scholarly journals Metabolism of Poly (A) Sequences of Maternal mRNA during Progesterone-Induced Maturation of Xenopus laevis Oocytes

1980 ◽  
Vol 5 (1) ◽  
pp. 67-79 ◽  
Author(s):  
Noriyuki Sagata ◽  
Koichiro Shiokawa ◽  
Kiyotaka Yamana
Keyword(s):  
Xenopus Laevis ◽  
Xenopus Laevis Oocytes ◽  
Maternal Mrna

scholarly journals A new system to evaluate characteristics of Niemann-Pick C1 Like 1-mediated cholesterol transport using Xenopus laevis oocytes

2021 ◽  
Vol 1863 (2) ◽  
pp. 183508
Author(s):  
Shunsuke Nashimoto ◽  
Saori Yagi ◽  
Naoki Takeda ◽  
Miku Nonaka ◽  
Yoh Takekuma ◽  
...  
Keyword(s):  
Xenopus Laevis ◽  
Cholesterol Transport ◽  
Xenopus Laevis Oocytes ◽  
Niemann Pick ◽  
New System

Characterization of Transport Activity of SLC11 Transporters in Xenopus laevis Oocytes by Fluorophore Quenching

2021 ◽  
pp. 247255522110041
Author(s):  
Raffaella Cinquetti ◽  
Francesca Guia Imperiali ◽  
Salvatore Bozzaro ◽  
Daniele Zanella ◽  
Francesca Vacca ◽  
...  
Keyword(s):  
Xenopus Laevis ◽  
Voltage Clamp ◽  
Expression System ◽  
Peptide Transporter ◽  
Xenopus Laevis Oocytes ◽  
Substrate Uptake ◽  
Transport Activity ◽  
Experimental Conditions ◽  
Metal Transporters ◽  
Electrode Voltage

Membrane proteins are involved in different physiological functions and are the target of pharmaceutical and abuse drugs. Xenopus laevis oocytes provide a powerful heterologous expression system for functional studies of these proteins. Typical experiments investigate transport using electrophysiology and radiolabeled uptake. A two-electrode voltage clamp is suitable only for electrogenic proteins, and uptake measurements require the existence of radiolabeled substrates and adequate laboratory facilities. Recently, Dictyostelium discoideum Nramp1 and NrampB were characterized using multidisciplinary approaches. NrampB showed no measurable electrogenic activity, and it was investigated in Xenopus oocytes by acquiring confocal images of the quenching of injected fluorophore calcein. This method is adequate to measure the variation in emitted fluorescence, and thus transporter activity indirectly, but requires long experimental procedures to collect statistically consistent data. Considering that optimal expression of heterologous proteins lasts for 48–72 h, a slow acquiring process requires the use of more than one batch of oocytes to complete the experiments. Here, a novel approach to measure substrate uptake is reported. Upon injection of a fluorophore, oocytes were incubated with the substrate and the transport activity measured, evaluating fluorescence quenching in a microplate reader. The technique permits the testing of tens of oocytes in different experimental conditions simultaneously, and thus the collection of significant statistical data for each batch, saving time and animals. The method was tested with different metal transporters (SLC11), DMT1, DdNramp1, and DdNrampB, and verified with the peptide transporter PepT1 (SLC15). Comparison with traditional methods (uptake, two-electrode voltage clamp) and with quenching images acquired by fluorescence microscopy confirmed its efficacy.

scholarly journals The Molecular Physiology of Sodium- and Proton-Coupled Solute Transporters

Physiology ◽  
1998 ◽  
Vol 13 (3) ◽  
pp. 123-131 ◽  
Author(s):  
Angela Steel ◽  
Matthias A. Hediger
Keyword(s):  
Xenopus Laevis ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Xenopus Laevis Oocytes ◽  
Missense Mutations ◽  
Molecular Physiology ◽  
Inherited Diseases ◽  
Biophysical Analysis ◽  
Solute Transporters

The expression of cloned Na+- and H+-coupled solute transporters in Xenopus laevis oocytes has permitted detailed molecular and biophysical analysis and illuminated unique mechanistic features. The identification of missense mutations in inherited diseases and site-directed mutagenesis studies have enhanced our understanding of their roles in physiological and pathological processes.

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