Detergent-resistant membrane fractions contribute to the total 1H NMR-visible lipid signal in cells.

Leukocytes and other cells show an enhanced intensity of mobile lipid in their 1H NMR spectra under a variety of conditions. Such conditions include stimulation, which has recently been shown to involve detergent-resistant, plasma membrane domains (DRMs) often called lipid rafts. As there is much speculation surrounding the origin of cellular NMR-visible lipid, we analysed subcellular fractions, including DRMs, by NMR spectroscopy. We demonstrated that DRMs isolated by density gradient centrifugation from lymphoid (CEM-T4, stimulated Jurkat cells), and monocytoid (THP-1) cells produced NMR-visible, lipid signals. Large scale subfractionation of THP-1 cells determined that while cytoplasmic lipid droplets constituted much of the total NMR-visible lipid, the contribution of DRMs was significant. Qualitative and quantitative lipid analyses revealed that DRMs and lipid droplets differed in their lipid composition. DRMs were enriched in cholesterol and ganglioside GM1, and contained relatively unsaturated fatty acids compared with the lipid droplets. Both lipid droplets and DRMs contained neutral lipids (triacylgycerols, cholesterol ester, fatty acids in THP-1 cells) that could, in addition to phospholipids, contribute to the NMR-visible lipid. The lipid droplets also exhibited different protein profiles and contained 500-fold less protein than DRMs, confirming that DRMs and droplets were fractionated as separate entities. The NMR-visible lipid in DRMs is therefore unlikely to be a contaminant from lipid droplets. We propose a micropartitioning of the NMR-visible mobile lipid of whole cells between intracellular lipid droplets, where most of this lipid resides, and detergent-resistant plasma membrane domains.     

Inter and intra‐population variation in shoaling and boldness in the zebrafish (Danio rerio)

Population differences in anti‐predator behaviour have been demonstrated in several species, although less is known about the genetic basis of these traits. To determine the extent of genetic differences in boldness (defined as exploration of a novel object) and shoaling within and between zebrafish ( Danio rerio ) populations, and to examine the genetic basis of shoaling behaviour in general, we carried out a study that involved laboratory‐raised fish derived from four wild‐caught populations. Controlling for differences in rearing environment, significant inter‐population differences were found in boldness but not shoaling. A larger shoaling experiment was also performed using one of the populations as the basis of a North Carolina type II breeding design (174 fish in total) to estimate heritability of shoaling tendency. A narrow‐sense heritability estimate of 0·40 was obtained, with no apparent dominance effects.     

EFFECT OF LIGHT WAVELENGTH ON SUPPRESSION AND PHASE DELAY OF THE MELATONIN RHYTHM

Different wavelengths of light were compared for melatonin suppression and phase shifting of the salivary melatonin rhythm. The wavelengths compared were 660 nm (red), 595 nm (amber), 525 nm (green), 497 nm (blue/green), and 470 nm (blue). They were administered with light-emitting diodes equated for irradiance of 130 μW/cm². Fifteen volunteers participated in all five wavelength conditions and a no light control condition, with each condition conducted over two consecutive evenings. Half-hourly saliva samples were collected from 19:00 to 02:00 on night 1 and until 01:00 on night 2. Light was administered for the experimental conditions on the first night only from midnight to 02:00. Percentage melatonin suppression on night 1 and dim light melatonin onset (DLMO) for each night were calculated. The shorter wavelengths of 470, 497, and 525 nm showed the greatest melatonin suppression, 65% to 81%. The shorter wavelengths also showed the greatest DLMO delay on night 2, ranging from 27 to 36 min. The results were consistent with the involvement of a scotopic mechanism in the regulation of circadian phase. (Chronobiology International, 18(5), 801-808, 2001)     

Breeding systems and genome evolution

The profound effects of inbreeding and other non-recombining breeding systems on genetic variability and molecular evolution are now beginning to be understood. Theoretical models predict how such populations are expected to differ from outcrossed populations, and DNA sequence data are being collected and used to test the predictions.     

A comparative histological and histochemical study of the post-gastric alimentary canal from three species of pleuronectid, the Atlantic halibut, the yellowtail flounder and the winter flounder

The histology and mucus histochemistry of the pleuronectid post-gastric alimentary canal was examined using light and electron microscopy. Distinct differences in goblet cell mucus histochemistry were observed between species, with the two closest taxonomic species, the winter flounder and the yellowtail flounder showing the most diversity and the halibut showing regional variation. Numbers of goblet cells within post-gastric regions did not differ significantly between species, but were significantly different between regions within species increasing toward the rectum. The post-gastric region was divisible into two areas based upon the ultrastructural features of lipid digestion and absorption in the intestine and pyloric caeca, and of exogenous protein in the rectum. The combination of species-specific histochemical differences in mucus and general histological and ultrastructural differences within the post-gastric regions between these species suggest a correlation between lumenal environmental conditions/histology and natural prey preference.