Role of prostaglandins in spinal transmission of the exercise pressor reflex in decerebrated rats.

Previous studies found that prostaglandins in skeletal muscle play a role in evoking the exercise pressor reflex; however the role played by prostaglandins in the spinal transmission of the reflex is not known. We determined, therefore, whether or not spinal blockade of cyclooxygenase (COX) activity and/or spinal blockade of endoperoxide (EP) 2 or 4 receptors attenuated the exercise pressor reflex in decerebrated rats. We first established that intrathecal doses of a non-specific COX inhibitor Ketorolac (100 µg in 10 µl), a COX-2-specific inhibitor Celecoxib (100 µg in 10 µl), an EP2 antagonist PF-04418948 (10 µg in 10 µl), and an EP4 antagonist L-161,982 (4 µg in 10 µl) effectively attenuated the pressor responses to intrathecal injections of arachidonic acid (100 µg in 10 µl), EP2 agonist Butaprost (4 ng in 10 µl), and EP4 agonist TCS 2510 (6.25 µg in 2.5 µl), respectively. Once effective doses were established, we statically contracted the hind limb before and after intrathecal injections of Ketorolac, Celecoxib, the EP2 antagonist and the EP4 antagonist. We found that Ketorolac significantly attenuated the pressor response to static contraction (before Ketorolac: 23 ± 5 mmHg, after Ketorolac 14 ± 5 mmHg; p<0.05) whereas Celecoxib had no effect. We also found that 8 µg of L-161,982, but not 4 µg of L-161,982, significantly attenuated the pressor response to static contraction (before L-161,982: 21 ± 4 mmHg, after L-161,982 12 ± 3 mmHg; p<0.05), whereas PF-04418948 (10 µg) had no effect. We conclude that spinal COX-1, but not COX-2, plays a role in evoking the exercise pressor reflex, and that the spinal prostaglandins produced by this enzyme are most likely activating spinal EP4 receptors, but not EP2 receptors.

Dynamics of muscle microcirculatory and blood-myocyte O(2) flux during contractions.

The O(2) requirements of contracting skeletal muscle may increase 100-fold above rest. In 1919, August Krogh's brilliant insights recognized the capillary as the principal site for this increased blood-myocyte O(2) flux. Based on the premise that most capillaries did not sustain RBC flux at rest, Krogh proposed that capillary recruitment [i.e. initiation of red blood cell (RBC) flux in previously non-flowing capillaries] increased the capillary surface area available for O(2) flux and reduced mean capillary-to-mitochondrial diffusion distances. More modern experimental approaches reveal that most muscle capillaries may support RBC flux at rest. Thus, rather than contraction-induced capillary recruitment per se, increased RBC flux and haematocrit within already-flowing capillaries probably elevate perfusive and diffusive O(2) conductances and hence blood-myocyte O(2) flux. Additional surface area for O(2) exchange is recruited but, crucially, this may occur along the length of already-flowing capillaries (i.e. longitudinal recruitment). Today, the capillary is still considered the principal site for O(2) and substrate delivery to contracting skeletal muscle. Indeed, the presence of very low intramyocyte O(2) partial pressures (PO(2)s) and the absence of intramyocyte PO(2) gradients, whilst refuting the relevance of diffusion distances, place an even greater importance on capillary hemodynamics. This emergent picture calls for a paradigm-shift in our understanding of the function of capillaries by de-emphasizing de novo'capillary recruitment'. Diseases such as heart failure impair blood-myocyte O(2) flux, in part, by decreasing the proportion of RBC-flowing capillaries. Knowledge of capillary function in healthy muscle is requisite for identification of pathology and efficient design of therapeutic treatments.

Nitric oxide bioavailability modulates the dynamics of microvascular oxygen exchange during recovery from contractions.

AIM: lowered microvascular PO(2) (PO(2) mv) during the exercise off-transient likely impairs muscle metabolic recovery and limits the capacity to perform repetitive tasks. The current investigation explored the impact of altered nitric oxide (NO) bioavailability on PO(2) mv during recovery from contractions in healthy skeletal muscle. We hypothesized that increased NO bioavailability (sodium nitroprusside: SNP) would enhance PO(2) mv and speed its recovery kinetics while decreased NO bioavailability (l-nitro arginine methyl ester: l-NAME) would reduce PO(2) mv and slow its recovery kinetics. METHODS: PO(2) mv was measured by phosphorescence quenching during transitions (rest-1 Hz twitch-contractions for 3 min-recovery) in the spinotrapezius muscle of Sprague-Dawley rats under SNP (300 microm), Krebs-Henseleit (CONTROL) and l-NAME (1.5 mm) superfusion conditions. RESULTS: relative to recovery in CONTROL, SNP resulted in greater overall microvascular oxygenation as assessed by the area under the PO(2) mv curve (PO(2 AREA) ; CONTROL: 3471 ± 292 mmHg s; SNP: 4307 ± 282 mmHg s; P < 0.05) and faster off-kinetics as evidenced by the mean response time (MRToff; CONTROL: 60.2 ± 6.9 s; SNP: 34.8 ± 5.7 s; P < 0.05), whereas l-NAME produced lower PO(2 AREA) (2339 ± 444 mmHg s; P < 0.05) and slower MRToff (86.6 ± 14.5s; P < 0.05). CONCLUSION: no bioavailability plays a key role in determining the matching of O(2) delivery-to-O(2) uptake and thus the upstream O(2) pressure driving capillary-myocyte O(2) flux (i.e. PO(2) mv) following cessation of contractions in healthy skeletal muscle. Additionally, these data support a mechanistic link between reduced NO bioavailability and prolonged muscle metabolic recovery commonly observed in ageing and diseased populations.