约氏疟原虫卵囊黑化与卵囊内游离钙关系的研究

中国寄生虫学与寄生虫病杂志 ›› 2003, Vol. 21 ›› Issue (2): 6-86.

约氏疟原虫卵囊黑化与卵囊内游离钙关系的研究

张锡林^*,徐文岳,王英,段建华,况明书

第三军医大学病原生物学教研室,重庆 400038

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2003-04-30 发布日期:2003-04-30

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

ZHANG Xi-lin,XU Wen-yue,WANG Ying,DUAN Jian-hua,KUANG Ming-shu

Department of Pathabiology,the Third Military Medical University,Chongqing 400038

Received:1900-01-01 Revised:1900-01-01 Online:2003-04-30 Published:2003-04-30

摘要/Abstract

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/LFluo-3/Am同时加入1μl/ml25%PluronicF-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。

关键词: 约氏疟原虫, 卵囊, 黑化, Ca~(2+), 共聚焦激光扫描显微镜(CLSM)

Abstract: 　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.

Key words: Plasmodium yoelii, oocyst, nnelanization, Ga2+, CLSM

张锡林;徐文岳;王英;段建华;况明书. 约氏疟原虫卵囊黑化与卵囊内游离钙关系的研究[J]. 中国寄生虫学与寄生虫病杂志, 2003, 21(2): 6-86.

ZHANGXi-lin;XUWen-yue;WANGYing;DUANJian-hua;KUANGMing-shu. Study on the Relationship between Intracellular Free Calcium and Melanization in Oocysts of Plasmodium yoelii[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2003, 21(2): 6-86.

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