应激对大鼠海马突触可塑性和学习记忆的影响

突触可塑性参与学习记忆，是学习记忆过程中必不可少的重要环节。应激能损伤或提高学习记忆、抑制长时程增强（Long一termPotentiation，LTP）、易化长时程抑制（Long一termdePressfon，LTD）。然而，应激经历依赖的长时程抑制（StressexperiencedePendentLTD，SLTD）在学习记忆中有什么意义还不清楚，这正是本文首先要探讨问题。其次，尽管我们在不同年龄时期的学习记忆能力不同，但是对所经历的重大应激事件仍然能产生牢固的记忆，这就有必要进一步通过阐明应激对突触可塑性的影响是否有年龄差异来解释SLTD的意义。再者，我们甚至不清楚为什么有的时候应激损伤学习记忆而有的时候又提高学习记忆。为了阐明这个问题，本文在最后一部分研究了哪些因素决定应激损伤还是提高学习记忆。为了探讨SLTD与学习记忆的关系，我们在第一部分实验中研究急性或亚急性的高台（ElevatedPlatform，EP）或足部电击（Footshock，Fs）对4周龄诚star大鼠海马CAI区LTD诱导的影响，以及这些应激条件对大鼠空间学习记忆的影响。结果显示：（l）急性EP易化LTD，亚急性EP不易化LTD；急性和亚急性FS都易化了LTD。（2）急性EP损伤空间记忆，亚急性EP对空间记忆没有影响；急性FS不影响空间记忆，亚急性FS却提高了空间记忆。这部分的实验结果表明：sLTD存在与否与应激对空间记忆的影响不是一一对应的关系，提示sLTD可能参与了应激记忆（对应激经历产生的记忆）。如果SLTD参与应激记忆，那么，研究应激对突触可塑性的影响是否有年龄依赖性将有助于解释不同年龄时期的学习记忆能力不同但是都能牢固记忆应激事件这一现象。为此，我们在第二部分实验中进一步研究EP对幼年、中年和老年动物（4，10和74周龄的Wistar大鼠）海马CAI区LTP／LTD诱导的影响。结果表明：（1）没有应激的情况下，LTP的大小随动物年龄的增加呈明显的倒U型关系，而低频刺激都不能诱导出LTD。（2）EP易化了这些年龄段大鼠的LTD，并完全阻断它们的LTP。值得注意的是，在三个年龄段大鼠中，EP易化的LTD大小一样。该部分实验结果提示：SLTD没有年龄依赖性，这与人和动物在不同年龄时期对所受到的重大应激事件都能产生牢固记忆这一现象相一致。所以，该部分结果进一步加强了第一部分结果的推测：SLTD参与应激记忆。应激有的时候提高动物的空间记忆能力，有的时候又损伤其空间记忆。为了阐明是什么因素决定应激损伤还是提高学习记忆，我们在第三部分实验中研究EP、FS和水迷宫游泳应激经历对六个年龄段动物（4，10，13，20，67，76周龄）在水迷宫任务中的影响。结果显示：（l）EP应激损伤无水迷宫游泳应激经历大鼠的空间记忆。（2）EP应激提高有游泳应激经历幼年大鼠的空间记忆；训练前外源性补充皮质酮或FS都抑制皮质酮或FS导致的空间记忆受损。（3）糖皮质激素受体的拮抗剂RU38486阻断上述的空间记忆成绩提高。所以，不同的应激经历和动物年龄两个因素共同决定了应激损伤还是提高学习记忆。

情绪应激对大鼠体液免疫功能的影响及其机制

This study was undertaken to investigate the effect of emotional stress on humoral immunoactivity and to examine whether the sympathetic nervous system was involved in the immunomodulation. In the present study, two types of emotional stressors were used. One was footshock apparatus used to cause the rats which were given footshock before, emotional stressed; the other was an empty water bottle used to cause the rats which were trained to drink water at two set times each day, emotional stressed. The effect of emotional stress on the primary immune function (anti-ovallum antibody level and spleen index), the endocrine response (corticosterone level, epinephrine and norepinephrine level), the behavioral changes (freezing, defecation, grooming and attacking behavior) were investigated. The main results were: 1. Two types of emotional stress significantly increased the level of plasma corticosterone, norepinephrine and epinephrine, as well as freezing, defecation and attacking behavior. 2. Two types of emotional stress significantly decreased the level of anti-ovallum antibody. A negative correlation between catecholamine level (epinephrine and norepinephrine) and antibody level or spleen index was found. 3. β-adrenergic receptor antagonist propranolol could reverse the immunomodulation induced by emotional stress. 4. After two types of emotional stress, c-fos expression was observed in the following brain areas or nucleus; arcuate nucleus, anterior commissure nucleus, diffuse part of dorsalmedial nucleus hypothalamus, lateral dorsal nucleus thalamus, medial nucleus amygdala, solitary nucleus, frontal cortex and cingulum. These brain areas and nucleus are involved in the central modulation of the autonomic nervous system. Taken together, these findings demonstrate that emotional stress can suppress humoral immunity and the activation of the sympathetic nervous system is involved in the humoral immunomodulation induced by emotional stress.

Corticosterone plasma concentrations in Carioca High- and Low-Conditioned freezing rats after a fear conditioned task

Our group in the Psychology Department at Pontifícia Universidade Católica do Rio de Janeiro (PUC-Rio) developed a rat genetic model of extreme freezing in response to contextual cues in an experimental chamber previously associated with footshock. One of the lines, Carioca High Freezing (CHF), exhibits an enhanced conditioned freezing response, whereas the other line, Carioca Low Freezing (CLF), shows the opposite response. The present study investigated corticosterone concentration between these two lines of animals and a random (RND) line of rats both under basal conditions and test condition after an emotional challenge using a contextual fear conditioning protocol. Comparisons between basal and test plasma corticosterone concentrations suggested differential basal and fear-induced differences between the two lines. The differences between basal conditions is an important and relevant aspect to be considered in behavioral experiments using or assessing stress and could help to understand variability in naïve populations.

Involvement of the insular cortex in the consolidation and expression of contextual fear conditioning

The insular cortex (IC) has been reported to be involved in the modulation of memory and autonomic and defensive responses. However, there is conflicting evidence about the role of the IC in fear conditioning. To explore the IC involvement in both behavioral and autonomic responses induced by contextual fear conditioning, we evaluated the effects of the reversible inhibition of the IC neurotransmission through bilateral microinjections of the non-selective synapse blocker CoCl2 (1 mm) 10 min before or immediately after the conditioning session or 10 min before re-exposure to the aversive context. In the conditioning session, rats were exposed to a footshock chamber (context) and footshocks were used as the unconditioned stimulus. Forty-eight hours later, the animals were re-exposed to the aversive context for 10 min, but no shock was given. Behavioral (freezing) as well as cardiovascular (arterial pressure and heart rate increases) responses induced by re-exposure to the aversive context were analysed. It was observed that the local IC neurotransmission inhibition attenuated freezing and the mean arterial pressure and heart rate increase of the groups that received the CoCl2 either immediately after conditioning or 10 min before re-exposure to the aversive context, but not when the CoCl2 was injected before the conditioning session. These findings suggest the involvement of the IC in the consolidation and expression of contextual aversive memory. However, the IC does not seem to be essential for the acquisition of memory associated with aversive context. © 2013 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

Novelty, but Not Operant Aversive Learning, Enhances Fos and Egr-1 Expression in the Medial Prefrontal Cortex and Hippocampal Areas of Rats

Immediate early genes (IEG) are presumed to be activated in response to stress, novelty, and learning. Evidence supports the involvement of prefrontal and hippocampal areas in stress and learning, but also in the detection of novel events. This study examined whether a previous experience with shocks changes the pattern of Fos and Egr-1 expression in the medial prefrontal cortex (mPFC), the hippocampal cornus ammonis 1 (CA1), and dentate gyrus (DG) of adult male Wistar rats that learned to escape in an operant aversive test. Subjects previously exposed to inescapable footshocks that learned to escape from Shocks were assigned to the treated group (EXP). Subjects from Group Novelty (NOV) rested undisturbed during treatment and also learned to escape in the test. The nonshock group (NSH) rested undisturbed in both sessions. Standard immunohistochemistry procedures were used to detect the proteins in brain sections. The results show that a previous experience with shocks changed the pattern of IEG expression, then demonstrating c-fos and egr-1 induction as experience-dependent events. Compared with NSH and EXP an enhanced Fos expression was detected in the mPFC and CA1 subfield of Group NOV, which also exhibited increased Egr-1 expression in the mPFC and DG in comparison to NSH. No differences were found in the DG for Fos, or in the CA1 for Egr-1. Novelty, and not the operant aversive escape learning, seems to have generated IEG induction. The results suggest novel stimuli as a possible confounding factor in studies on Fos and/or Egr-1 expression in aversive conditions.

Distinct mechanisms underlie activation of hypothalamic neurosecretory neurons and their medullary catecholaminergic afferents in categorically different stress paradigms.

Intermittent electrical footshock induces c-fos expression in parvocellular neurosecretory neurons expressing corticotropin-releasing factor and in other visceromotor cell types of the paraventricular hypothalamic nucleus (PVH). Since catecholaminergic neurons of the nucleus of the solitary tract and ventrolateral medulla make up the dominant loci of footshock-responsive cells that project to the PVH, these were evaluated as candidate afferent mediators of hypothalamic neuroendocrine responses. Rats bearing discrete unilateral transections of this projection system were exposed to a single 30-min footshock session and sacrificed 2 hr later. Despite depletion of the aminergic innervation on the ipsilateral side, shock-induced up-regulation of Fos protein and corticotropin-releasing factor mRNA were comparable in strength and distribution in the PVH on both sides of the brain. This lesion did, however, result in a substantial reduction of Fos expression in medullary aminergic neurons on the ipsilateral side. These results contrast diametrically with those obtained in a systemic cytokine (interleukin 1) challenge paradigm, where similar cuts ablated the Fos response in the ipsilateral PVH but left intact the induction seen in the ipsilateral medulla. We conclude that (i) footshock-induced activation of medullary aminergic neurons is a secondary consequence of stress, mediated via a descending projection transected by our ablation, (ii) stress-induced activation of medullary aminergic neurons is not necessarily predictive of an involvement of these cell groups in driving hypothalamic visceromotor responses to a given stressor, and (iii) despite striking similarities in the complement of hypothalamic effector neurons and their afferents that may be activated by stresses of different types, distinct mechanisms may underlie adaptive hypothalamic responses in each.