Journal of ultrastructure research最新文献

The functionally integratedTamarix salt gland consists of four piled pairs of cells, the three upper pairs of which constitute the main secretory cells while the lower pair corresponds to the so-called collecting cells. The whole gland is surrounded by the cuticle except for an area in the common wall between the inner secretory cells and the collecting cells (transfusion field). Each salt gland originates from a single protodermal cell with typical ultrastructure of a meristematic cell. In the progress of gland development no remarkable ultrastructural alterations occur in the secretory cells up to the stage of secretion. Since the secreted salt does not react with osmium tetroxide, the stage of secretion was estimated mainly on the basis of the mode of development of the subcellular structures considered to play an important role in the secretory process (wall protuberances, transfusion field, subcuticular space, small vacuoles, and mitochondria).

Clathrin assemblies were adsorbed to mica and freeze-dried by a new procedure that yields 3-D images with much topological detail. These permitted renewed inquiry into how clathrin trimers (i.e., “triskelions”) assemble into polygonal coats or baskets. Freeze-drying revealed unsuspected differences in the relative shapes and dimensions of individual trimer building blocks, as compared with the completed polygonal networks, which indicate that the assembly scheme first proposed by Crowther and Pearse (1) requires modification. Specifically, the freeze-etch images display the following new features: (1) Trimer arms possess terminal scroll-shaped hooks that can open or close and thus determine their lengths. (2) When extended, trimer arms are sufficiently long to pass around three sides of the final polygonal facet. Since current views hold that the arms pass around only two sides, the remaining length, including the terminal hook, must point into the basket interior. (3) Freeze-dried trimers display bends in their arms at specific loci that determine their final distribution in the completed baskets. (4) The completed struts of the final assemblies are uniform in calibre, cylindrical in profile, and travel directly between the vertices of each polygon, without any sign of the slew or width-variation that is predicted by the Crowther and Pearse model. Based on this direct comparison of protomer vs product, by a single technique that can image both, we offer a modified scheme for clathrin coat assembly, in which we predict that the individual arms in each clathrin triskelion emanate from its center in a slewed manner, but the final assembled struts of the basket need not be slewed. Attempts were made to capture assembly intermediates on mica to obtain support for the new scheme, but these unfortunately yielded ambiguous images of incomplete polygons with blunt projections, rather than the expected “halo” of uncommitted trimer arms. These we interpret to be “dead ends” that failed to polymerize further because they included proteolyzed components. Further assembly experiments, avoiding such hazards, are indicated.

Diplozoon is known to display an exceptional biology of reproduction: the hermaphroditic adults are permanently fused together and their genital ducts communicate. In contrast to all other polyopisthocotylean monogeneans in which the spermatozoa show an homogeneous biflagellate structure, the spermatozoon ofDiplozoon is aflagellate. It is filiform, and composed of a cytoplasmic region and a nuclear region. The cytoplasmic region exhibits mitochondria, a well-developed smooth endoplasmic reticulum, and up to 450 longitudinal singlet microtubules. The microtubules show links between them; seen in cross section, they are arranged as rows or polygons. The spermatozoon nuclear region contains the nucleus surrounded by cortical longitudinal microtubules. The spermiogenesis shows no zone of differentiation, a typical structure found in all other parasitic Platyhelminthes.Diplozoon is the first case of aflagellate spermatozoon found in the parasitic Platyhelminthes. The atypical sperm structure is not linked with phylogeny, but is well correlated with the atypical biology of reproduction.

Fine structural features of the murine myeloma MHFP-1 and two heterohybridomas secreting human IgM monoclonal antibody were examined. Intracisternal type-A retrovirus particles were found in both MHFP-1 and the heterohybridomas constructed by fusing MHFP-1 and human peripheral blood lymphocytes. The implications of this finding for the purification of human monoclonal antibody for therapeutic applications is discussed.

The ability of type A and type S synoviocytes to absorb horseradish peroxidase (HRP) and the intracellular fate of this tracer were studied by electron microscopic cytochemistry. Different concentrations of HRP (0.1–5 mg/ml) were injected into the left knee joint of rats and at intervals ranging from 1 min to 24 hr after injection the synovial membrane was fixed and incubated for HRP. Type A synoviocytes showed a striking ability to absorb HRP at low concentrations. At 1 and 5 min after injection reaction product was localized in coated pits and coated vesicles (110 nm) as well as in smooth-walled vesicles, vacuoles, and tubules. At 15 min to 4 hr postinjection the lysosomal system became increasingly loaded with reaction product. At 24 h after injection reaction product had disappeared. At higher concentrations of HRP similar observations were made in the A cells, but reaction product was still apparent in lysosomes at 24 hr postinjection. With respect to type S synoviocytes no reaction product was detected within these cells at any time interval after injection of low concentrations of HRP. However, at 5 min after injection of higher concentrations of HRP reaction product was localized in smooth vesicles and vacuoles mainly restricted to the large cytoplasmic processes facing the joint cavity. At 30 min to 4 hr postinjection the lysosomal system became progressively more loaded with HRP reaction product. At 24 hr after injection reaction product still remained in the lysosomal system.

The present findings that type A and type S synoviocytes showed major differences with respect to endocytic capacity and cellular structures involved in absorption of HRP support the interpretation that the A and S cells represent two distinct types of cells and further suggest that endocytosis in these two types of cells serve different functions.

The structure of two-dimensional crystals of membrane-bound Na,K-ATPase from rabbit kidney has been analyzed with a correlation averaging procedure. Two principally different crystal forms are observed with p1 and p21 symmetry, respectively. In the p1 form the averaged projection structure shows a triangular shaped protein domain interpreted as a protomer (αβ-unit) of Na,K-ATPase. In the p21-form the stain-deficient area is extended toward a twofold symmetry axis. The results are in good agreement with a previous analysis where Fourier methods were applied to well ordered crystals of pig kidney Na,K-ATPase and illustrate that the correlation averaging procedure can be used for the analysis of membrane crystals of Na,K-ATPase showing curved lattice lines.

The structural responses of cells in the distal convoluted, connecting, and collecting tubule to acute acid/base changes were investigated by electron microscopy. Acute metabolic acidosis was induced by administration of ammonium chloride, and acute metabolic alkalosis by potassium or sodium bicarbonate. Morphometric analyses were performed on micrographs of randomly selected distal nephron cells. No structural responses were found in distal convoluted tubule cells, connecting tubule cells, or principal cells but prominent changes were observed in intercalated cells (I cells). Thus, the surface density of the luminal membrane in I cells was significantly higher in acidotic animals and lower in KHCO₃ alkalotic animals than in controls. On the contrary, the surface density of the membrane that bounds apical vesicles was higher in KHCO₃ alkalotic and lower in acidotic animals than in controls. These results suggest that the luminal membrane is internalized during alkalosis and that the membrane that bounds apical vesicles is transferred to the luminal membrane during acidosis. Since a proton translocating ATPase may be present in the luminal membrane the observations are consistent with the possibility that cortical I cells participate in the maintenance of acid/base homeostasis.