Sex differences in the effect of amphetamine on immediate early gene expression in the rat dorsal striatum

Amphetamine (AMPH)-induced dopamine release in the striatum and AMPH-induced behavior in the rat have been demonstrated to be influenced by sex and hormonal status. The experiments reported here were conducted, therefore, to examine sex differences, hormonal influences and estrous cycle-dependent changes in AMPH-induced immediate early gene expression in the dorsal striatum. Cell counts were taken at three rostrocaudal levels from three to four regions of the dorsal striatum at each level (ventromedial, dorsomedial, dorsolateral, ventrolateral). The immunohistochemical localization of calbindin was used as a control. We report here that females on the afternoon of proestrus had a significantly greater percent of Fos-positive neurons after AMPH across the dorsolateral region of the middle and caudal striatum and in the ventrolateral region of the caudal striatum compared to females in diestrus, ovariectomized (OVX) females, castrated (CAST) males and intact males. There was no difference in AMPH-induced immediate early gene expression between OVX and diestrous rats. There were also no significant differences between CAST and intact males in AMPH-induced Fos expression, with the exception of the ventrolateral caudal striatum. In sum, the present findings indicate that AMPH-induced Fos expression is sexually dimorphic and modulated by gonadal hormones in lateral regions of the rat dorsal striatum.     

Fos expression induced by changes in arterial pressure is localized in distinct,longitudinally organized columns of neurons in the rat midbrain periaqueductal gray

The distribution of neurons expressing Fos within the periaqueductal gray (FAG) following pharmacologically induced high or low blood pressure was examined to determine (1) if PAG neurons are responsive to changes in arterial pressure (AP) and (2) the relationship of these cells to the functionally defined hypertensive and hypotensive columns in PAG. Changes in AP differentially induced robust Fos expression in neurons confined to discrete, longitudinally organized columns within PAG. Increased AP produced extensive Fos-like immunoreactivity within the lateral PAG, beginning at the level of the oculomotor nucleus. At the level of the dorsal raphe, Fos expression induced by increased AP shifted dorsally, into the dorsolateral division of PAG; this pattern of Fos labeling was maintained throughout the caudal one-third of PAG. Double-labeling for Fos and nicotinamide adenine dinucleotide phosphate diaphorase confirmed that Fos-positive cells induced by increased AP were located in the dorsolateral division of PAG at these caudal levels. Fos positive cells were codistributed, but not colocalized, with nicotinamide adenine dinucleotide phosphate diaphorase-positive cells. Decreased AP evoked a completely different pattern of Fos expression. Fos-positive cells were predominantly located within the ventrolateral PAG region, extending from the level of the trochlear nucleus through the level of the caudal dorsal raphe. Double-labeling studies for Fos and serotonin indicated that only 1–2 double-labeled cells per section were present. Saline infusion resulted in very few Foslike immunoreactive cells, indicating that volume receptor activation does not account for Fos expression in PAG evoked by changes in AP. These results indicate that (1) substantial numbers of PAG neurons are excited by pharmacologically induced changes in AP and (2) excitatory barosensitive PAG neurons are anatomically segregated based on their responsiveness to a specific directional change in AP. © 1995 Wiley-Liss, Inc.     

Predatory hunting and exposure to a live predator induce opposite patterns of Fos immunoreactivity in the PAG

Considering the periaqueductal gray's (PAG) general roles in mediating motivational responses, in the present study, we compared the Fos expression pattern in the PAG induced by innate behaviors underlain by opposite motivational drivers, in rats, namely, insect predation and defensive behavior evoked by the confrontation with a live predator (a cat). Exposure to the predator was associated with a striking Fos expression in the PAG, where, at rostral levels, an intense Fos expression was found largely distributed in the dorsomedial and dorsolateral regions, whereas, at caudal levels, Fos-labeled cells tended to be mostly found in the lateral and ventrolateral columns, as well as in the dorsal raphe nucleus. Quite the opposite, insect predation was associated with increased Fos expression predominantly in the rostral two thirds of the lateral PAG, where the majority of the Fos-immunoreactive cells were found at the oculomotor nucleus levels. Remarkably, both exposure to the cat and insect predation upregulated Fos expression in the supraoculomotor region and the laterodorsal tegmental nucleus. Overall, the present results clearly suggest that the PAG activation pattern appears to reflect, at least partly, the animal's motivational status. It is well established that the PAG is critical for the expression of defensive responses, and, considering the present findings, it will be important to investigate how the PAG contributes to the expression of the predatory behavior, as well.     

Fos-like immunoreactivity in the periaqueductal gray of rats exposed to a natural predator

In the present study we examined, in rats exposed to a predator (cat), the distribution of neurons expressing Fos along the continuum formed by the central gray surrounding the caudal pole of the third ventricle and the periaqueductal gray (PAG). After the predatory encounter, a distinct cluster of Fos-immunoreactive cells was observed in the precommissural nucleus. In the rostral two-thirds of the PAG, Fos expression was mostly seen in the dorsomedial and dorsolateral regions. In contrast, at caudal levels of the PAG, most of the Fos-labelled neurons were distributed in the lateral and ventrolateral PAG. These results are discussed and compared with the pattern of the PAG activation after fear conditioned to a context or elicited by aversive foot shock.     

Induction of c-Fos expression in specific areas of the fear circuitry in rat forebrain by anxiogenic drugs.

BACKGROUND: The fact that induction of anxiety- and panic-related symptoms is a property common to a range of drugs suggests that common neural substrates underlie their behavioral effects. METHODS: We used Fos immunocytochemistry to test the effects of four anxiogenic drugs (FG-7142, yohimbine, m-chlorophenylpiperazine [mCPP], and caffeine) on anxiety-related circuitry in rat forebrain. RESULTS: All four drugs commonly increased Fos-like immunoreactivity in 7 of 41 brain areas investigated, namely, central nucleus of the amygdala, bed nucleus of the stria terminalis, lateral septum, paraventricular nucleus of the hypothalamus, lateral hypothalamus, infralimbic and prelimbic cortex. All drugs but one (mCPP) also increased Fos expression in the basolateral and medial amygdala, the dorsomedial hypothalamus, cingulate cortex, and parts of the motor cortex. CONCLUSIONS: The results suggest that the anxiogenic drugs selected activate a restricted set of forebrain areas. Most of these areas have previously been shown to be activated by environmentally evoked anxiety and to have anatomic connections with hindbrain regions that are activated by the same drugs and by environmentally evoked anxiety. Together, these data are consistent with the theory of an integrated forebrain and hindbrain neuronal system that is important for anxiety states evoked by both drug and environmental manipulations.     

Laryngeal afferent stimulation enhances Fos immunoreactivity in periaqueductal gray in the cat.

The main functions of the larynx are protection of the airways, respiration, and vocalization. Previous studies have suggested a link between the mechanisms controlling vocalization and afferent feedback from the larynx. We inquired whether stimulation of the laryngeal afferents that run in the internal branch of the superior laryngeal nerve (ISLN) activates neurons of the periaqueductal gray (PAG), a midbrain region implicated in vocalization. We counted the number of neurons expressing Fos, the protein product of the immediate early gene c-fos, in the PAG. The counts were done both in experimental cats after electrical stimulation of the ISLN and nonstimulated controls. We also investigated the possible presence of nitric oxide synthase, an enzyme that synthesizes nitric oxide, in PAG neurons that respond to laryngeal afferent stimulation by double labeling for reduced nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase and Fos. Fos expression was significantly greater (P < or = 0.00714) in the lateral and dorsolateral regions of the PAG in the experimental group than in the controls. The Fos-immunoreactive neurons did not contain NADPH-diaphorase, a marker for nitric oxide synthase. Our study suggests that laryngeal afferent stimulation activates neurons in discrete longitudinal columns of the PAG including the regions that have previously been shown to be involved in vocalization, and that these neurons do not contain nitric oxide synthase.     

The cortical endogenous late potential in response to ultrasonic signals in the estrous rat

The evoked responses to 100-ms tone signals of 8, 22, 50 or 70 kHz were recorded from the frontal cortex in the conscious female rat throughout the estrous cycle. The stimulus-bound evoked potentials did not vary with the estrous cycle. At estrus, 50 kHz sounds produced a negative potential peaking at 200 ms in the cortex. In other combinations of sound frequency and estrous stage no late potentials were detected. This finding demonstrates the presence of human 'Event-related potential'-like endogenous potentials in the rat that responds to ultrasonic signals which mimic those used in intraspecific communication.     

Effects of acute and chronic fluoxetine treatments on restraint stress-induced Fos expression

Chronic treatment with antidepressants has been shown to attenuate behavioral changes induced by uncontrollable stress. The mechanisms and brain sites of this effect, however, remain controversial. The objective of the present work was to investigate the effects of chronic and acute treatment with fluoxetine (FLX), a selective serotonin reuptake blocker, on Fos expression in animals submitted to restraint stress. Male Wistar rats (n = 3-9/group) received, during 1 or 21 days, intraperitoneal. Injections of vehicle (saline + 0.2% Tween-80, 1 ml/kg) or FLX (10 mg/kg). One hour after the last injection they were forced restrained for 2 h and sacrificed immediately after. Non-stressed animals were sacrificed 2 h after the last injection. The brains were removed and processed for immunohistochemistry. Fos-like immunoreactivity (FLI) was quantified by a computer system. In acutely treated animals FLX decreased stress-induced FLI in the medial amygdala (MeA), bed nucleus of the stria terminalis (BNST), ventrolateral part, and dorsolateral periaqueductal gray (PAG). After chronic treatment, however, the drug induced a significant increase in FLI in the BNST (ventrolateral and medial parts), lateral septal nucleus (LSN, dorsal part), dorsal raphe nucleus (DRN), and locus coeruleus in restrained group. In non-restrained animals chronic treatment with FLX increased FLI in the MeA, BNST (ventrolateral and dorsolateral parts), LSN (dorsal and intermediate parts), dorsolateral and dorsomedial PAG and in the DRN. The results suggest that chronic fluoxetine treatment induce plastic changes that result in a different regional pattern of Fos expression.     

Disruption of maternal behaviour by acute conspecific interaction induces selective activation of the lateral periaqueductal grey

Maternal behaviour is sensitive to stress and opioidergic activation. The periaqueductal grey (PAG) is involved in coping strategies to stress, whereas morphine inhibition of maternal behaviour depends on the activation of the PAG. The aim of this study was to investigate whether the PAG is activated by disrupting maternal behaviour. Lactating Wistar rats were assigned to four groups: C (control); E1 (acute exposure to a male rat); E2 (daily 2-h exposure to another lactating female and a male rat from Day 3 to 6 of lactation); and E1 + 2 (treated first as E2 and, on Day 9, as E1). Maternal behaviour was recorded on Day 9 of lactation and analysed for 1 h. The E1 group spent more time retrieving their pups, took longer to initiate nursing, had shorter nursing bouts and spent more time in non-maternal activities compared with control. Rats submitted to E2 or E1 + 2 did not differ from the control. In another experiment, lactating rats were treated as above, except that 90 min after the end of the observation period the rats were killed and their brains were processed for immunohistochemical detection of Fos protein in the PAG. Fos increased in the lateral PAG only in the E1 group. We also observed that neurons activated by acute conspecific interaction in the PAG could be responsible for an opioid-dependent decrease in maternal behaviour as this effect was reversed by a microinjection of naltrexone, nor-binaltorphimine or naloxonazine into the lateral PAG. Chronic conspecific interaction alters the way this circuitry responds to acute conspecific interaction.     

Functional specializations within the tectum defense systems of the rat

Here we review the differential contribution of the periaqueductal gray matter (PAG) and superior colliculus (SC) to the generation of rat defensive behaviors. The results of studies involving sine-wave and rectangular pulse electrical stimulation and chemical (NMDA) stimulation are summarized. Stimulation of SC and PAG produced freezing and flight behaviors along with exophthalmus (fully opened bulged eyes), micturition and defecation. The columnar organization of the PAG was evident in the results obtained. Defecation was elicited primarily by lateral PAG stimulation, while the remaining defensive behaviors were similarly elicited by lateral and dorsolateral PAG stimulation, although with the lowest thresholds in the dorsolateral column. Conversely, the ventrolateral PAG did not appear to participate in unconditioned defensive behaviors, which were only elicited by high intensity stimulation likely to encroach on adjacent regions. In the SC, the most important differences relative to the PAG were the lack of stimulation-evoked jumping in both intermediate and deep layers, and of NMDA-evoked galloping in intermediate layers. Therefore, we conclude that the SC may be only involved in the increased attentiveness (exophthalmus, immobility) and restlessness (trotting) of prey species exposed to the cues of a nearby predator. These responses may be distinct from the full-blown flight reaction that is mediated by the dorsolateral and lateral PAG. However, other evidences suggest the possible influences of stimulation schedule, environment dimensions and rat strain in determining outcomes. Overall our results suggest a dynamically organized representation of defensive behaviors in the midbrain tectum.     

Ovarian steroids regulate neuropeptides in the trigeminal ganglion

Women are more than three times as likely as men to experience migraine headaches and temporomandibular joint pain, and painful episodes are often linked to the menstrual cycle. To understand how hormone levels may influence head and face pain, we assessed expression of pain-associated neuropeptides and estrogen receptor alpha (ERalpha) during the natural estrous cycle in mice. Gene expression was analyzed in the trigeminal ganglia of cycling female mice at proestrus, estrus and diestrus using RT-PCR. Peptide/protein expression in trigeminal neurons was analyzed using immunohistochemistry. ERalpha mRNA was present at all stages and highest at estrus. ERalpha protein was present in the cytoplasm of medium-sized and small trigeminal neurons. ERalpha immunoreactive neurons were most common at diestrus. CGRP and ANP mRNAs did not change across the estrous cycle, while expression of galanin and NPY mRNAs were strongly linked to the estrous cycle. Galanin mRNA levels peaked at proestrus, when expression was 8.7-fold higher than the diestrus levels. Galanin immunoreactivity also peaked at proestrus. At proestrus, 7.5% of trigeminal neurons contained galanin, while at estrus, 6.2% of trigeminal neurons contained galanin, and at diestrus, 4.9% of trigeminal neurons contained galanin. NPY mRNA peaked at estrus, when levels were 4.7-fold higher than at diestrus. Our findings suggest that estrogen receptors in trigeminal neurons modulate nociceptive responses through effects on galanin and NPY. Variations in neuropeptide content in trigeminal neurons across the natural estrous cycle may contribute to increases in painful episodes at particular phases of the menstrual cycle.     

Phosphorylated cyclic AMP response element binding protein expression induced in the periaqueductal gray by predator stress: its relationship to the stress experience, behavior and limbic neural plasticity

Electrophysiological studies in cats and recently in rats implicate neuroplasticity in the periaqueductal gray (PAG) and its afferents in stressor-induced increases in fearful behavior and anxiety-like behavior (ALB). Such increases may model aspects of affective changes following traumatic stress in humans. The present study explored the role of neuroplasticity in PAG and its connection with the central nucleus of the amygdala (ACE) in male rodent anxiety-like response to predator stress. In the first of two studies, the effects of predator stress on the induction of phosphorylated cyclic AMP response element binding protein (pCREB) were investigated. pCREB expression in the PAG and ventromedial hypothalamus (VMH) was examined immunohistochemically. Predator stress increased the degree of pCREB expression in PAG cells (measured densitometrically) but did not increase the number of cells expressing pCREB (measured stereologically). Moreover, predator stress-specific increase in pCREB-like immunoreactivity (lir) was restricted to the right lateral column of the PAG. In addition, pCREB lir in the right lateral column likely reflects aspects of the stress experience because the stressor (cat behavior) and the response to the stressor (rat defensive behavior) are highly predictive of degree of pCREB expression. There was no effect of predator stress on pCREB lir in the VMH.

Because pCREB expression has been associated with long-lasting potentiation (LLP) of neural transmission, we examined the effects of predator stress on transmission in the ACE-PAG pathway in a second study. Predator stress elevated evoked potential measures of ACE-PAG transmission in the right hemisphere but not in the left hemisphere 11–12 days after predator stress. This finding is consistent with the longer-lived effects of pharmacological stress on amygdalo-PAG transmission in the right hemisphere but not in the left hemisphere in cats. Of interest is the fact that the same aspects of the stressor experience and reaction to it, which are predictive of the degree of pCREB expression, are also highly predictive of the degree of potentiation of measures of ACE-PAG transmission. Behavioral analyses revealed that the most consistent effects of predator stress are on behavior in the plus maze (open arm exploration and risk assessment) and on startle. In addition, covariance analysis suggests that ACE-PAG potentiation mediates some but not all of the changes in ALB produced by predator stress. Because pCREB expression may be a precursor to neuroplastic changes in certain forms of memory and LLP, the present findings complement studies in the cat, showing that neuroplastic changes in the PAG underlie changes in affect following stress. Furthermore, these findings suggest that neuroplastic changes in PAG may be important mediators of predator stress-induced changes in affective behavior in rodents. Finally, consistent with cat and human studies, the right hemisphere appears particularly important in long-term response to stress.     

Effect of the Estrous Cycle on Olfactory Bulb Response to Vaginocervical Stimulation in the Rat: Results from Electrophysiology and Fos Immunocytochemistry Experiments

To determine whether the stage of the estrous cycle modified the response of olfactory bulb neurons to vaginocervical stimulation, (1) vaginocervical stimulation was applied to animals in proestrus-estrus and metestrus-diestrus and the extracellular electrophysiological response of units in the mitral cell layer of the main olfactory bulb was compared, and (2) the effect of vaginocervical or sham stimulation and the effect of the estrous cycle on the number of neurons stained immunocytochemically for Fos in the main and accessory olfactory bulb was examined. Animals in proestrus-estrus had basal firing rates of 21.8 ± 1.8 spikes per 5 s and vaginocervical stimulation produced an increase in firing rate. In contrast, animals in metestrus-diestrus had a slower basal firing rate (14.3 ± 2.3 spikes per 5 s) and vaginocervical stimulation produced a decrease in the firing rate. For animals in proestrus-estrus, vaginocervical stimulation increased the number of Fos-stained cells in the granular cell layer of the accessory olfactory bulb, and in the glomerular and in external plexiform layers of the main olfactory bulb. In contrast, the number of Fos-stained cells decreased in the granular cell layer of the main olfactory bulb after stimulation was applied to animals in proestrus-estrus. The number of Fos-stained cells in the granular layer of the accessory olfactory bulb and the granular and glomerular cell layers of the main olfactory bulb was modulated by the estrous cycle. Therefore, olfactory bulb activity, measured both electrophysiologically and by Fos staining, was affected by the estrous cycle and vaginocervical stimulation, and the two variables interacted. It is likely that integration of interoceptive and environmental stimulation is important for the normal expression of sexual behavior in the female rat.     

The estrous cycle affects pseudorabies virus (PRV) infection of the CNS

Previous work had suggested that mucosal immunity may be affected by the stage of the estrous cycle. Here, susceptibility to a neurotropic virus infection at different stages of the estrous cycle was assessed in a rodent model after direct injection of the virus into visceral organs. In the first two experiments, female Sprague-Dawley rats were infected with pseudorabies virus (PRV, Bartha's K-strain) by injection into either the cervix or the kidney after monitoring their estrous cycle. After either 4- or 5-day survival period post-infection, the rats were euthanized by transcardially perfusion and peripheral and central nervous system tissues were removed for immunocytochemical staining. The number of infected neurons was counted in various regions. Statistical analysis revealed that: (1) the number of infected cells in the sympathetic or parasympathetic ganglion, or the dorsal root ganglia was not affected regardless of the stage of the estrous cycle after cervix injection with PRV; (2) in contrast, the number of infected neurons in the spinal cord was affected significantly by the stage of the estrous cycle during viral infection of the cervix; (3) after kidney infection, the number of infected neurons found within the spinal cord or dorsal root ganglia varied significantly across the estrous cycle. In both cases, animals infected in proestrus or estrus had fewer infected neurons than animals infected in diestrus I or diestrus II (proestrous and estrous animals had less than 20% of infected cells found in diestrus I or diestrus II rats). In the third experiment, older, persistent estrous or persistent diestrous rats were infected by kidney injection and given a 4-day survival period, prior to virus isolation from lower thoracic spinal cord. Animals in persistent estrous had significantly less virus per gram of tissue than the persistent diestrous rats. These data suggest that the CNS of animals in proestrus or estrus is less susceptible to PRV infection compared to animals in either diestrus I or diestrus II. Because estrogen replacement therapy is known to restore some immune functions during reproductive ageing, it is speculated that plasma estrogen levels modulate the central nervous system's susceptibility to viral infections.     

Endogenous neurotensin is involved in estrous cycle related alterations in prepulse inhibition of the acoustic startle reflex in female rats

Ovarian hormones regulate prepulse inhibition (PPI) of the acoustic startle reflex. Results from studies in intact female rodents investigating sex, estrous cycle and ovarian hormone regulation of PPI are inconsistent. In experiment #1, we investigated whether PPI in female rats is influenced by the time of day of testing and the estrous cycle stage of the rat. PPI was examined across the day of proestrus (P) and diestrus 1 (D1) in female rats and compared to males. PPI in males and P females was significantly higher than in D1 females. PPI in males and D1 females was significantly affected by the time of day of testing with PPI being reduced in the afternoon and evening compared to morning. PPI in P females was not significantly affected by the time of day of testing. Previous studies have demonstrated estrous cycle regulation of central nervous system neurotensin (NT) neurons and peripherally administered NT receptor agonists regulate PPI in a manner similar to antipsychotic drugs. Experiment #2 of this study was designed to examine whether endogenous NT is involved in estrous cycle regulation of PPI. The NT receptor antagonist SR 142948A reduced the high levels of PPI during D1 and P. In contrast, when tested at a time of day in which PPI was low in D1 females, administration of both the typical antipsychotic drug haloperidol and the NT receptor antagonist significantly increased PPI. These data support an effect of time of day and estrous cycle stage on PPI in female rats. The estrous cycle variations in PPI are mediated in part by endogenous NT.     

大鼠导水管周围灰质注射谷氨酸可使Barrington核神经元表达Fos

形态学研究已发现大鼠内侧视前区（ＭＰＯ）和导水管周围灰质（ＰＡＧ）发出大量轴突,投射至与排尿反射密切相关的Ｂａｒｒｉｎｇｔｏｎ 核。本研究试图通过注射谷氨酸钠到ＭＰＯ或ＰＡＧ后,观察Ｂａｒｒｉｎｇｔｏｎ 核内的Ｆｏｓ表达情况,来了解以上两通路的性质。将谷氨酸钠注射到ＭＰＯ后,只有少量Ｆｏｓ阳性神经元出现在Ｂａｒｒｉｎｇ－ｔｏｎ 核。而将谷氨酸钠注射到ＰＡＧ后,Ｂａｒｒｉｎｇｔｏｎ 核内出现大量的Ｆｏｓ阳性神经元。此结果提示,ＰＡＧ可能对大鼠脑桥排尿反射活动具有兴奋性调节作用。     

Variations in opioid peptide levels during the estrous cycle in Sprague-Dawley rats

The estrous cycle, with its various hormonal conditions, may provide us with the means of understanding how endocrine states relate to opioid mechanisms. There has been increasing experimental support for interaction between sex steroids and opioid peptides in the central nervous system. Here, we describe fluctuations in endogenous brain immunoreactive (ir) peptide levels during various phases of the estrous cycle in the female Sprague-Dawley rat. Ir levels of dynorphin A, dynorphin B, Leu-enkephalin-Arg(6), Met-enkephalin-Arg(6)Phe(7) and nociceptin/orphanin FQ were measured in the pituitary gland and in 10 areas of the brain during the diestrus, proestrus and estrus phase. In several areas of the brain, basal levels of endogenous opioid peptides showed variation during the course of the estrous cycle. Significant differences were found between the diestrus state and the proestrus and/or estrus conditions, particularly in the nucleus accumbens, caudate putamen and the substantia nigra. The ir levels of the endogenous peptide nociceptin/orphanin FQ became altered in only one of the areas measured, indicating less variance during the estrous cycle. Correlation analyses revealed that significant associations between dynorphin A or dynorphin B and Leu-enkephalin-Arg(6) were found more often during estrus than during the diestrus and proestrus conditions. The ratio between the ir levels of Leu-enkephalin-Arg(6), a cleavage product of the enzymatic conversion of dynorphin peptides into shorter peptides in vivo, and dynorphin peptides was calculated. The significantly lower ratio between Leu-enkephalin-Arg(6) and dynorphin B in diestrus than in proestrus and estrus also indicates cyclic fluctuations in the enzymatic cleavage of dynorphin. These findings are discussed in relation to the possible role of interactions between sex steroids and opioid peptide mechanisms during the normal estrous cycle.     

Kappa opioid receptors in rat spinal cord vary across the estrous cycle

Kappa opioid receptors (KORs) were immunocytochemically localized in the lumbosacral spinal cord of female rats in different stages of the estrous cycle to examine the influence of hormonal status on receptor density. KOR labeling was primarily in fine processes and a few neuronal cell bodies in the superficial dorsal horn and the dorsolateral funiculus. Quantitative light microscopic densitometry of the superficial dorsal horn revealed that rats in diestrus had significantly lower KOR densities than those in proestrus or estrus. This suggests that female reproductive hormones regulate spinal KOR levels, which may contribute to variations in analgesic effectiveness of KOR agonists across the estrous cycle.     

Antidepressant treatment reduces Fos-like immunoreactivity induced by swim stress in different columns of the periaqueductal gray matter

Antidepressant treatment attenuates behavioral changes induced by uncontrollable stress. The periaqueductal gray matter (PAG) is proposed to be a brain site involved in the behavioral responses to uncontrollable stress and antidepressant effects. The main goal of the present study was to investigate the effect of antidepressant treatment on the pattern of neural activation of the PAG along its mediolateral and rostrocaudal subregions after a forced swim stress episode. Male Wistar rats were sub-acutely treated with desipramine (a selective noradrenaline re-uptake blocker, three injections of 10 mg/kg in 24 h) or clomipramine (a non-selective serotonin and noradrenaline re-uptake blocker, three injections of 10 mg/kg in 24 h) and submitted to the forced swimming test (FST). Two hours after the test their brain were removed for Fos immunohistochemistry. Fos-like immunoreactivity (FLI) in rostral, intermediate and caudal portions of dorsomedial (dmPAG), dorsolateral (dlPAG), lateral (lPAG) and ventrolateral (vlPAG) PAG were quantified by a computerized system. The FST session increased FLI in most parts of the PAG. Previous treatment with desipramine or clomipramine reduced FLI in all columns of the PAG. FLI in the PAG correlated positively with to the immobility time and negatively with to climbing behavior scored during the test. These results indicate that neurons in the PAG are activated by uncontrollable stress. Moreover, inhibitory action of antidepressants on this activity may be associated with the anti-immobility effects of these drugs in the FST.     

The effects of septal stimulation on spontaneous and tail-shock evoked neuronal activity in the brainstem of the rat

The effects of stimulation of the lateral septum (LS) and the medial septum (MS) with single pulses of current were assessed on the neural activity evoked by a noxious tail shock as well as on spontaneous activity in 72 cells located in the periventricular gray (PVG), periaqueductal gray (PAG), dorsal raphe (DR) and nucleus raphe magnus (NRM) of the anesthetized rat. Pronounced inhibitory effects of evoked activity, occurring with a 10–15 ms latency and lasting 20–100 ms, were found primarily in the DR and ventrolateral PAG. Inhibition was not confined to nociceptive cells, and could also be observed in some units unresponsive to noxious stimulation. Short-lived excitatory effects were encountered mostly in the PVG and dorsolateral PAG units. The results were interpreted as support for the notion of septal participation in pain inhibition, and the hypothesis was advanced that its mode of action may involve suppression of aspects of nociception at the supraspinal level.