O'Connell D et al. (2011), Xenopus laevis oocytes infected with multi-drug...

XB-ART-43593

J Gen Physiol 2011 Aug 01;1382:271-7. doi: 10.1085/jgp.201110661.

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Xenopus laevis oocytes infected with multi-drug-resistant bacteria: implications for electrical recordings.

O'Connell D , Mruk K , Rocheleau JM , Kobertz WR .

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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.

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Species referenced: Xenopus laevis

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References [+] :

Bement, Wound-induced assembly and closure of an actomyosin purse string in Xenopus oocytes. 1999, Pubmed, Xenbase

	Figure 1. Xenopus laevis oocytes infected with multi-drug–resistant bacteria. (A) Bright field micrograph of extracted oocytes showing the characteristic black foci and unpigmented halo. Bar, 0.5 mm. (B) Transmission electron micrograph of P. fluorescens cultured from compromised oocytes. (C) Scanning electron micrograph of bacteria on the surface of compromised oocytes.
	Figure 2. Infected oocytes exhibit reduced viability, which can be prevented with amikacin/ciprofloxacin supplementation. Histographs of the absolute change in leak current of oocytes incubated for 4 d in gentamicin/tetracycline (ND96-GT)- or amikacin/ciprofloxacin (ND96-ACT)-supplemented media. Change in leak current at –80 mV was calculated by subtracting the initial current value upon clamping from the value at 2 min. Data are from 2–5 batches of oocytes.
	Figure 3. Amikacin/ciprofloxacin supplementation prevents black foci formation. Bright-field micrographs of extracted oocytes incubated for 4 d in tetracycline/gentamicin-supplemented (ND96-GT; A) or amikacin/ciprofloxacin-supplemented media (ND96-ACT; B). Bars, 0.5 mm. Histopathology slices of a compromised oocyte (C) and a healthy oocyte (D), magnified in E and F, respectively. Brackets denote areas of increased pigmentation; arrows indicate areas devoid of pigment molecules, both of which were only observed in infected oocytes.