Measurements of (Na+,K+)ATPase after in vitro hypoxia and reoxygenation are affected by methods of membrane preparation

Journal of Neuroscience Methods
M BalestrinoIsabella Panfoli

Abstract

(Na+,K+ )ATPase activity was evaluated in membranes from rat hippocampal slices after in vitro hypoxia and reoxygenation. Membranes were prepared with two different methods, one using an isotonic medium and another using a hypotonic one. The changes that were found after hypoxia went into opposite directions in the two cases. Membranes prepared in a hypotonic medium are probably more suitable for these measurements. Using these membranes, hypoxia results in a slight decrease of (Na+,K+)ATPase activity and in a further decrease after reoxygenation. We also found that expressing (Na+,K+)ATPase activity as a percent of total ATPase activity is appropriate for membranes prepared under hypotonic conditions and can unveil (by reducing variability between experiments) significant changes that may be masked in small samples like ours.

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Citations

Feb 8, 2003·Brain Research. Molecular Brain Research·Kosuke KasaiMasaya Tohyama
Apr 30, 2009·The International Journal of Biochemistry & Cell Biology·Silvia RaveraAlessandro Morelli
May 11, 2013·Biology of the Cell·Daniela CalziaIsabella Panfoli
Aug 30, 2005·Archives of Biochemistry and Biophysics·A MorelliI M Pepe
Jun 3, 2006·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Matti Vornanen, Vesa Paajanen

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