Novel individualized power training protocol preserves physical function in adult and older mice

Sarcopenia, the age-related loss of muscle mass and strength, contributes to frailty, functional decline, and reduced quality of life in older adults. Exercise is a recognized therapy for sarcopenia and muscle dysfunction, though not a cure. Muscle power declines at an increased rate compared to force, and force output declines earlier than mass. Thus, there is a need for research of exercise focusing on improving power output and functionality in older adults. Our primary purpose was proof-of-concept that a novel individualized power exercise modality would induce positive adaptations in adult mice, before the exercise program was applied to an aged cohort. We hypothesized that after following our protocol, both adult and older mice would show improved function, though there would be evidence of anabolic resistance in the older mice. Male C57BL/6 mice (12 months of age at study conclusion) were randomized into control (n = 9) and exercise (n = 6) groups. The trained group used progressive resistance (with a weighted harness) and intensity (~ 4-10 rpm) on a custom motorized running wheel. The mice trained similarly to a human workout regimen (4-5 sets/session, 3 sessions/week, for 12 weeks). We determined significant (p < 0.05) positive adaptations post-intervention, including: neuromuscular function (rotarod), strength/endurance (inverted cling grip test), training physiology (force/power output per session), muscle size (soleus mass), and power/velocity of contraction (in vitro physiology). Secondly, we trained a cohort of older male mice (28 months old at conclusion): control (n = 12) and exercised (n = 8). While the older exercised mice did preserve function and gain benefits, they also demonstrated evidence of anabolic resistance.F31 AG044108 - NIA NIH HHS; R01 AG017768 - NIA NIH HHS; TL1 TR001440 - Institute for Translational Sciences, University of Texas Medical BranchAccepted manuscrip

Two Stable, Conducting Conformations of the Selectivity Filter in Shaker K+ Channels

We have examined the voltage dependence of external TEA block of Shaker K+ channels over a range of internal K+ concentrations from 2 to 135 mM. We found that the concentration dependence of external TEA block in low internal K+ solutions could not be described by a single TEA binding affinity. The deviation from a single TEA binding isotherm was increased at more depolarized membrane voltages. The data were well described by a two-component binding scheme representing two, relatively stable populations of conducting channels that differ in their affinity for external TEA. The relative proportion of these two populations was not much affected by membrane voltage but did depend on the internal K+ concentration. Low internal K+ promoted an increase in the fraction of channels with a low TEA affinity. The voltage dependence of the apparent high-affinity TEA binding constant depended on the internal K+ concentration, becoming almost voltage independent in 5 mM. The K+ sensitivity of these low- and high-affinity TEA states suggests that they may represent one- and two-ion occupancy states of the selectivity filter, consistent with recent crystallographic results from the bacterial KcsA K+ channel. We therefore analyzed these data in terms of such a model and found a large (almost 14-fold) difference between the intrinsic TEA affinity of the one-ion and two-ion modes. According to this analysis, the single ion in the one-ion mode (at 0 mV) prefers the inner end of the selectivity filter twofold more than the outer end. This distribution does not change with internal K+. The two ions in the two-ion mode prefer to occupy the inner end of the selectivity filter at low K+, but high internal K+ promotes increased occupancy of the outer sites. Our analysis further suggests that the four K+ sites in the selectivity filter are spaced between 20 and 25% of the membrane electric field

Increasing myosin light chain 3f (MLC3f) protects against a decline in contractile velocity

Disuse induces adaptations in skeletal muscle, which lead to muscle deterioration. Hindlimb-unloading (HU) is a well-established model to investigate cellular mechanisms responsible for disuse-induced skeletal muscle dysfunction. In myosin heavy chain (MHC) type IIB fibers HU induces a reduction in contraction speed (Vo) and a reduction in the relative myosin light chain 3f (MLC3f) protein content compared with myosin light chain 1f (MLC1f) protein. This study tested the hypothesis that increasing the relative MLC3f protein content via rAd-MLC3f vector delivery would attenuate the HU-induced decline in Vo in single MHC type IIB fibers. Fischer-344 rats were randomly assigned to one of three groups: control, HU for 7 days, and HU for 7 days plus rAd-MLC3f. The semimembranosus muscles were injected with rAd-MLC3f (3.75 x 1011-5 x 1011 ifu/ml) at four days after the initiation of HU. In single MHC type IIB fibers the relative MLC3f content decreased by 25% (12.00±0.60% to 9.06±0.66%) and Vo was reduced by 29% (3.22±0.14fl/s vs. 2.27±0.08fl/s) with HU compared to the control group. The rAd-MLC3f injection resulted in an increase in the relative MLC3f content (12.26±1.19%) and a concomitant increase in Vo (2.90±0.15fl/s) of MHC type IIB fibers. A positive relationship was observed between the percent of MLC3f content and Vo. Maximal isometric force and specific tension were reduced with HU by 49% (741.45±44.24μN to 379.09±23.77μN) and 33% (97.58±4.25kN/m2 to 65.05±2.71kN/m2), respectively compared to the control group. The rAd-MLC3f injection did not change the HU-induced decline in force or specific tension. Collectively, these results indicate that rAd-MLC3f injection rescues hindlimb unloading-induced decline in Vo in MHC type IIB single muscle fibers.Published versio

Membrane-delimited Inhibition of Maxi-K Channel Activity by the Intermediate Conductance Ca2+-activated K Channel

The complexity of mammalian physiology requires a diverse array of ion channel proteins. This diversity extends even to a single family of channels. For example, the family of Ca2+-activated K channels contains three structural subfamilies characterized by small, intermediate, and large single channel conductances. Many cells and tissues, including neurons, vascular smooth muscle, endothelial cells, macrophages, and salivary glands express more than a single class of these channels, raising questions about their specific physiological roles. We demonstrate here a novel interaction between two types of Ca2+-activated K channels: maxi-K channels, encoded by the KCa1.1 gene, and IK1 channels (KCa3.1). In both native parotid acinar cells and in a heterologous expression system, activation of IK1 channels inhibits maxi-K activity. This interaction was independent of the mode of activation of the IK1 channels: direct application of Ca2+, muscarinic receptor stimulation, or by direct chemical activation of the IK1 channels. The IK1-induced inhibition of maxi-K activity occurred in small, cell-free membrane patches and was due to a reduction in the maxi-K channel open probability and not to a change in the single channel current level. These data suggest that IK1 channels inhibit maxi-K channel activity via a direct, membrane-delimited interaction between the channel proteins. A quantitative analysis indicates that each maxi-K channel may be surrounded by four IK1 channels and will be inhibited if any one of these IK1 channels opens. This novel, regulated inhibition of maxi-K channels by activation of IK1 adds to the complexity of the properties of these Ca2+-activated K channels and likely contributes to the diversity of their functional roles

An Improve Naked Mole Rat Trap, Group III

This project attempts to improve on Dr. Stan Braude’s current Naked Mole Rat trap design. Dr. Braude currently uses his trap to catch and tag naked mole rats for his research. While Dr. Braude is happy with his current trap, he designed them himself over twenty years ago, and believes that they can be improved and modernized for increased ease and convenience. The key project goal is to deliver an improved trap to Dr. Braude that will meet his needs. The modifications that this project will attempt to deliver are a repeatable, solar powered trap that is compatible with his current design

Discovery of Photospheric Calcium Line Strength Variations in the DAZd White Dwarf G29-38

Metals in the photospheres of white dwarfs with Teff between 12,000 and 25,000 K should gravitationally settle out of these atmospheres in 1-2 weeks. Temporal variations in the line strengths of these metals could provide a direct measurement of episodic metal accretion. Using archival VLT and Keck spectroscopy, we find evidence that the DAZd white dwarf G29-38 shows significant changes in its Ca II K line strength. At the two best-observed epochs, we find that the Ca line equivalent width (EW) = 165 +- 4 mA (in 1996.885) and 280 +- 8 mA (in 1999.653), which is an increase of 70%. We consider the effect that pulsation has on the Ca EWs for this known variable star, and find that it adds an error of < 1 mA to these measurements. Calcium line strengths at other observational epochs support variations with timescales as short as two weeks. These Ca EW variations indicate that the metal accretion process in G29-38, presumably from its debris disk, is episodic on timescales of a few weeks or less, and thus the accretion is not dominated by Poynting-Robertson drag from an optically thin, continuous disk, which has a timescale of ~1 year.Comment: 15 pages, 4 figures, ApJ accepte

C57BL/6 life span study: age-related declines in muscle power production and contractile velocity

Quantification of key outcome measures in animal models of aging is an important step preceding intervention testing. One such measurement, skeletal muscle power generation (force * velocity), is critical for dynamic movement. Prior research focused on maximum power (P max), which occurs around 30-40 % of maximum load. However, movement occurs over the entire load range. Thus, the primary purpose of this study was to determine the effect of age on power generation during concentric contractions in the extensor digitorum longus (EDL) and soleus muscles over the load range from 10 to 90 % of peak isometric tetanic force (P 0). Adult, old, and elderly male C57BL/6 mice were examined for contractile function (6-7 months old, 100 % survival; ~24 months, 75 %; and ~28 months, 50 % P 0). The shape of the force-velocity curve also changed with age (a/P 0 increased). In addition, there were prolonged contraction times to maximum force and shifts in the distribution of the myosin light and heavy chain isoforms in the EDL. The results demonstrate that age-associated difficulty in movement during challenging tasks is likely due, in addition to overall reduced force output, to an accelerated deterioration of power production and contractile velocity under heavily loaded conditions.R01 AG017768 - NIA NIH HHS; F31 AG044108 - NIA NIH HHS; T32 AG029796 - NIA NIH HHS; R01 EY15313 - NEI NIH HHS; R01 EY015313 - NEI NIH HH

Pulsational Mapping of Calcium Across the Surface of a White Dwarf

We constrain the distribution of calcium across the surface of the white dwarf star G29-38 by combining time-series spectroscopy from Gemini-North with global time-series photometry from the Whole Earth Telescope. G29-38 is actively accreting metals from a known debris disk. Since the metals sink significantly faster than they mix across the surface, any inhomogeneity in the accretion process will appear as an inhomogeneity of the metals on the surface of the star. We measure the flux amplitudes and the calcium equivalent width amplitudes for two large pulsations excited on G29-38 in 2008. The ratio of these amplitudes best fits a model for polar accretion of calcium and rules out equatorial accretion

Infrared Spectral Observations While Drilling into a Frozen Lunar Simulant

Past and continuing observations indicate an enrichment of volatile materials in lunar polar regions. While these volatiles may be located near the surface, access to them will likely require subsurface sampling, during which it is desirable to monitor the volatile content. In a simulation of such activities, a multilayer lunar simulant was prepared with differing water content, and placed inside a thermal vacuum chamber at Glenn Research Center (GRC). The soil profile was cooled using liquid nitrogen. In addition to the soil, a drill and infrared (IR) spectrometer (~1600-3400 nm) were also located in the GRC chamber. We report the spectral observations obtained during a sequence where the drill was repeatedly inserted and extracted, to different depths, at the same location. We observe an overall increase in the spectral signature of water ice over the duration of the test. Additionally, we observe variations in the water ice spectral signature as the drill encounters different layers