CO₂ permeability and carbonic anhydrase activity of rat cardiomyocytes

To determine the CO₂ permeability (P_CO2 ) of plasma membranes of cardiomyocytes. These cells were chosen because heart possesses the highest rate of O₂ consumption/CO₂ production in the body.

Cardiomyocytes were isolated from rat hearts using the Langendorff technique. Cardiomyocyte suspensions exhibited a vitality of 2-14% and were studied by the previously described mass spectrometric ¹⁸ O-exchange technique deriving P_CO2 . We showed by mass spectrometry and by carbonic anhydrase (CA) staining that non-vital cardiomyocytes are free of CA and thus do not contribute to the mass spectrometric signal, which is determined exclusively by the fully functional vital cardiomyocytes.

Lysed cardiomyocytes yielded an intracellular CA activity for vital cells of 5070; that is, the rate of CO₂ hydration inside the cell is accelerated 5071-fold. Using this number, analyses of the mass spectrometric recordings from cardiomyocyte suspensions yield a P_CO2 of 0.10 cm s^-1 (SD ± 0.06, n = 15) at 37 °C.

In comparison with the P_CO2 of other cells, this value is quite high and about identical to that of the human red cell membrane. As no major protein CO₂ channels such as aquaporins 1 and 4 are present in rat cardiac sarcolemma, the high P_CO2 of this membrane is likely due to its low cholesterol content of about 0.2 (mol cholesterol)·(mol total membrane lipids)^-1 . Previous work predicted a P_CO2 of ≥0.1 cm s^-1 from this level of cholesterol. We conclude that the low cholesterol establishes a P_CO2 high enough to render the membrane resistance to CO₂ diffusion almost negligible, even under conditions of maximal O₂ consumption of the heart.