[Show abstract][Hide abstract] ABSTRACT: Delta opioid receptor (DOR) activation protects the adult mammalian brain during oxygen-glucose deprivation (OGD), but it is not known whether neonatal spinal motor circuits are also protected. Also, it is unclear whether the timing of spinal DOR activation relative to spinal OGD is important for neuroprotection. Thus, a split-bath in vitro neonatal rat brainstem/spinal cord preparation was used to record spontaneous respiratory motor output from cervical (C4-C5) and thoracic (T5-T6) ventral spinal roots while exposing only the spinal cord to OGD solution (0 mM glucose, bubbled with 95% N(2)/5% CO(2)) or DOR agonist drugs (DADLE, DPDPE). Spinal OGD solution application caused respiratory motor output frequency and amplitude to decrease until all activity was abolished (i.e. end-point times) after 25.9±1.4 min (cervical) and 25.2±1.4 min (thoracic). Spinal DOR activation via DPDPE (1.0 μM) prior-to and during spinal OGD increased cervical and thoracic end-point times to 35-48 min. Spinal DADLE or DPDPE (1.0 μM) application 15 min following spinal OGD onset increased cervical and thoracic end-point times to 36-45 min. Brief spinal DPDPE (1.0 μM) application for 10 min at 25 min before spinal OGD onset increased cervical and thoracic end-point times to 41-46 min. Overall, the selective DOR agonist, DPDPE, was more effective at increasing end-point times than DADLE. Naltrindole (DOR antagonist; 10 μM) pretreatment blocked DPDPE-dependent increase in end-point times, suggesting that DOR activation was required. Spinal naloxone (1.0 μM) application before and during spinal OGD also increased end-point times to 31-33 min, but end-point times were not altered by Mu opioid receptor (MOR) activation or DOR activation/MOR blockade, indicating that there are complex interactions between OGD and opioid signaling pathways. These data suggest DOR activation before, during, and after spinal OGD protects central motor networks and may provide neuroprotection during unpredictable perinatal ischemic events.
[Show abstract][Hide abstract] ABSTRACT: Intermittent hypoxia induces 5-HT-dependent, pattern-sensitive long-term facilitation (LTF) of spinal respiratory motor output. We used a split-bath in vitro neonatal rat brainstem-spinal cord preparation to test whether: 1) intermittent spinal 5-HT exposure (without hypoxia) is sufficient to induce LTF in phrenic and intercostal inspiratory motor outputs; 2) LTF magnitude is greater in intercostal versus phrenic activity; and 3) phrenic and intercostal motor output exhibits differential pattern sensitivity to 5-HT application. With a barrier at spinal segment C1, 5-HT (5 muM) was applied episodically (3 min 5-HT, 5 min wash, x3) to the spinal cord (C2-L1) while recording inspiratory bursts in cervical (C4 or C5) and thoracic (T5 or T6) ventral roots. Episodic 5-HT application increased cervical and thoracic burst amplitudes to 136+/-22% and 150+/-22% of baseline, respectively, at 120 min post-drug (P<0.01). Continuous 5-HT application (5 muM, 9 min) had no effect on cervical burst amplitude at 120 min post-drug, but increased thoracic burst amplitude to 142+/-11% of baseline at 120 min post-drug (P<0.001). Methysergide pretreatment abolished both cervical and thoracic 5-HT-induced LTF. Quantitative reverse transcriptase-polymerase chain reaction and immunocytochemistry revealed that 5-HT(2A) and 5-HT(7) receptor subtypes (receptors known to influence LTF expression in adult rats) are expressed in ventral cervical and thoracic spinal cord with no differences in expression levels due to spinal segment or age. Thus, 5-HT is sufficient to induce spinal LTF in neonatal rats and differences in pattern sensitivity suggest heterogeneity in underlying mechanisms.
[Show abstract][Hide abstract] ABSTRACT: Our goal was to determine whether time-dependent changes in respiratory motor output in vitro could be minimized by altering bath solution composition. Adult turtle brainstems were bathed in standard solution, nutrient-rich Dulbecco's Eagle media (100 or 25% concentration), or standard solution with phenylbiguanide (PBG, 5-HT3 agonist which increases respiratory drive). Except for a 63% frequency increase in PBG solution, hypoglossal bursts were unaltered within 100 min of observation. Respiratory activity was abolished within 7 h in 100% Dulbecco's compared with a mean of 24-31 h in other test solutions. At 12 h, burst frequency decreased faster in standard solution and 25% Dulbecco's (-0.28+/-0.07 and -0.13+/-0.05 bursts/h, respectively) compared with PBG solution (-0.09+/-0.04 bursts/h); amplitude declined at approximately 2%/h in all solutions. The tendency for episodic discharge decreased gradually in standard solution, but was eliminated in 25% Dulbecco's and PBG solution. Certain bath solutions may minimize time-dependent frequency reductions but may also cause breathing pattern changes.
[Show abstract][Hide abstract] ABSTRACT: Brain stem preparations from adult turtles were used to determine how bath-applied serotonin (5-HT) alters respiration-related hypoglossal activity in a mature vertebrate. 5-HT (5-20 microM) reversibly decreased integrated burst amplitude by approximately 45% (P < 0.05); burst frequency decreased in a dose-dependent manner with 20 microM abolishing bursts in 9 of 13 preparations (P < 0.05). These 5-HT-dependent effects were mimicked by application of a 5-HT(1A) agonist, but not a 5-HT(1B) agonist, and were abolished by the broad-spectrum 5-HT antagonist, methiothepin. During 5-HT (20 microM) washout, frequency rebounded to levels above the original baseline for 40 min (P < 0.05) and remained above baseline for 2 h. A 5-HT(3) antagonist (tropesitron) blocked the post-5-HT rebound and persistent frequency increase. A 5-HT(3) agonist (phenylbiguanide) increased frequency during and after bath application (P < 0.05). When phenylbiguanide was applied to the brain stem of brain stem/spinal cord preparations, there was a persistent frequency increase (P < 0.05), but neither spinal-expiratory nor -inspiratory burst amplitude were altered. The 5-HT(3) receptor-dependent persistent frequency increase represents a unique model of plasticity in vertebrate rhythm generation.
Journal of Applied Physiology 12/2001; 91(6):2703-12. · 3.06 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: An in vitro brainstem-spinal cord preparation from adult turtles was used to test the hypothesis that descending synaptic inputs to multifunctional spinal motoneurons (i.e., involved in respiration and locomotion) express activity-dependent depression or potentiation. The tissue was placed in a chamber that allowed for separate superfusion of the brainstem, spinal segments C(2)-C(4), and C(5)-D(1). Action potential conduction between the brainstem and spinal segments C(5)-D(1) was blocked by superfusing C(2)-C(4) with Na(+)-free solution. With C(5)-D(1) at [K(+)] = 10 mM, electrical stimulation at C(5) every 2 min evoked potentials in intact pectoralis (expiratory, inward rotation of shoulder) and serratus (inspiratory, outward rotation of shoulder) nerves that were stable for at least 2 hr. Application of conditioning stimulation (900 pulses at 1 or 10 Hz) at C(5) decreased pectoralis evoked potential amplitudes by approximately 40% initially and by 20% after 90 min; serratus evoked potentials were unaltered. Conditioning stimulation (100 Hz, 900 pulses) transiently depressed pectoralis evoked potential amplitude by <20% but produced a delayed 72% increase in serratus evoked potential amplitude after approximately 80 min. Conditioning stimulation (10 Hz) at C(5) also reduced the amplitude of sensory afferent evoked potentials in pectoralis produced by stimulating ipsilateral dorsal roots at C(8). Thus, long-lasting changes in descending synaptic inputs to multifunctional spinal motoneurons were frequency-dependent and heterosynaptic. We hypothesize that activity-dependent plasticity may modulate descending synaptic drive to spinal motoneurons involved in both respiration and locomotion.
The Journal of Neuroscience : The Official Journal of the Society for Neuroscience 05/2000; 20(9):3487-95. · 6.34 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A split-bath in vitro brainstem-spinal cord preparation from adult turtles (Pseudemys scripta) was used to test the hypotheses that: (1) bulbospinal respiratory synaptic transmission is mediated, at least in part, by N-methyl-D-aspartate (NMDA) glutamatergic receptors, and (2) this transmission is suppressed by low spinal pH (induced by hypercapnea). Recordings from intact pectoralis (expiratory) and serratus (inspiratory) nerves showed that the in vitro turtle brainstem-spinal cord preparation produces phasic expiratory and inspiratory bursts of activity similar to that produced by intact turtles. Bath application of AP-5 [(+/-)-2-amino-5-phosphonopentanoic acid, a noncompetitive NMDA receptor antagonist] to the spinal cord reversibly reduced pectoralis and serratus burst amplitude. In contrast, lowering the pH from 8.04 to 6.94 did not alter burst amplitude in either pectoralis or serratus nerves. These data suggest that spinal NMDA receptors: (1) mediate part of the bulbospinal respiratory synaptic transmission, and (2) are pH/P(CO2)-insensitive. The pH/P(CO2)-insensitivity of NMDA-dependent bulbospinal respiratory synaptic transmission may represent an important adaptation in turtles.
[Show abstract][Hide abstract] ABSTRACT: An in vitro brain stem preparation from adult turtles was used to determine effects of dopamine (DA) and norepinephrine (NE) on the pattern of respiratory motor output recorded from hypoglossal nerve roots (XII). Bath-applied DA (10-200 microM) increased the frequency of respiratory bursts (peaks) from 0.9 +/- 0.2 to 2.4 +/- 0.3 (SE) peaks/min, resulting in a 99 +/- 9% increase in neural minute activity. R[+]-SCH-23390 (10 microM, D1 antagonist) and eticlopride (20 microM, D2 antagonist) attenuated the DA-mediated increase in peak frequency by 52 and 59%, respectively. On the other hand, the DA-receptor agonists apomorphine (D1, D2), quinelorane (D2), and SKF-38393 (D1) had no effect on peak frequency. Prazosin, an alpha1-adrenergic antagonist (250 nM) abolished the DA-mediated frequency increase. Although NE (10-200 microM) and phenylephrine (10-200 microM, alpha1-adrenergic agonist) increased peak frequency from 0.5 +/- 0.1 to 1.2 +/- 0.3 peaks/min and from 0.6 +/- 0.1 to 1. 0 +/- 0.2 peaks/min, respectively, these effects were not as large as that with DA alone. The data suggest that both dopaminergic and adrenergic receptor activation in the brain stem increase respiratory frequency in turtles, but the DA receptor-mediated increase is dependent on coactivation of alpha1-adrenergic receptors.
Journal of Applied Physiology 08/1998; 85(1):105-14. · 3.06 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: An in vitro brain stem preparation from adult turtles (Chrysemys picta) was used to examine the effects of anoxia and increased temperature and pH/CO2 on respiration-related motor output. At pH approximately 7.45, hypoglossal (XII) nerve roots produced patterns of rhythmic bursts (peaks) of discharge (O.74 +/- 0.07 peaks/min 10.0 +/- 0.6 s duration) that were quantitatively similar to literature reports of respiratory activity in conscious, vagotomized turtles. Respiratory discharge was stable for 6 h at 22 degrees C; at 32 degrees C, peak amplitude and frequency progressively and reversibly decreased with time. Two hours of hypoxia had no effect on respiratory discharge. Acutely increasing bath temperature from 22 to 32 degrees C decreased episode and peak duration and increased peak frequency. Changes in pH/CO2 increased peak frequency from zero at pH 8.00-8.10 to maxima of 0.81 +/- 0.01 and 1.44 +/- 0.02 peaks/min at 22 degrees C (pH 7.32) and 32 degrees C (pH 7.46), respectively; pH/CO2 sensitivity was similar at both temperatures. We conclude that 1) insensitivity to hypoxia indicates that rhythmic discharge does not reflect gasping behavior, 2) increased temperature alters respiratory discharge, and 3) central pH/CO2 sensitivity is unaffected by temperature in this preparation (i.e., Q10 approximately 1.0).
Journal of Applied Physiology 03/1998; 84(2):649-60. · 3.06 Impact Factor