A Scaffold for Basal Body Patterning Revealed by a Monoclonal Antibody in the Hypotrich Ciliate Paraurostyla weissei

Fleury, Anne; Guyader, Hervé Le; Iftode, Francine; Laurent, Michel; Bornens, Michel

doi:10.1006/dbio.1993.1135

Cited by 37 publications

(38 citation statements)

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“…Immunodetection was conducted using a method modified from Fleury et al (1993). The cells were permeabilized in PHEM buffer (PIPES 60 mM, HEPES 25 mM, EGTA 10 mM, MgCl 2 2 mM) with 0.5% saponin for two minutes, followed by one-hour fixation in 4% paraformaldehyde in PHEM buffer at RT.…”

Section: Immunocytochemistrymentioning

confidence: 99%

Localization of α-, γ-, and δ-tubulin in the hypotrich ciliateStylonychia pustulata(Hyportrichida, Ciliophora)

Yin

Sheng

Gao

et al. 2013

Animal Cells and Systems

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Localization of α-, γ-, and δ-tubulin in the hypotrich ciliate Stylonychiapustulata (Hyportrichida, Ciliophora), Animal Cells and Systems, 17:4, 259-268, DOI: 10.1080/19768354.2013.828654 To link to this article: https://doi.org/10. 1080/19768354.2013 Ciliature microtubular organelles, which organize into ciliary units, are highly complex microtubular systems. To investigate the formation of ciliature microtubular organelles and the role that different tubulins play in this process, we examined the distribution and function of a-, g-, and d-tubulins in the hypotrich ciliate Stylonychia pustulata. Immunofluorescence analysis demonstrated that a-tubulin localized to all microtubular organelles, while g-and dtubulins colocalized at basal bodies. Immunogold labeling revealed that g-and d-tubulins colocalize to the proximal part of basal bodies, suggesting that g-and d-tubulins probably interact and may function together. In addition, the permanent localization of g-tubulin to the adoral zone of membranelles suggests that g-tubulin play a role in the maintenance of oral ciliature base-associated microtubules, but does not play a role in the biogenesis of these ciliature. We observe that d-tubulin is restricted to the basal body and the connective fibers of the basal body, suggesting this protein may be required for the extension and stability of the microtubules in these structures. Dramatically different concentrations of these three tubulins were observed in the cytoskeleton and the insoluble and soluble fractions of the cell, suggesting that these highly related proteins perform different functions in S. pustulata.

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Section: Immunocytochemistrymentioning

confidence: 99%

Localization of α-, γ-, and δ-tubulin in the hypotrich ciliateStylonychia pustulata(Hyportrichida, Ciliophora)

Yin

Sheng

Gao

et al. 2013

Animal Cells and Systems

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“…They are large enough to image easily with light microscopy and are amenable to routine cytological staining [13,18,19,20]. Staining of surface features for confocal microscopy is a known art, and a functional method was published by Ann Fleury [14,15], Fleury and Laurent [6] and Fleury et al [17].…”

Section: Ciliated Protistsmentioning

confidence: 99%

An inquiry into the morphology of Ciliate Protozoa using an engineering design approach

Benjamin¹,

Collins²,

Roberts³

2006

WIT Transactions on Ecology and the Environment, Vol 87

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Although cellular morphogenesis is an area of microbiology that is widely studied, very little is understood about the manner in which cells shape themselves or how they optimise their form to their environment. There exists an extensive literature on body shape, anatomy and life cycle of numerous single-celled micro-organisms including both prokaryotes and eukaryotes. However, with regards to morphology itself, there is a shortage of general empirical relationships to enable the interknitting of specific features that could lead to a biologically justifiable generic form. This paper comprises a concise review of the subject together with an assessment of the experimental approach which could be used, within an intended overall context of Engineering Design.

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“…Paraurostyla raises interesting problems in cellular differentiation mechanisms because this species undergoes a cycle of dedifferentiation [14,15]. Its thousands of cilia are grouped into clusters which define a highly asymmetric ciliature.…”

Section: Confocal Microscopy Bridges the Gap Between Light And Electrmentioning

confidence: 99%

“…Cells were processed for immunolabelling according to [15]. The issue is especially important in confocal microscopy since the optical slices are performed sequentially.…”

Section: Photodestruction Of Fluorophoresmentioning

confidence: 99%

Power and limits of laser scanning confocal microscopy

LAURENT¹

1994

Biology of the Cell

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In confocal microscopy, the object is illuminated and observed so as to rid the resulting image of the light from out-offocus planes. Imaging may be performed in the reflective or in the fluorescence mode. Confocal microscopy allows accurate and nondestructive optical sectioning in a plane perpendicular or parallel to the optical axis of the microscope. Further digital three-dimensional treatments of the data may be performed so as to visualize the specimen from a variety of angles. Several examples illustrating each of these possibilities are given. Three-dimensional reconstitution of nuclear components using a cubic representation and a raytracing based method are also given. Instrumental and experimental factors can introduce some bias into the acquisition of the 3-D data set: self-shadowing effects of thick specimens, spherical aberrations due to the sub-optimum use of the objective lenses and photobleaching processes. This last phenomenon is the one that most heavily hampers the quantitative analysis needed for 3-D reconstruction. We delineate each of these problems and indicate to what extent they can be solved. Some tips are given for the practice of confocal microscope and image recovery: how to determine empirically the thickness of the optical slices, how to deal with extreme contrasts in an image, how to prevent artificial flattening of the specimens. Finally, future prospects in the field are outlined. Particular mention of the use of pulsed lasers is made as they may be an alternative to UV-lasers and a possible means to attenuate photodamage to biological specimens. confocal microscopy / fluorescence microscopy / reflectance microscopy / photobleaching / three-dimensional imaging 13th I n t e r n a t i o n a l C o n g r e s s on E l e c t r o n M i c r o s c o p y

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A Scaffold for Basal Body Patterning Revealed by a Monoclonal Antibody in the Hypotrich Ciliate Paraurostyla weissei

Cited by 37 publications

References 0 publications

Localization of α-, γ-, and δ-tubulin in the hypotrich ciliateStylonychia pustulata(Hyportrichida, Ciliophora)

Localization of α-, γ-, and δ-tubulin in the hypotrich ciliateStylonychia pustulata(Hyportrichida, Ciliophora)

An inquiry into the morphology of Ciliate Protozoa using an engineering design approach

Power and limits of laser scanning confocal microscopy

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