The thermal transition in crude myelin proteolipid has a lipid rather than protein origin

Myelin proteolipid has been isolated from bovine brain and purified using organic solvents according to conventional procedures. The protein content of the purified sample, or crude proteolipid, contains a minimum of 75% w/w of proteolipid, with DM-20, a proteolipid molecule with an internal deletion of 35 out of 276 amino acid residues, as the only other component. Biochemical analysis has shown the differences in lipid composition between brain white matter, myelin and crude proteolipid preparations. The latter contained practically no cholesterol, while the other two samples had about 22-23% w/w. High-sensitivity differential scanning calorimetry experiments with both crude proteolipid and its extracted pool of lipids have shown similar reversible thermal transitions at 52 degrees C and 48 degrees C. The effect of increasing amounts of cholesterol on the two calorimetric transitions led in both cases to a continuous decrease in the melting temperature and in the transition enthalpy. Parallel Fourier-transform infrared spectroscopy studies of crude proteolipid have detected a reversible, co-operative lipid transition centred at 49 degrees C, with no detectable change in the amide region between 20 degrees C and 60 degrees C. Once more an increase in cholesterol content led to a decrease in the sharpness of this transition. It is concluded that the thermal transition detected in crude proteolipid, which has in the past been attributed to proteolipid thermal denaturation (Mateo et al. 1986), actually corresponds to a thermotropic phase transition of the lipids included in the crude proteolipid sample.     

Ca-controlled,reversible structural transition in myelin

When excised goldfish spinal cord is kept in physiological saline at room temperature, the myelin multilayers swell. As studied by X-ray diffraction, the original repeating distance of 150 Å (AS myelin) swells to 177 Å (AL myelin); no intermediate distances are seen. At least 70% of the myelin can undergo this gradual conversion. Omitting glucose from the saline, or substituting 2-deoxy-d-glucose for the glucose, or adding NaCN to the saline all promote conversion. Cooling retards the rate but does not prevent conversion. Omitting Ca does prevent conversion, however, and substituting Mg, Sr, or Ba for Ca also does so. Moreover, agents that increase the rate of conversion in Ca-containing saline by up to 5 fold (NaCN, colchicine, A23187) fail to convert myelin in Ca-free saline. We then converted AL myelin back to AS by withdrawing Ca. After converting in NaCN-containing saline, up to 3/4 of the AL myelin recompacts in Ca-free saline; however, none of the myelin recompacts if NaCN is present in the Ca-free saline. Little or no recompaction occurs after conversion in saline without NaCN. Based on our results, we suggest that the oligodendrocytes may maintain AS myelin in vivo by pumping Ca out of the myelin sheath. The myelin in a human PNS nerve has been induced to undergo a similar cycle of swelling and recompaction.     

Surprising cofactors in metalloenzymes

Transition metal complexes are located at the active sites of a number of enzymes involved in intriguing biochemical reactions. These complexes can exhibit a wide variety of chemical reactivity due to the ease at which transition metals can adopt different coordination environments and oxidation states. Crystallography has been a powerful technique for examining the structure and conformational variability of complex biological metallocenters. In particular, the past ten years have provided a wealth of structural information and several surprises concerning the metallocenters at the active sites of nitrogenase, hydrogenase and carbon monoxide dehydrogenase/acetyl-coenzyme A synthase.     

The thermal dependence of fast-start performance in fish

Many fish species use fast-starts to escape predators and capture prey. There is evidence for changes in fast-start behaviour with temperature, over acute, seasonal, developmental and evolutionary time scales. Maximum velocity often increases with acute temperature changes. Thermal acclimation can improve fast-start performance, although responses appear to be reduced in more eurythermal species. Changes in performance with thermal acclimation are often reflected at the molecular, biochemical and cellular levels of organisation. There appears to be little compensation in fast-start performance in Antarctic fish compared to warmer water species.     

Surprising functions of IAPs

The destruction of proteins in proteasomes is regulated at various levels. One of the early rate-limiting steps is "tagging" of proteins with ubiquitin, an 8 kD molecule that, when present in chains of four residues or more, serves as a signal recognized by proteasomal subunits. Ubiquitin is attached to target proteins by a series of enzymatic steps involving an El (ubiquitin activating enzyme), E2 (ubiquitin conjugating enzyme), and E3 (ubiquitin protein ligase). It is the last enzyme that confers target protein specificity. We have found that members of a family of RING domain containing proteins with anti- apoptotic activity, IAPs,     

Influences of thermal environment on fish growth

Thermoregulation in ectothermic animals is influenced by the ability to effectively respond to thermal variations. While it is known that ectotherms are affected by thermal changes, it remains unknown whether physiological and/or metabolic traits are impacted by modifications to the thermal environment. Our research provides key evidence that fish ectotherms are highly influenced by thermal variability during development, which leads to important modifications at several metabolic levels (e.g., growth trajectories, microstructural alterations, muscle injuries, and molecular mechanisms). In Atlantic salmon (Salmo salar), a wide thermal range (Δ_T 6.4°C) during development (posthatch larvae to juveniles) was associated with increases in key thermal performance measures for survival and growth trajectory. Other metabolic traits were also significantly influenced, such as size, muscle cellularity, and molecular growth regulators possibly affected by adaptive processes. In contrast, a restricted thermal range (Δ_T 1.4°C) was detrimental to growth, survival, and cellular microstructure as muscle growth could not keep pace with increased metabolic demands. These findings provide a possible basic explanation for the effects of thermal environment during growth. In conclusion, our results highlight the key role of thermal range amplitude on survival and on interactions with major metabolism‐regulating processes that have positive adaptive effects for organisms.     

Dependence on lipid structure of the coil-to-helix transition of bovine myelin basic protein

In aqueous solution bovine myelin basic protein has a close-to-random coil structure that is partially transformed to helix on interaction with lipids. Circular dichroism spectra have been used to follow this conformational transition which, with phospholipids, decreases in the order phosphatidylglycerol, phosphatidic acid approximately equal to phospholipids, decreases in the order phosphatidylethanolamine. There appears to be a strong correlation between the extent of alpha-helix formation and the degree of penetration of the hydrophobic region of the bilayer, as assessed by other methods. Cholesterol mixed in bilayers with phosphatidylserine has little effect on the protein secondary structure. Although basic protein binds strongly to cerebroside and to cerebroside sulphate, two of the other major myelin lipids, the intrinsic chirality of these lipids precludes assessment of their effect on the protein conformation. No significant changes in the circular dichroism spectra accompany the protein association with either of the zwitterionic bilayer-forming lipids, phosphatidylethanolamine and phosphatidylcholine. This seems to exclude extensive penetration into bilayers of these lipids and hence to exclude appreciable hydrophobic interactions; on the other hand, it is argued that little evidence exists for ionic attractions to these lipids. The optical activity of peptides derived from the basic protein by cleavage at the 42-43 and 88-89 peptides bonds (with cathepsin D) and at the 115-116 bond (with a skatole derivative) has also been measured in an attempt to locate the helix-forming regions within the primary structure.     

Ganglioside pattern and thermal tolerance of fish species

The brain ganglioside pattern of taxonomically close and distant related fish species living at different temperatures and the mammalian pattern were compared. The results demonstrate correlations between body temperature and brain ganglioside pattern: the higher the body temperature of animals, the lower is the relative proportion of polysialogangliosides in their brain.     

Size‐dependent thermal preferences in a pelagic fish

Large fish often inhabit colder waters than small fish. Using a simple bioenergetic model, we found that the optimal temperature for growth should decrease with increasing body size. We predicted that this mechanism would produce an ontogenetic change in thermal preference and then tested our predictions with Pacific salmon, Oncorhynchus spp. In a laboratory experiment, the slope of a regression of growth increment on initial size became steeper with increasing temperature, so that the optimal temperature for growth decreased with increasing body size. In field observations, larger and older salmon inhabited cooler areas, whereas smaller and younger salmon inhabited warmer areas. These patterns were consistent with a size‐dependent effect of temperature on condition factor, a parameter shown experimentally to be a measure of the most recent growth performance. Temperatures for maximising condition factor were lower for larger fish. Thus, an ontogenetic change in individual thermal preference toward cooler areas maximises the growth performance of fish, and the negative effects of climate warming on growth are hypothesised to be more severe for larger fish.     

A tropical fish community in thermal waters of southern Tuscany

In a small stream of southern Tuscany (Fossa Calda), fed by hot springs, we discovered a fish community dominated by tropical species, some of which have never previously been reported in Euro-Mediterranean natural freshwater environments. The aim of our research was to ascertain whether the three most abundant and widespread species (Amatitlania nigrofasciata, Hemichromis sp. and Oreochromis niloticus) have become established. Analysis of size class distribution and growth curves showed that the populations of these species are quite large and made up of juveniles, subadults and adults, as is the case in self-sustaining populations. These fishes were probably released intentionally, since they are widely used in aquariums and aquaculture and their survival in Fossa Calda was possible because of the constant high temperature of thermal waters. Spread to other streams seems unlikely. In fact, at some distance from the hot springs, where water temperatures are lower, populations of the tropical species were small.     

Comparison of fish impingement by a thermal power station with fish populations in the Ems Estuary

On an annual basis >10 million individuals of c . 35 fish species are impinged at the EC20 unit of the Eems power station located on the Ems Estuary. The most abundant are: herring Clupea harengus , gobies ( Gobiidae ), Nilsson's pipefish Sygnathus rostellatus , three-spined stickleback Gasterosteus aculeatus and sprat Sprattus sprattus . The impingement shows a seasonal pattern which reflects the presence of the fishes in the estuary. The results are compared with other impingement studies at power stations on the Belgian and Dutch estuaries.
In 1995, the cooling water entrance was displaced from the shoreline to 300 m off the coast at the edge of the tidal channel. The effect of this relocation on the level of fish impingement, determined by comparing the ratio of fish density in the cooling water and in the estuary before (1992–1993) and after (1996–1997) the relocation of the intake canal is discussed.     

Surprising developments in the legume root nodule

Root nodule morphogenesis involves the induction of mitotic activity in otherwise quiescent root cortical cells, giving rise to the nodule primordium. One gene expressed during nodule initiation, ENOD40, has been implicated in nodule growth and/or differentiation^(1,2). Interestingly, although the nucleotide sequence of ENOD40 genes from soybean^(3,4) and Medicago^(1,2) are highly homologous, they are unlikely to encode a similar protein product. In fact, a remarkable feature of these genes is their apparent lack of protein coding potential. Thus, ENOD40 is a member of the growing list of eukaryotic genes whose RNA product is implicated in control of cell growth and differentiation, the so called riboregulators⁽⁵⁾.     

Surprising Effects of Synaptic Excitation

Typically, excitatory synaptic coupling is thought of as an influence that accelerates and propagates firing in neuronal networks. This paper reviews recent results explaining how, contrary to these expectations, the presence of excitatory synaptic coupling can drastically slow oscillations in a network and how localized, sustained activity can arise in a network with purely excitatory coupling, without sustained inputs. These two effects stem from interactions of excitatory coupling with two different forms of intrinsic neuronal dynamics, and both serve to highlight the fact that the influence of synaptic coupling in a network depends strongly on the intrinsic properties of cells in the network.This work was partially supported by the National Science Foundation, under award DMS-0414023     

Surprising twists to exocyst function

What do neurons use the exocyst complex for? In this issue of Neuron, using mutations in one exocyst component, Mehta et al. reach the surprising conclusion that exocyst function is divisible: different components play distinct roles. These studies also suggest that the exocyst may regulate membrane insertion of cell adhesion molecules required for synaptic partner choice.     

The effect of bovine myelin basic protein on the phase transition properties of sphingomyelin

The basic protein of myelin can spontaneously associate with the synthetic phospholipid N-palmitoylsphingosinephosphatidylcholine. The protein alters the phase transition properties of the lipid from a single transition at 41.5°C to two overlapping transitions, one being slightly above and the other slightly below the transition temperature of the pure lipid. The effect was not seen upon the addition of poly(l-lysine) to this lipid nor does the myelin basic protein alter the phase transition properties of dimyristoylphosphatidylcholine. The results thus demonstrate that the myelin basic protein can interact with a major zwitterionic lipid component of myelin in addition to acidic phospholipids.