Ultrastructural localization of oxidative and peroxidative activities in a carrot suspension cell culture

Summary Oxidative and peroxidative activities were localized at the ultrastructural level in suspension cells of an anthocyanin-producing strain of carrot after treatment with dihydroxyphenylalanine (DOPA) and diaminobenzidine (DAB). In DOPA-treated cells a reaction ascribed to polyphenoloxidase (PPO) occurred in the thylakoids of plastids. After DAB treatment at pH 9.0 reactions occurred in microbodies and plastid thylakoids; after treatment at pH 6.8 additional reactions occurred in the mitochondrial cristae and cytoplasmic ground substance. No reaction occurred in the cell walls at either pH. A reaction could not be unequivocally detected in the vacuoles because of the natural occurrence of osmiophilic material. Application of peroxidase and PPO inhibitors indicated that four distinct systems were involved in the DAB reactions: catalase was correlated with the reaction in the microbodies, peroxidase with the reaction in the cytoplasmic ground substance, cytochromes with the mitochondrial reaction, and PPO with the reaction in the thylakoids of the plastids.Contribution No. 1964 of the Rhode Island Agricultural Experiment Station.     

Regulation of Muscarinic Receptor-Mediated Cyclic GMP Synthesis by Cultured Mouse Neuroblastoma Cells

Mouse neuroblastoma clone N1E-115 has muscarinic acetylcholine receptors that mediate cyclic GMP synthesis. This receptor-mediated response is not significantly higher than background until the cells have been maintained in the stationary phase for at least 1 week. The basis of the influence of time in culture on the cyclic GMP response was investigated. The relative amount of cyclic GMP synthesized by intact cells was measured by radioactively labeling the GTP pool with [³H]guanine, incubating cells with agonists, and then chromatographically isolating [³H]cyclic GMP. Carbamylcholine-, ionophore X-537A-, and sodium azide-induced cyclic GMP formation increased with time in culture to a maximum of 13-, 9-, and 2.5-fold above basal, respectively. There was no change in the number or the apparent affinity of the muscarinic receptors as measured by [³H]quinuclidinyl benzylate ([³H]QNB) binding. In addition, there was no change in the apparent affinity of the receptors for agonist as measured by the ability of carbamylcholine to displace the specific binding of [³H]QNB. Guanylate cyclase activity per milligram protein and per cell in-creased six- and sevenfold, respectively, from day 0 to day 22. However, this increase in guanylate cyclase appeared to precede the marked increase in sensitivity of the cells to agonists. These data suggest that, in addition to guanylate cyclase and muscarinic receptors, there is another factor which is responsible for the development of this muscarinic receptor-mediated response.     

CALCIFICATION BY COCCOLITHOPHORIDS: EFFECTS OF pH AND Sr1

Cells of Coccolithus huxleyi which fail to deposit CaCO₃ and form coccoliths often occur as unwanted components in cultures used for studies of calcification. Non-calcified cells generally cannot be made to recalcify, but they can be removed from cultures by treatment at elevated pH or by a method based on faster sinking of calcified cells. Lowering the concentrations of nitrate, phosphate, or trace metals in the medium did not restore calcifying ability of non-calcified cells. However, addition of strontium did promote recalcification of decalcified Cricosphaera carterae grown under calcium limitation. Strontium seemed to promote coccolith attachment to cells rather than to affect calcium uptake or coccolith formation itself.     

Observations on the structure of epidermal cells, particularly the cork and silica cells, from the flowering stem internode of Lolium temulentum L. (Gramineae)

Features of the epidermis such as stomata, hairs, cork and silica cells are described from both light and electron microscope studies. The stomatal complex consists of two guard cells and two subsidiary cells. After division of the guard mother cell a pore is left at each end of the dividing wall. The cork and silica cells arise from a single another cell and develop differentially. The silica cell enlarges more than the cork cell and finally becomes filled with solidified silica. The outer tangential and radial walls of the cork cells become very thick-walled, whereas the inner tangential and radial walls of the silica cells become thickened. The outer tangential wall of the silica cell remains thin and is covered with a thin layer- of cuticle. This wall frequently collapses in old cells leaving a depression in the surface of the stem. The change in the ultrastructure of the cork and silica cells are described and the possible functions of these cells discussed.     

Morphogenesis of mechanoreceptor and epidermal cells of crickets during the last instar,and its relation to molting-hormone level

Summary (1) The fine structure of the cercal campaniform sensilla and epidermal cells of Gryllus bimaculatus Deg. (Saltatoria, Gryllidae) was examined, and the ecdysteroid level was monitored throughout the last larval instar. (2) The epidermal cells show changes in shape, cytoplasmic inclusions and differentiation of the apical cell membrane, coupled to the phases of buildup and breakdown of the (cercus) cuticle. (3) The imaginal epicuticle of the epidermal cells begins to form later (by about approximately 6 h) than that of the campaniform sensilla. (4) The campaniform sensilla were studied with respect to (a) the morphogenesis of the cuticular apparatus, (b) the inclusion of phenol oxidases in the cuticular apparatus, and (c) changes in the sensory apparatus preparatory to molting. (5) After apolysis the folding of the tormogen-cell wall into microvilli transiently disappears. Microvilli re-form shortly before imaginai ecdysis, and at the same time an outer receptor-lymph space develops. The role of the tormogen-cell plaques is discussed. (6) The levels of - and -ecdysone were determined separately by radioimmunoassay. (7) At the beginning of the instar the hormone level, especially that of -ecdysone, falls. Prior to apolysis, the concentration of -ecdysone rises, reaching an intermediate peak after apolysis is complete. The maximum hormone concentration (approximately 2,000 ng/g) is reached after the cuticulin layer is deposited, primarily due to the increase in -ecdysone. While the proecdysial cuticle is forming, the hormone titer is reduced; at this time -ecdysone is its chief component. (8) The identification of the ecdysteroids monitored by radioimmunoassay was confirmed by gas chromatography.This paper is dedicated to Professor H. Risler on the occasion of his 65th birthdaySupported by the Deutsche Forschungsgemeinschaft     

Monoaminergic neurons of the mudpuppy retina

Summary The mudpuppy retina was investigated with the histofluorescence method of Falck and Hillarp in normal animals and in animals injected intraocularly with -methylnoradrenaline, 5,6-dihydroxytryptamine, or a combination of the two drugs. Catecholaminergic amacrine cells were found to form a thin layer of terminals at the border between the inner nuclear and the inner plexiform layers. Catecholaminergic interplexiform cells were not found. Indoleamine-accumulating amacrine cells were also observed. They are fifteen to twenty times more numerous than the catecholaminergic cells, and their terminals occur diffusely throughout the inner plexiform layer. In a number of eyes the majority of the indoleamine-accumulating terminals were eliminated with intraocular injections of the neurotoxin, 5,7-dihydroxytryptamine, but the reproducibility of this effect was not consistent. Intravitreal injections of 5,6-dihydroxytryptamine were used to label both types of neurons for electron microscopy. They were found to make conventional type synapses on amacrine cells and, less frequently, on bipolar cells.     

Long cytoplasmic processes in pancreatic polypeptide cells

Pancreatic polypeptide (PP) cells were studied in human endocrine pancreatic tumours and in normal human pancreata by immunohistochemical techniques and electron miscroscopy. The existence of long cytoplasmic processes was demonstrated both in tumours and normal tissue. These processes are in close contact with other endocrine cells or with acinar cells. This particular morphological aspect suggests that PP cells may control the function of other cells via paracrine secretion.     

Do neural crest cells in the pancreas differentiate into somatostatin-containing cells?

Summary The possibility that the somatostatin cells are derived from the neurectoderm has been questioned in avian embryos. Isotopic and isochronic transplantations of the neural primordium from quail into chick embryos were made at the vagal level (somites 1 to 7). Quail and chick cells can be distinguished by the structure of their nucleus. The somatostatin cells were characterized immunocytochemically. In no case did quail cells showing the immunological reaction originate from the neural crest.     

A new type of cell in the pulmonary epithelium of the newt Triturus alpestris Laur

Summary Single cells of a new type appear scattered among pneumocytes in the pulmonary epithelium. The surfaces of these cells communicate with the air space and display numerous finger-like microvilli. In comparison to pneumocytes, these cells have a more lucid cytoplasm and their apical parts contain large amounts of electron-lucent vesicles and electron-dense granules, which are probably released into the lumen of the lung. These secretory cells exhibit a yellow formaldehyde-induced fluorescence, which suggests that they belong to the class of APUD cells.     

Scanning and transmission electron microscopy of the squamose gill-filament epithelium from fresh- and seawater adaptedTilapia

Synopsis The architecture of the gill structure of variousTilapia species was studied in relation to their adaptability to hypersaline media. Using SEM and EM, it was shown that the squamose epithelial cells of the gills have species-typical patterns of ridges on their outer surfaces. These have previously been misinterpreted by other authors as microvilli or stereocillia. The ridges are more dense and better developed in euryhaline species, likeT. zillii, and less so in stenohaline species likeSarotherodon niloticus. Comparing freshwater and seawater-adapted individuals ofT. zillii, S. niloticus, S. galflaeus, andTristramella sacra, it was shown that in fresh water the surface cells are slightly swollen, extending over the openings of the chloride cells. During adaptation to sea water, these ridges become higher and denser and the cell surface shrinks, exposing the underlying orifices of the apical crypts of the chloride cells. The more euryhaline the species, the less change there is in the ridge pattern of the cells during passage from fresh to sea water. This evidence implicates the gill epithelium, together with the chloride cells, in the process of osmoregulation.