scholarly journals 3P133 ATP quantification and live-imaging in Xenopus laevis oocyte(09. Development & Differentation,Poster)

2013 ◽  
Vol 53 (supplement1-2) ◽  
pp. S233
Author(s):  
Takashi W. Ijiri ◽  
Jun-ichi Kishikawa ◽  
Hiromi Imamura ◽  
Maho Sakiie ◽  
Shuichi Ueno ◽  
...  
Keyword(s):  
Xenopus Laevis ◽  
Live Imaging ◽  
Xenopus Laevis Oocyte

scholarly journals Involvement of single-stranded tails in homologous recombination of DNA injected into Xenopus laevis oocyte nuclei.

1991 ◽  
Vol 11 (6) ◽  
pp. 3268-3277 ◽  
Author(s):  
E Maryon ◽  
D Carroll
Keyword(s):  
Homologous Recombination ◽  
Xenopus Laevis ◽  
Gel Electrophoresis ◽  
Xenopus Laevis Oocyte ◽  
Molecular Models ◽  
Mixed Substrates ◽  
Two Dimensional Gel Electrophoresis ◽  
Recombination Pathway ◽  
Made In

Homologous recombination of DNA molecules injected into Xenopus laevis oocyte nuclei is extremely efficient when those molecules are linear and have overlapping homologous ends. It was previously shown that a 5'----3' exonuclease activity in oocytes attacks injected linear DNAs and leaves them with single-stranded 3' tails. We tested the hypothesis that such tailed molecules are early intermediates on the pathway to recombination products. Substrates with 3' tails were made in vitro and injected into oocytes, where they recombined rapidly and efficiently. In experiments with mixed substrates, molecules with 3' tails entered recombination intermediates and products more rapidly than did molecules with flush ends. Molecules endowed in vitro with 5' tails also recombined efficiently in oocytes, but their rate was not faster than for flush-ended substrates. In most cases, the 5' tails served as templates for resynthesis of the 3' strands, regenerating duplex ends which then entered the normal recombination pathway. In oocytes from one animal, some of the 5' tails were removed, and this was exacerbated when resynthesis was partially blocked. Analysis by two-dimensional gel electrophoresis of recombination intermediates from 5'-tailed substrates confirmed that they had acquired 3' tails as a result of the action of the 5'----3' exonuclease. These results demonstrate that homologous recombination in oocytes proceeds via a pathway that involves single-stranded 3' tails. Molecular models incorporating this feature are discussed.

Imaging of Xenopus laevis Oocyte Plasma Membrane in Physiological-Like Conditions by Atomic Force Microscopy

2013 ◽  
Vol 19 (5) ◽  
pp. 1358-1363 ◽  
Author(s):  
Massimo Santacroce ◽  
Federica Daniele ◽  
Andrea Cremona ◽  
Diletta Scaccabarozzi ◽  
Michela Castagna ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Plasma Membrane ◽  
High Resolution ◽  
Membrane Proteins ◽  
Xenopus Laevis ◽  
Functional Study ◽  
Xenopus Laevis Oocyte ◽  
Force Microscopy ◽  
Atomic Force ◽  
Afm Imaging

AbstractXenopus laevis oocytes are an interesting model for the study of many developmental mechanisms because of their dimensions and the ease with which they can be manipulated. In addition, they are widely employed systems for the expression and functional study of heterologous proteins, which can be expressed with high efficiency on their plasma membrane. Here we applied atomic force microscopy (AFM) to the study of the plasma membrane of X. laevis oocytes. In particular, we developed and optimized a new sample preparation protocol, based on the purification of plasma membranes by ultracentrifugation on a sucrose gradient, to perform a high-resolution AFM imaging of X. laevis oocyte plasma membrane in physiological-like conditions. Reproducible AFM topographs allowed visualization and dimensional characterization of membrane patches, whose height corresponds to a single lipid bilayer, as well as the presence of nanometer structures embedded in the plasma membrane and identified as native membrane proteins. The described method appears to be an applicable tool for performing high-resolution AFM imaging of X. laevis oocyte plasma membrane in a physiological-like environment, thus opening promising perspectives for studying in situ cloned membrane proteins of relevant biomedical/pharmacological interest expressed in this biological system.

scholarly journals The mRNA encoding a β subunit of heterotrimeric GTP-binding proteins is localized to the animal pole of Xenopus laevis oocyte and embryos

1996 ◽  
Vol 59 (2) ◽  
pp. 141-151 ◽  
Author(s):  
Eric Devic ◽  
Laurent Paquereau ◽  
Karine Rizzoti ◽  
Armelle Monier ◽  
Bernard Knibiehler ◽  
...  
Keyword(s):  
Xenopus Laevis ◽  
Binding Proteins ◽  
Xenopus Laevis Oocyte ◽  
Β Subunit ◽  
Animal Pole ◽  
Gtp Binding Proteins ◽  
Gtp Binding

scholarly journals Nuclear hourglass technique: An approach that detects electrically open nuclear pores in Xenopus laevis oocyte

1999 ◽  
Vol 96 (23) ◽  
pp. 13530-13535 ◽  
Author(s):  
T. Danker ◽  
H. Schillers ◽  
J. Storck ◽  
V. Shahin ◽  
B. Kramer ◽  
...  
Keyword(s):  
Xenopus Laevis ◽  
Xenopus Laevis Oocyte ◽  
Nuclear Pores

Formation of branched DNA structures by Xenopus laevis oocyte extract

Nature ◽  
1977 ◽  
Vol 270 (5639) ◽  
pp. 754-756 ◽  
Author(s):  
DOMENICA GANDINI ATTARDI ◽  
EMILIO MATTOCCIA ◽  
GLAUCO P. TOCCHINI-VALENTINI
Keyword(s):  
Xenopus Laevis ◽  
Xenopus Laevis Oocyte ◽  
Dna Structures ◽  
Branched Dna

scholarly journals Xenopus laevis oocytes infected with multi-drug–resistant bacteria: implications for electrical recordings

2011 ◽  
Vol 138 (2) ◽  
pp. 271-277 ◽  
Author(s):  
Denice O'Connell ◽  
Karen Mruk ◽  
Jessica M. Rocheleau ◽  
William R. Kobertz
Keyword(s):  
Xenopus Laevis ◽  
Mammalian Cells ◽  
Resistant Bacteria ◽  
Xenopus Laevis Oocytes ◽  
Xenopus Laevis Oocyte ◽  
Drug Resistant ◽  
Animal Pole ◽  
Antibiotic Cocktail ◽  
Storage Media ◽  
Drug Resistant Bacteria

The Xenopus laevis oocyte has been the workhorse for the investigation of ion transport proteins. These large cells have spawned a multitude of novel techniques that are unfathomable in mammalian cells, yet the fickleness of the oocyte has driven many researchers to use other membrane protein expression systems. Here, we show that some colonies of Xenopus laevis are infected with three multi-drug–resistant bacteria: Pseudomonas fluorescens, Pseudomonas putida, and Stenotrophomonas maltophilia. Oocytes extracted from infected frogs quickly (3–4 d) develop multiple black foci on the animal pole, similar to microinjection scars, which render the extracted eggs useless for electrical recordings. Although multi-drug resistant, the bacteria were susceptible to amikacin and ciprofloxacin in growth assays. Supplementing the oocyte storage media with these two antibiotics prevented the appearance of the black foci and afforded oocytes suitable for whole-cell recordings. Given that P. fluorescens associated with X. laevis has become rapidly drug resistant, it is imperative that researchers store the extracted oocytes in the antibiotic cocktail and not treat the animals harboring the multi-drug–resistant bacteria.

scholarly journals Nuclear transport of U1 snRNP in somatic cells: differences in signal requirement compared with Xenopus laevis oocytes.

1994 ◽  
Vol 125 (5) ◽  
pp. 971-980 ◽  
Author(s):  
U Fischer ◽  
J Heinrich ◽  
K van Zee ◽  
E Fanning ◽  
R Lührmann
Keyword(s):  
Xenopus Laevis ◽  
Nuclear Import ◽  
Nuclear Transport ◽  
Somatic Cells ◽  
Xenopus Laevis Oocytes ◽  
Xenopus Laevis Oocyte ◽  
Nuclear Targeting ◽  
Transport Pathway ◽  
U1 Snrnp

The signal requirement for the nuclear import of U1 RNA in somatic cells from different species was investigated by microinjection of both digoxygenin-labeled wild type and mutant U1 RNA molecules and in vitro reconstituted U1 snRNPs. U1 RNA was shown to be targeted to the nucleus by a temperature-dependent process that requires the prior assembly of RNPs from the common proteins and the microinjected RNA. Competition in the cell between immunoaffinity-purified U1 snRNPs and digoxygenin-labeled U1 snRNPs reconstituted in vitro showed that the transport is saturable and should therefore be a mediated process. The transport of a karyophilic protein under the same conditions was not affected, indicating the existence of a U snRNP-specific transport pathway in somatic cells, as already seen in the Xenopus laevis oocyte system. Surprisingly, the signal requirement for nuclear transport of U1 snRNP was found to differ between oocytes and somatic cells from mouse, monkey and Xenopus, in that the m3GGpppG-cap is no longer an essential signaling component in somatic cells. However, as shown by investigation of the transport kinetics of m3GpppG- and ApppG-capped U1 snRNPs, the m3GpppG-cap accelerates the rate of U1 snRNP import significantly indicating that it has retained a signaling role for nuclear targeting of U1 snRNP in somatic cells. Moreover, our data strongly suggest that cell specific rather than species specific differences account for the differential m3G-cap requirement in nuclear import of U1 snRNPs.

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