Blast exposure causes dynamic microglial/macrophage responses and microdomains of brain microvessel dysfunction.

Exposure to blast overpressure (BOP) is associated with behavioral, cognitive, and neuroimaging abnormalities. We investigated the dynamic responses of cortical vasculature and its relation to microglia/macrophage activation in mice using intravital two-photon microscopy following mild blast exposure. We found that blast caused vascular dysfunction evidenced by microdomains of aberrant vascular permeability. Microglial/macrophage activation was specifically associated with these restricted microdomains, as evidenced by rapid microglial process retraction, increased ameboid morphology, and escape of blood-borne Q-dot tracers that were internalized in microglial/macrophage cell bodies and phagosome-like compartments. Microdomains of cortical vascular disruption and microglial/macrophage activation were also associated with aberrant tight junction morphology that was more prominent after repetitive (3×) blast exposure. Repetitive, but not single, BOPs also caused TNFα elevation two weeks post-blast. In addition, following a single BOP we found that aberrantly phosphorylated tau rapidly accumulated in perivascular domains, but cleared within four hours, suggesting it was removed from the perivascular area, degraded, and/or dephosphorylated. Taken together these findings argue that mild blast exposure causes an evolving CNS insult that is initiated by discrete disturbances of vascular function, thereby setting the stage for more protracted and more widespread neuroinflammatory responses.

Characterization of verbal and spatial memory changes from moderate to supraphysiological increases in serum testosterone in healthy older men.

BACKGROUND: It has been suggested that cognitive changes in response to T supplementation may occur within an ideal range. The objective of this study was to compare the cognitive responses of older, eugonadal men in whom moderate or large increases in serum testosterone levels was induced by exogenous testosterone supplementation. DESIGN: Randomized, double-blind, placebo-controlled study with subsequent grouping of participants according to average increase in circulating T from baseline. SETTING: Community dwelling participants. PARTICIPANTS: Fifty-seven healthy, eugonadal, community dwelling male volunteers, mean age 67 years (+/-11 years). INTERVENTIONS: Participants were randomized to receive weekly intramuscular (i.m.) injections of either 50, 100 or 300 mg T enanthate or placebo (saline) injection for 6 weeks. Cognitive evaluations using a battery of neuropsychological tests were conducted at baseline, weeks 3 and 6 of treatment and after 6 weeks of wash-out. MAIN OUTCOME MEASURES: Performance on cognitive tests of verbal and spatial memory. RESULTS: Men with moderate increases in serum T and/or its metabolites demonstrated significant improvements in verbal and spatial memory. In contrast, men with large or low increases in circulating T levels, failed to demonstrate significant changes in memory. CONCLUSION: These results suggest that in healthy older men, beneficial changes in cognitive function induced by T supplementation are most evident with moderate changes in cognition from moderate to high T supplementation increases in T levels. Large or no to low increases in T levels do not appear to appreciably effect cognition.

Testosterone improves spatial memory in men with Alzheimer disease and mild cognitive impairment.

OBJECTIVE: To determine the efficacy of testosterone (T) supplementation on cognition in a sample of men with Alzheimer disease (AD) or mild cognitive impairment (MCI). METHODS: Fifteen patients with AD and 17 patients with MCI aged 63 to 85 years completed a randomized, double-blind, placebo-controlled study. Nineteen participants received weekly intramuscular (IM) injections of 100 mg T enanthate and 13 participants received weekly injections of placebo (saline) for 6 weeks. Cognitive evaluations using a battery of neuropsychological tests were conducted at baseline, week 3, and week 6 of treatment and again after 6 weeks of washout. RESULTS: Peak serum total T levels were raised from baseline an average of 295% in the active treatment group. Improvements in spatial memory (p < 0.05) and constructional abilities (p < 0.05) and verbal memory were evident in the T group. No changes were noted for selective and divided attention or language. Prostate specific antigen did not significantly change during this brief treatment. CONCLUSION: Testosterone supplementation may benefit selective cognitive functions in men with Alzheimer disease and mild cognitive impairment.

The role of aromatization in testosterone supplementation: effects on cognition in older men.

OBJECTIVE: To determine the contribution of conversion of testosterone (T) to estradiol on cognitive processing in a population of healthy older men who received T supplementation. METHODS: Sixty healthy, community-dwelling volunteers aged 50 to 90 years completed a randomized, double-blind, placebo-controlled study. Participants were randomized to receive weekly IM injections of 100 mg T enanthate plus daily oral placebo pill (T group, n = 20), 100 mg testosterone enanthate plus 1 mg daily of anastrozole, an aromatase inhibitor (oral pill), to block the conversion of T to estradiol (AT group, n = 19), or saline injection and placebo pill (placebo group, n = 21) for 6 weeks. Cognitive evaluations using a battery of neuropsychological tests were conducted at baseline, week 3 and week 6 of treatment, and after 6 weeks of washout. RESULTS: Circulating total T was increased from baseline an average of 238% in the T and AT treatment groups. Estradiol increased an average of 81% in the T group and decreased 50% in the AT group during treatment. Significant improvements in spatial memory were evident in the AT and T treatment groups. However, only the group with elevated estradiol levels (T group) demonstrated significant verbal memory improvement. CONCLUSION: In healthy older men, improvement in verbal memory induced by testosterone administration depends on aromatization of testosterone to estradiol, whereas improvement in spatial memory occurs in the absence of increases in estradiol.

Chronic cortisol suppresses pituitary and hypothalamic peptide message expression in pigtailed macaques.

The effects of chronic elevations in circulating glucocorticoids on the expression of peptides and peptide receptors of the hypothalamic-pituitary-adrenal (HPA) axis have been studied extensively in rodents, but they have not been examined in primates. To determine the responses of the HPA axis in primates to elevated cortisol, hypothalamic and pituitary tissue from normal older pigtailed macaques (Macaca nemestrina) that had received daily oral administration of cortisol or placebo for 1 year were studied. Pro-opiomelanocortin in the anterior pituitary and corticotropin-releasing factor (CRF) mRNA expression in the hypothalamic paraventricular nucleus (PVN) were significantly reduced in cortisol-treated monkeys in comparison with controls. CRF receptor 1 (CRF-R1) expression in the anterior pituitary and arginine vasopressin mRNA expression in the PVN were unchanged by chronic cortisol administration. Sustained elevation of circulating glucocorticoids results in suppression of HPA peptide and peptide receptor expression in the PVN and anterior pituitary similar to those found in rodents. Chronic therapeutic administration of glucocorticoids in humans may have unintended consequences for hypothalamic and pituitary function.

High-dose estradiol improves cognition for women with AD: results of a randomized study.

OBJECTIVE: To characterize the cognitive and neuroendocrine response to treatment with a high dose of estrogen for postmenopausal women with AD. METHODS: Twenty postmenopausal women with AD were randomized to receive either 0.10 mg/day of 17 beta-estradiol by skin patch or a placebo patch for 8 weeks. Subjects were evaluated at baseline, at weeks 3, 5, and 8 during treatment, and again 8 weeks after treatment termination. During each visit, cognition was assessed with a battery of neuropsychological tests, and blood samples were collected to measure plasma estradiol as well as several other neuroendocrine markers of interest. RESULTS: Significant effects of estrogen treatment were observed on attention (Stroop Color Word Interference Test), verbal memory (Buschke Selective Reminding Test), and visual memory (Figure Copy/Memory). In addition, women treated with estrogen demonstrated improved performance on a test of semantic memory (Boston Naming Test) compared with subjects who received a placebo. Estrogen appeared to have a suppressive effect on the insulin-like growth factor (IGF) system such that plasma concentration of IGF binding protein-3 was significantly reduced and plasma levels of estradiol and IGF-I were negatively correlated during estrogen treatment. CONCLUSIONS: Administration of a higher dose of estrogen may enhance attention and memory for postmenopausal women with AD. Although these findings provide further clinical evidence to support a cognitive benefit of estrogen for women with AD, studies evaluating the effect of estradiol administration, in particular, using larger sample sizes and for longer treatment durations are warranted before the therapeutic potential of estrogen replacement for women with AD can be firmly established.

The alpha1-adrenergic antagonist prazosin ameliorates combat trauma nightmares in veterans with posttraumatic stress disorder: a report of 4 cases.

BACKGROUND: Central nervous system (CNS) adrenergic hyperresponsiveness may be involved in the pathophysiology of posttraumatic stress disorder (PTSD). Two Vietnam combat veterans with PTSD prescribed the centrally active alpha1-adrenergic antagonist prazosin for symptoms of benign prostatic hypertrophy unexpectedly reported elimination of combat trauma nightmares. This observation prompted an open-label feasibility trial of prazosin for combat trauma nightmares in chronic combat-induced PTSD. METHOD: Four consecutively identified combat veterans with chronic DSM-IV PTSD and severe intractable combat trauma nightmares participated in an 8-week open trial of escalating-dose prazosin. Nightmare severity response was rated using the nightmare item of the Clinician Administered PTSD Scale and the Clinical Global Impressions-Change scale. RESULTS: The 2 patients who achieved a daily prazosin dose of at least 5 mg were markedly improved, with complete elimination of trauma nightmares and resumption of normal dreaming. The 2 subjects limited to 2 mg of prazosin to avoid excessive blood pressure reduction were moderately improved with at least 50% reduction in nightmare severity. CONCLUSION: These clinical observations, together with neurobiological evidence for alpha1-adrenergic regulation of CNS neurobiological systems relevant to PTSD, provide rationale for placebo-controlled trials of prazosin for PTSD combat trauma nightmares.

Cognitive and neuroendocrine response to transdermal estrogen in postmenopausal women with Alzheimer's disease: results of a placebo-controlled, double-blind, pilot study.

Preliminary evidence from clinical studies indicates that treatment with estrogen augments cognitive function for women with Alzheimer's disease (AD). The neurobiology of estrogen, particularly its neuromodulatory and neuroprotective actions, provide a viable basis to support such cognition-enhancing effects. We conducted a placebo-controlled, double-blind, parallel-group design pilot clinical study to evaluate the cognitive and neuroendocrine response to estrogen administration for postmenopausal women with AD. Twelve women with probably AD of mild-moderate severity completed the study. During an eight week treatment period, six women received 0.05 mg/day dosage of 17 beta-estradiol via a skin patch and the remaining six wore a placebo skin patch. Subjects were randomized to equal distribution, and evaluated at baseline, at weeks 1, 3, 5, and 8 on treatment, and at weeks 9, 10, 11, and 13 off treatment. On each day of evaluation, cognition was assessed using a battery of neuropsychological tests, and blood samples were collected to measure plasma concentrations of estradiol and estrone. In addition, several neuroendocrine markers were measured in plasma to evaluate the relationship between estrogen-induced cognitive effects and fluctuations in the catecholaminergic and insulin-like growth factor systems. Significant effects of estrogen treatment were observed on attention (i.e. Stroop: number of self-corrections in the Interference condition, F[1,8] = 8.22, P < 0.03) and verbal memory (i.e., Buschke: delayed cued recall, F[3,30] = 4.31, P < 0.02). The salutary effects of estrogen on cognition were observed after the first week of treatment, and started to diminish when treatment was terminated. For women treated with estrogen, enhancement in verbal memory was positively correlated with plasma levels of estradiol (r = 0.96, P < 0.02) and negatively correlated with concentrations of insulin-like growth factor binding protein-3 (IGFBP-3) in plasma (r = -0.92, P < 0.03). Furthermore, a trend in the data was evident to suggest a negative relationship between plasma levels of insulin-like growth factor-1 (IGF-1) and verbal memory (r = -0.86, P = 0.06). Estrogen administration suppressed peripheral markers of the IGF system, as evidenced by a negative correlation between plasma concentration of estradiol and IGF-1 (r = -0.93, P < 0.03), and a trend for a similar relationship between plasma levels of estradiol and IGFBP-3 (r = -0.86, P = 0.06). With respect to the catecholamines assayed, norepinephrine was positively correlated with verbal memory (r = 0.95, P < 0.02) for women who were treated with estrogen. Furthermore, there was a trend to suggest a negative relationship between plasma epinephrine levels and the number of errors committed on a test of attention (r = -0.84, P = 0.07). In the placebo group, no significant effects of estrogen replacement were evident either on measures of cognition or on any of the neuroendocrine markers. The results of this study suggest that estrogen replacement may enhance cognition for postmenopausal women with AD. Furthermore, several markers of neuroendocrine activity may serve to index the magnitude of estrogen-induced facilitation on cognition. In addition, research findings from the present study will provide important information for the design of larger prospective clinical studies that are essential to definitively establish the therapeutic role of estrogen replacement for postmenopausal women with AD.

Estrogen regulates galanin but not tyrosine hydroxylase gene expression in the rat locus ceruleus.

The neuropeptide galanin (GAL) is coexpressed by the majority of noradrenergic neurons in the rat locus ceruleus (LC) and may function as an inhibitory modulator of noradrenergic transmission. Because estrogen has been shown to induce GAL expression in other brain regions and modulate noradrenergic transmission, we used in situ hybridization histochemistry to assess the effects of chronic estrogen treatment on GAL and tyrosine hydroxylase (TH) gene expression in the LC of ovariectomized female rats. We found that GAL mRNA levels were significantly elevated in rats implanted with a Silastic capsule containing estradiol compared to sham-implanted controls. Both the average optical density (P < or = 0.05) and the labelling area (P < or = 0.007) differed significantly between the groups. In contrast, TH gene expression measured in alternate brain sections did not differ between the groups. If GAL functions as an inhibitory modulator of noradrenergic transmission as postulated, these findings suggest that chronic estrogen treatment could reduce the noradrenergic tone of the brain in the absence of significant alterations in TH expression by enhancing the level of cosecreted GAL.

Nerve growth factor induces galanin gene expression in the rat basal forebrain: implications for the treatment of cholinergic dysfunction.

Nerve growth factor (NGF) is a potential treatment for cholinergic dysfunction associated with Alzheimer's disease (AD). In rats, NGF activates gene expression of the acetylcholine synthetic enzyme choline acetyltransferase (ChAT) and prevents age- and lesion-induced degeneration of basal forebrain (BF) cholinergic neurons. Cholinergic neurons in the BF coexpress galanin (GAL), a neuropeptide that has been shown to impair performance on memory tasks possibly through the inhibition of cholinergic memory pathways. NGF up-regulates both ChAT and GAL gene expression in cultured pheochromocytoma cells; however, the effect of chronic in vivo NGF administration on GAL gene expression within the BF has not been studied. We used in situ hybridization and quantitative autoradiography to assess GAL and ChAT gene expression within the BF of adult male rats following chronic intracerebroventricular infusion of NGF or cytochrome c. We now report that, in addition to stimulating ChAT gene expression, NGF strongly up-regulated the GAL gene in the rat cholinergic BF. NGF had no effect on GAL gene expression in other noncholinergic forebrain regions. NGF induction of GAL gene expression in the BF was specific, because gene expression for another neuropeptide, neurotensin, present within noncholinergic BF neurons was unchanged. Our data provide the first evidence that in vivo NGF administration up-regulates GAL gene expression in the cholinergic BF. These results suggest that the concurrent induction of GAL in the BF could limit the ameliorating actions of NGF on cholinergic dysfunction.

Galanin-binding sites in the female rat brain are regulated across puberty yet similar to the male pattern in adulthood.

The neuropeptide galanin (GAL) has been implicated in a variety of neuroendocrine functions and has been shown to be regulated by gonadal hormones in several brain regions. We have used slice binding and quantitative autoradiography techniques to determine whether the activation of GAL pathways across puberty in female rats is associated with changes in the density of GAL binding in telencephalic and diencephalic regions as we previously observed in male rats. We have also asked whether sex differences in GAL immunoreactivity and GAL gene expression detected in some brain regions would be paralleled by sex differences in 125I-GAL-binding density in adult male and female rat brains. To control for intrinsic differences in the level of endogenous GAL synthesis and release, brain slices from prepubertal female and adult male and female rats were treated with guanosine 5'-triphosphate (GTP) to induce dissociation of endogenous GAL from its binding sites prior to incubation with radiolabeled ligand. 125I-GAL binding was significantly reduced in seven brain regions of adult compared with prepubertal female rats. These regions included the islands of Calleja (p < or = 0.03), the medial amygdaloid nucleus, posterodorsal division (p < or = 0.05), median eminence (p < or = 0.02), medial habenular nucleus (p < or = 0.05), rhomboid thalamic nucleus (p < or = 0.05), and paraventricular (p < or = 0.05) and intermediodorsal (p < or = 0.02) thalamic nuclei. Only one region, the lateral preoptic area, exhibited significantly enhanced 125I-GAL binding in adult female (p < or = 0.04) compared with prepubertal animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Sex difference in coexpression by galanin neurons accounts for sexual dimorphism of vasopressin in the bed nucleus of the stria terminalis.

Vasopressin (VP) neurons in the bed nucleus of the stria terminalis (BNST) are steroid sensitive and sexually dimorphic. The number of VP messenger RNA (mRNA)-expressing neurons is larger in male than in female rats. This initial observation suggested that sexual dimorphism resulted from enhanced proliferation and/or survival of VP neurons after gonadal hormone exposure during the critical perinatal period. However, galanin (GAL) and VP mRNAs were recently reported to be coexpressed in the BNST of adult male rats, and GAL gene expression, unlike VP gene expression, is not sexually dimorphic. These findings are consistent with the hypothesis that the sex difference in VP cell number in the BNST results from a sex difference in the number of GAL neurons dedicated to express the VP gene. To test this hypothesis, double in situ hybridization histochemistry was performed for GAL and VP mRNAs in the BNST of adult male and female rats. For quantification, the posterior BNST was divided into its two anatomical regions: medial (BSTM) and lateral (BSTL) divisions. Extending previous results for the whole BNST, the number of GAL-expressing cells in either the BSTM or the BSTL was not sexually dimorphic. A significant sex difference was found in the number of GAL cells coexpressing VP in the BSTM (mean +/- SE, male, 124 +/- 8; female, 56 +/- 6; P < or = 0.0001), but not in the BSTL (male, 80 +/- 9; female, 83 +/- 15). Accordingly, the number of cells expressing GAL mRNA only was significantly lower (P < or = 0.002) in the BSTM of male (43 +/- 5) than in female (85 +/- 9) rats. Evidence is provided that the reduced incidence of coexpression of VP by GAL neurons in the BSTM of female rats may account for the reported sex difference in VP cell number in the entire BNST. The results suggest that gonadal hormones in the perinatal period may not influence the proliferation and/or survival of VP neurons in the BNST per se but influence, instead, the capacity of GAL neurons to synthesize VP.

Activation of galanin pathways across puberty in the male rat: galanin gene expression in the bed nucleus of the stria terminalis and medial amygdala.

Galanin and vasopressin are coexpressed in the bed nucleus of the stria terminalis and medial amygdala of the male rat. In adult males, the level of gene expression for both peptides in these regions is dependent on circulating levels of testosterone. We hypothesized that galanin messenger RNA levels would be enhanced in adult males compared with prepubertal males due to the rise in plasma testosterone levels. We used in situ hybridization and quantitative autoradiography to measure galanin messenger RNA in cells of the bed nucleus of the stria terminalis and medial amygdala of prepubertal and adult male rats. Our results show that significantly (P < or = 0.05) more galanin messenger RNA expressing neurons are detectable in the bed nucleus of the stria terminalis of adult compared with prepubertal male rats. In contrast, no differences were observed between the groups in the number of labeled neurons detected within the medial amygdala. However, the average labeling intensity was significantly enhanced in both the bed nucleus of the stria terminalis (P < or = 0.001) and medial amygdala (P < or = 0.001) of adult compared with prepubertal animals. The present findings are consistent with the hypothesis that gonadal hormones regulate galanin gene expression in some brain regions and suggest that the activation of the hypothalamic-pituitary-gonadal axis which occurs naturally with puberty is associated with activation of galanin pathways in the bed nucleus of the stria terminalis and medial amygdala.

Activation of galanin pathways across puberty in the male rat: assessment of regional densities of galanin binding sites.

Galanin-like immunoreactivity and galanin messenger RNA levels increase across puberty in neurons of gonadal steroid-dependent brain nuclei. We hypothesized that this activation and the associated increase in endogenous galanin release would result in changes across puberty in both galanin binding density and the level of receptor occupancy. Here we have assessed the density of galanin binding sites in several brain regions of prepubertal and adult male rats with or without GTP to induce dissociation of endogenous galanin from its binding sites. The developmental changes in the level of receptor occupancy were used as an indirect measure of changes in neuropeptide release from galanin expressing neurons. In standard binding conditions (buffer preincubation), 125I-labeled galanin binding showed a generalized decline in adult brains (34-68%) compared with prepubertal levels in most regions of the telencephalon and diencephalon. Following preincubation with 10(-5) M GTP, galanin binding showed a dramatic increase in most regions of the adult (152-504%) and several regions of the prepubertal brain (132-245%) over their standard binding levels. However, this increase was greatest in adult animals. Finally, although preincubation of brain slices with GTP eliminated most of the apparent age-related differences observed in standard binding conditions, several brain regions of the adult brain continued to show a significant reduction (38-76%) in 125I-labeled galanin binding compared with prepubertal animals. Only one region, the lateral preoptic area, exhibited enhanced 125I-labeled galanin binding in adult (160%) compared with prepubertal brain after GTP preincubation.(ABSTRACT TRUNCATED AT 250 WORDS)

Estrogen receptor and neurotensin/neuromedin-N gene expression in the preoptic area are unaltered with age in Fischer 344 female rats.

The sensitivity of hypothalamic centers to estrogenic regulation may be impaired with age and contribute to the loss of reproductive function in female rats. Here, we have tested the hypothesis that aging is associated with alterations in the level of expression of the estrogen receptor (ER) gene and/or the neurotensin/neuromedin-N (NT/N) gene in the preoptic area (POA) of female rats. We have used in situ hybridization histochemistry and quantitative autoradiography to compare ER gene expression and NT/N gene expression in the POA of ovariectomized and ovariectomized/estradiol-treated female rats at 3, 11, and 20 months of age. We found no evidence for an age-related impairment of either ER or NT/N gene expression in two subdivisions of the POA: the anterior medial preoptic nucleus and the medial preoptic nucleus. Likewise, estrogenic regulation of both ER messenger RNA levels and NT/N messenger RNA levels did not differ across age groups. These results indicate that transcription of the ER gene within the POA is not reduced with age and suggest that the receptor translated within the POA functions normally in old female rats. Our observations do not support a role for impaired expression of the ER gene or impaired estrogenic induction of NT/N gene expression by preoptic neurons in the development of reproductive acyclicity with aging.

Galanin in the bed nucleus of the stria terminalis and medial amygdala of the rat: lack of sexual dimorphism despite regulation of gene expression across puberty.

Neurons in the bed nucleus of the stria terminalis (BNST) and the medial amygdala (AMe) coexpress vasopressin and galanin (GAL) in the adult male rat. Here, we have asked whether GAL gene expression, like vasopressin gene expression in these same neurons, exhibits sexual dimorphism and whether GAL pathways in the BNST and AMe are activated with puberty in female rats as we have previously observed in male rats. In Exp 1, in situ hybridization histochemistry and quantitative autoradiography were used to compare GAL gene expression in the BNST and AMe of prepubertal (24-day-old) and adult (90-day-old) male and female rats. In the BNST, both the number of GAL mRNA-expressing neurons (F = 41.98; P < or = 0.0001; males, P < or = 0.007; females, P < or = 0.001) and the intensity of labeling (F = 40.35; P < or = 0.0001; males, P < or = 0.004; females, P < or = 0.002) were significantly increased in adult compared to prepubertal animals of both sexes. In the AMe of both males (P < or = 0.001) and females (P < or = 0.001), the intensity of labeling was significantly enhanced across puberty (F = 66.29; P < or = 0.0001); however, the number of GAL mRNA-expressing neurons in this region did not change. We found no evidence for sexual dimorphism of GAL gene expression in either brain region. In Exp 2, we replicated our observations of a lack of sexual dimorphism of GAL gene expression in the BNST of adult male and female rats. These findings are consistent with the hypothesis that GAL neurons in the BNST and AMe are steroid sensitive in both sexes. However, our failure to detect any differences in either the number of GAL mRNA-expressing neurons or the level of expression between male and female rats at either age indicates that these pathways do not exhibit sexual dimorphism.

Testosterone regulates galanin gene expression in the bed nucleus of the stria terminalis.

The regulation of galanin (GAL) gene expression in the bed nucleus of the stria terminalis (BNST) by testosterone (T) was investigated using in situ hybridization histochemistry. Castration of adult male rats significantly reduced both the number of cells which expressed GAL mRNA and the average number of grains per cell. These effects were reversed by testosterone treatment. Testosterone stimulates GAL gene expression in the same neurons that have previously been shown to exhibit steroid regulation of vasopressin gene expression.

Extra-hypothalamic vasopressin neurons coexpress galanin messenger RNA as shown by double in situ hybridization histochemistry.

Vasopressin (VP) neurons in the bed nucleus of the stria terminalis (BNST) and medial amygdala (AMe) exhibit sexual dimorphism and steroid dependency. VP neurons in the supraoptic nucleus and paraventricular nucleus have been shown to coexpress other transmitters including galanin (GAL). However, little is known about what other neurotransmitters may be colocalized with VP in the BNST and AMe. Here, we have used radio-labeled and digoxigenin-labeled cRNA probes to perform double in situ hybridization histochemistry for VP and GAL in the BNST and AMe of intact, adult male rats. We provide evidence that in the basal state, the majority of VP-synthesizing cells in the BNST and AMe of the adult male rat also express galanin mRNA. Likewise, the majority of GAL-expressing neurons in these regions also contain VP mRNA. These findings give further evidence for the similarity of the BNST and AMe and provide a rationale for studies investigating the role of GAL in functions involving extrahypothalamic VP pathways.

Polysymptomatic complaints and Briquet's syndrome in polycystic ovary disease.

To test the hypothesis that there is an association between polycystic ovary disease and Briquet's syndrome, the authors administered a health questionnaire to infertile women with polycystic ovary disease, infertile women with tubal disease, and normal women. The patients with polycystic ovary disease endorsed significantly more physical and psychological complaints than either control group. Structured interviews revealed that five of the 39 (13%) met diagnostic criteria for definite or probable Briquet's syndrome. This study gives support to an association between polysymptomatic complaints, Briquet's syndrome, and polycystic ovary disease.