The NAD+/NADH redox state in astrocytes: Independent control of the NAD+ and NADH content

The intracellular redox state is established by several redox pairs, such as NAD(+) /NADH and NADP(+) /NADPH and glutathione. This redox state is a crucial determinant of cellular metabolism and function. Astrocytes are an important cell population contributing to brain metabolism and brain energy supply, so a careful control of these redox pairs is essential for proper brain function. Despite this, little is known about control of the NAD(+) and NADH content within the brain or in astrocytes. Therefore, we here analyzed the NAD(+) and NADH content of mouse tissue and cultured cortical astrocytes. The NAD(+) /NADH ratio increased in most tissues during development from newborn to adult mice. The basal redox ratio of cultured astrocytes was about 3.8 and similar to the redox ratio of the cortex of newborn mice. Although the NADH content of these cells was highly sensitive to the concentration of energy substrates and to modulation of energy metabolism, the NAD(+) content was surprisingly constant under these conditions. In contrast, application of nicotine amide or nicotinamide mononucleotide, which are precursors for NAD(+) biosynthesis, slowly increased NAD(+) content while leaving NADH levels unaffected. Finally, inhibiting the NAD(+) -degrading enzyme poly-(ADP-ribose)-polymerase increased NAD(+) content slightly without affecting NADH levels, whereas inhibition of sirtuins had no effect. These results indicate that, in addition to converting NAD(+) to NADH and vice versa during redox reactions, the content of both partners of this redox pair is additionally controlled by other mechanisms.