Expression of endothelial nitric oxide synthase in ciliated epithelia of rats.

Endothelial nitric oxide synthase (eNOS), originally found in the endothelium of vascular tissue, also exists in other cell types, including ciliated epithelia of airways. The eNOS is ultrastructurally localized to the basal body of the microtubules of the cilia, and nitric oxide (NO) stimulates ciliary beat frequency (CBF). We examined whether the expression of eNOS is present in ciliated cells of other organs. Western blotting analysis revealed that eNOS was expressed in the rat cerebrum, lung, trachea, testis, and oviduct. Immunohistochemical staining showed that eNOS was localized in the ciliated epithelia of airways, oviduct, testis, and ependymal cells of brain in addition to the endothelium and smooth muscle of the vasculature. To confirm the activation of eNOS in the ciliated epithelia, we examined the effect of L-arginine (L-Arg), the substrate of NOS, on the production of nitrite and nitrate (NOx) in the cultured explants of rat trachea. L-Arg (100 microM) increased NOx levels significantly (p<0.05). In explants exposed to inhibitors of NOS, the effect of l-Arg on the production of NOx was blocked. These findings suggest that epithelial NO plays an important role in signal transduction associated with ciliary functions.     

Absence of Direct Delivery for Single Transmembrane Apical Proteins or Their “Secretory” Forms in Polarized Hepatic Cells

The absence of a direct route to the apical plasma membrane (PM) for single transmembrane domain (TMD) proteins in polarized hepatic cells has been inferred but never directly demonstrated. The genes encoding three pairs of apical PM proteins, whose extracellular domains are targeted exclusively to the apical milieu in Madin-Darby canine kidney cells, were packaged into recombinant adenovirus and delivered to WIF-B cells in vitro and liver hepatocytes in vivo. By immunofluorescence and pulse-chase metabolic labeling, we found that the soluble constructs were overwhelmingly secreted into the basolateral milieu, which in vivo is the blood and in vitro is the culture medium. The full-length proteins were first delivered to the basolateral surface but then concentrated in the apical PM. Our results imply that hepatic cells lack trans-Golgi network (TGN)-based machinery for directly sorting single transmembrane domain apical proteins and raise interesting questions about current models of PM protein sorting in polarized and nonpolarized cells.     

Reef to basin sediment transport using Halimeda as a sediment tracer,Grand Cayman Island,West Indies

Fragments of the calcareous green alga Halimeda form a large part of the sediment in the fringing reef system and adjacent deep marine environments of Grand Cayman Island, West Indies. Nine species combine to form three depth-related assemblages that are characteristic of the major reef-related environments (lagoonpatch reef, reef terraces, and deep reef). These modern plant assemblages form the basis of the use of Halimeda as a sediment tracer. Halimeda-based tracer studies of Holocene sediments indicate that only sediments containing deep reef species of Halimeda are presently being transported through the reef system by sediment creep and being deposited at the juncture of the upper and lower island slope. Sediments containing shallow reef Halimeda are retained within the reef and lithified by marine carbonate cements. Tracer studies of Pleistocene sediment indicate large amounts of reef-derived carbonate sand containing deep water Halimeda were produced during interglacial high stands of sea level. Much of this material was removed by turbidity currents moving out of the reef system to the island slope down submarine channels perpendicular to the reef trend. These channels may still be identified on bathymetric profiles, but are no longer receiving coarse reef debris and are veneered with a blanket of pelagic carbonate mud.     

Alpha- and beta-adrenergic and muscarinic cholinergic binding sites in the bladder and urethra of the rabbit

The affinities of a number of alpha- and beta-adrenergic binding sites and muscarinic cholinergic binding sites in rabbit urethra and bladder have been determined, using specific radioligand receptor binding assays. There was a greater density of beta-binding sites than alpha-binding sites in the bladder, while, in the urethra, there was a greater density of alpha-binding sites than beta-binding sites. The number of alpha-binding sites was fourfold greater in the urethra, whereas there were fewer beta-binding sites in the urethra. There were fewer muscarinic binding sites in the urethra than in the bladder. The dissociation constant for [3H]dihydroalprenolol at the beta-binding site was 6.4 nM, for [3H]dihydroergocryptine at the alpha-binding site was 2.11 nM, and for 3H-labelled l-quinuclidinyl benzilate at the muscarinic binding site was 0.22 nM.