Sirotkin et al 2000) and total RNA was isolated using the Biotecx Ultraspec RNA Isolation System. RNA from each stage was subjected to relative RT-PCR analysis as described by Ambion using the primers n5 seawi (5Ј-ccgggatccgaacaagcacccccaccgatg-3Ј) and n3 seawi (5Ј-caccaccctcgagttaagacagcagcatcacgtcgtac-3Ј).…”

Section: Relative Rt-pcr Analysis Of Staged Embryosmentioning

confidence: 99%

“…The resulting cDNA was gel purified using a QIAquick gel extraction kit (Qiagen Inc.) and sequenced using a Perkin Elmer DNA sequencing kit, the GPEIV and MIIGcom primers, and an Applied Biosystems ABI Prism 373 automated sequencer. GPEIV and MIIGcom primers were used to prepare a DIG-labeled probe for screening a ZAP cDNA library as previously described (Sirotkin et al 2000). A second DIG-labeled probe was synthesized using PCR primers C-term (5Ј-cggacactctgactgat gatgaag-3Ј) and C-termcom (5Ј-caacgagaagtttcctgagcgtatc-3Ј) from the C-terminal portion of plasmid 6-2.…”

Section: Cdna Library Screeningmentioning

confidence: 99%

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Seawi—a sea urchin piwi/argonaute family member is a component of MT-RNP complexes

Rodríguez¹,

Seipel²,

Hamill³

et al. 2005

RNA

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The piwi/argonaute family of proteins is involved in key developmental processes such as stem cell maintenance and axis specification through molecular mechanisms that may involve RNA silencing. Here we report on the cloning and characterization of the sea urchin piwi/argonaute family member seawi. Seawi is a major component of microtubule-ribonucleoprotein (MT-RNP) complexes isolated from two different species of sea urchin, Strongylocentrotus purpuratus and Paracentrotus lividus. Seawi co-isolates with purified ribosomes, cosediments with 80S ribosomes in sucrose density gradients, and binds microtubules. Seawi possesses the RNA binding motif common to piwi family members and binds P. lividus bep4 mRNA, a transcript that co-isolates with MT-RNP complexes and whose translation product has been shown to play a role in patterning the animal-vegetal axis. Indirect immunofluorescence studies localized seawi to the cortex of unfertilized eggs within granule-like particles, the mitotic spindle during cell division, and the small micromeres where its levels were enriched during the early cleavage stage. Lastly, we discuss how seawi, as a piwi/argonaute family member, may play a fundamentally important role in sea urchin animal-vegetal axis formation and stem cell maintenance.

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Section: Relative Rt-pcr Analysis Of Staged Embryosmentioning

confidence: 99%

Section: Cdna Library Screeningmentioning

confidence: 99%

Seawi—a sea urchin piwi/argonaute family member is a component of MT-RNP complexes

Rodríguez¹,

Seipel²,

Hamill³

et al. 2005

RNA

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“…Initially, egg myosin II was characterized biochemically (Mabuchi, 1972;Kane, 1983), followed by the unconventional myosins I and VI (D'Andrea, 1993;Terasaki, 1997). Other myosins that have been partially sequenced include amoeboid-like myosin I, V, VI, VII, IX (Sirotkin, 2000). Of these, only myosins V and VI have been completely sequenced.…”

Section: Myosinsmentioning

confidence: 99%

Analysis of cytoskeletal and motility proteins in the sea urchin genome assembly

Morris

Hoffman

Obar

et al. 2006

Developmental Biology

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The sea urchin embryo is a classical model system for studying the role of the cytoskeleton in such events as fertilization, mitosis, cleavage, cell migration and gastrulation. We have conducted an analysis of gene models derived from the Strongylocentrotus purpuratus genome assembly and have gathered strong evidence for the existence of multiple gene families encoding cytoskeletal proteins and their regulators in sea urchin. While many cytoskeletal genes have been cloned from sea urchin with sequences already existing in public databases, genome analysis reveals a significantly higher degree of diversity within certain gene families. Furthermore, genes are described corresponding to homologs of cytoskeletal proteins not previously documented in sea urchins. To illustrate the varying degree of sequence diversity that exists within cytoskeletal gene families, we conducted an analysis of genes encoding actins, specific actin-binding proteins, myosins, tubulins, kinesins, dyneins, specific microtubule-associated proteins, and intermediate filaments. We conducted ontological analysis of select genes to better understand the relatedness of urchin cytoskeletal genes to those of other deuterostomes. We analyzed developmental expression (EST) data to confirm the existence of select gene models and to understand their differential expression during various stages of early development.

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“…1996). Analysis of expression of myosin families, which encode 11 different mRNA, revealed that the onset of gastrulation is a pivotal point in modulation of myosin mRNA expression (Sirotkin et al . 2000).…”

Section: Factors Involved In Gastrulationmentioning

confidence: 99%

Gastrulation in the sea urchin embryo: A model system for analyzing the morphogenesis of a monolayered epithelium

Kominami

Takata

2004

Dev Growth Differ

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Processes of gastrulation in the sea urchin embryo have been intensively studied to reveal the mechanisms involved in the invagination of a monolayered epithelium. It is widely accepted that the invagination proceeds in two steps (primary and secondary invagination) until the archenteron reaches the apical plate, and that the constituent cells of the resulting archenteron are exclusively derived from the veg2 tier of blastomeres formed at the 60-cell stage. However, recent studies have shown that the recruitment of the archenteron cells lasts as late as the late prism stage, and some descendants of veg1 blastomeres are also recruited into the archenteron. In this review, we first illustrate the current outline of sea urchin gastrulation. Second, several factors, such as cytoskeletons, cell contact and extracellular matrix, will be discussed in relation to the cellular and mechanical basis of gastrulation. Third, differences in the manner of gastrulation among sea urchin species will be described; in some species, the archenteron does not elongate stepwise but continuously. In those embryos, bottle cells are scarcely observed, and the archenteron cells are not rearranged during invagination unlike in typical sea urchins. Attention will be also paid to some other factors, such as the turgor pressure of blastocoele and the force generated by blastocoele wall. These factors, in spite of their significance, have been neglected in the analysis of sea urchin gastrulation. Lastly, we will discuss how behavior of pigment cells defines the manner of gastrulation, because pigment cells recently turned out to be the bottle cells that trigger the initial inward bending of the vegetal plate.

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Characterization of sea urchin unconventional myosins and analysis of their patterns of expression during early embryogenesis

Cited by 11 publications

References 70 publications

Seawi—a sea urchin piwi/argonaute family member is a component of MT-RNP complexes

Seawi—a sea urchin piwi/argonaute family member is a component of MT-RNP complexes

Analysis of cytoskeletal and motility proteins in the sea urchin genome assembly

Gastrulation in the sea urchin embryo: A model system for analyzing the morphogenesis of a monolayered epithelium

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