Circulating levels of glucocorticoid hormones in WHHL and NZW rabbits: circadian cycle and response to repeated social encounter.

Social environment influences the progression of atherosclerosis in an important experimental model of disease, the Watanabe Heritable Hyperlipidemic rabbit (WHHL). Although the hypothalamic-pituitary-adrenocortical (HPA) system is likely to play an important role in the behavioral modulation of disease, relatively little is known about the glucocorticoid responses in these animals, or in other strains of rabbits. The purpose of the present study was to: (1) evaluate the rabbit glucocorticoid circadian rhythm, (2) compare plasma cortisol and corticosterone responses to social stress, and (3) examine strain differences (i.e., WHHL vs. New Zealand White (NZW)) in rabbit glucocorticoid responses to assess whether WHHLs have an aberrant HPA system. It was found that male rabbits secrete both corticosterone and cortisol in a circadian rhythm that peaks in the afternoon and reaches a nadir at 0600 h, i.e., approximately 12 h out-of-phase with the human glucocorticoid rhythm. Both glucocorticoids responded similarly to social stress induced by repeated daily 4 h pairings with another male rabbit; after 10 days of pairings, glucorticoid values were significantly correlated with the amount of defensive agonistic behavior exhibited. Finally, there were no significant strain differences in glucocorticoid circadian rhythms, baselines, or responses to social stress. These data suggest that glucocorticoid responses (i.e., circadian rhythms, responses to social stress) in the WHHL are similar to glucocorticoid responses in standard laboratory white rabbits.

Afferents to the central nucleus of the amygdala and functional subdivisions of the periaqueductal gray: neuroanatomical substrates for affective behavior.

Evidence suggests the periaqueductal gray (PAG) is involved in the integration of behavioral and autonomic components of affective behavior. Our laboratory has shown that electrical stimulation of the ventrolateral periaqueductal gray (vl PAG) versus the dorsolateral periaqueductal gray (dl PAG), in the rabbit, elicits two distinct behavioral/cardiorespiratory response patterns. Furthermore, evidence suggests that the amygdaloid central nucleus (ACe) may influence cardiovascular activity during emotional states. The purpose of this study was to delineate the topography and determine the origin of forebrain projections to the PAG and the ACe, as well as commonalties and differences in the pattern of afferents. Examination of common afferents may lend insights into their function as components of a forebrain system regulating autonomic activity during emotional states. Separate retrograde tracers were injected into functional subdivisions of the PAG and the ACe in rabbits. PAG injections led to neuronal labeling in numerous cortical regions including the ipsilateral medial prefrontal and insular cortices. Additionally, bilateral labeling was observed in several hypothalamic nuclei including the paraventricular nucleus, the dorsomedial nucleus and the ventromedial nucleus as well as the region lateral to the descending column of the fornix. Sparse labeling was also seen in various basal forebrain regions, thalamic nuclei and amygdaloid nuclei. Many of these regions were also labeled following injections in the ACe. Although double-labeled cells were never observed, afferents to the ACe were often proximal to PAG afferents. Implications of these findings are discussed in terms of two functionally distinct behavioral/cardiovascular response patterns.

Secretion of cryparin, a fungal hydrophobin.

Cryparin is a cell-surface-associated hydrophobin of the filamentous ascomycete Cryphonectria parasitica. This protein contains a signal peptide that directs it to the vesicle-mediated secretory pathway. We detected a glycosylated form of cryparin in a secretory vesicle fraction, but secreted forms of this protein are not glycosylated. This glycosylation occurred in the proprotein region, which is cleaved during maturation by a Kex2-like serine protease, leaving a mature form of cryparin that could be isolated from both the cell wall and culture medium. Pulse-chase labeling experiments showed that cryparin was secreted through the cell wall, without being bound, into the culture medium. The secreted protein then binds to the cell walls of C. parasitica, where it remains. Binding of cryparin to the cell wall occurred in submerged culture, presumably because of the lectin-like properties unique to this hydrophobin. Thus, the binding of this hydrophobin to the cell wall is different from that of other hydrophobins which are reported to require a hydrophobic-hydrophilic interface for assembly.

Effects of combined postischemic hypothermia and delayed N-tert-butyl-alpha-pheylnitrone (PBN) administration on histopathologicaland behavioral deficits associated with transient global ischemia in rats.

Previous cerebral ischemia studies have reported the limitations of restricted periods of postischemic hypothermia in producing long-term neuroprotection. The present experiment attempts to determine whether delayed treatment with the free radical scavenger N-tert-butyl-a-phenylnitrone (PBN) is protective at 2 months following transient global forebrain ischemia, and whether additive effects can be observed when PBN is administered in combination with moderate hypothermia. For this aim rats were subjected to 10 min of two-vessel forebrain ischemia followed by (a) 3 h of postischemic normothermia (37 degrees C); (b) 3 h of postischemic hypothermia (30 degrees C); (c) normothermic procedures combined with delayed injections of PBN (100 mg/kg) on days 3, 5 and 7 post-insult; (d) postischemic hypothermia combined with delayed PBN treatment; or (e) sham procedures. Outcome measures included cognitive behavioral testing and quantitative histopathological analysis at 2 months. Postischemic PBN injections induced a systemic hypothermia (1.5 degrees C-2.0 degrees C) that lasted for 2-2.5 h. Water maze testing revealed significant performance deficits relative to shams in the normothermic ischemic group, with the postischemic hypothermia and PBN groups showing intermediate values. A significant attenuation of cognitive deficits was observed in the animal group receiving the combination postischemic hypothermia and delayed PBN treatment. Quantitative CA1 hippocampal cell counts indicated that each of the ischemia groups exhibited significantly fewer viable CA1 neurons compared to sham controls. However, in rats receiving either delayed PBN treatment or 3 h of postischemic hypothermia, significant sparing of CA1 neurons relative to the normothermic ischemia group was observed. These data indicate that hypothermia combined with PBN treatment provides long-term cognitive improvement compared to nontreatment groups. PBN-induced mild hypothermia could contribute to the neuroprotective effects of this pharmacological strategy.

The influence of dsRNA viruses on the biology of plant pathogenic fungi.

Double-stranded RNA viruses are ubiquitous in fungi. They are non-infective and, like most prokaryotic plasmids, are only transmitted to compatible strains via cell fusion. Most are cryptic, but some with an established phenotype, such as the hypoviruses of the chestnut-blight fungus, have been studied for their potential as biological control agents of fungi.

Contribution of NMDA and non-NMDA receptors to synaptic transmission from the brachium of the inferior colliculus to the medial subdivision of the medial geniculate nucleus in the rabbit.

Previous work from this laboratory has demonstrated that monosynaptic inputs from the brachium of the inferior colliculus (BIC) to the medial subdivision of the medial geniculate nucleus (mMG) strengthen as a result of associative conditioning with an acoustic conditioned stimulus (i.e., fear conditioning). One model that has been proposed to underlie certain types of neuronal plasticity involves the recruitment of N-methyl-D-aspartic acid (NMDA)-type glutamate receptors. The purpose of the present study was to examine the relative contributions of glutamatergic NMDA and non-NMDA receptors to synaptic transmission within this pathway. Individual contributions of the specific receptor types were assessed through the use of 2-amino-5-phosphonovaleric acid (AP5), a selective NMDA receptor antagonist, and 6-cyano-5-nitroquinoxaline-2,3-dione (CNQX), a non-NMDA receptor antagonist. Bipolar stimulating electrodes were stereotaxically implanted in BIC and recording electrodes (attached to dual 32-gauge cannulae for delivery of drug) were positioned in mMG of New Zealand albino rabbits. Single pulses (150 micros, 100-350 microA) delivered to BIC resulted in short-latency (<4 ms) responses in mMG. BIC-evoked single-unit activity was recorded from mMG before, during, and at several intervals after injection of AP5, CNQX, and/or artificial cerebrospinal fluid (ACSF). Injection of either AP5 or CNQX, but not ACSF, significantly attenuated the short-latency BIC-evoked responses in the vast majority of cells tested. These findings suggest that the monosynaptic pathway from BIC to mMG is glutamatergic and that this pathway frequently employs NMDA-type receptors during electrically stimulated synaptic transmission. Due to the NMDA receptors' proposed role in plasticity (e.g., long-term potentiation), these results may have implications for understanding the mechanisms of synaptic plasticity observed at this synapse during associative learning.

Cardiovascular and perceptual effects of reporting pain during the foot and forehead cold pressor tests.

In research involving the cold pressor test, a tacit presumption is often made that reporting pain during stimulation is not in itself reactive. This study examined whether, for the foot and forehead cold pressor tests, activities involved in reporting pain may affect (a) the evoked pattern of cardiovascular response, and (b) the magnitude of self-perceived pain. In 40 normotensive college men, increases in systolic blood pressure were greater during test sessions that included verbal ratings of pain, as compared to sessions in which pain was not reported. In contrast to its effect on physiological activation, reporting pain did not significantly alter the participant's perception of the painfulness of the lest, on recollection shortly after the test. We conclude, therefore, that reporting pain during the cold pressor test may impose significant additional demands on the cardiovascular system, but it does not interfere significantly with the processing of nociceptive information.

Functional relationship between the hypothalamic vigilance area and PAG vigilance area.

The vigilance reaction is characterized by a large bradycardia, a pressor response, and inspiratory apnea in anesthetized rabbits and the inhibition of movement in conscious rabbits. This affective response pattern can be elicited by electrical stimulation of the dorsolateral hypothalamus (the hypothalamic vigilance area) or the ventrolateral periaqueductal gray (the periaqueductal gray vigilance area). The present study sought to advance our understanding of the functional relationship between the hypothalamic vigilance area (HVA) and the periaqueductal gray vigilance area (PVA) by measuring the effects of transverse transections of the caudal portion of the ventrolateral PAG (vlPAG) upon the cardiovascular responses elicited from the dorsolateral hypothalamus and the rostral vlPAG. Selective transverse transections of the caudal vlPAG significantly reduced the magnitudes of the bradycardia and pressor response elicited by stimulation of the PVA rostral to the transection site, but had minimal impact on the cardiovascular responses evoked by stimulation of the HVA. These findings suggest that the cardiovascular responses elicited by stimulation of the vlPAG are mediated by a neural pathway that is parallel, at least in part, to the one that subserves the response elicited from the HVA. The results also provide support for the view that the PAG is not an essential structure in the mediation of the autonomic components of affective behaviors involving behavioral inhibition.

Effects of guided imagery and music (GIM) therapy on mood and cortisol in healthy adults.

Healthy adults (N = 28) participated in a randomized trial of Bonny Method of Guided Imagery and Music (GIM; a depth approach to music psychotherapy) sessions on mood and cortisol. Participants in both GIM and wait-list control conditions completed the Profile of Mood States (POMS) and donated 15 cc of blood before and after the 13-week intervention period and again at a 6-week follow-up. Split-plot factorial and post hoc analyses demonstrated that after 6 biweekly sessions GIM participants reported significant decreases between pre- and postsession depression, fatigue, and total mood disturbance and had significant decreases in cortisol level by follow-up. Pretest to follow-up decrease in cortisol was significantly associated with decrease in mood disturbance. A short series of GIM sessions may positively affect mood and reduce cortisol levels in healthy adults. Such changes in hormonal regulation may have health implications for chronically stressed people.

Cardiorespiratory components of defense reaction elicited from paraventricular nucleus.

The present study was conducted to test the hypothesis that the paraventricular nucleus of the hypothalamus (PVN) is involved in the mediation or modulation of the cardiorespiratory components of the defense reaction (DR) in rabbits. Electrical stimulation of the PVN elicited increases in blood pressure and heart rate, hyperventilation, decreased blood flow to the visceral organs, and an increase in blood flow to the hindlimbs that was mediated by an atropine-sensitive vasodilation system. This response pattern is nearly identical to the one that is elicited by electrical stimulation of the hypothalamic defense area. In addition, the cardiomotor component of the baroreceptor reflex was observed to be suppressed during electrical stimulation of the PVN. Previous studies have shown that electrical stimulation of the hypothalamic defense area also leads to inhibition of the cardiomotor component of the baroreceptor reflex. The results of the present study provide evidence that the PVN is involved in the mediation or modulation of the defense reaction.

Behavioral characteristics of defense and vigilance reactions elicited by electrical stimulation of the hypothalamus in rabbits.

An automated tracking system was used to assess the behavioral changes elicited by electrical stimulation of the hypothalamic sites that yield the cardiorespiratory components of defense reaction and vigilance reaction in rabbits. Electrical stimulation of the hypothalamic defense area (HDA) at intensities near threshold led to cessation of body movements coupled with head movements suggesting increased attention to the environment. HDA stimulation at higher intensities evoked agitated running and hindlimb thumping; the amount of running was proportional to stimulus intensity. Electrical stimulation of the hypothalamic vigilance area (HVA) at intensities near threshold elicited orienting behaviors that were similar to those elicited by stimulating the HDA at low suprathreshold current intensities. Stimulation of the hypothalamic vigilance area (HVA) at higher intensities elicited phasic immobility, increased extensor muscle tension, and head tremor. The behavioral changes elicited by HDA and HVA stimulation were accompanied by pupil dilation and exophthalmos.

Modulation of the baroreceptor reflex by stimulation of the hypothalamic defense and vigilance areas.

Electrical stimulation of the hypothalamic defense area (HDA) elicits a pressor/tachycardia response that is believed to prepare an animal for fight or flight. In contrast, electrical stimulation of the hypothalamic vigilance area (HVA) evokes a pressor/bradycardia response that is associated with the inhibition of movement. The differences in the behavioral components of these two affective response patterns suggest differential modulation of the baroreceptor reflex. The present study tested this idea by assessing the effects of electrical stimulation of the HDA and the HVA upon the bradycardia/depressor response elicited by stimulation of the aortic nerve (AN) in rabbits. Concurrent HDA and AN stimulation was observed to attenuate the AN-elicited bradycardia but enhanced the depressor response elicited by AN stimulation. In contrast, concurrent stimulation of the HVA and AN enhanced the bradycardia elicited by AN stimulation but reduced the magnitude of the AN-elicited depressor response. These results provide evidence for differential modulation of the baroreceptor reflex during the defense and vigilance reactions.

Changes of synaptic efficacy in the medial geniculate nucleus as a result of auditory classical conditioning.

In this study we examined inputs to neurons in the medial subnucleus of the medial geniculate nucleus (mMG) for changes of synaptic efficacy associated with heart-rate conditioning to an auditory conditioned stimulus (CS). Conditioning-related changes of synaptic efficacy were measured in awake animals by examining mMG single-unit responses evoked by stimulation of one of two areas that send auditory CS and nonauditory information monosynaptically to the mMG, the brachium of the inferior colliculus (BlC) and the superior colliculus (SC). Synaptic efficacy was measured before, immediately after, and 1 hr after one session of classical conditioning with a tone CS and a corneal airpuff unconditioned stimulus. To determine whether conditioning produced changes of synaptic efficacy on the auditory BlC inputs to mMG cells and not general changes of cellular excitability, analyses of synaptic efficacy were performed on the mMG units that exhibited short-latency evoked responses (< 3.5 msec) to both BlC and SC stimulation. Analyses revealed that the BlC but not the SC test stimulus-evoked unit activity from the same neurons exhibited the following changes immediately after conditioning: decreases in unit response latency, increases in unit response reliability, and increases in spike frequency. BlC-evoked unit responses after pseudoconditioning did not exhibit these changes in unit responding. These results suggest that the synapses carrying auditory CS information to mMG neurons increase in strength as the result of associative conditioning with an acoustic CS. Some of these changes of synaptic efficacy remained 1 hr after training.

Immunohistochemical expression of the c-Fos protein in the spinal trigeminal nucleus following presentation of a corneal airpuff stimulus.

This study examined the expression of the c-Fos protein in the rabbit's central nervous system to determine which areas are activated by the presentation of a corneal airpuff. Previous work has shown that pairing a corneal airpuff unconditioned stimulus (US) with a tone conditioned stimulus (CS) produces reliable heart rate (HR) conditioning. In this study restrained awake rabbits received 100 corneal airpuffs. Brains were then processed immunohistochemically for the c-Fos protein. In animals that received the airpuff the ventral portion of the ipsilateral spinal trigeminal subnucleus caudalis (SVc) and interpolaris (SVi), and the dorsal raphe nucleus exhibited a greater number of c-Fos labeled cells compared to control animals. Another group of animals was given microinjections of WGA-HRP in the medial nucleus of the medial geniculate (mMG) to determine if this critical auditory area of the HR conditioning circuitry receives projections from SVc and SVi. These injections produced retrograde labeling in the same areas of SVc and SVi activated by the airpuff. Thus, a corneal airpuff activates neurons in SVc and SVi which could then activate neurons in mMG. This provides additional evidence that CS and US information converge in mMG.

Combined postischemic hypothermia and delayed MK-801 treatment attenuates neurobehavioral deficits associated with transient global ischemia in rats.

The present study was designed to determine whether postischemic hypothermia, delayed MK-801 (dizocilpine) administration, or a combination of these treatments can provide lasting neurobehavioral protection following transient global ischemia in rats. Rats were subjected to 10 min of normothermic (37 degrees C) ischemia induced by 2-vessel occlusion and hypotension (50 mmHg) or sham procedures. Ischemia was followed by either: (a) 3 h at normothermic brain temperatures, (b) 3 h of postischemic brain hypothermia at 30 degrees C, (c) hypothermia coupled with MK-801 (4 mg/kg, i.p.) on postischemic days 3, 5 and 7, or (d) postischemic MK-801 treatment alone. Neurobehavioral evaluation 6-8 weeks following surgery showed that normothermic ischemia (NI) was associated with water maze navigational deficits, including performance on a simple place task involving finding a hidden platform maintained in one position for 6 days, and a learning set task in which the platform was moved to a different location each day (both P's < 0.02 vs. sham). NI was also associated with increased locomotion in an open field (P < 0.01 vs. sham). A combination of postischemic hypothermia and delayed MK-801 injections provided partial protection from ischemic-associated hyperactivity in the open field (P < 0.02 vs. NI), and robust protection from simple place task deficits (P < 0.02 vs. NI). Evidence for significant protective effects of MK-801 or hypothermia alone was observed in the learning set, during the final trial blocks each day. These results provide further evidence for neuroprotective effects of these treatments at chronic survival intervals, and indicate that the therapeutic window for attenuating ischemic damage is considerably longer than has heretofore been appreciated.

Recovery of vibrissae-dependent behavioral responses following barrelfield damage is not dependent upon the remaining somatosensory cortical tissue.

Previous work has demonstrated that damage to the primary somatosensory cortex produces substantial deficits in a vibrissal cue-dependent discrimination task which recover gradually over the course of post-injury testing. The present study was designed to evaluate the possible site(s) and mechanisms underlying behavioral recovery in this task. Wistar rats were trained under red light in a T-maze to produce ipsilateral turns depending upon the presence of a vibrissal cue. Animals were then subjected to photothrombotic infarctions of either the ipsilateral medial parietal cortex, the ipsilateral primary and secondary somatosensory cortex (SI/SII), the primary and secondary somatosensory cortices of both hemispheres (bilateral SI/SII) or sham surgical procedures. Behavioral testing resumed 24 hours following surgery, and continued for a total of 60 days. The performance of animals with infarcts restricted to the medial parietal cortex did not differ from that of sham-operated controls. Animals with either unilateral or bilateral SI/SII infarcts exhibited a significant decrease in percent correct responding as compared to shams and rats in the medial parietal group. These deficits recovered to pre-infarct levels over approximately 35-40 days. This rate of recovery was slower than the recovery exhibited by animals given medial parietal infarcts which spared the primary barrelfield cortex. The results of this study suggest that neither the contralateral somatosensory cortex nor the vibrissal representation within ipsilateral SII cortex play a critical role in the recovery process. The possibility that subcortical structures underlie the deficits observed following barrelfield cortical damage is discussed.

Simultaneous single unit recording in the medial nucleus of the medial geniculate nucleus and amygdaloid central nucleus throughout habituation, acquisition, and extinction of the rabbit's classically conditioned heart rate.

The present study examined single neuron activity in the medial nucleus of the medial geniculate (mMG) and amygdaloid central nucleus (ACe) simultaneously across several phases of differential heart rate conditioning (habituation, acquisition, and extinction). Within the same animals, the magnitude of mMG and ACe unit responses to two tone conditioned stimuli (CS) exhibited habituation, differential acquisition, and extinction. Neurons in each area developed a differential response latency to the CSs during acquisition, suggesting that mMG and ACe may be involved in changes of synaptic efficacy. Units in both areas rapidly developed a differential response magnitude to the CSs (< 6 acquisition trials), however, mMG units responded to the CSs with a shorter latency than ACe units across all phases of training. This suggests that unlearned and learned CS information may access mMG before ACe. These results are consistent with the notion that conditioning-induced plasticity which occurs in mMG may influence the conditioning-induced plasticity that occurs further downstream in the amygdala.

Destruction of neurons in the VPM thalamus prevents rabbit heart rate conditioning.

The present study examined the role of the ventral posterior medial nucleus of the thalamus (VPM) in classical heart rate (HR) conditioning using an acoustic conditioned stimulus (CS) and a corneal air puff unconditioned stimulus (US). Previous research suggests that VPM neurons are activated during the presentation of a corneal air puff US. Rabbits were given ibotenic acid lesions in the VPM and subjected to one Pavlovian HR conditioning session. The results of the present study demonstrate that destruction of cell bodies in the VPM reduces HR conditioning to the level of a pseudoconditioning control without affecting HR baseline, or orienting responses to the CS. Lesions of the VPM also significantly augment the tachycardiac unconditioned response, suggesting that VPM lesions alter the somatosensory processing of the US.

Comparison of peripheral blood flow patterns associated with the defense reaction and the vigilance reaction in rabbits.

The present study compared the skeletal muscle and visceral blood flow patterns elicited by electrical stimulation of the hypothalamic defense area (HDA) and the hypothalamic vigilance area (HVA) of the rabbit. Electrical stimulation of the HDA evoked a pressor response, tachycardia, hyperventilation, an increase in blood flow to the skeletal muscles and decreased blood flow to visceral organs. Stimulation of the HVA yielded a pressor response, bradycardia, inspiratory apnea and decreased blood flow to both the skeletal muscles and the viscera. Intravenous injections of atropine methyl nitrate significantly reduced the HVA-elicited bradycardia and the HDA-elicited increase in blood flow to the skeletal muscles, thereby providing evidence that the bradycardia was mediated by vagal efferents and that the rabbit has an atropine-sensitive cholinergic vasodilation system. The decrease of blood flow to the visceral organs associated with the defense reaction and vigilance reaction was reversed by intravenous injections of the alpha-1 receptor blocker prazosin.