Jay P. Farber

Maternal nicotine administration and fetal brain stem damage: A rat model with implications for sudden infant death syndrome

Several recent studies have established maternal smoking as a significant risk factor for sudden infant death syndrome (SIDS). To test our hypothesis that nicotine, a component of cigarette smoke, may exert an injurious effect on the fetal brain stem, we administered nicotine in drinking water to Sprague-Dawley rats prior to and throughout the first 20 days of gestation. The nicotine dose and schedule of consumption in this experiment paralleled that of human usage. Fetuses from nicotine-treated mothers had significantly more dead cells in a standard section of the medulla than controls, but little difference was observed in the postnatal respiratory responses of treated and control animals to the inhalation of various gas mixtures. The birth weights of nicotine-exposed fetuses were significantly less than those of controls. Although the degree of fetal brain stem injury produced by nicotine in this experiment is small, it is our concern that maternal smoking might cause a more severe lesion in the human fetus and thereby increase the risk of SIDS by contributing to aberrant postnatal respiratory responses to noxious stimuli.

Sensory fibers containing vanilloid receptor-1 (VR-1) mediate spinal cord stimulation-induced vasodilation

BACKGROUND AND AIMS Spinal cord stimulation (SCS) is used to improve peripheral blood flow in selected populations of patients with ischemia of the extremities. Previous studies show that antidromic activation of sensory fibers is an important mechanism that contributes to SCS-induced vasodilation. However, the characteristics of sensory fibers involved in vasodilation are not fully known. This study investigated the contribution of vanilloid receptor type 1 (VR-1) containing fibers to SCS-induced vasodilation. METHODS A unipolar ball electrode was placed on the left dorsal column at the lumbar 2-3 spinal cord segments (L2-L3) in sodium pentobarbital anesthetized, paralyzed and ventilated rats. Cutaneous blood flows from both ipsilateral (left) and contralateral (right) hind foot pads were recorded with laser Doppler flow perfusion monitors. SCS (50 Hz; 0.2 ms) was applied through the ball electrode at 30%, 60%, 90% and 300% of motor threshold (MT). Resiniferatoxin (RTX), an ultra potent analog of capsaicin and VR-1 receptor agonist, was used to suppress the activities of VR-1 containing sensory fibers. RESULTS SCS at 30%, 60%, 90% and also at 300% of MT significantly increased cutaneous blood flow in the ipsilateral foot pad compared to that in the contralateral side. RTX (2 microg/kg, i.v.) significantly attenuated SCS-induced vasodilation of the ipsilateral side (P<0.05, n=7) compared with responses prior to RTX administration. A pledget of cotton soaked with RTX (2 microg/ml) placed on L2-L3 spinal cord significantly decreased SCS-induced vasodilation of the ipsilateral side at 30%, 60%, 90% and 300% of MT (P<0.05, n=7) compared with responses prior to RTX administration. Additionally, topical application of a pledget of cotton soaked with RTX (2 microg/ml) on the sciatic nerve at the middle level of the thigh or on the tibial nerve at the lower level of the lower hindlimb also decreased SCS-induced vasodilation (n=5). CONCLUSION SCS-induced vasodilation is predominantly mediated via VR-1 containing sensory fibers.

Effects of spinal cord stimulation with “standard clinical” and higher frequencies on peripheral blood flow in rats

BACKGROUND It is unclear whether spinal cord stimulation (SCS) at higher frequencies induces further increases in vasodilation and enhances clinical efficacy. OBJECTIVES This study investigated effects of SCS at both a normal frequency (as used clinically) and two higher frequencies on peripheral vasodilation. METHODS A unipolar ball electrode was placed on the left dorsal column at the lumbar 2-3 spinal cord segments (L2-L3) in sodium pentobarbital anesthetized, paralyzed, and artificially ventilated rats. Cutaneous blood flow recordings from both ipsilateral (left) and contralateral (right) hind foot pads were measured with laser Doppler flow perfusion monitors. SCS at frequencies of 50, 200, or 500 Hz was applied at 30%, 60%, and 90% of motor threshold (MT) using standard square waves. Resiniferatoxin (RTX: an ultrapotent analog of capsaicin) and a calcitonin gene-related peptide (CGRP) receptor blocker (CGRP(8-37)) was also used to elucidate mechanisms of SCS vasodilation at these higher frequencies. RESULTS SCS applied with the three frequencies produced similar MT (n=22). SCS at 500 Hz significantly increased cutaneous blood flow and decreased vascular resistance compared to changes induced by frequencies of 50 and 200 Hz (P<0.05, n=8). RTX (2 microg/kg, i.v.) as well as CGRP(8-37) (2.37 mg/kg, i.v.) significantly reduced SCS-induced vasodilation at 500 Hz (P<0.05, n=6) as compared to responses prior to administrations of these drugs. CONCLUSION SCS at 500 Hz significantly increased SCS-induced vasodilation without influencing MT. Furthermore, effects of SCS at 500 Hz are mediated via activation of TRPV1-containing fibers and a release of CGRP.

Extracellular signal-regulated kinase (ERK) and protein kinase B (AKT) pathways involved in spinal cord stimulation (SCS)-induced vasodilation

BACKGROUND AND AIMS SCS is used to improve peripheral circulation in selected patients with ischemia of the extremities. However the mechanisms are not fully understood. The present study investigated whether blockade of ERK and AKT activation modulated SCS-induced vasodilation. METHODS A unipolar ball electrode was placed on the left dorsal column at the lumbar 2-3 spinal segments in rats. Cutaneous blood flows from left and right hind foot pads were recorded with laser Doppler flow perfusion monitors. SCS was applied through a ball electrode at 60% or 90% of MT. U0126, an inhibitor of ERK kinase, or LY294002, an inhibitor of PI3K upstream of AKT, was applied to the lumbar 3-5 spinal segments (n=7, each group). RESULTS U0126 (100 nM, 5 microM and 250 microM) significantly attenuated SCS-induced vasodilation at 60% (100 nM: P<0.05; 5 microM and 250 microM: P<0.01, respectively) and 90% of MT (100 nM and 5 microM: P<0.05; 250 microM: P<0.01, respectively). LY294002 at 100 microM also attenuated SCS-induced vasodilation at 60% and 90% of MT (P<0.05). CONCLUSIONS These data suggest that ERK and AKT pathways are involved in SCS-induced vasodilation.

Comparison of Burst and Tonic Spinal Cord Stimulation on Spinal Neural Processing in an Animal Model

Spinal cord stimulation (SCS) using bursts of pulses suppressed neuropathic pain as well or better than tonic stimulation and limited the incidences of parasthesias. The present translational study explored possible differences in mechanisms of burst and tonic SCS on nociceptive spinal networks and/or the gracile nucleus supraspinal relay.

Pulmonary Petechiae: Ventilatory-Circulatory Interactions

Summary: Pulmonary petechial hemorrhages are typically observed in sudden infant death syndrome (SIDS), but their cause has not been determined. Some data suggest that SIDS infants die with an obstructed airway. Inspiratory efforts against the closed airway can decrease the efficiency of the cardiac pump by increasing the afterload on the left ventricle and/or by decreasing left ventricular compliance. If these effects are augmented by hypoxia-induced decreases in cardiac contractility as occlusion is maintained, the resultant failure of the left ventricle could produce a large increase in end-diastolic pressure. Increases in left ventricular end-diastolic pressure will then increase pressures in the pulmonary circuit, where rupture of small blood vessels gives rise to petechial hemorrhages.To evaluate this postulated mechanism of petechiae formation, lightly anesthetized adult rabbits were sacrificed using three 1-min end-expiratory airway occlusions plus a fourth occlusion maintained until death; systemic arterial, intratracheal, and pulmonary wedge pressures (the latter approximating left atrial pressure) were measured during inspiratory efforts and expiratory intervals. Those animals showing minimal evidence of left heart failure, as evidenced by a relatively small difference between wedge and intratracheal pressures during obstructed breathing, showed few pulmonary petechiae. When the above pressure difference was large (greater than 25 mmHg during inspiratory efforts and greater than 10 mmHg during expiratory intervals) numerous pulmonary petechiae (usually n > 40) could be observed. Progressive increases in pulmonary wedge pressure and bradycardia during occlusion suggested that direct and/or reflex hypoxic depression of the myocardium contributed to the observed effects. The occlusions produced little change in systemic arterial pressure. These results are consistent with the possibility that mechanical interactions between the heart and lung pumps, along with hypoxia-induced decreases in cardiac contractility accompanying airway obstruction, account for intrathoracic petechiae in SIDS.

Roles of dorsal column pathway and transient receptor potential vanilloid type 1 in augmentation of cerebral blood flow by upper cervical spinal cord stimulation in rats.

Clinical and basic studies have indicated that upper cervical spinal cord stimulation (cSCS) significantly increases cerebral blood flow (CBF), but the mechanisms are incompletely understood. This investigation was conducted to differentiate between stimulation of dorsal column fibers and upper cervical spinal cord cell bodies in cSCS-induced increases in CBF and decreases in cerebrovascular resistance (CVR). cSCS (50 Hz, 0.2 ms, 1 min) was applied on the left C1-C2 dorsal column of pentobarbital anesthetized, ventilated and paralyzed male rats. Laser Doppler flowmetry probes were placed bilaterally over the parietal cortex, and arterial pressure was monitored. cSCS at 30%, 60%, and 90% of motor threshold (MT) produced vasodilation bilaterally in cerebral cortices. Subsequently, cSCS was applied at 90% MT, and ipsilateral responses were recorded. Ibotenic acid (0.3 mg/ml, 0.1 ml) placed on dorsal surface of C1-C2 (n=7) to suppress cell body activity, did not affect cSCS-induced %DeltaCBF (42.5+/-8.1% vs. 36.8+/-7.1%, P>0.05) and %DeltaCVR (-19.4+/-4.2% vs. -15.2+/-5.6%, P>0.05). However, bilateral transection of the dorsal column at rostral C1 (n=8) abolished cSCS-induced changes in CBF and CVR. Also, rostral C1 transection (n=7) abolished cSCS-induced changes in CBF and CVR. Resinferatoxin (RTX), an ultrapotent transient receptor potential vanilloid type 1 (TRPV1) agonist, was used to inactivate TRPV1 containing nerve fibers/cell bodies. RTX (2 microg/ml, 0.1 ml) placed on the C1-C2 spinal cord (n=7) did not affect cSCS-induced %DeltaCBF (60.2+/-8.1% vs. 46.3+/-7.7%, P>0.05) and %DeltaCVR (-25.5+/-3.5% vs. -21.4+/-8.9%, P>0.05). However, i.v. RTX (2 microg/kg, n=9) decreased cSCS-induced %DeltaCBF from 65.0+/-9.5% to 27.4+/-7.2% (P<0.05) and %DeltaCVR from -28.0+/-7.6% to -14.8+/-4.2% (P<0.05). These results indicated that cSCS-increases in CBF and decreases in CVR occurred via rostral spinal dorsal column fibers and did not depend upon C1-C2 cell bodies. Also, our results suggested that cerebral but not spinal TRPV1 was involved in cSCS-induced cerebral vasodilation.

Convergent pathways for cardiac- and esophageal-somatic motor reflexes in rats

Chest pain of esophageal and cardiac origin is often difficult to distinguish due to similar sensations and localization. We have shown that spasm-like contractions of the spinotrapezius muscles evoked by noxious cardiac stimulation could potentially sensitize muscle afferent fibers and produce angina-like referred pain. In this study, we proposed that a similar type of spinotrapezius contraction evoked by esophageal stimulation could produce nociceptive responses with similar quality and localization as evoked by cardiac stimulation. An objective of this study was to show convergence of pathways to the spinotrapezius muscles by measuring electromyographic (EMG) activity between the cardiac- and esophageal-motor reflexes. We also investigated afferent pathways of esophageal-motor reflexes by disrupting or activating the left sympathetic chain and vagus nerves; these pathways form the afferent limbs of the cardiac-motor reflexes. Results showed that more than 95% of animals responding to noxious cardiac stimulation also responded to esophageal distension. Transection of the left sympathetic chain to reduce upper thoracic visceral afferent innervation significantly decreased cardiac-evoked EMG activity or total motor unit potentials (t-MUP). In contrast, however, the transection did not significantly decrease t-MUP evoked by esophageal distension. Bilateral vagotomy and vagal afferent stimulation increased and decreased the cardiac-evoked t-MUP, respectively. However, the same vagal manipulations did not influence t-MUP evoked by esophageal distension. This study demonstrated that the spinotrapezius muscle could be activated by noxious stimulation of two different visceral organs. The spinotrapezius muscle contractions evoked by esophageal distension are produced in part by activation of esophageal afferent fibers found in upper thoracic sympathetic nerves, but not by activation of the vagus nerves.

Short latency excitation of upper cervical respiratory neurons by vagal stimulation in the rat

Extracellular recordings were made from 29 respiration-phased neurons in the upper cervical spinal cord (C1-C3) in nine anesthetized rats while ipsilateral and contralateral vagi were stimulated via platinum hook electrodes. Neuronal responses to vagal stimulation were recorded using peristimulus histograms. An accumulation of spikes with an average latency of 4.0 +/- 0.8 (S.D.) ms occurred in 11 cells after ipsilateral stimulation. These results indicate that there are fibers in the vagus which oligosynaptically excite respiratory neurons in the upper cervical spinal cord.

Intraesophageal chemicals enhance responsiveness of upper thoracic spinal neurons to mechanical stimulation of esophagus in rats

Esophageal hypersensitivity is one of the most common causes of noncardiac chest pain in patients. In this study, we investigated whether exposure of the esophagus to acid and other chemical irritants affected activity of thoracic spinal neurons responding to esophageal distension (ED) in rats. Extracellular potentials of single thoracic (T3) spinal neurons were recorded in pentobarbital sodium-anesthetized, -paralyzed, and -ventilated male rats. ED (0.2 or 0.4 ml, 20 s) was produced by water inflation of a latex balloon placed orally into the middle thoracic region of the esophagus. The chemicals were administered via a tube that was passed through the stomach and placed in the thoracic esophagus. To irritate the esophagus, 0.2 ml of HCl (0.01 N), bradykinin (10 microg/ml), or capsaicin (10 microg/ml) were injected for 1-2 min. Only neurons excited by ED were included in this study. Results showed that intraesophageal instillation of HCl, bradykinin, and capsaicin increased activity in 3/20 (15%), 7/25 (28%), and 9/20 (45%) neurons but enhanced excitatory responses to ED in 9/17 (53%), 8/15 (53%), and 7/11 (64%) of the remaining spinal neurons, respectively. Furthermore, intraesophageal chemicals were more likely to enhance the responsiveness of low-threshold neurons than high-threshold neurons to the esophageal mechanical stimulus. Normal saline (pH 7.4, 0.2 ml) or vehicle instilled in the esophagus did not significantly affect activity or ED responses of neurons. We conclude that enhanced responses of thoracic spinal neurons to ED by the chemically challenged esophagus may provide a possible pathophysiological basis for visceral hypersensitivity in patients with gastroesophageal reflux and/or esophagitis.