Seasonality of the red blood cell stress response in rainbow trout (Oncorhynchus mykiss)

The β-adrenergic stress response in red blood cells (RBCs) of rainbow trout shows seasonal changes in expression. We have explored the mechanisms underpinning this response by following over a period of 27 months changes in β-adrenergic receptor (β-AR) binding characteristics, β-adrenergically stimulated RBC Na+/H+ exchanger (βNHE) activity, together with β-AR and βNHE mRNA levels and plasma steroid hormone and lactate levels. These parameters were measured at approximately monthly intervals in a single population of fish held under semi-natural conditions. Membrane-bound, high-affinity β-ARs were present in RBCs at all sampling times, varying from 668 ± 112 to 2654 ± 882 receptors cell-1 (mean ± SEM; n=8). βNHE activity, however, was reduced by 57 and 34% in December 1999 and February 2001, respectively, compared to an otherwise sustained influx that averaged 110.4 ± 2.3 mmol l-1 RBCs h-1 (n = 119). Only one reduction coincided with a spawning period but both were preceded by transient increases in circulating testosterone. βNHE activity measured under standard conditions was not correlated with the number or affinity of β-ARs nor with water temperature, but both β-AR numbers and βNHE activity were positively related to their respective mRNA levels (P = 0.005 and 0.038, respectively). Pharmaceutical intervention in the transduction cascade linking the β-AR and βNHE failed to indicate any failure of the transduction elements in RBCs displaying low βNHE activity. Similarly, we failed to demonstrate any link between seasonal cortisol fluctuations and seasonally reduced βNHE activity. However, the βNHE activity of age-separated RBC fractions showed that younger RBCs had a significantly higher βNHE response than older RBCs, consistent with the seasonal reductions in βNHE being linked to turnover of red cells and erythropoiesis. Testosterone is known to induce erythropoiesis and we conclude that seasonal reductions in βNHE are not caused by changes in β-AR numbers, but may be linked to testosterone-induced erythropoiesis

A rapid and massive gene expression shift marking adolescent transition in C. elegans

Organismal development is the most dynamic period of the life cycle, yet we have only a rough understanding of the dynamics of gene expression during adolescent transition. Here we show that adolescence in Caenorhabditis elegans is characterized by a spectacular expression shift of conserved and highly polymorphic genes. Using a high resolution time series we found that in adolescent worms over 10,000 genes changed their expression. These genes were clustered according to their expression patterns. One cluster involved in chromatin remodelling showed a brief up-regulation around 50 h post-hatch. At the same time a spectacular shift in expression was observed. Sequence comparisons for this cluster across many genotypes revealed diversifying selection. Strongly up-regulated genes showed signs of purifying selection in non-coding regions, indicating that adolescence-active genes are constrained on their regulatory properties. Our findings improve our understanding of adolescent transition and help to eliminate experimental artefacts due to incorrect developmental timing

Electrospray ionisation mass spectrometric analysis of lipid restructuring in the carp (Cyprinus carpio L.) during cold acclimation

Cold acclimation of carp from 30°C to 10°C causes a restructuring of liver microsomal phospholipids characterised by increased proportions of monounsaturated fatty acid in phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Here, we have used electrospray ionisation mass spectrometry (ESI-MS) to determine the patterns of alteration to individual molecular species compositions of PC, PE and phosphatidylinositol (PI) in response to gradually decreasing temperature.The results demonstrate that cold induces precise changes to a limited number of phospholipid species, and that these changes are distinct and different for each phospholipid class. The major change for PC was increased 16:1/22:6, but for PE the species that increased was 18:1/22:6. By contrast, the PI species that increased during cold acclimation were characterised by an sn-1 monounsaturated fatty acid in combination with arachidonoyl or eicosapentaenoyl fatty acid at the sn-2 position. Analysis of acyl distribution indicates that cold only caused the accumulation of monounsaturated fatty acids at the sn-1 and not at the sn-2 position of phospholipids. These results highlight the tight and restricted range of modifications that membranes make to their phospholipid composition in response to thermal stress. <br/