The Improvement Of The Method-FLUTAX And Visualization Of The Microtubular Cytoskeleton Of Euplotes

With special phenomena of encystment, excystment and binary fission, as well as complex cytoskeleton, the study of cytoskeleton of ciliates has been one of the important reaearch fields in protozoology and cell biology . To further investigate the characters and component of cytoskeleton of ciliates, the author found an valid method-FLUTAX ; With this method ,to observed the cytoskeleton of ciliates in different physiological states ; by means of SDS -PAGE, to investigate the componnt of cytoskeleton of ciliates in in different physiology.The results are showed as fellow:1. The improvement of method of FLUTAXBased on the method of Arregui(2002),the author improved the method to adapt to studying cytoskeleton of Hypotrich ciliates. The article expatiates the modification of the reagent of FLUTAX, demandment of the material, emphasizes the technique of pervasion and fixation. With the modified method of FLUTAX ,we can clearly observe microtubule organelles of the different Hypotrich ciliates. This paper affords an important valid approach to investigate the shape, assembly and function of the microtubules cytoskeleton.2. The shape aad the differentiation of microtubules cytoskeleton of euplotes encysticus in different physiological states.With the improved method provided by the author , the shape and differenation of the microtubule and microtubule-like structure of euplotes encysticus in different physiological statesman be visioned clearly. Durong the binary fission of the cell, the new cirri kinetosomes come into being in certain given position of the vental side, they develop into cirri and migrate and divide into two sets of cirris, at last take the place of the old ones .on the ventral side of cell, the adoral zone of membrane(AZM), the underlating membrane(UM), froto-ventral and transverse cirri(FVC,TC) and caudalcirri migrate and then are disintegrated . The mode of the ariseing of the new doral cirris are different form the ventral cirris's, which emerge closely behind the old cirri, and then the senond , the third ...at last ,those new cirris form two set cirri,and the old ones disintegrate and are absorbed. Just right before the encystmemnt of euplotes encysticus ,the ciliary shafts and some kinetosomes of the frotalal and ventral cirris are disintegrated and disappeard , the remaining ones aggregate togeher ,and then are folded with the cortical pellicle within the cyst, the cell become the round cyst, while the dorsal cirris remains intact without being absorbed. Right before the excystment of euplotes encysticus, the microtubule -like structures are differentiated into component cirris based on the remaining structures , the cell come out of the cyst as "amoeba"-like form , and turn to vegetative cell.Summarizeing the characters of the differentiation of the microtubulelike organelle of the euplotes encysticus,duoring theose process of fission ,encystment and excystment, the author speculate that: microtubuie-like structure is the pivotal structure in the ife activity,of the cell, which change continuously douring the life activity. The old cirris play very important role to the new cirri orgalles. It may contribute substance source to the new ones ,and induce the direct orientation of the new ones. In addition ,during the excystment ,the cell form amoeba-like , this may be caused by the special differentiated form of the microtubule. 3. Comparison of protein component between vegetative cells and resting cellsCompared with vegetative cells and the resting cells of euplotes encysticus by means of SDS-PAGE .The result showed as fellow: the resting cells of euplotes encysticus preserve some part of the protein bands: 95. 3. 82. 9, 77. 2. 65. 7s 62. 3. 58.3. 48.6. 40.4, 37.2. 35.2. 31.0. 25.1. 18.4. 16. 3 kD(whichare share with the vegetative cells, whereas lost some protein bands, meanwhile, produce some new particular proteins: 70.1. 56.3. 46.2. 28.2 26.2, 24.2 22.2 kD. The result shows that the protein composition has changed notably in the course of encystment, the change of the compoment of microtubu...