Neural mechanisms generating locomotion studied in mammalian brain stem‐spinal cord in vitro

1988 ◽  
Vol 2 (7) ◽  
pp. 2283-2288 ◽  
Author(s):  
Jeffrey C. Smith ◽  
Jack L. Feldman ◽  
Brian J. Schmidt
1999 ◽  
Vol 87 (3) ◽  
pp. 1066-1074 ◽  
Author(s):  
Chun-Kuei Su

To understand the origination of sympathetic nerve discharge (SND), I developed an in vitro brain stem-spinal cord preparation from neonatal rats. Ascorbic acid (3 mM) was added into the bath solution to increase the viability of preparations. At 24°C, rhythmic SND (recorded from the splanchnic nerve) was consistently observed, but it became quiescent at <16°C. Respiratory-related SND (rSND) was discernible and was well correlated with C4 root activity. Power spectral analysis of SND revealed a dominant 2-Hz oscillation. In most preparations (86%), such oscillation was persistent, whereas it only slightly reduced its magnitude after isolation from the brain stem. The removal of neural structures rostral to the superior cerebellar artery (equivalent to the level of facial nuclei) reduced rSND, increased tonic SND, but did not affect the temporal coupling between SND and C4 root activity. Our data suggest a prominent contribution of SND from the neural mechanisms confined within the neonatal rat spinal cord. This ascorbic acid-enhanced in vitro preparation is a very useful model to study neural mechanisms underlying sympathorespiratory integration.


1999 ◽  
Vol 82 (2) ◽  
pp. 1074-1077 ◽  
Author(s):  
Isabelle Delvolvé ◽  
Pascal Branchereau ◽  
Réjean Dubuc ◽  
Jean-Marie Cabelguen

An in vitro brain stem–spinal cord preparation from an adult urodele ( Pleurodeles waltl) was developed in which two fictive rhythmic motor patterns were evoked by bath application of N-methyl-d-aspartate (NMDA; 2.5–10 μM) with d-serine (10 μM). Both motor patterns displayed left-right alternation. The first pattern was characterized by cycle periods ranging between 2.4 and 9.0 s (4.9 ± 1.2 s, mean ± SD) and a rostrocaudal propagation of the activity in consecutive ventral roots. The second pattern displayed longer cycle periods (8.1–28.3 s; 14.2 ± 3.6 s) with a caudorostral propagation. The two patterns were inducible after a spinal transection at the first segment. Preliminary experiments on small pieces of spinal cord further suggested that the ability for rhythm generation is distributed along the spinal cord of this preparation. This study shows that the in vitro brain stem–spinal cord preparation from Pleurodeles waltl may be a useful model to study the mechanisms underlying the different axial motor patterns and the flexibility of the neural networks involved.


2011 ◽  
Vol 105 (6) ◽  
pp. 2818-2829 ◽  
Author(s):  
Eugene Zaporozhets ◽  
Kristine C. Cowley ◽  
Brian J. Schmidt

Previous studies of the in vitro neonatal rat brain stem-spinal cord showed that propriospinal relays contribute to descending transmission of a supraspinal command signal that is capable of activating locomotion. Using the same preparation, the present series examines whether enhanced excitation of thoracic propriospinal neurons facilitates propagation of the locomotor command signal in the lesioned spinal cord. First, we identified neurotransmitters contributing to normal endogenous propriospinal transmission of the locomotor command signal by testing the effect of receptor antagonists applied to cervicothoracic segments during brain stem-induced locomotor-like activity. Spinal cords were either intact or contained staggered bilateral hemisections located at right T1/T2 and left T10/T11 junctions designed to abolish direct long-projecting bulbospinal axons. Serotonergic, noradrenergic, dopaminergic, and glutamatergic, but not cholinergic, receptor antagonists blocked locomotor-like activity. Approximately 73% of preparations with staggered bilateral hemisections failed to generate locomotor-like activity in response to electrical stimulation of the brain stem alone; such preparations were used to test the effect of neuroactive substances applied to thoracic segments (bath barriers placed at T3 and T9) during brain stem stimulation. The percentage of preparations developing locomotor-like activity was as follows: 5-HT (43%), 5-HT/ N-methyl-d-aspartate (NMDA; 33%), quipazine (42%), 8-hydroxy-2-(di- n-propylamino)tetralin (20%), methoxamine (45%), and elevated bath K+ concentration (29%). Combined norepinephrine and dopamine increased the success rate (67%) compared with the use of either agent alone (4 and 7%, respectively). NMDA, Mg2+ ion removal, clonidine, and acetylcholine were ineffective. The results provide proof of principle that artificial excitation of thoracic propriospinal neurons can improve supraspinal control over hindlimb locomotor networks in the lesioned spinal cord.


1992 ◽  
Vol 67 (2) ◽  
pp. 373-388 ◽  
Author(s):  
T. Matsushima ◽  
S. Grillner

1. To elucidate the neural mechanisms responsible for coordinating undulatory locomotor movements, the intersegmental phase lag was analyzed from ventral roots along the spinal cord during fictive swimming. It was induced by bath application of N-methyl-D-aspartate (NMDA) in in vitro preparations of lamprey spinal cord, while the excitability of different segments were modified. The phase lag between consecutive segments during normal forward swimming is 1% of the cycle duration in a broad range of values. Rostral segments are activated before more caudal ones. 2. Under control conditions, whole preparations (12-24 segment long; n = 22) were perfused with NMDA solutions of the same concentration (100-150 microM). The intersegmental phase lag values varied in a continuous range with a single peak around a median value of forward +0.74% per segment (range: forward +2.23% to backward -0.97%). 3. To examine whether excitability differences along the spinal cord could modify the intersegmental phase lag, different levels of excitatory amino acids (NMDA) were applied to spinal cord preparations positioned in a partitioned chamber. Different portions of the cord could be perfused separately by NMDA solutions of different concentrations (50-150 microM). If rostral segments were perfused with the higher NMDA solution, the lag was inevitably in the forward direction. Conversely, if the caudal portion was perfused with the higher NMDA solution, caudally located ventral roots became activated before the rostral ventral roots in a caudorostral succession, thus reversing the direction of the fictive swimming wave to propagate as during backward swimming. If the middle portion was perfused by the highest NMDA solution, this portion instead became leading, and the activity propagated from this point in both the rostral and the caudal directions. The portion located in the pool with highest NMDA concentration always gave rise to a "leading" segment. 4. When a portion of the preparation was perfused with an NMDA solution of a high concentration (75-150 microM), the cycle duration was close to that recorded when the whole preparation was perfused with the same high NMDA solution. The ensemble cycle duration is, therefore, largely determined by the leading segment. 5. The phase lag changes were not restricted to the region around the barrier separating pools with different NMDA solutions.(ABSTRACT TRUNCATED AT 400 WORDS)


1994 ◽  
Vol 266 (3) ◽  
pp. R658-R667 ◽  
Author(s):  
K. Sugaya ◽  
W. C. De Groat

An in vitro neonatal (1-7 day) rat brain stem-spinal cord-bladder (BSB) preparation was used to examine the central control of micturition. Isovolumetric bladder contractions occurred spontaneously or were induced by electrical stimulation of the ventrolateral brain stem, spinal cord, bladder wall (ES-BW), or by perineal tactile stimulation (PS). Transection of the spinal cord at the L1 segment increased the amplitude of ES-BW- and PS-evoked contractions, and subsequent removal of the spinal cord further increased spontaneous and ES-BW-evoked contractions but abolished PS-evoked contractions. Hexamethonium (1 mM), a ganglionic blocking agent, mimicked the effect of cord extirpation. Tetrodotoxin (1 microM) blocked ES-BW- and PS-evoked contractions but enhanced spontaneous contractions. Bicuculline methiodide (10-50 microM), a gamma-aminobutyric acid A receptor antagonist, increased the amplitude of spontaneous, ES-BW- and PS-evoked contractions. These results indicate that PS-evoked contractions are mediated by spinal reflex pathways, whereas spontaneous and ES-BW-evoked contractions that are elicited by peripheral mechanisms are subject to a tonic inhibition dependent on an efferent outflow from the spinal cord. PS-evoked micturition is also subject to inhibitory modulation arising from sites rostral to the lumbosacral spinal cord. Although electrical stimulation of bulbospinal excitatory pathways can initiate bladder contractions in the neonatal rat, these pathways do not appear to have an important role in controlling micturition during the first postnatal week.


1993 ◽  
Vol 70 (6) ◽  
pp. 2241-2250 ◽  
Author(s):  
M. K. Floeter ◽  
A. Lev-Tov

1. The excitation of lumbar motoneurons by reticulospinal axons traveling in the medial longitudinal fasciculus (MLF) was investigated in the newborn rat using intracellular recordings from lumbar motoneurons in an in vitro preparation of the brain stem and spinal cord. The tracer DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine) was introduced into the MLF of 6-day-old littermate rats that had been fixed with paraformaldehyde to evaluate the anatomic extent of this developing pathway. 2. Fibers labeled from the MLF by DiI were present in the cervical ventral and lateral white matter and a smaller number of labeled fibers extended to the lumbar enlargement. Patches of sparse terminal labeling were seen in the lumbar ventral gray. 3. In the in vitro preparation of the brain stem and spinal cord, MLF stimulation excited motoneurons through long-latency pathways in most motoneurons and through both short-(< 40 ms) and long-latency connections in 16 of 40 motoneurons studied. Short- and longer-latency components of the excitatory response were evaluated using mephenesin to reduce activity in polysynaptic pathways. 4. Paired-pulse stimulation of the MLF revealed a modest temporal facilitation of the short-latency excitatory postsynaptic potential (EPSP) at short interstimulus intervals (20–200 ms). Trains of stimulation at longer interstimulus intervals (1–30 s) resulted in a depression of EPSP amplitude. The time course of the synaptic depression was compared with that found in EPSPs resulting from paired-pulse stimulation of the dorsal root and found to be comparable. 5. The short-latency MLF EPSP was reversibly blocked by 6-cyano-7-nitroquinoxaline (CNQX), an antagonist of non-N-methyl-D-aspartate glutamate receptors, with a small CNQX-resistant component. Longer-latency components of the MLF EPSP were also blocked by CNQX, and some late components of the PSP were sensitive to strychnine. MLF activation of multiple polysynaptic pathways in the spinal cord is discussed.


2006 ◽  
Vol 96 (4) ◽  
pp. 2042-2055 ◽  
Author(s):  
James F. Einum ◽  
James T. Buchanan

An in vitro preparation of the nervous system of the lamprey, a lower vertebrate, was used to characterize the properties of spinal neurons with axons projecting to the brain stem [i.e., spinobulbar (SB) neurons)]. To identify SB neurons, extracellular electrodes on each side of the spinal cord near the obex recorded the axonal spikes of neurons impaled with sharp intracellular microelectrodes in the rostral spinal cord. The ascending spinal neurons ( n = 144) included those with ipsilateral (iSB) (63/144), contralateral (cSB) (77/144), or bilateral (bSB) (4/144) axonal projections to the brain stem. Intracellular injection of biocytin revealed that the SB neurons had small- to medium-size somata and most had dendrites confined to the ipsilateral side of the cord, although about half of the cSB neurons also had contralateral dendrites. Most SB neurons had multiple axonal branches including descending axons. Electrophysiologically, the SB neurons were similar to other lamprey spinal neurons, firing spikes throughout long depolarizing pulses with some spike-frequency adaptation. Paired intracellular recordings between SB and reticulospinal (RS) neurons revealed that SB neurons made either excitatory or inhibitory synapses on RS neurons and the SB neurons received excitatory input from RS neurons. Mutual excitation and feedback inhibition between pairs of RS and SB neurons were observed. The SB neurons also received excitatory inputs from primary mechanosensory neurons (dorsal cells), and these same SB neurons were rhythmically active during fictive swimming, indicating that SB neurons convey both sensory and locomotor network information to the brain stem.


1985 ◽  
Vol 249 (3) ◽  
pp. R317-R322 ◽  
Author(s):  
D. R. Kostreva ◽  
J. A. Armour ◽  
Z. J. Bosnjak

The hypothesis tested was that some cardiac reflexes can be mediated by the neural mechanisms in thoracic sympathetic ganglia in addition to those existing in the spinal cord or brain stem. In mongrel dogs anesthetized with pentobarbital sodium the left stellate (SG) and middle cervical (MCG) ganglion were decentralized, and the left thoracic vagosympathetic trunk was cut cranial to the MCG and caudal to the MCG but above the heart. The left thoracic vagosympathetic trunk below the MCG was then stimulated afferently. A single bolus of 2-[14C]deoxyglucose was injected intravenously and the stimuli repeated periodically toward the decentralized ganglia for 45 min. The heart and ganglia were then removed, frozen, and sectioned for autoradiography. Significant increases in glucose utilization were found in both the SG and MCG compared with ganglia from nonstimulated control animals. Significant increases in the glucose utilization of the endocardial third of the left ventricle were also observed in the reflexly stimulated hearts. Neural mechanisms in the acutely decentralized SG and MCG could modify cardiac glucose utilization within specific areas of the heart. In addition, such ganglionic mechanisms may be concentrated within the caudal half of the SG. These data suggest that many neurons in the thoracic autonomic ganglia may be involved in local cardiac reflexes.


1992 ◽  
Vol 262 (4) ◽  
pp. R595-R603 ◽  
Author(s):  
Y. Xia ◽  
C. Jiang ◽  
G. G. Haddad

Using enzyme histochemistry and in vitro electrophysiological recordings in brain slices, we studied 1) the relative activity of cytochrome c oxidase (Cytox) and hexokinase (HK) and 2) cellular function by examining ionic homeostasis across cell membranes in the turtle and newborn (5 days old) and adult rat central nervous system. We found that Cytox was higher in the rostral than in the caudal brain regions of the adult rat and that the activity in the newborn is at least as high as in the adult rat. In contrast, adult turtles had very low Cytox activity throughout the central nervous system. Compared with that in the adult rat, HK activity in the newborn was generally lower in the rostral brain and cerebellum but similar or higher in the brain stem and spinal cord. In the turtle, HK activity was higher in the cerebellum, brain stem, and ventral horn of the spinal cord than in those in the rat. During anoxia, extracellular K+ increased by approximately 10-fold (from 3.2 to approximately 32 mM) in the adult brain stem but only by 2.6 mM in newborn rats. After glycolysis was blocked with iodoacetic acid (10-20 mM), extracellular K+ increased remarkably in both adult and newborn rats to approximately 35 mM. In contrast, the turtle brain tissue showed a slight and insignificant increase in extracellular K+ during complete anoxia or with iodoacetic acid; there was a modest increase in K+ when anoxia and iodoacetate were administered together. We conclude that 1) the newborn rat brain must rely either on higher glycolytic capacity or on a reduction of metabolic rate during O2 deprivation and 2) the turtle brain can subsist on nonglucose fuels or on fuels not requiring the citric acid cycle and the electron transfer chain.


2014 ◽  
Vol 117 (9) ◽  
pp. 1027-1036 ◽  
Author(s):  
Veronica Siljehav ◽  
Yuri Shvarev ◽  
Eric Herlenius

Prostaglandin E2 (PGE2) serves as a critical mediator of hypoxia, infection, and apnea in term and preterm babies. We hypothesized that the prostaglandin E receptor type 3 (EP3R) is the receptor responsible for PGE2-induced apneas. Plethysmographic recordings revealed that IL-1β (ip) attenuated the hypercapnic response in C57BL/6J wild-type (WT) but not in neonatal (P9) EP3R−/− mice ( P < 0.05). The hypercapnic responses in brain stem spinal cord en bloc preparations also differed depending on EP3R expression whereby the response was attenuated in EP3R−/− preparations ( P < 0.05). After severe hypoxic exposure in vivo, IL-1β prolonged time to autoresuscitation in WT but not in EP3R−/− mice. Moreover, during severe hypoxic stress EP3R−/− mice had an increased gasping duration ( P < 0.01) as well as number of gasps ( P < 0.01), irrespective of intraperitoneal treatment, compared with WT mice. Furthermore, EP3R−/− mice exhibited longer hyperpneic breathing efforts when exposed to severe hypoxia ( P < 0.01). This was then followed by a longer period of secondary apnea before autoresuscitation occurred in EP3R−/− mice ( P < 0.05). In vitro, EP3R−/− brain stem spinal cord preparations had a prolonged respiratory burst activity during severe hypoxia accompanied by a prolonged neuronal arrest during recovery in oxygenated medium ( P < 0.05). In conclusion, PGE2 exerts its effects on respiration via EP3R activation that attenuates the respiratory response to hypercapnia as well as severe hypoxia. Modulation of the EP3R may serve as a potential therapeutic target for treatment of inflammatory and hypoxic-induced detrimental apneas and respiratory disorders in neonates.


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