xenopus laevis oocytes

The females are killed by decapitation after 10–15 min of anesthesia in 0.2% MS222 (Sandoz, Basel, Switzerland). Not only has polyploidy been important in the evolution of the genus Xenopus itself, but it also appears to have played a major role in the evolution of species associated with it. Mediation ofanion transport in oocytes ofXenopus laevis by biosynthetically inserted band3 protein from mouse spleen erythroid cells M.Morgan. Ecocyte Bioscience. To verify research from this laboratory indicating that sheep omasal epithelium contains mRNA encoding for a peptide transporter(s) and to determine di- to octapeptide transport capability, we injected poly(A) + RNA isolated from sheep omasal epithelium into Xenopus laevis oocytes. Microinjection of Xenopus laevis oocytes is an excellent system for studying nuclear transport because of the large size of the oocyte and its high nuclear pore complex (NPC) density. 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. Dev Biol 36:669- cleavage furrow formation in Xenopus laevis oocytes and eggs. The coevolutionary influence of polyploidy goes a step further in this regard, because it appears that allopolyploidy may confer some resistance to certain species of these parasites (Jackson and Tinsley, 2003). Springer Nature is developing a new tool to find and evaluate Protocols. During the primary oocyte stage of oogenesis, the nucleus is called a germinal vesicle. A schematic representation is shown. The collagenase treatment should be terminated when about 50% of the oocytes are stripped of overlying follicle cells (1.5–3 hr). If healthy oocytes die quickly after only 1 day, it may help to switch antibiotics and use 0.1 mg/ml gentamycin. This helps to avoid overdigestion and, often, many of the nondenuded cells will become defolliculated later as the sack of follicle cells sticks to the bottom of the culture dish. Bioz Stars score: 90/100, based on 1 PubMed citations. Actinomycin or enucleation do not block progesterone action but cycloheximide inhibits maturation (3,5,6,7). (1996) Molecular cloning of a high-affinity receptor for the growth factor-like lipid mediator lysophosphatidic acid from. Recherche d'information médicale. In compliance with the UK Animals (Scientific Procedures) Act 1986, female Xenopus were anesthetized with benzocaine (ethyl 4-aminobenzoate) and minimum amounts of oocytes were used. D. Bertrand, ... M. Ballivet, in Methods in Neurosciences, 1991. 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. 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. (1984) Expression of functional acetylcholine receptor from cloned cDNAs. Females have large cloacal papillae, and males develop dark inner surfaces on their forearms (nuptial pads). These keywords were added by machine and not by the authors. bonjour j ai 14 tetard de xenopus albinos on ma donner de l aquatic code veggys comme nourriture est ce bon? 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). SURGICAL (SURVIVAL) OOCYTE COLLECTION FROM XENOUS LAEVIS PURPOSE: This document covers the procedures for the aseptic surgical (survival) collection of oocytes from Xenopus laevis following the UBC Policy 00x to standardize the procedures used to collect the oocytes and to maintain procedure and post‐procedure monitoring documents. A last and strong argument in favour of membrane as a target for triggering meiosis reinitiation has been brought about by a series of injection experiments. We have found it best to send oocytes out as dissected ovary lobes on wet ice but can provide defolliculated oocytes if necessary, albeit at extra cost. On the other hand, some inhibitors of progesterone-induced meiosis were also acting at membrane level, thus on the first step of the cascade of events triggered by the steroid (Table 1). pus (zĕn′ə-pəs) n. Any of various aquatic, tongueless, clawed frogs of the genus Xenopus of southern Africa, certain species of which, especially the African clawed frog, are widely used for biological research. Test potentials that ranged between -80 mV to +60 mV applied in 10mV increments were applied to injected oocytes. The search-image is essentially a uniform pigmentation of the animal pole of stage V and VI oocytes (9). Xrax, Xlhx2, and Xtbx3 are contained centrally within the Xsix3 expression domain (Fig. It is estimated that the protein content is about 500 ug in each oocyte and approximately 10-50 mg/mL in the extract. 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). In Xenopus, misexpression of a transcriptional activator form of XSix3 by fusion with the VP16 activation domain (XSix3–VP16) blocks Xrax and Xpax6 expression and dramatically reduces or even eliminates eye formation, suggesting that XSix3 normally acts as a repressor with respect to eye field and eye formation (Gestri et al., 2005). Oviductin is inhibited by DFP, aprotinin, soybean trypsin inhibitor, leupeptin, antipain EDTA, and EGTA, and also weakly by TLCK, TPCK, and 1,10-phenanthroline; pepstatin, E-64, and iodoacetamide do not inhibit [5]. Oocytes are not used for injection of mRNA for at least 2 hr after defolliculation. Xenopus laevis females, purchased from the African Xenopus Facility (Noordhoek, Republic of South Africa) (or Xenopus, Ltd., Nutfield Ridge, Surrey, England; Carolina Biological Supply Company, Burlington, NC; Xenopus 1, Ann Arbor, MI) are kept at 23°C in tanks with a circulating supply of water (low calcium, low chlorine). Researchers have found that the genetic ZZ/ZW sex determination system in this genus (females are heterozygous for dominant female-determining factors and for recessive male-determining factors, although no morphologically distinct sex chromosomes have been found) is disrupted by polyploidy (reviewed in Kobel and Du Pasquier, 1986). Xenopus laevis oocytes are arrested in prophase of the first meiotic division. 5 ). Oogenesis in the anuran Xenopus laevis can be divided into six stages based on the anatomy of the developing oocyte. Trotti, D., Peng, J. Methods Mol Biol. Xenopus laevis Six3 is expressed broadly (relative to the other EFTFs) in the developing neural plate at all stages (Ghanbari et al., 2001; Zhou et al., 2000; Zuber et al., 2003). Both Xenopus laevis oocytes and mammalian cells are widely used for heterologous expression of several classes of proteins, and membrane proteins especially, such as ion channels or receptors, have been extensively investigated in both cell types. The frog's ventral surface is solid yellowish white to spotted. However, there is a clear discrepancy between diploid X. tropicalis (2n = 20), tetraploid X. epitropicalis (4x = 40), and the other Xenopus polyploids (all 4x = 36 or multiples thereof). Furthermore, several pharmacological membrane active agents were able to mimick progesterone action. Sarah Cohen 1, Shelly Au 1, Nelly Pant é 1. The widespread use of the oocyte expression system is based upon the fact that oocytes can not only translate exogenous RNA from various sources efficiently and faithfully, but also carry out posttranslational modifications, such as precursor processing, phosphorylation, glycosylation, intracellular trafficking and protein targeting into the plasma membrane, complex subunit assembly, and even exporting secretory proteins. The skin secretions of Xenopus include thyrotropin releasing hormone, caerulein, and xenopsin. The lungs, heart, and liver of Xenopus are large, and the urinary bladder is spherical. 2.4E). Here we describe how to isolate Xenopus laevis oocytes, prepare capped sense RNA (cRNA) of the maize boron importer TASSEL‐LESS1 (TLS1) as an example, microinject the cRNA into the isolated oocytes, and functionally assess the boron import capabilities of TLS1 in an oocyte swelling assay. Xenopus laevis (African clawed frog) Status. The relatively broad expression domain and range of phenotypes observed following its inactivation indicate that XSix3 is required not only for eye field specification, but also for much of rostral brain formation. Not logged in We use cookies to help provide and enhance our service and tailor content and ads. The EXRC supplies oocytes to many UK laboratories regularly and also to some on mainland Europe. Specifically, W dominance is weakened with increasing dosages of male-determining factors, and genetic sex determination may give way to environmental sex determination at ploidies where the dosages are equal. [New Latin Xenopūs, genus name : xeno- + Greek pous, foot; see octopus.] Xenopus laevis oocytes. Gurdon, J. If the wash solution around the defolliculated oocytes is very cloudy even after several rinses and becomes even more cloudy after 30 min in culture, then the oocytes are probably unusable. Preparation. An extensive array of techniques is featured. N2 - The possibility that maternal RNA may be localized in specific oocyte regions was examined in fully grown oocytes of Xenopus laevis. Each region was analyzed to determine the amount of total cellular RNA, poly(A)+RNA, and newly synthesized ribosomal RNA. Xenopus is a genus of African frogs that are commonly known as the African clawed frogs. Web. The first step is followed by the appearance of a cytoplasmic factor, known as maturation promoting factor (MPF), which is formed after 3-5 h of external progesterone treatment. B., Dunlop, J, et al. (1971) Use of frog eggs and oocytes for the study of messenger RNA and its translation in living cells. (2001) Isotonic transport by the Na, Broer, S., Schneider, H. P., Broer, A., et al. Preparation. All frog surgery is done under anesthesia induced by 0.2% 3-aminobenzoic acid ethyl ester (tricaine methane sulfonate or MS-222). You-Jun Fei, ... Frederick H. Leibach, in Progress in Nucleic Acid Research and Molecular Biology, 1997. Emed Res 2: 100008. Agents interacting with meiosis in Xenopus laevis oocytes, Some steroids: progesterone, cortisol, testosterone, pregnenolone, Some organo-mercurials: parachloromercuribenzoate, mersalyl, Some ions: lanthanum, zinc, calcium*† magnesium*, Some antiarrythmic drugs: D-600(isoptin), D-200(verapamil), Some O-receptor adrenergic antagonists: isomers of propranolol, alprenolol, dichloroisoproterenol, Some local anesthetics: dibucaine, tetracaine, Some CNS acting drugs: barbital, imipramine, chlorpromazine, fluphenazine, fenfluramine, cycloheximide (inhibitor of protein synthesis), theophylline, IBMX (inhibitors of phosphodiesterases), dithiothreitol (protecting agent for SH groups), gammexane (inhibitor of phospholipid metabolism). (S.) tropicalis. Traditionally, Xenopus species with 20 or 36 chromosomes had been considered as diploids, those with 72 chromosomes as tetraploids, and the 108 chromosomes of X. ruwenzoriensis as a hexaploid (e.g., Bogart, 1980). Xenopus laevis oocytes. (We first tested rosiglitazone (RGZ), a TZD, on human ENaC activity in Xenopus laevis oocytes. Author information: (1)School of Biochemistry and Molecular Biology, Australian National University, Canberra, Australia. Table 1. Contrary to progesterone, its efficiency was linearly correlated with its time exposure, due to its different solubility properties. The same strategy was applied by different investigators to successfully clone rabbit PEPT1 and rabbit PEPT2 from the rabbit intestinal and renal cDNA libraries, respectively (29,30). Copyright © 2021 Elsevier B.V. or its licensors or contributors. 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). exocytosis in Xenopus laevis oocytes and eggs in response to cytosolic Ca ~÷. Females can be operated on several times but usually they are sacrificed on the second taking of 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. Phytochemistry 170:112187. For injection (11, 12), we use untreated females during most of the year. Page 3 of 12 Volume 2, Article ID: 100008 extracellular KCM-211 solution at a flow rate of 75 mL/hour. Sterile, 70% ethanol-treated surgical instruments are used for all procedures. Life span of X. laevis in the wild is reported to be greater than 10 years. BACKGROUND. Xenopus floats at the water's surface, and vision is directed upward; therefore, objects passing above will elicit a hiding response from the frog. XSix3 misexpression has dose-dependent effects. It is known that this ion does not penetrate the cells and interferes with Ca2+-movements from membrane sites and/or adenylate cyclase activity (15). L.L Lindsay, in Handbook of Proteolytic Enzymes (Third Edition), 2013. 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). Activity in Xenopus laevis Oocytes. John P. Leonard, in Methods in Neurosciences, 1989. Antimicrobial compounds (magainins) are also found in skin secretions (Kreil, 1996). 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. This subspecies has a fifth toe that is much longer than the tibia. Once a method is successfully developed, a great deal of data can be obtained for the transporter in question by bathing oocytes in different metal chelate solutions and/or adding various inhibitors. Female frogs usually produce healthy oocytes if certain measures are taken (5). (A) Development of ICSI embryos to each stage is summarized. Thus, OsYSL2 maintains a fairly narrow substrate specificity and is likely to function in phloem transport of Fe and Mn. XSix3 represses Xen2 expression while expanding Xpax6 and Xrax expression into the midbrain. The first step of this physiological process seems to involve a target site at the oocyte membrane, as shown by a variety of experimental data (4). Emed Res 2: 100008. INTRODUCTION The oocyte of Xenopus laevis has provided a … Summary. In vitro transcribed poly(A)-cRNA is microinjected into the cytoplasm of the oocyte and on the next day the function of the encoded protein can be measured. A 1–2 cm incision is made in the abdomen and several ovarian lobes removed (6). Since ATP synthase activity is redox regulated, we explored a regulatory role for reversible thiol oxidation. It is believed that the first duplication/hybridization event occurred at least 30 million years ago, with more recent events perhaps having been stimulated by environmental changes during the Pleistocene (Tymowska, 1991). Selection of oocytes is an often-overlooked important step. Please Specify Uncurling arrive around 38 NF

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