James W. Grau

Controllability, coping behavior, and stress-induced analgesia in the rat

Abstract Exposure to painful or stressful stimuli produces an analgesic reaction which can persist for 1–2 h post‐stress. In the typical stress‐induced analgesia study the subject is not permitted to alter or exert control over the aversive event to which it is exposed. That is, its behavior affects neither the duration or intensity of the event. The experiments reported here attempted to determine whether this inability of the subject to control the aversive event is an important determinant of stress‐induced analgesia, or whether simple exposure to painful events is a sufficient condition for its production. In the first experiment rats were given either escapable electric shocks (the subjects behavior could terminate the shock), equal amounts of inescapable shock, or no shock. Tail‐flick to radiant heat was assessed 30 min later. The group given inescapable shock was strongly analgesic, while the group given an equal amount of escapable shock was only mildly analgesic. Thus the controllability of the shock or the availability of a coping response determined the antinociceptive reaction which followed. The second experiment revealed that this differential effect of controllability on tail‐flick responding is masked, shortly after the end of the shock session, by a transient analgesic effect of shock which is not sensitive to the controllability dimension. The implications of these results for stress‐induced analgesia and the activation of opioid systems are discussed.

Instrumental learning within the spinal cord: I. Behavioral properties.

Four experiments are reported that explore whether spinal neurons can support instrumental learning. During training, one group of spinal rats (master) received legshock whenever one hindlimb was extended. Another group (yoked) received legshock independent of leg position. Master, but not yoked, rats learned to maintain their leg in a flexed position, exhibiting progressively longer flexions as a function of training (Experiment 1). All subjects were then tested by applying controllable shock to the same leg (Experiment 2). Master rats reacquired the instrumental response more rapidly (positive transfer), whereas yoked rats failed to learn (a learned helplessness-like effect). Disrupting response-outcome contiguity by delaying the onset and offset of shock by 100 ms eliminated learning (Experiment 3). Experiment 4 showed that shock onset contributes more to learning than does shock offset.

BDNF and learning: Evidence that instrumental training promotes learning within the spinal cord by up-regulating BDNF expression.

We have previously shown that the spinal cord is capable of learning a sensorimotor task in the absence of supraspinal input. Given the action of brain-derived neurotrophic factor (BDNF) on hippocampal learning, the current studies examined the role of BDNF in spinal learning. BDNF is a strong synaptic facilitator and, in association with other molecular signals (e.g. cAMP-response element binding protein (CREB), calcium/calmodulin activated protein kinase II (CaMKII) and synapsin I), important for learning. Spinally transected rats given shock to one hind leg when the leg extended beyond a selected threshold exhibited a progressive increase in flexion duration that minimized shock exposure, a simple form of instrumental learning. Instrumental learning resulted in elevated mRNA levels of BDNF, CaMKII, CREB, and synapsin I in the lumbar spinal cord region. The increases in BDNF, CREB, and CaMKII were proportional to the learning performance. Prior work has shown that instrumental training facilitates learning when subjects are tested on the contralateral leg with a higher response criterion. Pretreatment with the BDNF inhibitor TrkB-IgG blocked this facilitatory effect, as did the CaMKII inhibitor AIP. Intrathecal administration of BDNF facilitated learning when subjects were tested with a high response criterion. The findings indicate that instrumental training enables learning and elevates BDNF mRNA levels within the lumbar spinal cord. BDNF is both necessary, and sufficient, to produce the enabling effect.

Instrumental learning within the spinal cord II. Evidence for central mediation

Rats spinally transected at the second thoracic vertebra can learn to maintain their leg in a flexed position if they receive legshock for extending the limb. These rats display an increase in the duration of a flexion response that minimizes net shock exposure. The current set of experiments was designed to determine whether the acquisition of this behavioral response is mediated by the neurons of the spinal cord (i.e., is centrally mediated) or reflects a peripheral modification (e.g., a change in muscle tension). Experiment 1 found that preventing information from reaching the spinal cord by severing the sciatic nerve blocked the acquisition of this behavioral response. Spinalized rats also failed to learn if the spinal cord was anesthetized with lidocaine during exposure to response-contingent shock (Experiment 2). Experiment 3 demonstrated that prior exposure to response-contingent shock on one hindleg facilitated acquisition of the response when subjects were later tested on the opposite leg. These findings suggest that acquisition of the instrumental response depends on neurons within the spinal cord.

Extent and control of shock affects naltrexone sensitivity of stress-induced analgesia and reactivity to morphine

Opioid and nonopioid mediated changes in pain sensitivity have been observed after exposure to various stressful conditions. A series of inescapable shocks sequentially produces an early form of analgesia which is not affected by the opiate antagonist, naltrexone, and a late antinociceptive response which is sensitive to reversal by naltrexone. Here, this is shown to be true over a wide range of doses. In a further experiment subjects given either escapable or inescapable shock were analgesic immediately after the stress session. However, the analgesia of inescapably shocked subjects was more sensitive to reversal by naltrexone. A final experiment revealed that inescapably shocked subjects, but not escapably shocked subjects, were hyperreactive to the analgesic effects of morphine 24 hr after shock. These results suggest that activation of an opiate system occurs only after extended exposure to stress and that this activation is greater when the stress is inescapable. Implications for opioid versus nonopioid mechanisms of stress-induced analgesia are discussed.

Analysis of the unique cue in configural discriminations

Four experiments used an autoshaping procedure with pigeons to investigate the basis of configural discriminations. The elements of both a negative patterning (A+, B+, AB-) and a conditional discrimination (AC+, BD+, AD-, BC-) were paired, in a second-order procedure with two new key lights, X and Y. Responding was then tested to X and Y presented in compound with each other and with A and B. The pattern of responding to compounds containing X and Y was like the pattern of responding to compounds containing their associates, A and B. This suggests that A and B can be replaced by their associates without disrupting responding to their compounds. Because X and Y are physically different from A and B, this in turn suggests that any unique cue controlling responding to their compounds does not depend on the physical presence of the component stimuli. Instead the unique stimulus appears to arise from the joint activation of memory representations.

MICRORNA DYSREGULATION FOLLOWING SPINAL CORD CONTUSION: IMPLICATIONS FOR NEURAL PLASTICITY AND REPAIR

Spinal cord injury (SCI) is medically and socioeconomically debilitating. Currently, there is a paucity of effective therapies that promote regeneration at the injury site, and limited understanding of mechanisms that can be utilized to therapeutically manipulate spinal cord plasticity. MicroRNAs (miRNAs) constitute novel targets for therapeutic intervention to promote repair and regeneration. Microarray comparisons of the injury sites of contused and sham rat spinal cords, harvested 4 and 14 days following SCI, showed that 32 miRNAs, including miR124, miR129, and miR1, were significantly down-regulated, whereas SNORD2, a translation-initiation factor, was induced. Additionally, three miRNAs including miR21 were significantly induced, indicating adaptive induction of an anti-apoptotic response in the injured cord. Validation of miRNA expression by qRT-PCR and in situ hybridization assays revealed that the influence of SCI on miRNA expression persists up to 14 days and expands both anteriorly and caudally beyond the lesion site. Specifically, changes in miR129-2 and miR146a expression significantly explained the variability in initial injury severity, suggesting that these specific miRNAs may serve as biomarkers and therapeutic targets for SCI. Moreover, the pattern of miRNA changes coincided spatially and temporally with the appearance of SOX2, nestin, and REST immunoreactivity, suggesting that aberrant expression of these miRNAs may not only reflect the emergence of stem cell niches, but also the reemergence in surviving neurons of a pre-neuronal phenotype. Finally, bioinformatics analysis of validated miRNA-targeted genes indicates that miRNA dysregulation may explain apoptosis susceptibility and aberrant cell cycle associated with a loss of neuronal identity, which underlies the pathogenesis of secondary SCI.

Instrumental Learning Within the Spinal Cord: Underlying Mechanisms and Implications for Recovery After Injury:

Using spinally transected rats, research has shown that neurons within the L4-S2 spinal cord are sensitive to response-outcome (instrumental) relations. This learning depends on a form of N-methyl-D-aspartate (NMDA)-mediated plasticity. Instrumental training enables subsequent learning, and this effect has been linked to the expression of brain-derived neurotrophic factor. Rats given uncontrollable stimulation later exhibit impaired instrumental learning, and this deficit lasts up to 48 hr. The induction of the deficit can be blocked by prior training with controllable shock, the concurrent presentation of a tonic stimulus that induces antinociception, or pretreatment with an NMDA or gamma-aminobutyric acid-A antagonist. The expression of the deficit depends on a kappa opioid. Uncontrollable stimulation enhances mechanical reactivity (allodynia), and treatments that induce allodynia (e.g., inflammation) inhibit learning. In intact animals, descending serotonergic neurons exert a protective effect that blocks the adverse consequences of uncontrollable stimulation. Uncontrollable, but not controllable, stimulation impairs the recovery of function after a contusion injury.

The central representation of an aversive event maintains the opioid and nonopioid forms of analgesia.

Exposure to an aversive event, such as shock, can elicit either an opioid or nonopioid analgesia in rats. We suggest that the central representation of an aversive event in working memory activates both forms of analgesia. We formalize this basic hypothesis by coupling it with a current model of animal learning and memory, SOP (Wagner, 1981). SOP is designed to capture the standard operating procedures that govern memory systems. Our application of SOP suggests that manipulations which disrupt the maintenance of information in working memory should alter the magnitude and time course of analgesia. Three experiments are reported that support our proposal. Experiment 1 showed that analgesia decays more rapidly if the representation of the aversive event is displaced from working memory by presenting a postshock distractor. Experiment 2 demonstrated that the postshock distractor alters the magnitude and time course of both the opioid and nonopioid forms of analgesia. Experiment 3 demonstrated that pharmacologically disrupting working memory, by administering a high dose of pentobarbital, prevents mild shock from inducing a strong change in pain reactivity. Implications of the results are discussed.

Instrumental learning within the spinal cord: IV. Induction and retention of the behavioral deficit observed after noncontingent shock.

Spinalized rats given shock whenever 1 hind leg is extended learn to maintain that leg in a flexed position, a simple form of instrumental learning. Rats given shock independent of leg position do not exhibit an increase in flexion duration. Experiment 1 showed that 6 min of intermittent legshock can produce this deficit. Intermittent tailshock undermines learning (Experiments 2-3), and this effect lasts at least 2 days (Experiment 4). Exposure to continuous shock did not induce a deficit (Experiment 5) but did induce antinociception (Experiment 6). Intermittent shock did not induce antinociception (Experiment 6). Experiment 7 addressed an alternative interpretation of the results, and Experiment 8 showed that presenting a continuous tailshock while intermittent legshock is applied can prevent the deficit.