Inflammation enhances Y1 receptor signaling, neuropeptide Y-mediated inhibition of hyperalgesia, and substance P release from primary afferent neurons.

Neuropeptide Y (NPY) is present in the superficial laminae of the dorsal horn and inhibits spinal nociceptive processing, but the mechanisms underlying its anti-hyperalgesic actions are unclear. We hypothesized that NPY acts at neuropeptide Y1 receptors in the dorsal horn to decrease nociception by inhibiting substance P (SP) release, and that these effects are enhanced by inflammation. To evaluate SP release, we used microdialysis and neurokinin 1 receptor (NK1R) internalization in rat. NPY decreased capsaicin-evoked SP-like immunoreactivity in the microdialysate of the dorsal horn. NPY also decreased non-noxious stimulus (paw brush)-evoked NK1R internalization (as well as mechanical hyperalgesia and mechanical and cold allodynia) after intraplantar injection of carrageenan. Similarly, in rat spinal cord slices with dorsal root attached, [Leu(31), Pro(34)]-NPY inhibited dorsal root stimulus-evoked NK1R internalization. In rat dorsal root ganglion neurons, Y1 receptors colocalized extensively with calcitonin gene-related peptide (CGRP). In dorsal horn neurons, Y1 receptors were extensively expressed and this may have masked the detection of terminal co-localization with CGRP or SP. To determine whether the pain inhibitory actions of Y1 receptors are enhanced by inflammation, we administered [Leu(31), Pro(34)]-NPY after intraplantar injection of complete Freund's adjuvant (CFA) in rat. We found that [Leu(31), Pro(34)]-NPY reduced paw clamp-induced NK1R internalization in CFA rats but not uninjured controls. To determine the contribution of increased Y1 receptor-G protein coupling, we measured [(35)S]GTPγS binding simulated by [Leu(31), Pro(34)]-NPY in mouse dorsal horn. CFA inflammation increased the affinity of Y1 receptor G-protein coupling. We conclude that Y1 receptors contribute to the anti-hyperalgesic effects of NPY by mediating the inhibition of SP release, and that Y1 receptor signaling in the dorsal horn is enhanced during inflammatory nociception.

Y1 receptor knockout increases nociception and prevents the anti-allodynic actions of NPY.

OBJECTIVE: Recent pharmacologic studies in our laboratory have suggested that the spinal neuropeptide Y (NPY) Y1 receptor contributes to pain inhibition and to the analgesic effects of NPY. To rule out off-target effects, the present study used Y1-receptor-deficient (-/-) mice to further explore the contribution of Y1 receptors to pain modulation. METHODS AND RESULTS: Y1(-/-) mice exhibited reduced latency in the hotplate test of acute pain and a longer-lasting heat allodynia in the complete Freund's adjuvant (CFA) model of inflammatory pain. Y1 deletion did not change CFA-induced inflammation. Upon targeting the spinal NPY systems with intrathecal drug delivery, NPY reduced tactile and heat allodynia in the CFA model and the partial sciatic nerve ligation model of neuropathic pain. Importantly, we show for the first time that NPY does not exert these anti-allodynic effects in Y1(-/-) mice. Furthermore, in nerve-injured CD1 mice, concomitant injection of the potent Y1 antagonist BIBO3304 prevented the anti-allodynic actions of NPY. Neither NPY nor BIBO3304 altered performance on the Rotorod test, arguing against an indirect effect of motor function. CONCLUSION: The Y1 receptor contributes to pain inhibition and to the analgesic effects of NPY.

Validation of a single-day maximal lactate steady state assessment protocol.

AIM: The classical maximal lactate steady state (MLSS) assessment protocol takes multiple days to measure thus necessitates athletes to return to a laboratory for several visits. The purpose of this study was to assess the validity and reliability of a new protocol (Palmer protocol), which proposes to measure MLSS in a single-day. METHODS: Nine endurance-trained males (age 21.1 +/- 1.6 years, VO2max of 63.2 +/- 3.2 ml x kg(-1) x min(-1)) performed the Palmer protocol and the classical MLSS assessment protocol. The classical MLSS protocol consisted of several constant-velocity runs of increasing intensity. The MLSS was defined as the highest velocity associated with an increase in blood lactate concentration ([La-]) = or < 1.0 mmol x L (-1) during the final 20 min of a 30 min run. Concurrent validity was assessed by calculating a Pearson product correlation coefficient between the running velocity at MLSS from the classical protocol and from the single-day Palmer protocol. Test-retest reliability was assessed by calculating a Pearson product correlation coefficient between the running velocities from 2 separate trials of the single-day Palmer protocol. RESULTS: The velocity at MLSS from the single-day Palmer protocol (236.4 +/- 27.8 m x min(-1)) produced a strong correlation of 0.97 (p<0.001) with the velocity at MLSS from the classical protocol (226.3 +/- 22.6 m x min(-1)). An equally strong correlation was calculated from test-retest reliability of the single-day Palmer protocol (r=0.97), (p<0.001). CONCLUSION: These results suggest that the single-day Palmer protocol is valid and reliable in the estimation of MLSS.

The effects of supplemental carbohydrate ingestion on intermittent isokinetic leg exercise.

BACKGROUND: The purpose of this investigation was to examine the effects of carbohydrate (CHO) supplementation on isokinetic leg extension/flexion exercise performance, blood glucose responses, blood free fatty acid (FFA) responses, and blood lactate (La) responses. METHODS: Eight resistance trained males (mean+/-SEM, age: 23.7+/-1.3 yrs, height: 180.0+/-3.5 cm, bodymass: 94.9+/-4.9 kg) participated in a randomized, double blind protocol with testing sessions separated by 7-d. Subjects were given CHO or placebo (P) while performing 16 sets of 10 repetitions at 120 degrees x s(-1) on a Cybex isokinetic dynamometer. Performance variables measured were; total work (TW), average work (AW), peak torque (PT) and average torque (AT). Plasma glucose (PG), FFA, and La were measured prior to testing (PRE), after set 8 (MID), and 16 (POST). RESULTS: Results indicated that the CHO treatment elicited significantly (p<0.05) more TW (CHO: 41.1+/-3.9 kJ; P: 38.1+/-3.9 kJ) and AW (CHO: 2.6+/-0.2 kJ; P: 2.4+/-0.2 kJ). There were no differences (p<0.05) between treatments for PT of the hamstrings (CHO: 91.6+/-6.5 Nm; P: 87.4+/-8.5 Nm) and quadriceps (CHO: 129.7+/-9.5 Nm; P: 123.0+/-10.6 Nm). The AT of the hamstrings (CHO: 77.8+/-5.2 Nm; P: 75.7+/-8.7 Nm) and quadriceps (CHO: 116.9+/-8.9 Nm; P: 110.0+/-8.5 Nm) were not statistically different (p>0.05) between the treatments. PG was significantly higher at the POST blood draw in the CHO treatment. No significant differences (p>0.05) were observed between the treatments for FFA and La concentrations. CONCLUSIONS: The data from this investigation indicate that the use of CHO supplementation during isokinetic leg exercise allows for the performance of more work.

Carbohydrate supplementation attenuates muscle glycogen loss during acute bouts of resistance exercise.

The effects of carbohydrate (CHO) supplementation on muscle glycogen and resistance exercise performance were examined with eight highly resistance trained males (mean +/- SEM, age: 24.3 +/- 1.1 years, height: 171.9 +/- 2.0 cm, body mass: 85.7 +/- 3.5 kg; experience 9.9 +/- 2.0 years). Subjects participated in a randomized, double blind protocol with testing sessions separated by 7 days. Testing consisted of an initial isokinetic leg exercise before and after an isotonic resistance exercise (IRT) session consisting of 3 leg exercises lasting approximately 39 min. Subjects consumed a CHO (1.0 g CHO.kg body mass(-1)) or placebo treatment (PLC), prior to and every 10-min (0.5 g CHO.kg body mass(-1)) during the IRT. Muscle tissue was obtained from the m vastus lateralis after a supine rest (REST) immediately after the initial isokinetic test (POST-ISO) and immediately after the IRT (POST-IRT). The CHO treatment elicited significantly less muscle glycogen degradation from the POST-ISO to POST-IRT (126.9 +/- 6.5 to 109.7 +/- 7.1 mmol.kg wet weight(-1)) compared to PLC (121.4 +/- 8.1 to 88.3 +/- 6. 0 mmol.kg wet weight(-1)). There were no differences in isokinetic performance between the treatments. The results of this investigation indicate that the consumption of a CHO beverage can attenuate the decrease in muscle glycogen associated with isotonic resistance exercise but does not enhance the performance of isokinetic leg exercise.