Direct Fluorescent Labeling And Observation With Scanning Electronic Microscopy Of Uroleptus Sp.(Hypotrichia, Ciliophroa)

Hypotrichous ciliates are undoubtedly the most complicated and certainly the most evolved group among so many ciliated protozoa. The cells contain a variety of structural components forming the cortex, and studies on the cortex especially the microtubular organelles and microtubular cytoskeleton in it has become one of the hottest spot in cell biology. With the relative species-specific stability, Microtubular organelles and its associated microtubules has contributed to a better understanding on the protozoan ciliate evolutionary relationship between species and made the research on the regulation model of eukaryotic cell and cell diversity good sense as well.As a fascinating ciliate, the phylogenic status of the uroleptus ciliate has always been a huge controversy, and the introduction of a variety of modern biology experimental technology makes the controversy more complicated. In this study, the microtubular organelles and microtubular cytoskeleton in the cortex were discussed by using direct fluorescent labeling and scanning electronic microscopy. And the result can provide the information of the construction characteristics of microtubular organelles and other characteristics such as its organization, its location its orientation. The result also illustrated the specific characteristic of the morphology of the ciliates and provided many basic information about the diversity of the structure of the ciliates. The result can provide grate useful value about the classification and systematic study of uroleptus ciliates. The results are as follows:1 Observation of the cortex surface in the uroleptus ciliate with scanning electronic microscopyThe cortex surface in the uroleptus ciliate was analyzed with scanning electronic microscopy. The uroleptus ciliate is 160-180μm in. length and 30μm in width, which has a body with streamline shape and a distinct long tail. Its ventral side is flat and its dorsal side is distinctly vaulted. The cortex ciliature of the uroleptus ciliate consists of adoral zone of membranelles(AZM), undulating membranes(UM), front-ventral-transverse cirri(FVTC), left and right marginal cirri(L -and- RMC), dorsal kineties(DK). The AZM has a shape like a question mark and located in the left front of the ciliate, which accounts for 1/5 of the body length. The UM comprises a endoral membrane (EM) and a paroral membrane(PM) which both start at the base of the AZM. The UM is parallel to the PM, which both membrane locate at the left side of the AZM. The ciliate has three FT which has a distinct differentiation and were arranged in linearly at the front part of the ciliate. There exists two rows of midventral cirri which is arranged in a zigzag position. The right row is distinctly larger than the left one. The two rows both start at the middle part of the AZM and extent to the rear part of the ciliate. A LMC and a RMC are located in the ventral side of the ciliate. The LMC start at the middle part of the left AZM and the other one start at the second cirri of the MVC. THE two rows parallel to each other at the forepart of the ciliate and converge at the rear. The TC consists of four cirri which were arranged in a checkmark position. The CC is not recognizable and not distinct. More than five rows of DK locate in the dorsal side of the ciliate.The article put forward an improvement to this ciliate which is very long and easily deformed. It is an important reference to the preparation method with scanning electronic microscopy to similar ciliates.2 Microtubular organelles and microtubular cytoskeleton in uroleptus sp. revealed by direct fluorescent labeling.The microtubular organelles and microtubular cytoskeleton in uroleptus sp. were revealed by direct fluorescent labeling of FLUTAX. The results showed that the organelles consist of adoral zone of membranelles(AZM), undulating membranes(UM), front-ventral-transverse cirri(FVTC), left and right marginal cirri(L -and- RMC), dorsal kineties(DK). Beneath the cirri there exists microtubular cytoskeleton of different developed and different orientation. The microbubular cytoskeleton of AZM comprises membranelle brackets and its associated microtubules; the base associated microtubules consist of anterior longitudinal microtubules (ALM), posterior longitudinal microtubules (PLM), transverse microtubules(TM); the transverse microtubules of the left marginal cirri underdeveloped, the transverse microtubules of the right marginal cirri developed, the posterior longitudinal microtubules of both left and right marginal cirri underdeveloped. There are many rows of dorsal kineties on the dorsal surface. The result also indicated that the cytoskeleton of Uroleptus sp.is built up mainly with the microtubules as its major units, and the extent of development of the base associated microtubules beneath all of these cirri possibly is related with the phylogenic status of the ciliates. As a result, the microtubular cytoskeleton,which are a very important component of the cortex,can not only deepen the understanding of the origin, evolution, classification of the uroleptus ciliate, but also can contribute to the diversity of the ciliate phylogeny, the origin of the ciliates, the mechanism of eukaryotic cell.