Muñoz Yorka, Fuenzalida Karen, Bronfman Miguel, Gatica Arnaldo, Sepúlveda Marcelo, Bacigalupo Juan, Roth Alejandro D, Delgado Ricardo
Department of Biology, Faculty of Sciences, University of Chile, Santiago, Chile,
Faculty of Biological Sciences, Pontifical Catholic University of Chile, Santiago, Chile.
Biol Res. 2013;46(3):289-94. doi: 10.4067/S0716-97602013000300010.
Phototransduction, the mechanism underlying the electrical response to light in photoreceptor cells, has been thoroughly investigated in Drosophila melanogaster, an essential model in signal transduction research. These cells present a highly specialized photosensitive membrane consisting of thousands of microvilli forming a prominent structure termed a rhabdomere. These microvilli encompass the phototransduction proteins, most of which are transmembrane and exclusively rhabdomeric. Rhabdomere membrane lipids play a crucial role in the activation of the transient receptor potential ionic channels (TRP and TRPL) responsible for initiating the photoresponse. Despite its importance, rhabdomere lipid composition has not been established. We developed a novel preparation enriched in rhabdomere membranes to perform a thorough characterization of the lipidomics of Drosophila rhabdomeres. Isolated eyes (500) were homogenized and subjected to a differential centrifugation protocol that generates a fraction enriched in rhabdomere membrane. Lipids extracted from this preparation were identified and quantified by gas chromatography coupled to mass spectrometry. We found an abundance of low sterol esters (C16:0, C18:0), highly abundant and diverse triglycerides, free fatty acids, a moderate variety of mono and diacyglycerols (C:16:0, 18:0, C18:1) and abundant phospholipids (principally C18:2). This preparation opens a new avenue for investigating essential aspects of phototransduction.
光转导是光感受器细胞对光产生电反应的基础机制,在果蝇(信号转导研究中的重要模型)中已得到充分研究。这些细胞呈现出一种高度特化的感光膜,由数千个微绒毛组成,形成一个突出的结构,称为视杆。这些微绒毛包含光转导蛋白,其中大部分是跨膜蛋白且仅存在于视杆中。视杆膜脂质在激活负责启动光反应的瞬时受体电位离子通道(TRP和TRPL)中起关键作用。尽管其很重要,但视杆脂质组成尚未确定。我们开发了一种富含视杆膜的新型制剂,以全面表征果蝇视杆的脂质组学。将分离的眼睛(500个)匀浆,并进行差速离心方案,以产生富含视杆膜的级分。从该制剂中提取的脂质通过气相色谱-质谱联用进行鉴定和定量。我们发现了大量的低甾醇酯(C16:0、C18:0)、高度丰富且多样的甘油三酯、游离脂肪酸、种类适中的单酰甘油和二酰甘油(C:16:0、18:0、C18:1)以及丰富的磷脂(主要是C18:2)。这种制剂为研究光转导的基本方面开辟了一条新途径。