摘要:

Summary—We have developed an automated image analysis method to study the ciliary beat frequency of ciliated cells of the primary culture from rabbit trachea. The ciliated outgrowth image is digitized and the variation in optical density is automatically calculated for each selected area of interest. 32 measurements of ciliary beat frequency are, in this way, calculated simultaneously in 6 min. With this reliable device, some studies on baseline frequency of control culture have been carried out. There was no variation in the mean frequencies of ciliated cells of the primary culture of different tracheas in our culture conditions. Moreover, the values of ciliary beat frequency at the starting point of the outgrowth were similar to those at the periphery of the outgrowth. There is nevertheless a slow decrease in frequenciesversusthe duration of culture. We have also established that the frequency of ciliary beat of some cells fluctuates in a periodic pattern whereas the majority of the ciliated population beat in a stable way. The image analysis process allows us to perform a cartography of frequencies on the video display. It also allows us to have access to the frequency of one cilium. Our method therefore seems to be reliable and furthermore simple in the evaluation of the potential effect of inhaled toxic compounds on ciliated cells of mammalian respiratory trac

ISSN:0248-4900    

DOI:10.1016/0248-4900(91)90064-T

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

Summary—Monoclonal antibody CC‐248 labels cilia differentially on Triton X‐100 permeabilized ciliated epithelium of quail oviduct by indirect immunofluorescence. On isolated ciliated cells, a punctuated staining is seen at the distal region over the bend of cilia. Electron micrographs of immunoperoxidase and immunogold techniques showed that the punctuated fluorescence corresponds to a helical disposition of CC‐248 antigenic sites. This labeling was arranged on the axonemal distal region either as a simple or a double helix externally disposed around the nine microtubular doublets. These results suggest the existence of a detergent insoluble structure in the ciliary matrix that might concern the ciliary skeleton, probably acting as an elastic recoil that keeps the structural integrity of the axoneme during bending. The cross‐reactivity of CC‐248 MAb with the intermediate filament cytoskeleton of ciliated and smooth muscle cells indicates that this structure might be related to the intermediate fila

ISSN:0248-4900    

DOI:10.1016/0248-4900(91)90065-U

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

Summary—In ciliated cells of metazoa, striated rootlets associated with basal bodies anchor the ciliary apparatus to the cytoskeleton. We have used here a monoclonal antibody against a 175 kDa protein associated with the striated rootlets of quail ciliated cells [14], to study ciliated cells of different species. In mussel gill epithelium the antibody recognized a protein of 92 kDa which shows a periodic distribution along the striated rootlets. In frog ciliated palate epithelium, two different rootlets are associated with basal bodies, both are decorated and only one protein of 48 kDa is recognized on immunoblot. The antigen is arranged in a helix around the striated rootlets. In rabbit ciliated oviduct epithelium, we detected the presence of very small and thin rootlets which are weakly labeled. We have shown that an epitope associated with the striated rootlets is preserved through evolution although the molecular weight of the peptide varies. We have also observed the appearance of this epitope on protein associated with junctional complexes in rabbit and cytoskeleton component in quail oviduc

ISSN:0248-4900    

DOI:10.1016/0248-4900(91)90066-V

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

Summary—In this paper the authors describe the ultrastructure of the mature spermatozoon and the spermatid inStruthio camelusandDromaius novaehollandiae. The first species is characterized by a rod‐like foratorium within an endonuclear canal in the anterior third of the nucleus, while the second is characterized by an extremely reduced completely extranuclear perforatorium. Other differences are in the sperm dimensions, the number of mitochondria and the length of the axonemal accessory fibers. Considering both the present data and previous findings, Palaeognath birds appear to be a peculiar and monophyletic group, characterized by: 1), a conical acrosome surrounding the nucleus; 2), a fibrous sheath aroundmost of the axoneme; and 3), an elongated distal centriole occupying the entire midpiece. Within this group, Tinamiformes seem to be more primitive than Struthioniformes. In the latter orderDromaiusis distinctly different from the reducedStruthioandRheawhich are closely related to one another by the presence of a rod‐like endonuclear perforat

ISSN:0248-4900    

DOI:10.1016/0248-4900(91)90067-W

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

Summary—The infraciliary lattice (ICL) is the innermost cortical cytoskeletal network ofParamecium. Its meshes which run around the proximal end of basal bodies form a continuous contractile network beneath the cell surface. We had previously shown that the network, which could be recovered in a contracted form and selectively solubilized by EGTA from an ICL‐enriched cell fraction, was principally composed of 23–24 kDa polypeptides cross‐reacting with antibodies raised against the 22 kDa Ca2+‐binding proteins of the ecto‐endoplasmic boundary (EEB), a contractile cytoskeletal network of another ciliateIsotricha prostoma. We show here 1) that the ICL also comprises a 220 kDa polypeptide; 2) that the 23–24 kDa polypeptides are resolved in 2D gels into 11 spots of acidic pI, 7 of which are both Ca2+‐binding and cross‐reacting with the anti EEB polypeptides; 3) that the network displays a high Ca2+‐affinity as the treshold for solubilization/co‐precipitation of both high and low MW polypeptides is around 10−8M free Ca2+; 4) thatin vivocontraction of the network occurs upon physiological increase of internal calcium concentration. The likely phylogenetic relationships of the 23–24 kDa ICL polypeptides with the calmodulin related family of Ca2+‐modulated polypeptides and the functions of the ICL in cell contractility and Ca

ISSN:0248-4900    

DOI:10.1016/0248-4900(91)90068-X

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

Summary—The diversity of microtubular networks was analyzed in quail oviduct and inParameciumcells using conventional and confocal immunofluorescence as well as pre‐ and post‐embedding EM immunocytochemistry with a variety of anti‐tubulin antibodies. The 6‐11B‐1 monoclonal antibody, specific for the post‐translational acetylation of Lys 40 of α‐tubulin [40], and a polyclonal antibody raised againstParameciumaxonemal tubulin (anti‐PA tubulin antibody) [16] both decorated stable microtubular arrays inParamecium ieciliary axonemes and a set of microtubular bundles associated with the cortex, suggesting that the two antibodies may be directed against the same epitope. However, several differences in the immunocytological patterns yielded by each antibody on the two cell types were evident. For example, in quail, as in all other Metazoa, the anti‐PA tubulin antibody only decorated axonemes enclosed in normal ciliary membrane while it was unreactive on cytoplasmic tubulins. Immunoblotting of peptide maps of axonemal tubulins demonstrated that the epitopes of the two antibodies were indeed completely different. Double immunolabelling of dividing paramecia using a universal anti‐tubulin antibody and the anti‐PA tubulin one revealed that all newly assembled microtubular arrays were first detected by the universal antibody and, only shortly afterwards, by the anti‐PA tubulin one. This provided a strong indication that the anti‐PA tubulin antibody is directed against a post‐translational modification taking place on already assembled microtubules (MTs) (as previously known to be the case for acetylation and detyrosination). In taxol‐treated quail cells undergoing ciliogenesis, massive assembly of MTs and even axonemes occurred in the cytoplasm. These MTs were not decorated by the anti‐PA tubulin antibody however, suggesting that in Metazoa the post‐translational modification can only take place within the ciliary lumen. The present work provides one further mechanism for generating MT immunological and biochemical diversity post‐translationally; this may account for the high multiplicity of tubulin isoforms observed in ciliates which contain very little if

ISSN:0248-4900    

DOI:10.1016/0248-4900(91)90069-Y

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

ISSN:0248-4900    

DOI:10.1016/0248-4900(91)90043-M

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY