Static Contraction Of Skeletal Muscle Research Articles

Endothelial Nitric Oxide Synthase (eNOS) Blockade Within the Ventrolateral Medulla Differentially Modulates the Exercise Pressor Reflex in Stroked‐Rats

We have previously shown that blockade of endothelial nitric oxide synthase (eNOS) within the rostral (RVLM) and caudal ventrolateral medulla (CVLM) differentially modulates cardiovascular responses which was associated with changes in extracellular glutamate and GABA concentrations during static skeletal muscle contraction, the Exercise Pressor Reflex. In this study using left‐sided stroked‐rats we determined if microdialyzing a specific eNOS antagonist into the left RVLM and/or the left CVLM would alter cardiovascular responses during the Exercise Pressor Reflex. Rats were stroked by employing a transient (90‐minute) left‐sided middle cerebral artery occlusion (MCAO) followed by 24 hr reperfusion. In Protocol 1, microdialysis of a selective eNOS antagonist, L‐N(5)‐(1‐iminoethyl)ornithine (L‐NIO; 10 µM) for 120 minutes into the left RVLM significantly potentiated cardiovascular responses during a static muscle contraction compared to those observed in intact rats. In Protocol 2, application of L‐NIO into the left CVLM significantly attenuated the cardiovascular responses during muscle contraction. Finally, in Protocol 3, administration of L‐NIO simultaneously into both the left RVLM and left CVLM produced results similar to those observed with Protocol 1. These results demonstrate that blockade of eNOS within the two regions of the ventrolateral medulla differentially modulates cardiovascular responses during static exercise in stroked‐rats.

Modulation of inducible nitric oxide synthase (iNOS) expression and cardiovascular responses during static exercise following iNOS antagonism within the ventrolateral medulla

Previous reports indicate that inducible nitric oxide synthase (iNOS) blockade within the rostral ventrolateral medulla (RVLM) and caudal ventrolateral medulla (CVLM) differentially modulated cardiovascular responses, medullary glutamate, and GABA concentrations during static skeletal muscle contraction. In the current study, we determined the role of iNOS antagonism within the RVLM and CVLM on cardiovascular responses and iNOS protein expression during the exercise pressor reflex in anesthetized rats. Following 120 min of bilateral microdialysis of a selective iNOS antagonist, aminoguanidine (AGN; 10 µM), into the RVLM, the pressor responses were attenuated by 72 % and changes in heart rate were reduced by 38 % during a static muscle contraction. Furthermore, western blot analysis of iNOS protein abundance within the RVLM revealed a significant attenuation when compared to control animals. In contrast, bilateral administration of AGN (10 µM) into the CVLM augmented the increases in mean arterial pressure by 60 % and potentiated changes in heart rate by 61 % during muscle contractions, but did not alter expression of the iNOS protein within the CVLM. These results demonstrate that iNOS protein expression within the ventrolateral medulla is differentially regulated by iNOS blockade that may, in part, contribute to the modulation of cardiovascular responses during static exercise.

Inducible nitric oxide synthase blockade within the ventrolateral medulla differentially modulates the exercise pressor reflex in stroke rats (681.5)

We have previously shown that blockade of neuronal nitric oxide synthase (nNOS) within the rostral (RVLM) and caudal ventrolateral medulla (CVLM) differentially modulates cardiovascular responses via changes in glutamate and GABA concentrations during static skeletal muscle contraction. Using left‐sided stroked rats in this study, we determined if microdialyzing a specific iNOS antagonist into the left RVLM and/or the left CVLM would alter cardiovascular responses during static muscle contraction commonly known as the Exercise Pressor Reflex. Rats were stroked by employing a 90 minute left‐sided middle cerebral artery occlusion (MCAO) followed by 24 hours of reperfusion. In protocol 1, microdialysis of a selective iNOS antagonist, aminoguanidine (AGN; 10 µM) for 120 min into the left RVLM significantly attenuated cardiovascular responses during a static muscle contraction compared to those induced in intact rats. In Protocol 2, application of AGN into the left CVLM significantly potentiated the cardiovascular responses during muscle contractions. Finally, in Protocol 3, administration of AGN simultaneously into both the left RVLM and left CVLM produced results similar to those observed with Protocol 1. These results demonstrate that blockade of iNOS within the two regions of the ventrolateral medulla differentially modulates cardiovascular responses during static exercise in stroked rats.

Cardiovascular responses during static skeletal muscle contraction following neuronal NOS blockade within the ventrolateral medulla

We have previously shown that blockade of neuronal nitric oxide synthase (nNOS) within the rostral (RVLM) and caudal ventrolateral medulla (CVLM) differentially modulates cardiovascular responses via changes in glutamate and GABA concentrations during static skeletal muscle contraction (Brain Res. 977: 80–89, 2003). In this study, we determined if microdialyzing a specific nNOS antagonist into the left RVLM or the left CVLM and into both the left RVLM and CVLM would affect cardiovascular responses expression during the exercise pressor reflex in left‐sided stroked rats. Rats were stroked by employing a 90 minute left‐sided middle cerebral artery occlusion (MCAO) followed by 24 hours of reperfusion. In protocol 1, microdialysis of a selective nNOS antagonist, 1‐(2‐trifluoromethylphenyl)‐ imidazole (TFMP, 1.0 microM), for 120 min into the left RVLM significantly potentiated cardiovascular responses during a static muscle contraction compared to intact rats. In Protocol 2, application of TFMP into the left CVLM significantly attenuated cardiovascular responses during muscle contractions. Finally, administration of TFMP into both the left RVLM and left CVLM produced results similar to those observed with Protocol 1. These results demonstrate that blockade of nNOS within the ventrolateral medulla differentially modulates cardiovascular responses during static exercise in stroked rats.

Angiotensin II type-2 (AT2) receptor antagonism alters cardiovascular responses to static exercise and simultaneously changes glutamate/GABA levels within the ventrolateral medulla

Angiotensin II receptors (Ang II), classified into AT1 and AT2 subtypes, are located in different regions of the central nervous system, including the cardiovascular control centers in the medulla oblongata. We previously reported the role of Ang II AT1 receptors within the medulla on cardiovascular responses and glutamate/GABA neurotransmission during the exercise pressor reflex [Patel, D., Böhlke, M., Phattanarudee, S., Kabadi, S., Maher, T.J., Ally, A., 2008. Cardiovascular responses and neurotransmitter changes during blockade of angiotensin II receptors within the ventrolateral medulla. Neurosci. Res. 60 (3), 340–348]. In this study, we investigated the role of the AT2 receptor subtype within the ventrolateral medullary region (VLM) in modulating increases in mean arterial pressure (MAP) and heart rate (HR) in response to static skeletal muscle contraction. Methods Using microdialysis methods in anesthetized rats, we administered AR-AT2 antagonists into the rostral (RVLM) and caudal (CVLM) VLM and determined its effects on cardiovascular responses and glutamate/GABA neurotransmission following muscle contraction. Bilateral microdialysis of a selective AT2 antagonist, PD 123319 (10 μM), for 30 min into the RVLM augmented MAP and HR responses during a static muscle contraction. Simultaneously, the drug increased glutamate and decreased GABA levels within the RVLM. After 60 min of discontinuation of the drug, only MAP and HR values but not the neurotransmitter levels in response to a muscle contraction returned to baseline. In contrast, bilateral microdialysis of the drug into the CVLM attenuated cardiovascular responses during a static muscle contraction, decreased glutamate and increased GABA. However, only the cardiovascular responses recovered after 60 min of discontinuation of the drug. These results demonstrate that AT2 within both RVLM and CVLM plays important differential roles in modulating neurotransmission and cardiovascular function during the exercise pressor reflex.

Spinal P2X receptor modulates muscle pressor reflex via glutamate

Static contraction of skeletal muscle evokes reflex increases in blood pressure and heart rate. Previous studies showed that P2X receptors located at the dorsal horn of the spinal cord play a role in modulating the muscle pressor reflex. P2X stimulation can alter release of the excitatory amino acid, glutamate (Glu). In this report, we tested the hypothesis that stimulation of P2X receptors enhances the concentrations of Glu ([Glu]) in the dorsal horn, and that blocking P2X receptors attenuates contraction-induced Glu increases and the resultant reflex pressor response. Contraction was elicited by electrical stimulation of the L(7) and S(1) ventral roots of 14 cats. Glu samples were collected from microdialysis probes inserted in the L(7) level of the dorsal horn of the spinal cord, and dialysate [Glu] was determined using the HPLC method. First, microdialyzing alpha,beta-methylene ATP (0.4 mM) into the dorsal horn significantly increased [Glu]. In addition, contraction elevated [Glu] from baseline of 536 +/- 53 to 1,179 +/- 192 nM (P < 0.05 vs. baseline), and mean arterial pressure by 39 +/- 8 mmHg in the control experiment. Microdialyzing the P2X receptor antagonist pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid (10 mM) into the dorsal horn attenuated the contraction induced-Glu increase (610 +/- 128 to 759 +/- 147 nM; P > 0.05) and pressor response (16 +/- 3 mmHg, P < 0.05 vs. control). Our findings demonstrate that P2X modulates the cardiovascular responses to static muscle contraction by affecting the release of Glu in the dorsal horn of the spinal cord.

Neuronal Nitric Oxide Synthase (nNOS) blockade within the ventrolateral medulla differentially modulates cardiovascular responses and nNOS expression during static skeletal muscle contraction

Nitric oxide (NO) is synthesized from L-arginine through the activity of the enzyme, NO synthase (NOS). Previous studies have demonstrated the role of the 3 isoforms of NOS, namely endothelial NOS (eNOS), neuronal NOS (nNOS), and inducible NOS (iNOS) in cardiovascular regulation. Local blockade of nNOS in RVLM vs. CVLM differentially alters local glutamate and GABA release, and thereby results in opposite cardiovascular responses to static muscle contraction (Brain Res. 2003, 977, 80–89). In this study, we examined whether nNOS antagonism within the RVLM and CVLM affected cardiovascular responses during the exercise pressor reflex and simultaneously modulated medullary nNOS protein expression using anesthetized rats. Bilateral microdialysis of a selective nNOS antagonist, 1-(2-trifluoromethylphenyl)-imidazole (TRIM, 1.0 μM) for 120 min into the RVLM, potentiated cardiovascular responses during a static muscle contraction. Western blot analysis of nNOS expression within the RVLM showed significant attenuation of the protein when compared to the data obtained from control animals microdialyzed with vehicle. In contrast, bilateral application of TRIM into the CVLM attenuated cardiovascular responses during muscle contractions and increased nNOS protein expression within the CVLM. These results demonstrated that nNOS protein expression within the brainstem was pharmacologically altered by nNOS blockade within the RVLM or CVLM, which in turn might have contributed to the augmentation or attenuation of cardiovascular responses, respectively, during static exercise.

Cardiovascular responses and neurotransmitter changes during static muscle contraction following blockade of inducible nitric oxide synthase (iNOS) within the ventrolateral medulla

The enzyme nitric oxide synthase (NOS) which is necessary for the production of nitric oxide from l-arginine exists in three isoforms: neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS). Our previous studies have demonstrated the roles of nNOS and eNOS within the rostral (RVLM) and caudal ventrolateral medulla (CVLM) in modulating cardiovascular responses during static skeletal muscle contraction via altering localized glutamate and GABA levels ( Brain Res. 977 (2003) 80–89; Neuroscience Res. 52 (2005) 21–30). In this study, we investigated the role of iNOS within the RVLM and CVLM on cardiovascular responses and glutamatergic/GABAergic neurotransmission during the exercise pressor reflex. Bilateral microdialysis of a selective iNOS antagonist, aminoguanidine (AGN; 1.0 μM), for 60 min into the RVLM attenuated increases in mean arterial pressure (MAP), heart rate (HR), and extracellular glutamate levels during a static muscle contraction. Levels of GABA within the RVLM were increased. After 120 min of discontinuation of the drug, MAP and HR responses and glutamate/GABA concentrations recovered to baseline values during a subsequent muscle contraction. In contrast, bilateral application of AGN (1.0 μM) into CVLM potentiated cardiovascular responses and glutamate concentration while attenuating levels of GABA during a static muscle contraction. All values recovered after 120 min of discontinuation of the drug. These results demonstrate that iNOS within the ventrolateral medulla plays an important role in modulating cardiovascular responses and glutamatergic/GABAergic neurotransmission that regulates the exercise pressor reflex.

Spinal P2X receptor modulates reflex pressor response to activation of muscle afferents

Static contraction of skeletal muscle evokes increases in blood pressure and heart rate. Previous studies suggested that the dorsal horn of the spinal cord is the first synaptic site responsible for those cardiovascular responses. In this study, we examined the role of ATP-sensitive P2X receptors in the cardiovascular responses to contraction by microdialyzing the P2X receptor antagonist pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) into the L7 level of the dorsal horn of nine anesthetized cats. Contraction was elicited by electrical stimulation of the L7 and S1 ventral roots. Blockade of P2X receptor attenuated the contraction induced-pressor response [change in mean arterial pressure (delta MAP): 16 +/- 4 mmHg after 10 mM PPADS vs. 42 +/- 8 mmHg in control; P < 0.05]. In addition, the pressor response to muscle stretch was also blunted by PPADS (delta MAP: 27 +/- 5 mmHg after PPADS vs. 49 +/- 8 mmHg in control; P < 0.05). Finally, activation of P2X receptor by microdialyzing 0.5 mM alpha,beta-methylene into the dorsal horn significantly augmented the pressor response to contraction. This effect was antagonized by prior PPADS dialysis. These data demonstrate that blockade of P2X receptors in the dorsal horn attenuates the pressor response to activation of muscle afferents and that stimulation of P2X receptors enhances the reflex response, indicating that P2X receptors play a role in mediating the muscle pressor reflex at the first synaptic site of this reflex.

Cardiovascular responses and neurotransmitter changes following blockade of nNOS within the ventrolateral medulla during static muscle contraction

Nitric oxide (NO) is synthesized from l-arginine through the activity of the synthetic enzyme, NO synthase (NOS). Previous studies have demonstrated the roles of the three isoforms of NOS, namely endothelial NOS (eNOS), neuronal NOS (nNOS), and inducible NOS (iNOS) in cardiovascular regulation. However, no investigation has been done to study their individual role in modulating cardiovascular responses during static skeletal muscle contraction. In this study, we determined the effects of microdialyzing a specific nNOS antagonist into the rostral (RVLM) and caudal ventrolateral medulla (CVLM) on cardiovascular responses and glutamatergic/GABAergic neurotransmission during the exercise pressor reflex using rats. We hypothesized that the NO modulation of the exercise pressor reflex was largely influenced by specific nNOS activity within the ventrolateral medulla. Bilateral microdialysis of a selective nNOS antagonist, 1-(2-trifluoromethylphenyl)-imidazole (1.0 μM), for 30 or 60 min into the RVLM potentiated cardiovascular responses and glutamate release during a static muscle contraction. Levels of GABA within the RVLM were decreased. The cardiovascular responses and neurochemical changes to muscle contraction recovered following discontinuation of the drug. In contrast, bilateral application of the nNOS antagonist into CVLM attenuated cardiovascular responses and glutamate release during a static muscle contraction, but augmented GABA release. These results demonstrate that nNOS in the ventrolateral medulla plays an important role in modulating glutamatergic/GABAergic neurotransmission that regulates the exercise pressor reflex, and contributes to the sympathoexcitatory and sympathoinhibitory actions of NO within the RVLM and CVLM, respectively.

Cardiovascular responses during stimulation of hindlimb skeletal muscle nerves in anaesthetized rats.

1. Cardiovascular responses during static skeletal muscle contraction in anaesthetized rats appear to be contradictory. The present study attempted to explain such variations by stimulating different peripheral nerves supplying the hindlimb skeletal muscles using anaesthetized Sprague-Dawley rats. 2. Muscle contractions were evoked by a 30 s stimulation of the sciatic, tibial, peroneal nerves or the sciatic nerve with transected peroneal branch at threefold the motor threshold, 0.1 msec duration and 40 Hz frequency. 3. Contractions during stimulation of the tibial or the sciatic nerve with severed peroneal branch evoked similar increases in arterial pressure and heart rate. Following stimulation of the tibial nerve, blood pressure, heart rate and muscle tension increased by 23 +/- 3 mmHg, 31 +/- 5 b.p.m. and 789 +/- 34 g, respectively. For the sciatic nerve with transected peroneal branch, increases the respective increases were 27 +/- 5 mmHg, 32 +/- 6 b.p.m. and 802 +/- 43 g. In contrast, peroneal nerve stimulation produced depressor and bradycardic responses of -22 +/- 5 mmHg and -40 +/- 9 b.p.m., respectively. Interestingly, intact sciatic nerve stimulation elicited pressor, depressor or no responses (average being -10 +/- 8 mmHg), along with a consistent increase in heart rate of 24 +/- 7 b.p.m. 4. The results demonstrate that static muscle contraction following stimulation of the tibial or sciatic nerve with transected peroneal branch, elicits consistent increases in blood pressure and heart rate. Furthermore, stimulation of the peroneal nerve elicits a depressor response, while stimulation of the intact sciatic nerve evokes variable cardiovascular responses. Overall, anaesthetized rats can be excellent models to study the variable cardiovascular responses during activation of group III and/or group IV muscle afferents.

Effects of opioid receptor activation on cardiovascular responses and extracellular monoamines within the rostral ventrolateral medulla during static contraction of skeletal muscle.

During static muscle contraction, activation of opioid receptors alters the extracellular glutamate concentrations within the rostral ventrolateral medulla (RVLM). In addition, microdialysis of glutamate in the ventrolateral medulla (VLM) increases the release of norepinephrine (NE), dopamine (DA), and serotonin (5-HT). Therefore, we hypothesized that extracellular concentrations of these monoamines as well as cardiovascular responses during static skeletal muscle contraction would be modulated following administration of [D-Ala(2)]methionine enkephalinamide (DAME), an opioid receptor agonist, into the RVLM. Microdialysis of 100 microM DAME into the RVLM of 10 rats significantly (P<0.01) decreased extracellular levels (in pg/10 microl) of NE (from 3.3+/-0.3 to 1.9+/-0.3), DA (from 5.5+/-0.2 to 3.7+/-0.3), and 5-HT (from 6.1+/-0.8 to 3.6+/-0.2) during static exercise. After microdialysis of DAME, the exercise pressor reflex also significantly (P<0.01) decreased mean arterial pressure (MAP) by 13+/-3 mmHg and heart rate (HR) by 16+/-6 bpm, compared with control (MAP=22+/-4 mmHg and HR=31+/-7 bpm). Subsequently, after 30 min microdialysis of naloxone, an opioid receptor antagonist, muscle contraction increased the extracellular monoamine levels (in pg/10 microl, 3.8+/-0.3 NE; 5.2+/-0.3 DA; and 5.5+/-0.4 5-HT) similar to the control groups and evoked a reversal of cardiovascular responses. Similarly, 30 min of microdialyzing naloxone, added to the perfusing medium containing DAME, reversed the attenuating effects of DAME on monoamines, MAP, and HR during a muscle contraction. Furthermore, microdialysis of 100 microM naloxone alone for 30 min potentiated cardiovascular responses and monoamine levels during a muscle contraction. In summary, the present data demonstrates that microdialysis of DAME into RVLM attenuates the exercise pressor reflex mediated increases in MAP, HR and extracellular levels of biogenic monoamines. A subsequent microdialysis of naloxone reversed the effects suggesting that an opioidergic mechanism within RVLM modulates the exercise pressor reflex. Overall, the present study provides further insights into the opioidergic modulation of the exercise pressor reflex.

Simultaneous glutamate and γ-aminobutyric acid release within ventrolateral medulla during skeletal muscle contraction in intact and barodenervated rats

The purpose of this study was to determine if baroreflex modulates cardiovascular responses and neurotransmitter release within rostral (RVLM) and caudal (CVLM) ventrolateral medulla during static contraction of skeletal muscle using anesthetized rats. We evoked cardiovascular responses by a static muscle contraction and measured simultaneous release of glutamate and γ-aminobutyric acid (GABA) in both the RVLM and CVLM using microdialysis probes, two inserted bilaterally into the RVLM and two into the CVLM. In intact anesthetized rats, a muscle contraction increased release of glutamate concomitantly in both the RVLM and CVLM along with significant increases in heart rate and arterial blood pressure. In contrast, concentrations of GABA increased within the RVLM, but decreased significantly within the CVLM during the pressor response. These changes were due to contraction-evoked activation of muscle afferents since tibial nerve stimulation following muscle paralysis failed to evoke glutamate, GABA, or any cardiovascular changes. On the other hand, static muscle contractions in baroreceptor denervated rats augmented the increases in heart rate and blood pressure. Furthermore, muscle contraction significantly enhanced the release of glutamate in the RVLM but attenuated its release in the CVLM. In addition, concentrations of GABA within the RVLM were attenuated following a muscle contraction in denervated rats without any changes in GABA within the CVLM. These results demonstrate that the baroreceptors influence cardiovascular responses to static muscle contraction associated with dynamic changes in glutamate and GABA release within the RVLM and CVLM.

Neuronal application of capsaicin modulates somatic pressor reflexes.

Static contraction of skeletal muscle elicits a reflex increase in cardiovascular function. Likewise, noxious stimuli activate somatic nociceptors eliciting a reflex increase in cardiovascular function. On the basis of recent work involving spinothalamic cells in the dorsal horn, we hypothesized that the dorsal horn cells involved in the aforementioned reflexes would be sensitized by applying capsaicin (Cap) to a peripheral nerve. If correct, then Cap would enhance the cardiovascular increases that occur when these reflexes are evoked. Cats were anesthetized, and the popliteal fossa was exposed. Static contraction was induced by electrical stimulation of the tibial nerve at an intensity that did not directly activate small-diameter muscle afferent fibers, whereas nociceptors were stimulated by high-intensity stimulation (after muscle paralysis) of either the saphenous nerve (cutaneous nociceptors) or a muscular branch of the tibial nerve (muscle nociceptors). The reflex cardiovascular responses to these perturbations (contraction or nociceptor stimulation) were determined before and after direct application of Cap (3%) onto the common peroneal nerve, using a separate group of cats for each reflex. Compared with control, application of Cap attenuated the peak change in mean arterial pressure (MAP) evoked by static contraction (DeltaMAP in mmHg: 38 +/- 10 before and 24 +/- 8 after ipsilateral Cap; 47 +/- 10 before and 33 +/- 10 after contralateral Cap). On the other hand, Cap increased the peak change in MAP evoked by stimulation of the saphenous nerve from 57 +/- 8 to 77 +/- 9 mmHg, as well as the peak change in MAP elicited by activation of muscle nociceptors (36 +/- 9 vs. 56 +/- 14 mmHg). These results show that the reflex cardiovascular increases evoked by static muscle contraction and noxious input are differentially affected by Cap application to the common peroneal nerve. We hypothesize that a Cap-induced alteration in dorsal horn processing is the locus for this divergent effect on these reflexes.

Reflex responses in plasma catecholamines caused by static contraction of skeletal muscle.

To examine a hypothesis of whether static muscle contraction produces a release of catecholamines from the adrenal medulla via reflex stimulation of preganglionic adrenal sympathetic nerve activity induced by receptors in the contracting muscle, we compared the reflex responses in a concentration of epinephrine (Ep) and norepinephrine (NEp) in arterial plasma during static contraction and during a mechanical stretch of the hindlimb triceps surae muscle in anesthetized cats. Static contraction was evoked by electrically stimulating the peripheral ends of the cut L(7) and S(1) ventral roots at 20 or 40 Hz. Mean arterial pressure (MAP) and heart rate (HR) increased 23 +/- 3.1 mmHg and 19 +/- 4.3 beats/min during static contraction. Ep in arterial plasma increased 0.18 +/- 0.072 ng/ml over the control of 0.14 +/- 0.051 ng/ml within 1 min from the onset of static contraction, and NEp increased 0.47 +/- 0.087 ng/ml over the control of 0.71 +/- 0.108 ng/ml. Following a neuromuscular blockade, although the same ventral root stimulation failed to produce the cardiovascular and plasma catecholamine responses, the mechanical stretch of the muscle increased MAP, HR, and plasma Ep, but not plasma NEp. With bilateral adrenalectomy, the baseline Ep became negligible (0.012 +/- 0.001 ng/ml) and the baseline NEp was lowered to 0.52 +/- 0.109 ng/ml. Neither static contraction nor mechanical stretch produced significant responses in plasma Ep and NEp following the adrenalectomy. These results suggest that static muscle contraction augments preganglionic adrenal sympathetic nerve activity, which in turn secretes epinephrine from the adrenal medulla into plasma. A muscle mechanoreflex from the contracting muscle may play a role in stimulation of the adrenal sympathetic nerve activity.

Dorsal horn administration of l-arginine accentuates the pressor response evoked by activation of muscle mechanoreceptors

Recent work from our laboratory suggests that nitric oxide production in the dorsal horn has a modulatory influence on the pressor reflex evoked by static contraction of skeletal muscle. In this study, we tested the hypothesis that nitric oxide production in the dorsal horn is involved in producing the pressor reflex elicited by activation of skeletal muscle mechanoreceptors. Cats were anesthetized with α-chloralose (80 mg/kg) and urethane (100 mg/kg) and a laminectomy was performed. With the exception of the L 7 dorsal root, the dorsal and ventral roots from L 5 to S 2 were sectioned on one side. Muscle mechanoreceptors were activated by manually stretching the ipsilateral triceps surae muscle 1.5 cm. To block nitric oxide synthase, a 50 mM solution of nt-nitro- l-argenine methyl ester ( l-NAME) (a dose that altered the pressor reflex to static contraction) was microdialyzed into the dorsal horn at L 6 and S 1. Dialysis of l-NAME failed to attenuate the peak change in mean arterial pressure evoked by muscle stretch (45±6 mmHg before and 44±9 mmHg after 2 h of l-NAME dialysis). On the other hand, 2 h of l-arginine dialysis (50 mM) increased the peak pressor response to muscle stretch from 43±3 to 57±5 mmHg. These data suggest that administration of l-arginine enhances the excitability of dorsal horn cells receiving input from muscle mechanoreceptors, thus increasing the pressor response evoked by activation of this type of muscle afferent neuron.

Spinal modulation of the muscle pressor reflex by nitric oxide and acetylcholine

Static contraction of skeletal muscle activates the sympathetic nervous system, which in turn increases cardiovascular function. These changes are mediated, in part, by a reflex arising from the contracting muscle. This reflex is termed the exercise pressor reflex or, more simply, the muscle pressor reflex (MPR). Over the past few years, studies have been performed investigating the sensory processing that occurs in the dorsal horn of the spinal cord as it pertains to the MPR. Several putative neurotransmitters and receptors have been implicated in mediating the MPR at the level of the dorsal horn. In addition, several receptor systems have been shown to modulate the MPR at the dorsal horn. We have recently performed studies investigating the potential modulatory role of dorsal horn nitric oxide (NO) and acetylcholine (ACH) on the MPR. Along these lines, our experiments suggest that NO enhances the excitability of dorsal horn cells receiving input from muscle afferent neurons, while ACH decreases the MPR when its concentration in the dorsal horn is elevated. The purpose of this manuscript is to review recently published findings from our laboratory and apply this information in an effort to better understand the integration of sensory input that occurs in the dorsal horn as it pertains to cardiovascular regulation. This review is also designed to stimulate questions as to how these two neurochemicals exert their actions and whether or not they represent or can represent important physiological mechanisms involved in regulating the dorsal horn integration of the MPR.

Pressor reflex evoked by static muscle contraction: role of nitric oxide in the dorsal horn.

In this study, we tested the hypothesis that nitric oxide (NO) production in the dorsal horn is involved in producing the pressor reflex elicited by static contraction of skeletal muscle. Cats were anesthetized with alpha-chloralose (80 mg/kg) and urethane (100 mg/kg), and a laminectomy was performed. With the exception of the L7 dorsal root, the dorsal and ventral roots from L5 to S2 were sectioned on one side and static contraction of the ipsilateral triceps surae muscle was evoked by electrically stimulating the peripheral ends of the L7 and S1 ventral roots. Dialysis of the NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME; 50 mmol/l syringe concentration, based upon dose-response data) into the dorsal horn at L6 and S1 failed to attenuate the peak change in mean arterial pressure (MAP) evoked by static contraction (DeltaMAP in mmHg: 57 +/- 5 before and 50 +/- 6 after 2 h of L-NAME). However, this dialysis of L-NAME reduced the magnitude of the initial pressor response as the MAP at 10 s of the contraction fell from 27 +/- 4 to 17 +/- 4 mmHg. On the other hand, 2 h of L-arginine dialysis (50 mmol/l) shifted the curve representing the time course of the pressor response upward and increased the peak pressor response to static contraction from 51 +/- 9 to 68 +/- 9 mmHg. A 2-h dialysis of D-NAME (50 mmol/l), the inactive enantiomer of L-NAME, had no effect on the time course or the peak pressor response (DeltaMAP in mmHg: 78 +/- 12 before and 72 +/- 15 after). These data suggest that NO production in the dorsal horn has a modulatory influence on the pressor reflex evoked by static contraction of skeletal muscle and that increasing the level of NO in the dorsal horn enhances the excitability of dorsal horn cells to muscle afferent input.

Segmental effect of NMDA block in the dorsal horn on the pressor reflex

The purpose of this study was to examine the influence of NMDA receptor blockade in the dorsal horn of adjacent spinal segments as it pertains to the pressor reflex evoked by static contraction and stretch of skeletal muscle. In this preparation, cats were anesthetized and the afferent fibers mediating the pressor reflex entered the spinal cord via the L 7 dorsal root. Blockade of dorsal horn NMDA receptors at L 6 and L 7 attenuated the pressor reflex evoked by static contraction and muscle stretch. However, NMDA block in the L 6 dorsal horn alone failed to alter the peak increase in MAP produced by static contraction and muscle stretch, but the initial pressor response evoked by static contraction was attenuated. These data support the hypothesis that the pressor reflex is partially mediated by activation of NMDA receptors in the dorsal horn, and this occurs at multiple spinal segments. Further, these data suggest that activation of NMDA receptors plays an important role in initiating the rise in arterial pressure produced by static contraction of skeletal muscle.

Segmental effect of spinal NK-1 receptor blockade on the pressor reflex.

The physiological effects of substance P (SP) are mediated via activation of neurokinin-1 (NK-1) receptors. The purpose of this study was to test the hypothesis that blockade of NK-1 receptors in the dorsal horn, both at the site of entry for the primary afferent neurons and adjacent spinal segments, attenuates the pressor reflex evoked by static contraction and stretch of skeletal muscle. Cats were anesthetized with alpha-chloralose and urethan, and a laminectomy was performed. With the exception of the L7 dorsal root, the dorsal and ventral roots from L5 to S2 were sectioned on one side of the spinal cord. Thus the primary afferent fibers mediating the pressor reflex enter the spinal cord via the L7 dorsal root in these experiments. Based on dose-response data, dialysis of the NK-1 receptor antagonist CP-96,345 (5 mM for 2 h) into the L7 dorsal horn ipsilateral to the contracting muscle attenuated the pressor response to static contraction (75 +/- 15 vs. 46 +/- 7 mmHg; n = 5 cats) but not muscle stretch (60 +/- 12 vs. 50 +/- 8 mmHg). Administration of the inactive enantiomer of CP-96,345, CP-96,344 (5 mM for 2 h), into the L7 dorsal horn failed to alter the cardiovascular changes elicited by contraction (45 +/- 7 vs. 43 +/- 6 mmHg) and stretch (31 +/- 8 vs. 32 +/- 11). Dialysis of 5 mM CP-96, 345 into the dorsal horn at the L6 and S1 segments for 2 h decreased the peak pressor response to static contraction (58 +/- 9 vs. 31 +/- 6 mmHg; n = 7) and muscle stretch (61 +/- 6 vs. 44 +/- 8 mmHg). These data suggest that the activation of NK-1 receptors, both at the site of entry and in regions outside of the entry site for afferent neurons, is involved in the spinal processing that produces the pressor reflex evoked by static contraction of skeletal muscle.