牡蛎寄生虫——原生动物马里努斯派金虫的脂质类组成及其对荧光磷脂酰胆碱类似物的代谢。

Lipid class composition of the protozoan Perkinsus marinus, an oyster parasite, and its metabolism of a fluorescent phosphatidylcholine analog.

作者信息

Soudant P, Chu F L, Marty Y

机构信息

Virginia Institute of Marine Science, College of William and Mary, Gloucester Point 23062, USA.

出版信息

Lipids. 2000 Dec;35(12):1387-95. doi: 10.1007/s11745-000-0656-1.

DOI:10.1007/s11745-000-0656-1

PMID:11202001

Abstract

Perkinsus marinus is one of two important protozoan parasites of the eastern oyster, Crassostrea virginica. The other is Haplosporidium nelsoni. Lipids extracted from 7-d-old in vitro cultured P. marinus meronts, incubated with fluorescent-labeled phosphatidylcholine (FL PC) and nonincubated P. marinus meronts, were analyzed by a high-performance liquid chromatography (HPLC) system equipped with a diol phase column, in combination with thin-layer chromatography coupled with a flame-ionization detector (TLC/FID), and high-performance thin-layer chromatography (HPTLC). Various polar and neutral lipid classes were separated by HPLC using a two-gradient solvent system. Five polar lipid classes--phosphatidylcholine (PC), phosphatidylethanolamine (PE), cardiolipin (CL), sphingomyelin (SM), and phosphatidylserine (PS)--were identified from P. marinus extracts. Four neutral lipid classes--triacylglycerol (TAG), steryl ester (SE), cholesterol (CHO), and fatty alcohol--were distinguished. TLC/FID analysis of meront lipids showed that the weight percentages of PC, PE, CL, SM, PS/PI, TAG, SE, and CHO were 21, 10.7, 4, 2.3, 4.3, 48.7, 7.8, and 1.2%, respectively. HPLC and HPTLC analyses revealed the presence of two SM and PS isomers in P. marinus extracts. Perkinsus marinus effectively incorporated FL PC acquired from the medium and metabolized it to various components (i.e., free fatty acid, monoacylglycerol, diacylglycerol, TAG, PE, and CL). Uptake and interconversion of FL PC in P. marinus meronts increased with time. After 48 h the total uptake of fluorescence (FL) was 28.9% of the FL PC added to the medium, and 43% of the incorporated FL resided in TAG.

摘要

马里努斯派金虫是美国东海岸牡蛎（弗吉尼亚牡蛎）的两种重要原生动物寄生虫之一。另一种是尼尔森单孢子虫。从体外培养7天的马里努斯派金虫裂殖体中提取脂质，将其与荧光标记的磷脂酰胆碱（FL PC）一起孵育，以及未孵育的马里努斯派金虫裂殖体，通过配备二醇相柱的高效液相色谱（HPLC）系统进行分析，并结合薄层色谱与火焰离子化检测器（TLC/FID）以及高效薄层色谱（HPTLC）。使用双梯度溶剂系统通过HPLC分离各种极性和中性脂质类别。从马里努斯派金虫提取物中鉴定出五种极性脂质类别——磷脂酰胆碱（PC）、磷脂酰乙醇胺（PE）、心磷脂（CL）、鞘磷脂（SM）和磷脂酰丝氨酸（PS）。区分出四种中性脂质类别——三酰甘油（TAG）、甾醇酯（SE）、胆固醇（CHO）和脂肪醇。裂殖体脂质的TLC/FID分析表明，PC、PE、CL、SM、PS/PI、TAG、SE和CHO的重量百分比分别为21%、10.7%、4%、2.3%、4.3%、48.7%、7.8%和1.2%。HPLC和HPTLC分析显示马里努斯派金虫提取物中存在两种SM和PS异构体。马里努斯派金虫有效地摄取了从培养基中获得的FL PC并将其代谢为各种成分（即游离脂肪酸、单酰甘油、二酰甘油、TAG、PE和CL）。马里努斯派金虫裂殖体中FL PC的摄取和相互转化随时间增加。48小时后，荧光（FL）的总摄取量为添加到培养基中的FL PC的28.9%，并且掺入的FL中有43%存在于TAG中。