Cystic Fibrosis Methods and Protocols (Methods in Molecular by William R. Skach

By William R. Skach

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Channel-generated currents show Lorentzian noise in the spectrum described by a distribution of frequencies according to S(f) = S0/[1 + (f/fc)2] (4) Up to the corner frequency (fc) Lorentzian noise shows a constant amplitude (S0) and then rolls off with a slope of –2 in a double-logarithmic plot (see Note 2). The number (n) of observed Lorentzians in the spectrum predicts n + 1 states in a steady-state gating scheme. Spectra recorded from CFTR-mediated currents contain two distinct Lorentzian noise components (Fig.

J. (1994) Cystic fibrosis heterozygote resistance to cholera toxin in the cystic fibrosis mouse model. Science 266, 107–109. 33. Cuthbert, A. , Colledge, W. , and Evans, M. J. (1995) The genetic advantage hypothesis in cystic fibrosis heterozygotes: a murine study. J. Physiol. (Lond) 482, 449–454. 34. George, J. W. and Lerche, N. W. (1990) Electrolyte abnormalities associated with diarrhea in rhesus monkeys: 100 cases (1986-1987). J. Am. Vet. Med. Assoc. 196, 1654–1658. 35. Elmore, D. , Anderson, J.

To do this a glass micropipet that is filled with salt solution is placed onto a single cell on the stage of a microscope. A tight seal between the glass pipet and the cell membrane is achieved by applying negative pressure to the pipet interior. The seal electrically and mechanically isolates a small membrane patch inside the pipet. The sealing procedure is a critical step during patch clamping of CFTR. The quality of the seal is measured by its electrical resistance. In order to measure CFTR, single-channel currents that are <1 picoampere (pA = 10–12 A or ~6 million Cl– ions per second) seals >10 gigaohms (GΩ) are necessary.

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