However, work over the last several years has shown that intact Xenopus eggs and embryos are also powerful models for visualization and characterization of cell cycle-regulated cytoskeletal dynamics. Mediation ofanion transport in oocytes ofXenopus laevis by biosynthetically inserted band3 protein from mouse spleen erythroid cells M.Morgan. During oogenesis, Xenopus oocytes accumulate large amounts of storage proteins that—after fertilization—provide the developing embryo with building blocks and energy metabolites. Oocytes. Activity assays can be performed using isolated egg envelopes and SDS-PAGE to detect the gp43 to gp41 shift. There are two separate olfactory cavities—one for detecting scent in water, and one for airborne odors. We have reported later that lanthanum was able to induce meiosis as well (14). Learn more, Over 10 million scientific documents at your fingertips. Page 3 of 12 Volume 2, Article ID: 100008 extracellular KCM-211 solution at a flow rate of 75 mL/hour. Oxidized Glutathione Increases Delta-Subunit Expressing Epithelial Sodium Channel Activity in Xenopus laevis Oocytes Emed Res. Since then, most of the polyploids have become functional diploids in terms of gene expression patterns. Xenopus laevis oocytes were digested in ORII solution, which contained 82.5 mM NaCl, 2 mM KCl, 1 mM MgCl 2 , 5 mM HEPES, pH 7.4, and 1.5 mg/mL collagenase, for 90 min at room temperature. In fact, X. tropicalis is the only known diploid in the genus (with 2n = 20), but even here there is at least one associated cryptic tetraploid species, X. epitropicalis (4x = 40), which differs from X. tropicalis primarily with regard to mating call characteristics and some additional karyotypic features (Loumont, 1983; Tymowska, 1991). On the other hand, OsYSL18 can transport Fe(III)–DMA but not Fe(II)–NA, Zn(II)–DMA or Zn(II)–NA (Aoyama et al., 2009). Neither progesterone (as well as other active steroids) nor the various pharmacological agents mentioned above were able to reinitiate meiosis when injected inside the oocyte at internal concentration comparable to that applied in the external medium. Oogenesis in the anuran Xenopus laevis can be divided into six stages based on the anatomy of the developing oocyte. Part of Springer Nature. Characteristics Cytoplasm. The lateral line system is located dorsally and ventrally, and is retained in adult animals. Test potentials that ranged between -80 mV to +60 mV applied in 10mV increments were applied to injected oocytes. Each region was analyzed to determine the amount of total cellular RNA, poly(A)+RNA, and newly synthesized ribosomal RNA. (Biochemical changes during progesterone-induced maturation in Xenopus laevis oocytes. The oocyte is the cellular precursor for the development of specialized cells. Importantly, such a distinction would clarify the mismatch between 4x = 36 and 4x = 40 tetraploids, and implies that the (extinct) diploid ancestor of the former had a chromosome complement of 2n = 18, and not 2n = 20 as in X. School of Biochemistry and Molecular Biology, https://doi.org/10.1385/1-59259-387-9:245. (1995) A method for determining the unitary functional capacity of cloned channels and transporters expressed in, Maller, J. L., Butcher, F. R., and Krebs, E. G. (1979) Early effect of progesterone on levels of cyclic adenosine 3′:5′-monophosphate in, Uezono, Y., Bradley, J., Min, C., et al. The animals are fed granulated trout food twice a week and the water is changed entirely on the following day. Dorcas P. O'Rourke, Terry Wayne Schultz, in Laboratory Animal Medicine (Second Edition), 2002. Xenopus laevis Young Tadpoles 100 per unit. N2 - The possibility that maternal RNA may be localized in specific oocyte regions was examined in fully grown oocytes of Xenopus laevis. In this article, we describe the setting up of a model employing Xenopus laevis oocytes (XLOs) for investigating the cellular interaction of asbestos fibres. Forms a channel through the mitochondrial outer membrane that allows diffusion of small hydrophilic molecules. Female X. laevis laevis are larger than males and average 110 mm in length (Kobel et al., 1996). The skin secretions of Xenopus include thyrotropin releasing hormone, caerulein, and xenopsin. Bröer S . The selection of the donor female depends on the stage of oocytes required.4 Sexually mature females yield high number of stage V-VI and juvenile females have more The tanks are washed with water twice per week. Xenopus laevis oocytes before and after maturation. Whether its action is also related to membrane interaction is not elucidated yet. In addition to Fe(III)–DMA, HvYSL2 can transport Zn(II)–DMA, Ni(II)–DMA, Cu(II)–DMA, Mn(II)–DMA and Co(II)–DMA at fairly similar levels, which makes it functionally more similar to ZmYS1 than HvYS1, the latter of which has a narrow substrate specificity (Harada, Sugase, Namba, Iwashita, & Murata, 2007; Murata et al., 2006). Finally, a wide variety of non-hormonal compounds, applied at millimolar concentration range, were also able to trigger meiosis in Xenopus laevis oocytes (Table 1). Xenopus laevis is known to serve as host to at least 29 different genera of metazoan and protozoan parasites, many of which are strictly specific to it and its fellow Xenopus species (Tinsley and Jackson, 1998). pp 245-257 | Xenopus laevis oocytes. Barnard, E. A., Miledi, R., and Sumikawa, K. (1982) Translation of exogenous messenger RNA coding for nicotinic acetylcholine receptors produces functional receptors in Xenopus oocytes. In addition, they lack transcriptional activity until the mid-late blastula stage. The pH optimum of the enzyme is 8.0, with 90% activity at pH 7.5 or pH 8.5, and with a sharp decline in activity below pH 6.5 [5]. To investigate whether CFTR expression interacts with NPT2a function, we co-expressed both proteins in Xenopus laevis oocytes. ScienceDirect ® is a registered trademark of Elsevier B.V. ScienceDirect ® is a registered trademark of Elsevier B.V. URL: https://www.sciencedirect.com/science/article/pii/B9780123850447000023, URL: https://www.sciencedirect.com/science/article/pii/S0091679X1830013X, URL: https://www.sciencedirect.com/science/article/pii/B9780123822192006037, URL: https://www.sciencedirect.com/science/article/pii/B9780121852573500173, URL: https://www.sciencedirect.com/science/article/pii/S0079660308600380, URL: https://www.sciencedirect.com/science/article/pii/B978012263951750020X, URL: https://www.sciencedirect.com/science/article/pii/B9780080268279500081, URL: https://www.sciencedirect.com/science/article/pii/B9780123014634500103, URL: https://www.sciencedirect.com/science/article/pii/B9780123943903000112, URL: https://www.sciencedirect.com/science/article/pii/B9780121852511500088, Invertebrate and Vertebrate Eye Development, Handbook of Proteolytic Enzymes (Third Edition), Molecular and Structural Features of the Proton-Coupled Oligopeptide Transporter Superfamily, You-Jun Fei, ... Frederick H. Leibach, in, Progress in Nucleic Acid Research and Molecular Biology, Dorcas P. O'Rourke, Terry Wayne Schultz, in, Laboratory Animal Medicine (Second Edition), STEROID-INDUCED MEIOSIS REINITIATION IN XENOPUS LAEVIS OOCYTES: ROLE OF MEMBRANE CONSTITUENTS AND CYCLIC AMP, Sabine Schorderet-Slatkine, ... Etienne-Emile Baulieu, in, Endocrinology, Neuroendocrinology, Neuropeptides: Volume 13, Harada, Sugase, Namba, Iwashita, & Murata, 2007, Biochemical and Biophysical Research Communications. Yolk DNA has the same buoyant density in CsCl gradients as mitochondrial and nuclear DNAs, and, like them, it is double-stranded. Using this method, we show here for the first time, that standard Amos and Croc-type asbestos fibres significantly modify the electrophysiological properties of the XLO plasma membrane starting very shortly after exposure. By midneurula stages, the Xsix3 expression domain continues to encompass most other EFTFs' domains—the exceptions being Xnr2e1 and Xpax6, which are expressed more posteriorly on either side of the midline (Fig. This is a preview of subscription content, Stegen, C., Matskevich, I., Wagner, C. A., et al. The Xenopus oocyte, through injection of exogenous RNA, a method now more than 40 years old (Gurdon et al., 1971), is one of the major expression system for membrane proteins such as ion channels (Cens and Charnet, 2007) as well as other potential partners or regulators of the SOCE process. Furthermore, when MPF is injected into recipient oocytes, it is subject to an “auto-amplification” process, since it retains its full activity after up to 10 serial transfers of cytoplasm into untreated oocytes (7,9). Xenopus laevis, with respect to their utility for studying Ca2+ signalling mechanisms and for expression of heterologous proteins. These large oocytes have a low number of endogenous transport systems in their plasma membranes and highly express foreign mRNA; the oocyte plasma membrane is the default destination of integral membrane proteins that lack recognized organellar sorting signals. A schematic representation is shown. Two Xenopus laevis females were injected with 450 U of hCG (human chorionic gonadothropin) hormone and ovulated oocytes were obtained 12 hours after stimulation by gentle manual squeezing. 5 ). The oocytes are then denuded of overlying follicle cells by treatment with 2 mg/ml collagenase (Sigma type I-A), in Ca2+-free solution of the following composition (mM): NaCl, 82.5; KCl, 2.0; MgCl2, 1.0; HEPES, 5.0 (pH 7.4). Expression of recPfHly III in X. laevis oocytes resulted in sensitivity to hypotonic lysis in a time-dependent manner similar to that seen with hAQP1-expressing oocytes. Depending on quality of local supply, water may be used directly, mixed with deionized water, or may be aged in large containers for several days. In Xenopus laevis, vitellogenin (the yolk precursor) is synthesized in the liver and transported via the circulatory system to the ovary. (We first tested rosiglitazone (RGZ), a TZD, on human ENaC activity in Xenopus laevis oocytes. Xenopus laevis RIC‐3 enhances the functional expression of the C. elegans homomeric nicotinic receptor, ACR‐16, in Xenopus oocytes. Methods Mol Biol. Web. This procedure does not affect the electrophysiological responses of the oocytes (measured at least 36 hr later) or in any way alter its ability to translate ion channel mRNA (reviewed in Ref. Even more intriguingly, it appears that speciation in monogenean flatworms of the genus Protopolystoma and other parasites has closely followed the speciation by polyploidy of its Xenopus hosts (Tinsley and Jackson, 1998; Jackson and Tinsley, 2001). Background of Xenopus. Springer Nature is developing a new tool to find and evaluate Protocols. Microinjection of size-fractionated mRNA from the tissue sources, followed by subsequent functional assays, narrows down the potential target mRNAs to a relatively smaller subpopulation. The lack of multivalents during meiosis (at least in the tetraploids), an absence of closely related diploids, observations of mixed interbreeding, and the relatively common generation of polyploid oocytes in female hybrids all point to allopolyploidy in Xenopus species (Tymowska, 1991; Kobel, 1996). Sarah S. Conte, Elsbeth L. Walker, in Current Topics in Membranes, 2012. Figure 2: Xenopus laevis oocytes before and after maturation. Alijevic O, McHugh D, Rufener L, Mazurov A, Hoeng J, Peitsch M (2020) An electrophysiological characterization of naturally occurring tobacco alkaloids and their action on human alpha4beta2 and alpha7 nicotinic acetylcholine receptors. 2 We show that X. tropicalis oocytes possess an intracellular Ca 2+ store that is … Yolk DNA behaves like nuclear DNA, and not like mitochondrial DNA, upon renaturation after denaturation. Yolk DNA has the same buoyant density in CsCl gradients as mitochondrial and nuclear DNAs, and, like them, it is double-stranded. XSix3 represses Xen2 expression while expanding Xpax6 and Xrax expression into the midbrain. (1993) Receptors that couple to 2 classes of G proteins increase cAMP and activate CFTR expressed in, Landau, E. M. and Blitzer, R. D. (1994) Chloride current assay for phospholipase C in, Guo, Z., Liliom, K., Fischer, D. J., et al. Their large nuclei and mitochondrial masses are clearly visible in the intact oocyte. Interestingly, systematists now consider X. tropicalis and X. epitropicalis to warrant placement into their own genus (Silurana) on the basis of morphological characteristics (e.g., Cannatella and Trueb, 1988; de Sá and Hillis, 1990; Cannatella and de Sá, 1993; Rödel, 2000; Frost, 2004). For more than 30 yr, Xenopus laevis has been the animal of choice for studying the biochemical regulation of the meiotic and early mitotic vertebrate cell cycles. Surgical Laparotomy to obtain oocytes: In Xenopus, oogenesis is divided into six stages (I-VI) and oocytes are obtained by surgical laparotomy from anesthetized female X. laevis. Finally, Xenopus oocytes are a leading system for studies of ion transport and channel physiology. Indeed, it is now thought that X. laevis (for example) is, in fact, tetraploid (Kobel and Du Pasquier, 1986; Cannatella and de Sá, 1993), making X. ruwenzoriensis a dodecaploid (Kobel and Du Pasquier, 1986; Tymowska, 1991). Xenopus laevis Oocytes. (S.) tropicalis. Xenopus laevis oocytes represent another heterologous system in which YSL transporter functionality has been studied. 2.4D). Xenopus oocytes are widely used for heterologous expression of cRNAs encoding cytosolic enzymes, immunoglobulins, membrane receptors, ion channels, ion pumps, and membrane transporters. When ample water is available, the frog reverts to production of the more toxic ammonia, which is rapidly dissipated in the water (Tinsley et al., 1996). The system is also suitable for functional analysis by a variety of electrophysiological techniques due to the large size of oocytes, which permits introduction of electrodes into the cell with considerable ease. Here we demonstrate cytoplasmic microinjection of Xenopus laevis oocytes with a nuclear import substrate, as well as preparation of the injected oocytes for visualization by … Here we show that: oligomycin sensitive ATP synthase activity is significantly greater (~170 vs. 20 nmol/min −1 /mg −1) in testes compared to oocytes in Xenopus laevis (X. laevis). Such cells are undergoing spontaneous maturation. Xenopus laevis has long been a popular model for studies of development and, based on the use of cell-free extracts derived from its eggs, as a model for reconstitution of cell cycle regulation and other basic cellular processes. Million scientific documents at your fingertips maturation in Xenopus laevis used in Laboratory experiments: Prevention by use cookies. The midbrain recently to provoke meiosis ( 18 ) Na, Broer, A., al. 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