Meacci Elisabetta, Bini Francesca, Battistini Chiara
Department of Biochemical Sciences, University of Florence, Florence, Italy.
Methods Mol Biol. 2012;874:155-65. doi: 10.1007/978-1-61779-800-9_12.
Sphingosine 1-phosphate (S1P) is a bioactive sphingolipid regulator of numerous important physiological and pathological processes in mammalian and nonmammalian cells. There are emerging evidence that many cell types can produce and release S1P; therefore, the quantification of its intracellular and extracellular content as well as the activity of sphingosine kinase (SphK), the enzyme responsible of S1P synthesis, is crucial to attribute to the SphK/S1P axis a functional significance in response to many different stimuli and in physiopathological conditions.This chapter describes experimental procedures to measure intracellular S1P formation in skeletal muscle cells and skeletal muscle fibers by using sphingolipid precursors. It also underlines the relevance of measuring S1P production in specific cellular compartments in order to attribute to S1P signaling a role in the biology of skeletal muscle cells.
1-磷酸鞘氨醇(S1P)是一种生物活性鞘脂调节剂,对哺乳动物和非哺乳动物细胞中的众多重要生理和病理过程发挥作用。越来越多的证据表明,许多细胞类型都可以产生并释放S1P;因此,对其细胞内和细胞外含量以及鞘氨醇激酶(SphK,负责S1P合成的酶)的活性进行定量,对于确定SphK/S1P轴在应对多种不同刺激和生理病理状况时的功能意义至关重要。本章介绍了使用鞘脂前体来测量骨骼肌细胞和骨骼肌纤维中细胞内S1P形成的实验方法。同时强调了在特定细胞区室中测量S1P产生的相关性,以便确定S1P信号在骨骼肌细胞生物学中的作用。