K. Sue Hageman

Effects of Type II diabetes on muscle microvascular oxygen pressures

We tested the hypothesis that muscle microvascular O2 pressure (PmvO2; reflecting the O2 delivery (QO2) to O2 uptake (VO2) ratio) would be lowered in the spinotrapezius muscle of Goto-Kakizaki (GK) Type II diabetic rats (n=7) at rest and during twitch contractions when compared to control (CON; n=5) rats. At rest, PmvO2 was lower in GK versus CON rats (CON: 29+/-2; GK: 18+/-2Torr; P<0.05). At the onset of contractions, GK rats evidenced a faster change in PmvO2 than CON (i.e., time constant (tau); CON: 16+/-4; GK: 6+/-2s; P<0.05). In contrast to the monoexponential fall in PmvO2 to the steady-state level seen in CON, GK rats exhibited a biphasic PmvO2 response that included a blunted (or non-existent) PmvO2 decrease followed by recovery to a steady-state PmvO2 that was at, or slightly above, resting values. Compared with CON, this decreased PmvO2 across the transition to a higher metabolic rate in Type II diabetes would be expected to impair blood-muscle O2 exchange and contractile function.

Effects of nitric oxide synthase inhibition on vascular conductance during high speed treadmill exercise in rats

To determine the functional role of nitric oxide (NO) in regulating vascular conductance during high intensity dynamic exercise in skeletal muscles composed of all major fibre types, female Wistar rats (277 ± 4 g; n= 7) were run on a motor‐driven treadmill at a speed and gradient (60 m min‐1, 10% gradient) established to yield maximal oxygen uptake (Vo2,max). Vascular conductance (ml min‐1 (100g)‐1 mmHg‐1), defined as blood flow normalised to mean arterial pressure (MAP), was determined using radiolabelled microspheres during exercise before and after NO synthase (NOS) inhibition with NG‐nitro‐L‐arginine methyl ester (l‐NAME; 10 mg kg‐1, i.a.). The administration of l‐NAME increased MAP from pre‐l‐NAME baseline values, demonstrating that NOS activity is reduced. The administration of l‐NAME also reduced vascular conductance in 20 of the 28 individual hindlimb muscles or muscle parts examined during high speed treadmill exercise. These reductions in vascular conductance correlated linearly with the estimated sum of the percentage of slow twitch oxidative (SO) and fast twitch oxidative glycolytic (FOG) types of fibres in each muscle (Δconductance = ‐0.0082(%SO +%FOG) ‐ 0.0105; r= 0.66; P < 0.001). However, if the reduction in vascular conductance found in the individual hindquarter muscles or muscle parts was expressed as a percentage decrease from the pre‐l‐NAME value (%Δ= (pre‐l‐NAME conductance — post‐l‐NAME conductance)/pre‐l‐NAME conductance × 100), then the reduction in vascular conductance was similar in all muscles examined (average %Δ= ‐23 ± 2%). These results suggest that NO contributes substantially to the regulation of vascular conductance within and among muscles of the rat hindquarter during high intensity exercise. When expressed in absolute terms, the results suggest that the contribution of NO to the regulation of vascular conductance during high intensity exercise is greater in muscles that possess a high oxidative capacity. In contrast, if results are expressed in relative terms, then the contribution of NO to the regulation of vascular conductance during high intensity exercise is similar across the different locomotor muscles located in the rat hindlimb and independent of the fibre type composition.

Aging alters the contribution of nitric oxide to regional muscle hemodynamic control at rest and during exercise in rats

Advanced age is associated with altered skeletal muscle hemodynamic control during the transition from rest to exercise. This study investigated the effects of aging on the functional role of nitric oxide (NO) in regulating total, inter-, and intramuscular hindlimb hemodynamic control at rest and during submaximal whole body exercise. We tested the hypothesis that NO synthase inhibition (N(G)-nitro-l-arginine methyl ester, l-NAME; 10 mg/kg) would result in attenuated reductions in vascular conductance (VC) primarily in oxidative muscles in old compared with young rats. Total and regional hindlimb muscle VCs were determined via radiolabeled microspheres at rest and during treadmill running (20 m/min, 5% grade) in nine young (6-8 mo) and seven old (27-29 mo) male Fisher 344 × Brown Norway rats. At rest, l-NAME increased mean arterial pressure (MAP) significantly by ∼17% and 21% in young and old rats, respectively. During exercise, l-NAME increased MAP significantly by ∼13% and 19% in young and old rats, respectively. Compared with young rats, l-NAME administration in old rats evoked attenuated reductions in 1) total hindlimb VC during exercise (i.e., down by ∼23% in old vs. 43% in young rats; P < 0.05), and 2) VC in predominantly oxidative muscles both at rest and during exercise (P < 0.05). Our results indicate that the dependency of highly oxidative muscles on NO-mediated vasodilation is markedly diminished, and therefore mechanisms other than NO-mediated vasodilation control the bulk of the increase in skeletal muscle VC during the transition from rest to exercise in old rats. Reduced NO contribution to vasomotor control with advanced age is associated with blood flow redistribution from highly oxidative to glycolytic muscles during exercise.

Effects of Type II diabetes on exercising skeletal muscle blood flow in the rat

The purpose of the present investigation was to examine the muscle hyperemic response to steady-state submaximal running exercise in the Goto-Kakizaki (GK) Type II diabetic rat. Specifically, the hypothesis was tested that Type II diabetes would redistribute exercising blood flow toward less oxidative muscles and muscle portions of the hindlimb. GK diabetic (n = 10) and Wistar control (n = 8, blood glucose concentration, 13.7 ± 1.6 and 5.7 ± 0.2 mM, respectively, P < 0.05) rats were run at 20 m/min on a 10% grade. Blood flows to 28 hindlimb muscles and muscle portions as well as the abdominal organs and kidneys were measured in the steady state of exercise using radiolabeled 15-μm microspheres. Blood flow to the total hindlimb musculature did not differ between GK diabetic and control rats (161 ± 16 and 129 ± 15 ml·min(-1)·100 g(-1), respectively, P = 0.18). Moreover, there was no difference in blood flow between GK diabetic and control rats in 20 of the individual muscles or muscle parts examined. However, in the other eight muscles examined that typically are comprised of a majority of fast-twitch glycolytic (IIb/IIdx) fibers, blood flow was significantly greater (i.e., ↑31-119%, P < 0.05) in the GK diabetic rats. Despite previously documented impairments of several vasodilatory pathways in Type II diabetes these data provide the first demonstration that a reduction of exercising muscle blood flow during submaximal exercise is not an obligatory consequence of this condition in the GK diabetic rat.

Nitric oxide synthase inhibition during treadmill exercise reveals fiber-type specific vascular control in the rat hindlimb

The control of vascular tone during exercise is highly complex and integrated. Specifically, in regards to the contribution of nitric oxide (NO), the observed magnitude and muscle fiber-type dependency of the NO contribution to exercise hyperemia may differ depending on the timing of NO synthase (NOS) inhibition with respect to the exercise bout (i.e., administration prior to vs. during exercise). We tested the hypothesis that, in the presence of prior cyclooxygenase inhibition (indomethacin, 5 mg/kg(-1)), NOS inhibition (N(G)-nitro-L-arginine methyl ester, L-NAME; 10 mg/kg) administered during submaximal treadmill exercise would blunt blood flow and vascular conductance (VC) in the hindlimb muscle(s) of the rat with the greatest reductions in blood flow and VC occurring in the predominantly oxidative muscles. Adult female Wistar rats (n = 10, age: 3-4 mo) ran on a motor-driven treadmill (20 m/min, 10% grade). Total and regional hindlimb muscle blood flow and VC were determined via radiolabeled microspheres before (control) and after L-NAME administration during exercise. L-NAME reduced (P < 0.05) total hindlimb muscle blood flow (control: 123 + or - 10, L-NAME: 103 + or - 7 ml x min(-1) x 100g(-1)) and VC (control: 0.95 + or - 0.09, L-NAME: 0.63 + or - 0.05 ml x min(-1) x 100g(-1) x mmHg(-1)). There was a significant correlation (r = 0.51, P < 0.05) between the absolute reductions in VC after L-NAME and the percent sum of type I and IIa fibers in the individual muscles and muscle parts; however, there was no correlation (P = 0.62) when expressed as blood flow. Surprisingly, the highly oxidative muscles demonstrated a marked ability to maintain oxygen delivery, which differs substantially from previous reports of L-NAME infusion prior to exercise in these muscles. The demonstration that NO is an important regulator of blood flow and VC in the rat hindlimb during treadmill exercise, but that the fiber-type dependency of NO is altered markedly when NOS inhibition is performed during, vs. prior to, exercise, lends important insights into the integrated nature of vascular control during exercise.

Regional blood flow responses to acute ANG II infusion: Effects of nitric oxide synthase inhibition

We hypothesized that nitric oxide (NO) opposes regional vasoconstriction caused by acute angiotensin II (ANG II) infusion in conscious rats. Mean arterial pressure (MAP), blood flow, and vascular conductance (regional blood flow/ MAP; ml/min/100 g/mm Hg) were measured and/or calculated before and at 2 min of ANG II infusion (0.05 or 1 microg/kg/min, i.a.) in the absence and presence of NO synthase (NOS) inhibition [N(G)-nitro-L-arginine methyl ester (L-NAME), 0.25 or 1 mg/kg, i.a.]. ANG II reduced stomach and hindlimb conductance only after NOS inhibition. For example, whereas 0.05 microg/kg/min ANG II did not attenuate conductance in the stomach (i.e., 1.04+/-0.08 to 0.93+/-0.12 ml/min/100 g/mm Hg), this variable was reduced (i.e., 0.57+/-0.14 to 0.34-/+0.05 ml/min/100 g/mm Hg; p < 0.05) when ANG II was infused after 0.25 mg/kg L-NAME. In addition, whereas hindlimb conductance was similar before and after administering 1 microg/kg/min ANG II (i.e., 0.13+/-0.01 and 0.09+/-0.02, respectively), this variable was reduced (i.e., 0.07+/-0.01 and 0.02+/-0.00, respectively; p < 0.05) when ANG II was infused after 1 mg/kg L-NAME. These findings indicate that NO opposes ANG II-induced vasoconstriction in the stomach and hindlimb. In contrast, whereas both doses of ANG II decreased (p < 0.05) vascular conductance in the kidneys and small and large intestine regardless of whether NOS inhibition was present, absolute vascular conductance was lower (p < 0.05) after L-NAME. For example, 1 microg/kg ANG II reduced renal conductance from 3.34+/-0.31 to 1.22+/-0.14 (p < 0.05). After 1 mg/kg L-NAME, renal conductance decreased from 1.39+/-0.18 to 0.72+/-0.16 (p < 0.05) during ANG II administration. Therefore the constrictor effects of NOS inhibition and ANG II are additive in these circulations. Taken together, our results indicate that the ability of NO to oppose ANG II-induced constriction is not homogeneous among regional circulations.

Endogenous prostaglandins limit angiotensin-II induced regional vasoconstriction in conscious rats.

&NA; In conscious rats, we tested the hypothesis that prostaglandins attenuate regional vasoconstriction caused by acute infusion of angiotensin II. Mean arterial pressure, regional blood flow, and vascular conductance in response to 2‐minute infusions of 0.05 or 1 &mgr;g/kg/min Ang II were assessed before and during indomethacin treatment (5 mg/kg). Effects of the lower dose of Ang II (n = 8) on regional blood flow were not altered by indomethacin, but conductance in the kidney (2.98 ± 0.35 vs. 2.19 ± 0.32), stomach (1.15 ± 0.13 vs. 0.83 ± 0.13), and white gastrocnemius muscle (0.11 ± 0.02 vs. 0.07 ± 0.01 mL/min/100g/mm Hg) were reduced. Changes in conductance were not seen in the pancreas or spleen. In response to the higher dose of Ang II (n = 7), indomethacin reduced blood flow in the kidney, red and white gastrocnemius, and soleus muscles. Reductions in conductance were found in the kidney, stomach and small intestine, and in the red and white gastrocnemius, and soleus muscles (2.27 ± 0.9 vs. 1.79 ± 0.14, 0.44 ± 0.07 vs. 0.27 ± 0.03, 0.68 ± 0.11 vs. 0.60 ± 0.07, 0.43 ± 0.08 vs. 0.16 ± 0.03, 0.10 ± 0.02 vs. 0.05 ± 0.01, and 0.26 ± 0.03 vs. 0.15 ± 0.02 mL/min/100g, respectively). No changes occurred in the pancreas and spleen. Indomethacin had no effect on baseline blood flow or conductance in any of these organs. These results suggest that prostaglandins attenuate vasoconstriction caused by Ang II in a manner that is organ‐specific and dependent on the dose of Ang II. Consequently, prostaglandins may limit vasoconstriction and potential ischemia caused by elevated levels of this hormone.

Effect of chronic heart failure in older rats on respiratory muscle and hindlimb blood flow during submaximal exercise

Submaximal exercise diaphragm blood flow (BF) is elevated in young chronic heart failure (CHF) rats, while it is unknown if this occurs in older animals. Respiratory and hindlimb muscle BFs (radiolabeled microspheres) were measured at rest and during submaximal exercise (20m/min, 5% grade) in older healthy (n=7) and CHF (n=6) Fischer 344X Brown Norway rats (27-29 mo old). Older CHF, compared to healthy, rats had greater (p<0.01) left ventricular end-diastolic pressure and right ventricle and lung weight (normalized to body weight). During submaximal exercise, respiratory and hindlimb muscle BFs increased (p<0.02) in both groups, while diaphragm BF was higher (CHF: 257±32; healthy: 121±9mL/min/100g, p<0.01) and hindlimb BF lower (CHF: 111±10; healthy: 133±12mL/min/100g, p=0.04) in older CHF compared to healthy rats. Submaximal exercise hindlimb BF was negatively related (r=-0.93; p=0.03) to diaphragm BF in older CHF rats. During submaximal exercise, diaphragm BF is elevated in older CHF compared to healthy rats in proportion to the compromised hindlimb BF.

Respiratory muscle blood flow during exercise: effects of sex and ovarian cycle

Sex and ovarian cycle have been speculated to modify respiratory muscle blood flow control during exercise, but the findings are inconclusive. We tested the hypotheses that females would have higher respiratory muscle blood flow and vascular conductance (VC) compared with males during exercise and that this difference would be accentuated in proestrus vs. ovariectomized (OVA) females. Mean arterial pressure (carotid artery catheter) and respiratory muscle blood flow (radiolabeled microspheres) were measured during moderate-intensity (24 m/min, 10% grade) exercise in male (n = 9), female (n = 9), and OVA female (n = 7) rats and near-maximal (60 m/min, 5% grade) exercise in male (n = 5) and female (n = 7) rats. At rest, diaphragm, intercostal, and transversus abdominis blood flow were not different (P = 0.33) among groups. During moderate-intensity exercise, diaphragm (M: 124 ± 16; F: 140 ± 14; OVA: 140 ± 20 ml·min-1·100 g-1), intercostal (M: 33 ± 5; F: 34 ± 5; OVA: 30 ± 5 ml·min-1·100 g-1), and transversus abdominis blood flow (M: 24 ± 4; F: 35 ± 7; OVA: 35 ± 9 ml·min-1·100 g-1) significantly increased in all groups compared with rest but were not different (P = 0.12) among groups. From rest to moderate-intensity exercise, diaphragm (P < 0.03) and transversus abdominis (P < 0.04) VC increased in all groups, whereas intercostal VC increased only for males and females (P = 0.01). No differences (P > 0.13) existed in VC among groups. During near-maximal exercise, diaphragm (M: 304 ± 62; F: 283 ± 17 ml·min-1·100 g-1), intercostal (M: 29 ± 8; F: 40 ± 6 ml·min-1·100 g-1), and transversus abdominis (M: 85 ± 14; F: 86 ± 9 ml·min-1·100 g-1) blood flow and VC were not different (P > 0.27) between males and females. These data demonstrate that respiratory muscle blood flow and vascular conductance at rest and during exercise are not affected by sex or ovarian cycle in rats.NEW & NOTEWORTHY It has been proposed that sex and ovarian cycle modulate respiratory muscle blood flow control during exercise. We demonstrate herein that neither sex nor ovarian cycle influences respiratory muscle blood flow or vascular conductance at rest or during exercise in rats.

Effects of Altered Nitric Oxide Availability on Rat Muscle Microvascular Oxygenation During Contractions: 1628

AIM To explore the role of nitric oxide (NO) in controlling microvascular O2 pressure (P(O2)mv) at rest and during contractions (1 Hz). We hypothesized that at the onset of contractions sodium nitroprusside (SNP) would raise P(O2)mv and slow the kinetics of P(O2)mv change whereas l-nitro arginine methyl ester (L-NAME) would decrease P(O2)mv and speed its kinetics. METHODS We superfused the spinotrapezius muscle of female Sprague-Dawley rats (n = 7, body mass = 298 +/- 10 g) with SNP (300 microM) and L-NAME (1.5 mm) and measured P(O2)mv (phosphorescence quenching) during contractions. RESULTS SNP decreased mean arterial pressure (92 +/- 5 mmHg) below that of control (CON, 124 +/- 4 mmHg) and L-NAME (120 +/- 4 mmHg) conditions. SNP did not raise P(O2)mv at rest but it did elevate the P(O2)mv-to-MAP ratio (50% increase, P < 0.05) and slow the kinetics by lengthening the time-delay (TD, 14.0 +/- 5.0 s) and time constant (tau, 24.0 +/- 10.0 s) of the response compared with CON (TD, 8.4 +/- 3.3 s; tau, 16.0 +/- 4.5 s, P < 0.05 vs. SNP). L-NAME decreased P(O2)mv at rest and tended to speed tau (10.1 +/- 3.8 s, P = 0.1), while TD (8.1 +/- 1.0 s) was not significantly different. L-NAME also caused P(O2)mv to fall transiently below steady-state contracting values. CONCLUSIONS These results indicate that NO availability can significantly affect P(O2)mv at rest and during contractions and suggests that P(O2)mv derangements in ageing and chronic disease conditions may potentially result from impairments in NO availability.