Stian Ellefsen

Differential regulation of AMP-activated kinase and AKT kinase in response to oxygen availability in crucian carp (Carassius carassius)

We investigated whether two kinases critical for survival during periods of energy deficiency in anoxia-intolerant mammalian species, AMP-activated kinase (AMPK), and protein kinase B (AKT), are equally important for hypoxic/anoxic survival in the extremely anoxia-tolerant crucian carp (Carassius carassius). We report that phosphorylation of AMPK and AKT in heart and brain showed small changes after 10 days of severe hypoxia (0.3 mg O2/l at 9 degrees C). In contrast, anoxia exposure (0.01 mg O2/l at 8 degrees C) substantially increased AMPK phosphorylation but decreased AKT phosphorylation in carp heart and brain, indicating activation of AMPK and deactivation of AKT. In agreement, blocking the activity of AMPK in anoxic fish in vivo with 20 mg/kg Compound C resulted in an elevated metabolic rate (as indicated by increased ethanol production) and tended to reduce energy charge. This is the first in vivo experiment with Compound C in a nonmammalian vertebrate, and it appears that AMPK plays a role in mediating anoxic metabolic depression in crucian carp. Real-time RT-PCR analysis of the investigated AMPK subunit revealed that the most likely composition of subunits in the carp heart is alpha2, beta1B, gamma2a, whereas a more even expression of subunits was found in the brain. In the heart, expression of the regulatory gamma2-subunit increased in the heart during anoxia. In the brain, expression of the alpha1-, alpha2-, and gamma1-subunits decreased with anoxia exposure, but expression of the gamma2-subunit remained constant. Combined, our findings suggest that AMPK and AKT may play important, but opposing roles for hypoxic/anoxic survival in the anoxia-tolerant crucian carp.

Proteomic changes in the crucian carp brain during exposure to anoxia

During exposure to anoxia, the crucian carp brain is able to maintain normal overall protein synthesis rates. However, it is not known if there are alterations in the synthesis or expression of specific proteins. This investigation addresses this issue by comparing the normoxic and anoxic brain proteome. Nine proteins were found to be reduced by anoxia. Reductions in the glycolytic pathway proteins creatine kinase, fructose biphosphate aldolase, glyceraldehyde‐3‐phosphate dehydrogenase, triosephosphate isomerase and lactate dehydrogenase reflect the reduced production and requirement for adenosine tri‐phosphate during anoxia. In terms of neural protection, voltage‐dependent anion channel, a protein associated with neuronal apoptosis, was reduced, along with gefiltin, a protein associated with the subsequent need for neuronal repair. Additionally the expression of proteins associated with neural degeneration and impaired cognitive function also declined; dihydropyrimidinase‐like protein‐3 and vesicle amine transport protein‐1. One protein was found to be increased by anoxia; pre‐proependymin, the precursor to ependymin. Ependymin fulfils multiple roles in neural plasticity, memory formation and learning, neuron growth and regeneration, and is able to reverse the possibility of apoptosis, thus further protecting the anoxic brain.

Block periodization of high‐intensity aerobic intervals provides superior training effects in trained cyclists

The purpose of this study was to compare the effect of two different methods of organizing endurance training in trained cyclists. One group of cyclists performed block periodization, wherein the first week constituted five sessions of high‐intensity aerobic training (HIT), followed by 3 weeks of one weekly HIT session and focus on low‐intensity training (LIT) (BP; n = 10, VO2max = 62 ± 2 mL/kg/min). Another group of cyclists performed a more traditional organization, with 4 weeks of two weekly HIT sessions interspersed with LIT (TRAD; n = 9, VO2max = 63 ± 2 mL/kg/min). Similar volumes of both HIT and LIT was performed in the two groups. While BP increased VO2max, peak power output (Wmax) and power output at 2 mmol/L [la−] by 4.6 ± 3.7%, 2.1 ± 2.8%, and 10 ± 12%, respectively (P < 0.05), no changes occurred in TRAD. BP showed relative improvements in VO2max compared with TRAD (P < 0.05). Mean effect size (ES) of the relative improvement in VO2max, Wmax, and power output at 2 mmol/L [la−] revealed large to moderate effects of BP training compared with TRAD training (ES = 1.34, ES = 0.85, and ES = 0.71, respectively). The present study suggests that block periodization of training provides superior adaptations to traditional organization during a 4‐week endurance training period, despite similar training volume and intensity.

Expression of heat shock proteins in anoxic crucian carp (Carassius carassius): support for cold as a preparatory cue for anoxia

The crucian carp (Carassius carassius) tolerates anoxia for days to months depending on temperature. During episodes of stress, heat shock proteins (HSPs) are important for limiting cellular damage, mainly by ensuring protein function. Accordingly, we hypothesized that anoxia would change the expression of HSPs and that this response would be temperature dependent. Real-time RT-PCR was used to investigate the effects of 1 and 7 days anoxia (A1 and A7) on the expression of HSP70a, HSP70b, HSC70, HSP90, and HSP30 in the brain and heart of 8 degrees C- and 13 degrees C-acclimated crucian carp. In general, the expression of all HSPs changed in response to anoxia, although varying in size and direction, and with organ and temperature. HSP70a expression increased drastically (approximately 10-fold) in A7 brains and hearts at 13 degrees C but not at 8 degrees C. HSC70 and HSP90 expression decreased in A7 brains (by 60-70%), but not in A7 hearts. HSC70 expression increased in A1 brains and hearts at both temperatures (by 60-160%), and HSP30 expression decreased in A7 brains and hearts at both temperatures (by 50-80%). Notably, normoxic fish showed 7- and 11-fold higher HSP70a expression in the brain and heart at 8 degrees C compared with 13 degrees C. This difference disappeared during anoxia, suggesting that cold may function as a cue for preconditioning the crucian carps HSP70a expression to the approaching anoxic winter period.

Hypoxia inducible factor 1α links fast-patterned muscle activity and fast muscle phenotype in rats

Non‐technical summary  Muscle fibres change when they are used differently, such as by exercise. Genetic studies have shown that a hyperactive form of the gene regulatory protein hypoxia inducible factor 1α (HIF‐1α) occurs twice as often among strength‐trained athletes as in the normal population. HIF‐1α is ‘sensing’ the oxygen levels in cells, and the oxygen levels change by an order of magnitude in working muscle. We show that an ‘endurance’ type of activity reduces the level of HIF‐1α, while short intense bursts of activity increases it. When HIF‐1α was produced in higher quantities by introducing artificial genes in the muscle fibres they became larger and faster, and with a less oxidative metabolism. Thus, oxygen itself could be a trigger for changes in muscle. The composition of muscle fibres is strongly correlated to major lifestyle conditions such as diabetes and chronic obstructive pulmonary disease, and HIF‐1α might provide a new molecular link.

Irisin in Blood Increases Transiently after Single Sessions of Intense Endurance Exercise and Heavy Strength Training

Purpose Irisin is a recently identified exercise-induced hormone that increases energy expenditure, at least in rodents. The main purpose of this study was to test the hypothesis that Irisin increases acutely in blood after singular sessions of intense endurance exercise (END) and heavy strength training (STR). Secondary, we wanted to explore the relationship between body composition and exercise-induced effects on irisin, and the effect of END and STR on muscular expression of the irisin gene FNDC5. Methods Nine moderately trained healthy subjects performed three test days using a randomized and standardized crossover design: one day with 60 minutes of END, one day with 60 minutes of STR, and one day without exercise (CON). Venous blood was sampled over a period of 24h on the exercise days. Results Both END and STR led to transient increases in irisin concentrations in blood, peaking immediately after END and one hour after STR, before gradually returning to baseline. Irisin responses to STR, but not END, showed a consistently strong negative correlation with proportions of lean body mass. Neither END nor STR affected expression of FNDC5, measured 4h after training sessions, though both protocols led to pronounced increases in PGC-1α expression, which is involved in transcriptional control of FNDC5. Conclusion The results strongly suggest that single sessions of intense endurance exercise and heavy strength training lead to transient increases in irisin concentrations in blood. This was not accompanied by increased FNDC5 expression, measured 4h post-exercise. The results suggest that irisin responses to resistance exercise are higher in individuals with lower proportions of lean body mass.

Effects of 12 weeks of block periodization on performance and performance indices in well-trained cyclists.

The purpose of this study was to compare the effects of two different methods of organizing endurance training in trained cyclists during a 12‐week preparation period. One group of cyclists performed block periodization (BP; n = 8), wherein every fourth week constituted five sessions of high‐intensity aerobic training (HIT), followed by 3 weeks of one HIT session. Another group performed a more traditional organization (TRAD; n = 7), with 12 weeks of two weekly HIT sessions. The HIT was interspersed with low‐intensity training (LIT) so that similar total volumes of both HIT and LIT were performed in the two groups. BP achieved a larger relative improvement in VO2max than TRAD (8.8 ± 5.9% vs 3.7 ± 2.9%, respectively, P < 0.05) and a tendency toward larger increase in power output at 2 mmol/L [la−] (22 ± 14% vs 10 ± 7%, respectively, P = 0.054). Mean effect size (ES) of the relative improvement in VO2max, power output at 2 mmol/L [la−], hemoglobin mass, and mean power output during 40‐min all‐out trial revealed moderate superior effects of BP compared with TRAD training (ES range was 0.62–1.12). The present study suggests that BP of endurance training has superior effects on several endurance and performance indices compared with TRAD.

Expression of genes involved in GABAergic neurotransmission in anoxic crucian carp brain (Carassius carassius)

The crucian carp, Carassius carassius, survives days to months without oxygen, depending on temperature. In the anoxic crucian carp brain, increased GABAergic inhibition, mediated by increased extracellular levels of GABA, has been shown to suppress electric activity and ATP consumption. To investigate an involvement of gene expression in this response, we utilized real-time RT-PCR to test the effect of 1 and 7 days anoxia (8 degrees C) on the expression of 22 genes, including nine GABA(A) receptor subunits (alpha(1-6), beta(2), delta, and gamma(2)), three GABA(B) receptor subunits (G(B)1a-1b and G(B)2), three enzymes involved in GABA metabolism (GAD65 and GAD67, GABAT), four GABA transporters (GAT1, 2a-b and 3), two GABA(A) receptor-associated proteins (GABARAP 1 and 2), and the K(+)/Cl(-) cotransporter KCC2. While the expression of GABA(A) receptor subunits was dominated by alpha(4)-, alpha(6)-, and delta-subunits, all of which are located to extrasynaptic sites in mammalian brains and respond to elevations in extracellular levels of GABA by showing tonic activity patterns, the expression of GABA transporters was dominated by GAT2 (a and b) and GAT3, which also show extrasynaptic location in mammals. These expression patterns differ from those observed in mammals and may be a prerequisite for GABAergic inhibition of anoxic metabolic rate in crucian carp. Furthermore, while the expression of the majority of the genes was largely unaltered by anoxia, the expression of GAT2 and GAT3 decreased to 20%. This suggests impairment of GABA transport, which could be a mechanism behind the accumulation of extracellular GABA and the increased GABAergic inhibition.

The Effects of Adding Different Whole-Body Vibration Frequencies to Preconditioning Exercise on Subsequent Sprint Performance

Rønnestad, BR and Ellefsen, S. The effects of adding different whole-body vibration frequencies to preconditioning exercise on subsequent sprint performance. J Strength Cond Res 25(12): 3306–3310, 2011—The phenomenon postactivation potentiation can possibly be used to acutely improve sprint performance. The purpose of this study was to investigate the effect of adding whole-body vibration (WBV) to body-loaded half-squats, performed as preconditioning activity to the 40-m sprint test. Nine male amateur soccer players performed 1 familiarization session and 6 separate test sessions. Each session included a standardized warm-up followed by 1 of the after preconditioning exercises: 30-seconds of half-squats with WBV at either 50 or 30 Hz or half-squats without WBV. The 40-m sprint was performed 1 minute after the preconditioning exercise. For each subject, each of the 3 protocols was repeated twice on separate days in a randomized order. Mean values were used in the statistical analysis. Performing the preconditioning exercise with WBV at a frequency of 50 Hz resulted in a superior 40-m sprint performance compared to preconditioning exercise without WBV (5.48 ± 0.19 vs. 5.52 ± 0.21 seconds, respectively, p < 0.05). There was no difference between preconditioning exercise with WBV at a frequency of 30 Hz and the no-WBV condition. In conclusion, preconditioning exercise performed with WBV at 50 Hz seems to enhance 40-m sprint performance in recreationally trained soccer players. The present findings suggest that coaches can incorporate such exercise into the warm-up to improve sprint performance or the quality of the sprint training.

Strength training improves performance and pedaling characteristics in elite cyclists.

The purpose was to investigate the effect of 25 weeks heavy strength training in young elite cyclists. Nine cyclists performed endurance training and heavy strength training (ES) while seven cyclists performed endurance training only (E). ES, but not E, resulted in increases in isometric half squat performance, lean lower body mass, peak power output during Wingate test, peak aerobic power output (Wmax), power output at 4 mmol L−1 [la−], mean power output during 40‐min all‐out trial, and earlier occurrence of peak torque during the pedal stroke (P < 0.05). ES achieved superior improvements in Wmax and mean power output during 40‐min all‐out trial compared with E (P < 0.05). The improvement in 40‐min all‐out performance was associated with the change toward achieving peak torque earlier in the pedal stroke (r = 0.66, P < 0.01). Neither of the groups displayed alterations in VO2max or cycling economy. In conclusion, heavy strength training leads to improved cycling performance in elite cyclists as evidenced by a superior effect size of ES training vs E training on relative improvements in power output at 4 mmol L−1 [la−], peak power output during 30‐s Wingate test, Wmax, and mean power output during 40‐min all‐out trial.