约氏疟原虫卵囊黑化与卵囊内游离钙关系的研究  

Study on the Relationship between Intracellular Free Calcium and Melanization in Oocysts of Plasmodium yoelii

作　　者：张锡林[1] 徐文岳[1] 王英[1] 段建华[1] 况明书[1] 

机构地区：[1]第三军医大学病原生物学教研室,重庆400038

出　　处：《中国寄生虫学与寄生虫病杂志》2003年第2期84-86,F003,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No.30200237)

摘　　要：目的 研究大劣按蚊蚊胃的约氏疟原虫卵囊黑化时卵囊内Ca^(2+)的变化,探讨疟原虫感染不易感蚊种时卵囊黑化与囊内Ca^(2+)的相互关系。方法 以荧光染料Fluo-3/Am作卵囊内钙指示剂,应用共聚焦激光扫描显微镜(CLSM)观察不同时期约氏疟原虫卵囊内Ca^(2+)分布、变化以及检测的实验条件。结果 3μmol/L Fluo-3/Am同时加入1μl/ml 25%Pluronic F-127在37℃恒温下,孵育约氏疟原虫卵囊60min即可达到最佳的负载效果。Fluo-3/Am浓度的提高和孵育时间延长只能增加背景染色,若降低孵育浓度和缩短孵育时间则难于达到最佳的负载效果。约氏疟原虫成熟卵囊的囊内Ca^(2+)为(137.15±7.02)nmol/L,完全黑化卵囊为(18.44±1.75)nmol/L,并在囊壁出现明显的钙沉着。 结论 约氏疟原虫卵囊完全黑化时,囊内Ca^(2+)明显减少,并有外排现象。Objective To study the change of intracellular free Ca2+ in the oocyst when it melanized and to find out the relationship between the melanized oocyst and its intracellular level of free Ca2+ in a Plasmodium-refractory strain of Anopheles dirus. Methods The distribution and experimental condition of the intracellular free Ca2+ in oocyst of Plasmodium yoelii was measured with Ca2+ sensitive dye Fluo-3/AM and Plutonic F-127 under confocal laser scanning microscope (CLSM) at different time. Results The best load condition was that the oocysts were incubated in 3 μmol/ml Fluo-3/AM adding 1 μl/ml 25% Pluronic F-127 for 60 min at 37 ℃ . Fluorescent imaging of oocysts was affected by an increase or decrease of the concentration of Fluo-3/Am and incubation time. The distribution of intracellular free Ca2+ was heterogeneous in the oocysts. The mean value of Ca2 + in the mature oocysts was (137.15 ±7.02) nmol/L(X±S) but was (18.44± 1.75) nmol/L in melanized oocysts with Ca2+ sedimentation in the wall of oocyst. Conclusion The results suggest that the level of the intracellular free Ca2+ in oocyst decreased and excreted during its melanization in a Plasmodium-reiractory anopheline mosquito species.

关 键 词：约氏疟原虫 卵囊黑化 游离钙 共聚焦激光扫描显微镜 

分 类 号：R382.31[医药卫生—医学寄生虫学]