Biomedical subjects

Ole Kiehn

Publications and source records attributed to Ole Kiehn.

11 recordsLinked to original sources

PubMed2004

Central pattern generators deciphered by molecular genetics.

Central pattern generators (CPGs) are localized neuronal networks that have the ability to produce rhythmic movements even in the absence of movement-related sensory feedback. They are found in all animals, including man, and serve as informative model systems for understanding how neuronal networks produce behavior. Traditionally, CPGs have been investigated with electrophysiological techniques. Here we review recent molecular and genetic approaches for dissecting the organization and development of CPGs.

Animals↗

PubMed2003

Synaptic targets of commissural interneurons in the lumbar spinal cord of neonatal rats.

There is strong evidence that commissural interneurons, neurons with axons that extend to the contralateral side of the spinal cord, play an important role in the coordination of left/right alternation during locomotion. In this study we investigated the projections of commissural interneurons to motor neurons and other commissural interneurons on the other side of the spinal cord in neonatal rats. To establish whether there are direct contacts between axons of commissural interneurons and motor neurons, we carried out two series of experiments. In the first experiment we injected biotinylated dextran amine (BDA) into the lateral motor column to retrogradely label commissural interneurons that may have direct projections to motor neurons. Stained neurons were recovered in the ventromedial areas of the contralateral gray matter in substantial numbers. In the second experiment BDA was injected into the ventromedial gray matter on one side of the lumbar spinal cord, whereas motor neurons were simultaneously labeled on the opposite side by applying biocytin onto the ventral roots. BDA injections into the ventromedial gray matter labeled a strong axon bundle that arose from the site of injection, crossed the midline in the ventral commissure, and extensively arborized in the contralateral ventral gray matter. Many of these axons made close appositions with dendrites and somata of motor neurons and also with commissural interneurons retrogradely labeled with BDA. The results suggest that commissural interneurons may establish monosynaptic contacts with motor neurons on the opposite side of the spinal cord. Our findings also indicate that direct reciprocal connections between commissural interneurons on the two sides of the spinal cord may also exist.

Afferent Pathways↗

PubMed2003

Functional identification of interneurons responsible for left-right coordination of hindlimbs in mammals.

Local neuronal networks that are responsible for walking are poorly characterized in mammals. Using an innovative approach to identify interneuron inputs onto motorneuron populations in a neonatal rodent spinal cord preparation, we have investigated the network responsible for left-right coordination of the hindlimbs. We demonstrate how commissural interneurons (CINs), whose axons traverse the midline to innervate contralateral neurons, are organized such that distinct flexor and extensor centers in the rostral lumbar spinal cord define activity in both flexor and extensor caudal motor pools. In addition, the nature of some connections are reconfigured on switching from rest to locomotion via a mechanism that might be associated with synaptic plasticity in the spinal cord. These results from identified pattern-generating interneurons demonstrate how interneuron populations create an effective network to underlie behavior in mammals.

Animals↗

PubMed2003

Role of EphA4 and EphrinB3 in local neuronal circuits that control walking.

Local circuits in the spinal cord that generate locomotion are termed central pattern generators (CPGs). These provide coordinated bilateral control over the normal limb alternation that underlies walking. The molecules that organize the mammalian CPG are unknown. Isolated spinal cords from mice lacking either the EphA4 receptor or its ligand ephrinB3 have lost left-right limb alternation and instead exhibit synchrony. We identified EphA4-positive neurons as an excitatory component of the locomotor CPG. Our study shows that dramatic locomotor changes can occur as a consequence of local genetic rewiring and identifies genes required for the development of normal locomotor behavior.

Animals↗

PubMed2003

Physiological, anatomical and genetic identification of CPG neurons in the developing mammalian spinal cord.

The basic motor patterns underlying rhythmic limb movements during locomotion are generated by neuronal networks located within the spinal cord. These networks are called Central Pattern Generators (CPGs). Isolated spinal cord preparations from newborn rats and mice have become increasingly important for understanding the organization of the CPG in the mammalian spinal cord. Early studies using these preparations have focused on the overall network structure and the localization of the CPG. In this review we concentrate on recent experiments aimed at identifying and characterizing CPG-interneurons in the rodent. These experiments include the organization and function of descending commissural interneurons (dCINs) in the hindlimb CPG of the neonatal rat, as well as the role of Ephrin receptor A4 (EphA4) and its Ephrin ligand B3 (EphrinB3), in the construction of the mammalian locomotor network. These latter experiments have defined EphA4 as a molecular marker for mammalian excitatory hindlimb CPG neurons. We also review genetic approaches that can be applied to the mouse spinal cord. These include methods for identifying sub-populations of neurons by genetically encoded reporters, techniques to trace network connectivity with cell-specific genetically encoded tracers, and ways to selectively ablate or eliminate neuron populations from the CPG. We propose that by applying a multidisciplinary approach it will be possible to understand the network structure of the mammalian locomotor CPG. Such an understanding will be instrumental in devising new therapeutic strategies for patients with spinal cord injury.

Animals↗

PubMed2003

Mechanisms underlying the noradrenergic modulation of longitudinal coordination during swimming in Xenopus laevis tadpoles.

Noradrenaline (NA) is a potent modulator of locomotion in many vertebrate nervous systems. When Xenopus tadpoles swim, waves of motor neuron activity alternate across the body and propagate along it with a brief rostro-caudal delay (RC-delay) between segments. We have now investigated the mechanisms underlying the reduction of RC-delay s by NA. When recording from motor neurons caudal to the twelfth postotic cleft, the mid-cycle inhibition was weak and sometimes absent, compared to more rostral locations. NA enhanced and even unmasked inhibition in these caudal neurons and enhanced inhibition in rostral neurons, but to a lesser extent. Consequently, the relative increase in the amplitude of the inhibition was greater in caudal neurons, thus reducing the RC-inhibitory gradient. We next investigated whether NA might affect the electrical properties of neurons, such that enhanced inhibition under NA might promote postinhibitory rebound firing. The synaptic inputs during swimming were simulated using a sustained positive current, superimposed upon which were brief negative currents. When these conditions were held constant NA enhanced the probability of rebound firing--indicating a direct effect on membrane properties--in addition to any indirect effect of enhanced inhibition. We propose that NA preferentially enhances weak caudal inhibition, reducing the inhibitory gradient along the cord. This effect on inhibitory synaptic transmission, comprising parallel pre- and postsynaptic components, will preferentially facilitate rebound firing in caudal neurons, advancing their firing relative to more rostral neurons, whilst additionally increasing the networks ability to sustain the longer cycle periods under NA.

Adrenergic alpha-Agonists↗

PubMed2002

Firing properties of identified interneuron populations in the mammalian hindlimb central pattern generator.

Little is known about the network structure of the central pattern generator (CPG) controlling locomotor movements in mammals. The present experiments aim at providing such knowledge by focusing on commissural interneurons (CINs) involved in left-right coordination. During NMDA and 5-HT-initiated locomotor-like activity, we recorded intracellularly from caudally or descending projecting L2 and L3 CINs (dCINs) located in the ventromedial area of the lumbar spinal cord in newborn rats. This region is crucial for rhythmic motor output and left-right coordination. The overall sample of dCINs represented a heterogenous population with neurons that fired in all phases of the locomotor cycle and exhibited varying degrees of rhythmicity, from strongly rhythmic to nonrhythmic. Among the rhythmic, putative CPG dCINs were populations that fired in-phase with the ipsilateral or with the contralateral L2 locomotor-like activity. There was a high degree of organization in the dorsoventral location of rhythmic dCINs, with neurons in-phase with the ipsilateral L2 activity located more ventrally. Spikes of rhythmically active dCINs were superimposed on membrane oscillations that were generated predominantly by synaptic input, with little direct contribution from the intrinsic pacemaker hyperpolarization-activated inward current. For both ipsilaterally and contralaterally firing dCINs the dominant synaptic drive was in-phase with the ipsilateral L2 motor activity. This study provides the first characterization of putative CPG interneurons in the mammalian spinal cord. Our results suggest an anatomical and physiological separation of CPG commissural interneurons in the ventral horn and demonstrate that it is possible to target specific interneuron subpopulations in the mammalian locomotor network.

Action Potentials↗

PubMed2002

Synchronization of motor neurons during locomotion in the neonatal rat: predictors and mechanisms.

We describe here the robust synchronization of motor neurons at a millisecond time scale during locomotor activity in the neonatal rat. Action potential activity of motor neuron pairs was recorded extracellularly using tetrodes during locomotor activity in the in vitro neonatal rat spinal cord. Approximately 40% of motor neuron pairs recorded in the same spinal segment showed significant synchronization, with the duration of the central peak in cross-correlograms between motor neurons typically ranging between approximately 30 and 100 msec. The percentage of synchronized motor neuron pairs was considerably higher for pairs with similar locomotor-related activity and strong rhythmic modulation. We also found synchronization between the activities of different motor pools, even if located several segments apart. Such distant synchronization was abolished in the absence of chemical synapses, although local coupling between motor neurons persisted. On the other hand, both local and distant coupling between motor neurons were preserved after antagonism of gap junction coupling between motor neurons. These results demonstrate that motor neuron activity is strongly synchronized at a millisecond time scale during the production of locomotor activity in the neonatal rat. These results also demonstrate that chemical synaptic inputs, in addition to electrical synapses, contribute to this synchronization, suggesting the existence of multiple mechanisms underlying motor neuron synchronization in the neonatal rat. The fast synchronization described here might be involved in activity-dependent processes during development or in the coordination of individual motor neurons into a functional population underlying behavior.

Action Potentials↗

PubMed2002

Projection patterns of commissural interneurons in the lumbar spinal cord of the neonatal rat.

We have studied the axonal projection patterns of commissural interneurons (CINs) in the neonatal rat spinal cord. Some CINs are integral components of the neuronal networks in the vertebrate spinal cord that generate locomotor activity. By using differential retrograde labeling protocols with fluorescent dextran amines, we show that CINs with ascending axons (ascending CINs, or aCINs) and CINs with descending axons (descending CINs, or dCINs) constitute largely different populations. We show that aCINs and dCINs occupy partially overlapping domains in the transverse plane. The aCINs are located at the dorsal margin, within the dorsal horn, centrally within the intermediate zone, and in the medial region of the ventral horn, whereas the dCINs are located predominantly among the ventral and central aCINs and in smaller numbers within the dorsal horn. The labeled aCINs and dCINs project for at least one and a half segment rostrally or caudally and are present in roughly equal numbers. We also demonstrate the presence of a third, smaller population of CINs whose axons bifurcate to project for at least one and a half segment both rostrally and caudally (adCINs). The adCINs are located predominantly among the central and ventral groups of aCINs and dCINs. Finally, we demonstrate the presence of CINs with axons projecting for fewer than one and a half segment in either direction. These "short-range CINs" are intermingled with the aCINs, dCINs, and adCINs. Our results provide an anatomical framework for further electrophysiological studies aimed at identifying the CINs that participate in the mammalian locomotor central pattern generator.

Animals↗

PubMed2002

Organization of left-right coordination in the mammalian locomotor network.

Neuronal circuits involved in left-right coordination are a fundamental feature of rhythmic locomotor movements. These circuits necessarily include commissural interneurons (CINs) that have axons crossing the midline of the spinal cord. The properties of CINs have been described in some detail in the spinal cords of a number of aquatic vertebrates including the Xenopus tadpole and the lamprey. However, their function in left-right coordination of limb movements in mammals is poorly understood. In this review we describe the present understanding of commissural pathways in the functioning of spinal cord central pattern generators (CPGs). The means by which reciprocal inhibition and integration of sensory information are maintained in swimming vertebrates is described, with similarities between the three basic populations of commissural interneurons highlighted. The subsequent section concentrates on recent evidence from mammalian limbed preparations and specifically the isolated spinal cord of the neonatal rat. Studies into the role of CPG elements during drug-induced locomotor-like activity have afforded a better understanding of the location of commissural pathways, such that it is now possible, using whole cell patch clamp, to record from anatomically defined CINs located in the rhythm-generating region of the lumbar segments. Initial results would suggest that the firing pattern of these neurons shows a greater diversity than that previously described in swimming central pattern generators. Spinal CINs play an important role in the generation of locomotor output. Increased knowledge as to their function in producing locomotion is likely to provide valuable insights into the spinal networks required for postural control and walking.

Animals↗

PubMed2002

Gap junctions and motor behavior.

The production of any motor behavior requires coordinated activity in motor neurons and premotor networks. In vertebrates, this coordination is often assumed to take place through chemical synapses. Here we review recent data suggesting that electrical gap-junction coupling plays an important role in coordinating and generating motor outputs in embryonic and early postnatal life. Considering the recent demonstration of a prevalent expression of gap-junction proteins and gap-junction structures in the adult mammalian spinal cord, we suggest that neuronal gap-junction coupling might also contribute to the production of motor behavior in adult mammals.

Animals↗