In vivo detection of gene expression in liver by 31P nuclear magnetic resonance spectroscopy employing creatine kinase as a marker gene

In vivo assessment of gene expression is desirable to obtain information on the extent and duration of transduction of tissue after gene delivery. We have developed an in vivo, potentially noninvasive, method for detecting virally mediated gene transfer to the liver. The method employs an adenoviral vector carrying the gene for the brain isozyme of murine creatine kinase (CK-B), an ATP-buffering enzyme expressed mainly in muscle and brain but absent from liver, kidney, and pancreas. Gene expression was monitored by 31P magnetic resonance spectroscopy (MRS) using the product of the CK enzymatic reaction, phosphocreatine, as an indicator of transfection. The vector was administered into nude mice by tail vein injection, and exogenous creatine was administered in the drinking water and by i.p. injection of 2% creatine solution before 31P MRS examination, which was performed on surgically exposed livers. A phosphocreatine resonance was detected in livers of mice injected with the vector and was absent from livers of control animals. CK expression was confirmed in the injected animals by Western blot analysis, enzymatic assays, and immunofluorescence measurements. We conclude that the syngeneic enzyme CK can be used as a marker gene for in vivo monitoring of gene expression after virally mediated gene transfer to the liver.

Sea-surface temperature reconstruction in the Southwest Pacific

Uniquely in the Southern Hemisphere the New Zealand micro-continent spans the interface between a subtropical gyre and the Subantarctic Circumpolar Current. Its 20° latitudinal extent includes a complex of submerged plateaux, ridges, saddles and basins which, in the present interglacial, are partial barriers to circulation and steer the Subtropical (STF) and Subantarctic (SAF) fronts. This configuration offers a singular opportunity to assess the influence of bottom topography on oceanic circulation through Pleistocene glacial - interglacial (G/I) cycles, its effect on the location and strength of the fronts, and its ability to generate significant differences in mixed layer thermal history over short distances. For this study we use new planktic foraminiferal based sea-surface temperature (SST) estimates spanning the past 1 million years from a latitudinal transect of four deep ocean drilling sites. We conclude that: 1. the effect of the New Zealand landmass was to deflect the water masses south around the bathymetric impediments; 2. the effect of a shallow submerged ridge on the down-current side (Chatham Rise), was to dynamically trap the STF along its crest, in stark contrast to the usual glacial-interglacial (G-I) meridional migration that occurs in the open ocean; 3. the effect of more deeply submerged, downstream plateaux (Campbell, Bounty) was to dynamically trap the SAF along its steep southeastern margin; 4. the effects of saddles across the submarine plateaux was to facilitate the development of jets of subtropical and subantarctic surface water through the fronts, forming localized downstream gyres or eddies during different phases in the G-I climate cycles; 5. the deep Pukaki Saddle across the Campbell-Bounty Plateaux guided a branch of the SAF to flow northwards during each glacial, to form a strong gyre of circumpolar surface water in the Bounty Trough, especially during the mid-Pleistocene Climate Transition (MIS 22-16) when exceptionally high SST gradients existed across the STF; 6. the shallower Mernoo Saddle, at the western end of the Chatham Rise, provided a conduit for subtropical water to jet southwards across the STF in the warmest interglacial peaks (MIS 11, 5.5) and for subantarctic water to flow northwards during glacials; 7. although subtropical or subantarctic drivers can prevail at a particular phase of a G-I cycles, it appears that the Antarctic Circumpolar Current is the main influence on the regional hydrography. Thus complex submarine topography can affect distinct differences in the climate records over short distances with implications for using such records in interpreting global or regional trends. Conversely, the local topography can amplify the paleoclimate record in different ways in different places, thus enhancing its value for the study of more minor paleoceanographic influences that elsewhere are more difficult to detect. Such sites include DSDP 594, which like some other Southern Ocean sites, has the typical late Pleistocene asymmetrical saw-tooth G-I climate pattern transformed to a gap-tooth pattern of quasi-symmetrical interglacial spikes that interrupt extended periods of minimum glacial temperatures.