Ikaga 2015 Biochem Biophys Rep
|Ikaga R, Namekatac I, Kotiadisd VN, Ogawab H, Duchend MR, Tanakac H, Iida-Tanakab N (2015) Knockdown of aquaporin-8 induces mitochondrial dysfunction in 3T3-L1 cells. Biochem Biophys Rep 4:187–95.|
Abstract: Aquaporin-8 (AQP8), a member of the aquaporin water channel family, is expressed in various tissue and cells, including liver, testis, and pancreas. AQP8 appears to have functions on the plasma membrane and/or on the mitochondrial inner membrane. Mitochondrial AQP8 with permeability for water, H2O2 and NH3 has been expected to have important role in various cells, but its information is limited to a few tissues and cells including liver and kidney. In the present study, we found that AQP8 was expressed in the mitochondria in mouse adipose tissues and 3T3-L1 preadipocytes, and investigated its role by suppressing its gene expression.
AQP8-knocked down (shAQP8) cells were established using a vector expressing short hairpin RNA. Cellular localization of AQP8 was examined by western blotting and immunocytochemistry. Mitochondrial function was assessed by measuring mitochondrial membrane potential, oxygen consumption and ATP level measurements.
In 3T3-L1 cells, AQP8 was expressed in the mitochondria. In shAQP8 cells, mRNA and protein levels of AQP8 were decreased by about 75%. The shAQP8 showed reduced activities of complex IV and ATP synthase; it is probable that the impaired mitochondrial water handling in shAQP8 caused suppression of the electron transport and ADP phosphorylation through inhibition of the two steps which yield water. The reduced activities of the last two steps of oxidative phosphorylation in shAQP8 cause low routine and maximum capacity of respiration and mitochondrial hyperpolarization.
Mitochondrial AQP8 contributes to mitochondrial respiratory function probably through maintenance of water homeostasis.
The AQP8-knocked down cells we established provides a model system for the studies on the relationships between water homeostasis and mitochondrial function.
• Keywords: Aquaporin-8, Mitochondria, Adipocyte, Mouse 3T3-L1 preadipocytes
Labels: MiParea: Respiration, mt-Structure;fission;fusion, mtDNA;mt-genetics, Genetic knockout;overexpression
Organism: Mouse Tissue;cell: Fat, Other cell lines Preparation: Intact cells Enzyme: Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Complex V;ATP synthase
Coupling state: LEAK, ROUTINE, ET Pathway: CIV, ROX HRR: Oxygraph-2k