Alteration of epithelial cell lipid synthesis by n-nitrosonornicotine

Summary Changes in the lipid composition of a cell membrane due to the binding of one cell modulator may affect binding of a second modulator, whether that binding is receptor-mediated (specific) or non-receptor-mediated (nonspecific). Such altered binding interactions have been demonstrated in oral epithelial cells, wherein N-nitrosonornicotine (NNN), a nonspecific ligand, enhances phorbol ester binding. To characterize membrane changes that may be responsible for such an effect, the current study examined lipid changes in hamster oral epithelial (HCP) cells associated with NNN binding. HCP cultures at two cell densities, 5 × 10⁶ cells/100 mm plate (subconfluent cultures) or 10 × 10⁶ cells/100 mm plate (confluent cultures) were incubated in Keratinocyte-Serum-Free Medium and exposed to 10μM NNN or DMSO (solvent control) for 48 h. Lipids were labeled with¹⁴C-acetate, then extracted, separated by thin layer chromatography, and the¹⁴C-lipids located by autoradiography and counted. Exposure of subconfluent cultures to NNN for 48 h, with¹⁴C-acetate present during the final 24 h, resulted in altered phospholipid and fatty acid labeling. Phospholipid labeling increased slightly in the presence of NNN compared to controls, while fatty acid labeling showed a modest but significant decrease in the presence of NNN. Similar changes occurred in the confluent cultures. Prelabeling of lipids in subconfluent cultures, followed by exposure to NNN in the absence of radiolabel, resulted in significantly (P<0.05) greater phospholipid labeling in the presence of NNN compared to control cultures. At the same time, fatty acid labeling decreased significantly. The NNN caused the cells to have decreased levels of several long chain polyunsaturated fatty acids and an increased amount of one long chain saturated (24:0) fatty acid. Significant differences were detected in the synthesis of various lipid classes between subconfluent and confluent cultures, but the presence of the NNN did not affect these differences. These results suggest that there is a shift of fatty acid type in the cells’ phospholipids in the presence of NNN, a finding that may reflect the membrane’s adaptation to the modulator’s presence.     

Ciliary activity in the mouse oviduct as studied by transmission and scanning electron microscopy

The mouse oviduct is covered by dense tracts of ciliated cells interspersed at random with occasional non-ciliated cells. Correlation between scanning electron microscopy and thin section images indicates that in the isolated fimbria most cilia are short (5 µm) and inactive, resting at the end of a uterad-directed effective stroke. These cilia terminate in a 9S−2 tip, the microtubules ending in an electron-dense plaque underneath the cell membrane. At the tip of the cilium a crown of fine extracellular hairs is attached to the ciliary membrane. In the ampulla and isthmus the ciliated cells decrease progressively in number and appear to lie in crypts.     

Centriole and basal body formation during ciliogenesis revisited.

This review is concerned with the formation during ciliogenesis of centrioles and basal bodies, primarily in epithelial multi-ciliated cells from the developing vertebrate respiratory and reproductive tracts. During ciliated cell differentiation, in these as well as in other cell types, cilium formation is preceded by the formation of centrioles assembled from precursor structures having little resemblance to the mature organelle. The origin, composition and function of the centriole precursor structures in generating large numbers of centrioles in a short period of time during ciliogenesis is discussed. This review also focuses on the biochemistry of centrioles and basal bodies and on recent experimental evidence that DNA might be associated with these structures.     

Quantitative analysis of dynamic behavior of osteoblasts during in vitro formation of micro‐mass cell cultures

Improvements in bone tissue engineering require an understanding of cellular and tissue level behavior of osteoblast‐like cells. Experiments indicate that in the absence of an anchoring material, intercellular adhesion may be based on signals that promote cell activity resulting in the formation of a spheroid cell‐matrix. The aim of the present study is to investigate the formation of scaffold‐free three‐dimensional micro‐mass cell spheroids in vitro, and to characterize quantitatively the cell movement. A new correlation based automated tracking method is evaluated in order to optimize the processing parameters and to identify statistical parameters that characterize the cell behavior. Results suggest that the temporal development of the mean distance of the cells to the center of gravity may be described by an exponential function, thus providing a characteristic time constant as a quantitative measure of cell dynamics. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Intercellular signaling in glial cells: calcium waves and oscillations in response to mechanical stimulation and glutamate.

Intercellular Ca2+ signaling in primary cultures of glial cells was investigated with digital fluorescence video imaging. Mechanical stimulation of a single cell induced a wave of increased [Ca2+]i that was communicated to surrounding cells. This was followed by asynchronous Ca2+ oscillations in some cells. Similar communicated Ca2+ responses occurred in the absence of extracellular Ca2+, despite an initial decrease in [Ca2+]i in the stimulated cell. Mechanical stimulation in the presence of glutamate induced a typical communicated Ca2+ wave through cells undergoing asynchronous Ca2+ oscillations in response to glutamate. The coexistence of communicated Ca2+ waves and asynchronous Ca2+ oscillations suggests distinct mechanisms for intra- and intercellular Ca2+ signaling. This intercellular signaling may coordinate cooperative glial function.     

Changes in the adrenal cortex of the rat after chronic administration of the steroidogenic inhibitor U-8113

In concert with studies of the effects of various pharmacologic inhibitors of corticosteroidogenesis on adrenocortical morphology, U-8113, an analog of amphenone B, was administered daily to Sprague-Dawley rats for 7, 14, 21 or 30 day. The primary morphological responses involved increases in adrenal weight, width of zona fasciculata, width of zona reticularis, intracellular lipids, mitochondrial size, mitochondrial vacuolation and crystalline-like inclusions, small coated vesicles, lysosomes, autophagic vacuoles and cholesterol ester clefts. In particular, the increases in lysosomes, coated vesicles and autophagic vacuoles containing morphologically altered mitochondria were considered reflective of mechanisms designed to maintain cellular integrity amidst functional impairment. Lipid analysis revealed marked increases in cholesterol esters and phospholipids, supportive of morphological observations. When permitted a 14 day recovery period following either 14 or 30 days of inhibitor therapy, most fine structural alterations and lipid derangements were diminished, and the cells approximated normal parameters.