Research On The Microtubular Cytoskeleton Of Paraurostyla Weissei

With complex microtubular cytoskeleton which played important role in the maintaince of the figure and the function of the cell, the ciliates have been the ideal materials in the study of microtubular cytoskeleton which dedifferentiated and reclassifided in the cell division and reorganization. In the past, partial cytoskeleton of some ciliatures or some position of the cortex of few ciliates was obtained, and it aimed at the fibre but not the tubulin. In order to futher investigate on the characters of the microtubular cytoskeleton and the orientation, distribution of the tubulin of the ciliates, the microtubular cytoskeleton of Paraurostyla weissei which was the more complicated ciliates with complicated microtubular structures and tightly- assembled cortical ciliatures was analyzed with SEM, FLUTAX, a-tubulin antibody,SDS-PAGE and the distribution of tubulin was explored with SDS-PAGE. the results were as the following.1 Investigation on the development of oral, cirral, and dorsal primordial of Paraurostyla weissei fixed in different period with SEM.In the interphase, Paraurostyla weissei had " AZM—FC-VC-TC- L- & RMC " model in its ventral side. Six rows of kineties were labeled in the doral side. In dividing cells, part of the old AZM renewed, the primordium of the AZM of proter arised at the original area; the primordium of the new AZM of opisthe appeared posterior to the old AZM, the UM-primordium, FVTC-primordium appeared at the right area of it. and at this place the old ciliature disorganized, new ciliature grew on definite position of cell cortices. From the creation, disposition and movement of the corresponding ciliature anlagens, to finally getting the new ciliature of proter and the opisthe, the marginal cirri primordium appered in the forside and middle side of the old cirri, then extended and ciliated at the right area and replace the old ones finally. At the early stage of the development of the doral kinety, there are two sets of dorsal ciliary primordia on each side of the equator of the cell, and three rows of primordia in each side, then, the primordium ciliated and eloganted to the rearward of the cell. In this process, the third column formed two columns of kineties and part of the primordial formed the caudal cirri. Furthermore, there are two columns of primodia appears beside the new right marginal cirri and finally formed the mature doralkineties. The course of development of the ciliatures in reorganization process is similar with the proter in dividing cells, but part of the AZM renewed which was different with the proter in division. In summary, the old ciliature is the template of the new ones and devote themselves into the formation of the new with some substances.2 The morphology and morphogenesis of the ciliature microtubular organelles in the ventral cortex of Paraurostyla weissei analyzed with fluorescence labeling of FLUTAX.These organelles consisted of adoral zone of membranelles (AZM), undulating membranes (UM), frontal-ventral-transverse cirri (FVTC), left & right marginal cirri (L- & RMC) and the base associated microtubules of these ciliatures. The microtubular cytoskeleton of AZM comprised membranelle brackets; The base associated microtubules of FVTC, L- & RMC contained anterior longitudinal microtubules (ALM), posterior longitudinal microtubules (PLM) and transverse microtubules (TM), these microtubules sent forth from the ciliatures and extended to different orientation, formed the microtubule net which below the surface of ventral cortex. besides this, the constructive character of marginal cirri base-associated microtubules of P. weissei were different from those of other ciliates (e.g. Stylonychia mytilus), whose TM of left marginal cirri emitted from the ciliature and extended to the right area. During the process of the morphogenesis, part of the old AZM was renewed, together with the leftever constitute an intact AZM at the orginal area which belonged to the proter. The old marginal cirri played an important part in the orientation of the new marginal cirri and supplied its development with some substances. After that, the new marginal cirri formed at the right area but not the originally area of the old ones. In some cells, one row of cirri was appeared at the left of the new left marginal cirri, it may be formed because of the old left marginal cirri which degenerated incompletely. During morphogenesis of physiological reorganization. The evolvement of ciliatures were similar with the process of cell division. but one suit of ciliature was formed. And a new primordium of adoral zone of memebranelles always forms and connects with the old AZM to make a new one.but there was obvious clearance between them. At last it was filled up with the process of physiological reorganization. According to the differentidtion feature of the microtubule organelles in different physiological states, the writer assumes that microtubule organelle made a continuous change in different paste of Paraurostyla weissei's life cycle, and so it's the foremost important structure of cell. In the progress of shape creation and structure disposition, former microtubule structure may have physical contribution to the building of new construction, and it has the function of structure inducement and pattern location.3 research on the growing progress of microtubule organelles and the distribution of a-tubulin in Paraurostyla weissei in different period.By combining with a-tubulin in the organelles and its skeleton, it tracked and marked the creation, movement and placement of ciliature skeleton and organelle microtubules during the cell cycle. Regarding the interphase cell, besides the same results achieved by labeling of FLUTAX, some new configuration and phenomenawere obtained: at the base of the ventro-AZM, two bundles of base-associated microtubules crossed with each other and formed a "V" shape; at the base of the transverse cirri, besides the triangular microtubular layer which was assembled by its anterior longitudinal microtubules (ALM), there also existed obvious posterior longitudinal microtubules; six columns of kineties were observed at the doral of Paraurostyla weissei/, and several caudal cirri existed at the end of the cell. From these results we conclude that the a-tubulin expresses stably in Paraurostyla weissei in different period and exists in the whole ciliatures. During the morphogenesis of cell division and physiological reorganization. The ciliature base-associated microtubules were disappeared with the ciliature from left to right. The results indicate that the the morphogenesis of the ventral ciliature followed the same sequence. Besides this, during the growing of dorsal ciliatures, the new ciliate anlagens grow at the back of several old kineties,and finally becoming tow new set of dorsal kinety, the proter and the opisthe, and caudal cirri were formed from part of these new dorsal kineties-primordium. These results indicate the basal components of the ciliature cytoskeleton and the ciliature base-associated microtubules of P. weissei are tubulins, From these results we also conclude that the a-tubulin expresses stably in Paraurostyla weissei in different period and exists in the whole ciliatures. and In the progress of shape creation and structure disposition, a-tubulin do some contribution in the endenger of the new ciliature primordium and the former microtubule structure may play important role in the differentiation and development of new ones . 4 accomplishment on some comparison to the total protein, cytoskeleton protein and cytoskeleton protein in which the tubulin had been extracted.The tubulins had a high throughput expression in both the total protein and the cytoskeleton protein; the cytoskeleton protein and the total protein have many bands in common, the common protein has high expression. Then, writer assumes that cytoskeleon whose main element is microtubule cytoskeleton is of hypertextualization in protozoa. After extracting the tubulin from the cell. there was a band at 66KD around left, it may be responsible for the maintenance of the cell and the stability of the cytoskeletonConclusions: In this study , the constitution and distributing characteristics of microtubular organelles and base-associated microtubules of Paraurostyla weissei were observed, and the constitutions and model of microtubular cytoskeleton was revealed in detail. The results might not only be helpful in the understanding of microtubular cytoskeleton in ciliates, but also provide foundamental data to discuss the cell model and its forming and the evolution and differentiation of microtubules in eukaryotes. Furthermore, the primary explore of the function of some unclear protein was carried out in this paper, and it was worthwhile to make further researches on its frondose functions in the future.