Scott N. Currie

Spinal Motor Patterns in the Turtlea

Abstract: Rhythmic alternation between ipsilateral hip flexors and extensors occurs during the normal pattern of fictive rostral scratching in response to unilateral midbody stimulation in D3‐end turtles (complete spinal transection posterior to the forelimb enlargement). Unilateral midbody stimulation evokes rhythmic bursts of ipsilateral hip flexor activity with no hip extensor activity in D3‐end turtles with D6‐D7 contralateral hemisection (transverse hemisection anterior to the hindlimb enlargement). Bilateral midbody stimulation in these turtles evokes reconstruction of rhythmic alternation between intact side hip flexors and extensors. These normal motor patterns in response to two‐site stimulation are reconstructed because one‐site stimulation in this preparation activates only hip flexor rhythms (J. Neurosci. 18: 467).

NMDA receptors in layers II and III of rat cerebral cortex

Glutamate receptors are found in all layers of the cerebral cortex, but NMDA receptors are concentrated in layers II and III in the adult. We investigated the location of these receptors, and their contribution to the responses of cells in layers V and VI, by iontophoresing NMDA at various distances from the cell body along the apical dendrite of the cells, first in artificial CSF, then in TTX to abolish action potentials. Comparison of responses at various distances along the apical dendrite showed that the response generally increases as distance from the cell body decreases. Comparison of responses in layers II and III, before and after TTX, showed that TTX reduced the response considerably. We conclude first that NMDA receptors in layers II and III are located primarily on cells in layers II and III, rather than on the apical dendrites of cells in layers V and VI, and second that the contribution of NMDA receptors to the response of cells in layers V and VI comes primarily from receptors close to the cell body.

Fictive hindlimb motor patterns evoked by AMPA and NMDA in turtle spinal cord-hindlimb nerve preparations.

Application of the glutamate agonists alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA, 5-10 microM), or N-methyl-D-aspartate (NMDA, 50-100 microM) to the turtle spinal cord produced fictive hindlimb motor patterns in low-spinal immobilized animals (in vivo) and in isolated spinal cord-hindlimb nerve preparations (in vitro). For in vivo experiments, drugs were applied onto the dorsal surface of 2-4 adjacent spinal cord segments in and near the anterior hindlimb enlargement. Motor output was recorded unilaterally or bilaterally from hindlimb muscle nerves. AMPA elicited vigorous motor patterns in vivo that included strict hip flexor-extensor and right-left alternation. In most turtles, the monoarticular knee extensor nerve FT-KE was active during the HE phase of AMPA evoked burst cycles, similar to the timing of pocket scratch motor patterns. NMDA was less effective in vivo, typically producing only weak and irregular bursting from hip nerves and little or no knee extensor (KE) discharge. Sensory stimulation of a rostral scratch reflex in vivo could reset an ongoing AMPA-evoked motor rhythm, indicating that cutaneous reflex pathways interact centrally with the chemically activated rhythm generator. Most in vitro preparations consisted of six segments of spinal cord, including the entire 5-segment hindlimb enlargement (D8-S2) and the segment immediately anterior to the enlargement (D7), with attached hindlimb nerves. In contrast to in vivo experiments, in vitro preparations exhibited highly regular, long-lasting motor rhythms when NMDA was superfused over the spinal cord. AMPA also produced rhythmic motor patterns in vitro, but these lasted only a few minutes before they were replaced with tonic discharge. FT-KE timing during in vitro chemically elicited activity was similar to that of sensory-evoked pocket scratch motor patterns. Some NMDA-evoked rhythmicity persisted even in 3-segment (D6-D8) and 1-segment (D8) in vitro preparations, demonstrating that neural mechanisms for chemically activated rhythmogenesis reside even in a single segment of the hindlimb enlargement.

Glycinergic inhibition in the turtle spinal cord regulates the intensity and pattern of fictive flexion reflex motor output

Application of strychnine sulfate (10-50 mu M) to the anterior hindlimb enlargement of the turtle spinal cord increased the amplitude of the ipsilateral fictive flexion reflex and revealed a contralateral (crossed) fictive flexion reflex response to cutaneous stimulation of the foot. Strychnine abolished the crossed inhibition of fictive flexion reflex that was normally evoked by contralateral foot stimulation, unmasking the crossed excitation. Our observations are consistent with the hypothesis that intraspinal inhibition mediated by strychnine-sensitive glycine receptors regulates the amplitude of fictive flexion reflex motor output and confines the response to the appropriate neural pathways.

Location of Spinal Cord Pathways That Control Hindlimb Movement Amplitude and Interlimb Coordination During Voluntary Swimming in Turtles

We performed mechanical lesions of the midbody (D2-D3; second to third postcervical spinal segments) spinal cord in otherwise intact turtles to locate spinal cord pathways that 1) activate and control the amplitude of voluntary hindlimb swimming movements and 2) coordinate hindlimb swimming with the movement of other limbs. Pre- and postlesion turtles were held by a band clamp around the carapace just beneath the water surface in a clear Plexiglas tank and videotaped from below so that kinematic measurements could be made of voluntary forward swimming with motion analysis software. Movements of the forelimbs (wrists) and hindlimbs (knees and ankles) were tracked relative to stationary reference points on the plastron to obtain bilateral measurements of hip and forelimb angles as functions of time along with foot trajectories. We measured changes in limb movement amplitude, cycle period, and interlimb phase before and after spinal lesions. Our results indicate that locomotor command signals that activate and regulate the amplitude of voluntary hindlimb swimming travel primarily in the dorsolateral funiculus (DLF) at the D2-D3 level and cross over to drive contralateral hindlimb movements. This suggests that electrical stimulation of the D3 DLF, which was previously shown to evoke swimming movements in the contralateral hindlimb of low-spinal turtles, activated the same locomotor command pathways that the animal uses during voluntary behavior. We also show that forelimb-hindlimb coordination is maintained by longitudinal spinal pathways that are largely confined to the ventrolateral funiculus (VLF) and mediate phase coupling of ipsilateral limbs, presumably by interenlargement propriospinal fibers.

Electrically evoked locomotor activity in the turtle spinal cord hemi-enlargement preparation

We assessed the locomotor capacity of the left half of the spinal cord hindlimb enlargement in low-spinal turtles. Forward swimming was evoked in the left hindlimb by electrical stimulation of the right dorsolateral funiculus (DLF) at the anterior end of the third postcervical spinal segment (D3). Animals were held by a band-clamp in a water-filled tank so that hindlimb movements could be recorded from below with a digital video camera. Left hindlimb hip and knee movements were tracked while electromyograms (EMGs) were recorded from left hip and knee muscles. In turtles with intact spinal cords, electrical stimulation of the right D3 DLF evoked robust forward swimming movements of the left hindlimb, characterized by rhythmic alternation between hip flexor (HF) and hip extensor (HE) EMG discharge, with knee extensor (KE) bursts occurring during the latter part of each HE-off phase. After removing the right spinal hemi-enlargement (D8-S2), DLF stimulation still evoked rhythmic locomotor movements and EMG bursts in the left hindlimb that included HF-HE alternation and KE discharge. However, post-surgical movements and EMG bursts had longer cycle periods, and movements showed lower amplitudes compared to controls. These results show that (1) sufficient locomotor CPG circuitry resides within the turtle spinal hemi-enlargement to drive major components of the forward swim motor pattern, (2) contralateral circuitry contributes to the excitation of the locomotor CPG for a given limb, and (3) a sufficient portion of the descending DLF pathway crosses over to the contralateral cord anterior to the hindlimb enlargement to activate swimming.

SUCKLING-INDUCED BURST DISCHARGES OF SUPRAOPTIC OXYTOCIN NEURONS IN RATS: PROSTAGLANDIN MEDIATION OF OXYTOCIN ACTIONS

Yu-Feng Wang Pulsatile release of hormones and neurotransmitters is widespread in mammals and typical of oxytocin secretion during lactation. Sucklingevoked pulsatile release of oxytocin follows a transient activation of hypothalamic oxytocin neurons in a burst-like firing pattern and leads to a bolus release of oxytocin and milk ejections. Prostaglandin (PG), a nonhormonal messenger cytokine, has been implicated in the activation of oxytocin neurons. To clarify the role of PGs in the activation of oxytocin neurons, we made the following observations. In patch-clamp recordings of oxytocin neurons in the supraoptic nucleus of brain slices from lactating rats, bath application of PGE2 evoked a significantly higher incidence of bursts than oxytocin did. Indomethacin, an inhibitor of PG synthesis, totally blocked oxytocinbut not PGE2-evoked bursts. Moreover, suckling-induced milk ejections and increased molecular association between extracellular signal-regulated protein kinase (ERK) 1/2 and actin were blocked by intracerebroventricular application of indomethacin. Intracerebroventricular administration of PD98059, a blocker of ERK 1/2 phosphorylation/activation, also blocked suckling-induced cyclooxygenase 2 (Cox-2, a PG synthetase) in oxytocin neurons and suckling-increased molecular association between ERK 1/2 and Cox-2. These results indicate that PGs are essential signaling links in the suckling-evoked bursting of oxytocin neurons; however, to evoke full bursts, PGs need other signaling events downstream to oxytocin receptors. This finding highlights the mediator effect of PGs on oxytocin-evoked pulsatile oxytocin release, and provides a model of signaling process of hormonal actions and therapeutic target of hormone-related diseases.

Sensory‑evoked turning locomotion in red‑eared turtles: kinematic analysis and electromyography

We examined the limb kinematics and motor patterns that underlie sensory-evoked turning locomotion in red-eared turtles. Intact animals were held by a band-clamp in a water-filled tank. Turn-swimming was evoked by slowly rotating turtles to the right or left via a motor connected to the shaft of the band-clamp. Animals executed sustained forward turn-swimming against the direction of the imposed rotation. We recorded video of turn-swimming and computer-analyzed the limb and head movements. In a subset of turtles, we also recorded electromyograms from identified limb muscles. Turning exhibited a stereotyped pattern of (1) coordinated forward swimming in the hindlimb and forelimb on the outer side of the turn, (2) back-paddling in the hindlimb on the inner side, (3) a nearly stationary, “braking” forelimb on the inner side, and (4) neck bending toward the direction of the turn. Reversing the rotation caused animals to switch the direction of their turns and the asymmetric pattern of right and left limb activities. Preliminary evidence suggested that vestibular inputs were sufficient to drive the behavior. Sensory-evoked turning may provide a useful experimental platform to examine the brainstem commands and spinal neural networks that underlie the activation and switching of different locomotor forms.