Evolution of a small-muscle polymorphism in lines of house mice selected for high activity levels

To study the correlated evolution of locomotor behavior and exercise physiology, we conducted an artificial selection experiment. From the outbred Hsd:ICR strain of Mus domesticus, we began eight separate lines, each consisting of 10 breeding pairs. In four of the lines, we used within-family selection to increase voluntary wheel running. The remaining four lines were random-bred (within lines) to serve as controls. Various traits have been monitored to test for correlated responses. Here, we report on organ masses, with emphasis on the triceps surae muscle complex, an important extensor of the ankle. Mice from the selected lines exhibit reduced total body mass, increased relative (mass-corrected) kidney mass, and reduced relative triceps surae mass. In addition, a discrete muscle-mass polymorphism was observed: some individuals had triceps surae that were almost 50% lighter than normal for their body mass. This small-muscle phenotype was observed in only three of the eight lines: in one control line, it has fluctuated in frequency between zero and 10%, whereas in two of the selected lines it has increased in frequency to approximately 50% by generation 22. Data from a set of parents and offspring (generations 23 and 24) are consistent with inheritance as a single autosomal recessive allele. Evidence for the adaptive significance of the small-muscle allele was obtained by fitting multiple-generation data to hierarchical models that include effects of genetic drift and/or selection. The small-muscle allele is estimated to have been present at low frequency (approximately 7%) in the base population, and analysis indicates that strong selection favors the allele in the selected but not control lines. We hypothesize that the small muscles possess functional characteristics and/or that the underlying allele causes pleiotropic effects (e.g., reduced total body mass; increased relative heart, liver, and kidney mass) that facilitate high levels of wheel running. Nevertheless, at generation 22, wheel running of affected individuals did not differ significantly from those with normal-sized muscles, and the magnitude of response to selection has been similar in all four selected lines, indicating that multiple genetic "solutions" are possible in response to selection for high activity levels.     

Efficient activation of nucleoside phosphoramidites with 4,5-dicyanoimidazole during oligonucleotide synthesis.

A new activator for the coupling of phosphoramidites to the 5'-hydroxyl group during oligonucleotide synthesis is introduced. The observed time to complete coupling is twice as fast with 4, 5-dicyanoimidazole (DCI) as the activator, compared with 1 H -tetrazole. The effectiveness of DCI is thought to be based on its nucleophilicity. DCI is soluble in acetonitrile up to 1.1 M at room temperature and can be used as the sole coupling activator during routine automated solid phase synthesis of oligonucleotides. The addition of 0.1 M N -methylimidazole to 0.45 M 1 H -tetrazole also results in higher product yields during oligonucleotide synthesis than observed with 1 H -tetrazole alone.     

Stream position choice in salmon parr: maximisation of foraging efficiency

Juvenile Atlantic salmon or parr (Salmo salar L.) maintain station at certain locations in flowing stream water. This position choice is assumed to involve the maximization of energy intake, based upon food availability which is usually directly related to water flow rate. Conversely energy expenditure, including station holding behaviour, foraging and defending preferred sites, is inversely related to water flow rate. Adaptations of parr to life in fast flows implies that station holding is energetically inexpensive at water speeds up to the maximum sustained holding speed, which is fish specific, thus the most important energetic consideration for parr is the ability to maximize food intake. Ten groups of three parr were each observed for 60 min within an artificial stream tank over a heterogeneous substratum. Individual position choice and behaviour were recorded continuously. For each location chosen by the parr the potential upstream line‐of‐sight (LOS), defined as the maximum distance upstream that the water surface would be visible, was calculated. At those sites where foraging behaviour was observed, the mean upstream potential LOS was significantly greater than at sites where other behaviours were observed and at 400 randomly generated sites within the tank. When foraging, parr usually take food from the stream drift and there is a significant time expenditure on food location, identification and catching. Results presented here would seem to confirm that to maximize time available to make these decisions, a fish would be expected to maximize the distance over which it can observe potential food particles.     

A method for identification of inhibitors of the phosphorylation reactions of bacterial response regulator proteins using (31)P nuclear magnetic resonance spectroscopy.

Bacterial response regulators are attractive targets for antibacterial drug development, yet random screening against these targets has failed as yet to identify chemicals that constitute viable leads. Alternative methods to provide leads for drug development based on identification and optimization of low affinity ligands from NMR screens have been described. However, leads from these processes still require verification in a bioassay, which is often problematic if compounds have unfavorable optical and solubility properties. A simple method, based on using NMR to observe the activity of the target, is described. It has the advantages of being able to characterize both low affinity leads and a wider selection of compounds in a structure activity relationships series, without the problems affecting a fluorescence assay. In this example we use (31)P to monitor the turnover of a bacterial response regulator, but the generic approach could be applied to other nuclei and thus a range of biological systems.     

Hypohydration and prior heat stress exacerbates decreases in cerebral blood flow velocity during standing.

Hypohydration is associated with orthostatic intolerance; however, little is known about cerebrovascular mechanisms responsible. This study examined whether hypohydration reduces cerebral blood flow velocity (CBFV) in response to an orthostatic challenge. Eight subjects completed four orthostatic challenges (temperate conditions) twice before (Pre-EU and Pre-Hyp) and following recovery from passive heat stress ( approximately 3 h at 45 degrees C, 50% relative humidity, 1 m/s air speed) with (Post-EU) or without (Post-Hyp) fluid replacement of sweat losses (-3% body mass loss). Measurements included CBFV, mean arterial pressure (MAP), heart rate (HR), end-tidal CO(2), and core and skin temperatures. Test sessions included being seated (20 min) followed by standing (60 s) then resitting (60 s) with metronomic breathing (15 breaths/min). CBFV and MAP responses to standing were similar during Pre-EU and Pre-Hyp. Standing Post-Hyp exacerbated the magnitude (-28.0 +/- 1.4% of baseline) and duration (9.0 +/- 1.6 s) of CBFV reductions and increased cerebrovascular resistance (CVR) compared with Post-EU (-20.0 +/- 2.1% and 6.6 +/- 0.9 s). Standing Post-EU also resulted in a reduction in CBFV, and a smaller decrease in CVR compared with Pre-EU. MAP decreases were similar for Post-EU (-18 +/- 4 mmHg) and Post-Hyp (-21 +/- 5 mmHg) from seated to standing. These data demonstrate that despite similar MAP decreases, hypohydration, and prior heat stress (despite apparent recovery) produce greater CBFV reduction when standing. These observations suggest that hypohydration and prior heat stress are associated with greater reductions in CBFV with greater CVR, which likely contribute to orthostatic intolerance.     

The Cooperative Breeding System of the Red-cockaded Woodpecker

The breeding system of the red-cockaded woodpecker is described based on data collected over six years from a population of 500 marked individuals in the Sandhills of North Carolina. Male-female pairs were the most common social unit (59%), but 30% of social units contained one or more adult helpers, and 11% consisted of solitary males. Helpers were almost exclusively male: 27% of males remained in their natal group as helpers for at least one year, whereas only four (1%) females did. Most breeding females remained as breeders in the same group from one year to the next (56%), but a surprising number (12%) moved to another group. Many movements were related to incest avoidance or mate death, but 39% involved deserting a mate, usually following successful reproduction. We suggest that females sometimes are forced from groups by immigrants or other group members. The median distance of movements by adult females was only 1.3 km. In contrast to females, no breeding males switched groups. Survival of both breeding (76%) and helper (80%) males was higher than that of breeding females (69%). Males exhibited two distinct life-history strategies. Some remained as helpers on their natal territory for one or more years, and became breeders by inheriting breeding status in the natal group (17% per year) or by replacing a deceased breeder in a nearby group (13% per year, median distance moved 1.0 km). Other males dispersed from their natal group permanently during their first year. Some of these males were floaters at age one year, others were solitary, and a few became helpers in a non-natal group, but many were breeders. In contrast to males that first functioned as helpers, those that dispersed after fledging moved long distances (median dispersal distance 4.5 km), longer even than dispersing female fledglings moved (median distance 3.2 km). The habitat saturation model of the evolution of cooperative breeding is based on selection between the two life-history strategies exhibited by male red-cockaded woodpeckers. The model therefore may be tested directly with this species. Another indication that this model is appropriate for this species is the existence of a resource (cavity trees) that might provide an ecological basis for habitat saturation.     

VINE PHOTOSYNTHESIS AND RELATIONSHIPS TO CLIMBING MECHANICS IN A FOREST UNDERSTORY

Photosynthesis in a deciduous forest understory was studied for three exotic vine species (Pueraria lobata, Lonicera japonica, and Hedem helix) and five common native species (Rhus radicans, Clematis virginiana, Smilax rotundifolia, Vitis vulpina, and Parthenocissus quinquefolid) possessing a variety of climbing mechanisms. The adventitious-root climbers (H. helix and R. radicans) had the lowest maximum photosynthetic rates of all species (5.5 and 6.4 μmol m^–2 s^–1, respectively). The twining vine P. lobata was the most poorly adapted to the understory with a high light-compensation point (43 μmol m^–2 s^–1), low photosynthesis under low light (0.5 μmol m² s^–1 at 50 μmol m^–2 s^–1), and the highest light requirement for obtaining 90% of maximum photosynthesis (860 μmol m^–2 s^–1). Lonicera japonica, another twining vine, was better-adapted to low light conditions, but vines with tendril climbing mechanics were physiologically the best adapted to low light. The adhesive-tendril climber P. quinquefolia was the most highly adapted to shade, with a low light compensation point (20 μmol m^–2 s^–1), a high photosynthetic rate under low light (3.5 μmol m^–2 s^–1), and a low light saturation point (160 μmol m^–2 s^–1). Results suggest that physiological adaptability of vines to low-light environments may be related to climbing mechanics.     

Bone Load Estimation for the Proximal Femur Using Single Energy Quantitative CT Data

A density-based load estimation method was applied to determine femoral load patterns. Two-dimensional finite element models were constructed using single energy quantitative computed tomography (QCT) data from two femora. Basic load cases included parabolic pressure joint loads and constant tractions on the greater trochanter. An optimization procedure adjusted magnitudes of the basic load cases, such that the applied mechanical stimulus approached the ideal stimulus throughout each model. Dominant estimated load directions were generally consistent with published experimental data for gait. Other estimated loads suggested that loads at extreme joint orientations may be important to maintenance of bone structure. Remodeling simulations with the estimated loads produced density distributions qualitatively similar to the QCT data sets. Average nodal density errors between QCT data and predictions were 0.24 g/cm(3) and 0.28 g/cm(3). The results indicate that density-based load estimation could improve understanding of loading patterns on bones.     

Evidence for Phosphoprotein Microspheres in Bone

Bone sialoprotein and osteopontin are bone-specific phosphoproteins, but their function is uncertain and their ultrastructural associations remain unclear. Insight into their role was sought by special attention to their general distribution and specific morphology under the high-power optical microscope. Their extracellular staining characteristics were examined in cryosections of adult rat skeletal tissues using two immunohistochemical methods. The two proteins were clearly evident in immature woven bone of endochondral and intramembranous origin (although cartilage was negative, even when calcified). In mature lamellar bone, bone sialoprotein remained ubiquitous, while osteopontin was confined to cement lines and other relatively discrete sites of past and present resorption activity, particularly near blood vessels. In neither case was the distribution of the stain structureless and diffuse. Invariably (except when non-specific), it was sharply defined and had the form of microspheres measuring approximately 1 m in diameter. In both immature and mature regions, these objects appeared in sheets, chains or groups in a pattern that was evidently coincident with a similar structural arrangement found within the inorganic phase of bone. It was concluded that phosphoproteins are not randomly located throughout the collagenous matrix but are apparently integral to calcified microsphere populations, and it is suggested that these structures are well placed to control the chemical State of the mineral over their surfaces and influence remodelling.