Sex steroid hormones in depressive disorders as a basis for new potential treatment strategies.

The sex steroid hormones (SSHs) such as testosterone, estradiol, progesterone, and their metabolites have important organizational and activational impacts on the brain during critical periods of brain development and in adulthood. A variety of slow and rapid mechanisms mediate both organizational and activational processes via intracellular or membrane receptors for SSHs. Physiological concentrations and distribution of SSHs in the brain result in normal brain development. Nevertheless, dysregulation of hormonal equilibrium may result in several mood disorders, including depressive disorders, later in adolescence or adulthood. Gender differences in cognitive abilities, emotions as well as the 2-3 times higher prevalence of depressive disorders in females, were already described. This implies that SSHs may play a role in the development of depressive disorders. In this review, we discuss preclinical and clinical studies linked to SSHs and development of depressive disorders. Our secondary aim includes a review of up-to-date knowledge about molecular mechanisms in the pathogenesis of depressive disorders. Understanding these molecular mechanisms might lead to significant treatment adjustments for patients with depressive disorders and to an amelioration of clinical outcomes for these patients. Nevertheless, the impact of SSHs on the brain in the context of the development of depressive disorders, progression, and treatment responsiveness is complex in nature, and depends upon several factors in concert such as gender, age, comorbidities, and general health conditions.

Rapid actions of oestrogens and their receptors on memory acquisition and consolidation in females.

Increased attention has been paid in recent years to the ways in which oestrogens and oestrogen receptors rapidly affect learning and memory. These rapid effects occur within a timeframe that is too narrow for the classical genomic mode of action of oestrogen, thus suggesting nonclassical effects as underlying mechanisms. The present review examines recent developments in the study of the rapid effects of 17ß-oestradiol and oestrogen receptor (ER) agonists on learning and memory tasks in female rodents, including social recognition, object recognition, object placement (spatial memory) and social learning. By comparing studies utilising systemic or intracranial treatments, as well as pre- and post-acquisition administration of oestradiol or ER agonists, the respective contributions of individual ERs within specific brain regions to various forms of learning and memory can be determined. The first part of this review explores the effects of systemic administration of 17ß-oestradiol and ER agonists on memory when administered either pre- or post-acquisition. The second part not only focuses on the effects of pre- and post-acquisition infusions of 17ß-oestradiol or ER agonists into the dorsal hippocampus on memory, but also discusses the contributions of other brain regions, including the medial amygdala, medial prefrontal cortex and paraventricular nucleus of the hypothalamus. The cellular mechanisms mediating the rapid effects of 17ß-oestradiol on memory, including activation of intracellular signalling cascades and epigenetic processes, are discussed. Finally, the review concludes by comparing pre- and post-acquisition findings and effects of 17ß-oestradiol and ER agonists in different brain regions.

Epigenetics, oestradiol and hippocampal memory consolidation.

Epigenetic alterations of histone proteins and DNA are essential for hippocampal synaptic plasticity and cognitive function, and contribute to the aetiology of psychiatric disorders and neurodegenerative diseases. Hippocampal memory formation depends on histone alterations and DNA methylation, and increasing evidence suggests that the regulation of these epigenetic processes by modulatory factors, such as environmental enrichment, stress and hormones, substantially influences memory function. Recent work from our laboratory suggests that the ability of the sex-steroid hormone 17ß-oestradiol (E2 ) to enhance novel object recognition memory consolidation in young adult female mice is dependent on histone H3 acetylation and DNA methylation in the dorsal hippocampus. Our data also suggest that enzymes mediating DNA methylation and histone acetylation work in concert to regulate the effects of E2 on memory consolidation. These findings shed light on the epigenetic mechanisms that influence hormonal modulation of cognitive function, and may have important implications for understanding how hormones influence cognition in adulthood and ageing. The present review provides a brief overview of the literature on epigenetics and memory, describes in detail our findings demonstrating that epigenetic alterations regulate E2 -induced memory enhancement in female mice, and discusses future directions for research on the epigenetic regulation of E2 -induced memory enhancement.

Dose-dependent effects of post-training estradiol plus progesterone treatment on object memory consolidation and hippocampal extracellular signal-regulated kinase activation in young ovariectomized mice.

Previous work from our laboratory has shown that the ability of estradiol to enhance object memory consolidation in young ovariectomized mice is dependent on dorsal hippocampal activation of the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) signaling pathway [Fernandez SM, Lewis MC, Pechenino AS, Harburger LL, Orr PT, Gresack JE, Schafe GE, Frick KM (2008) Estradiol-induced enhancement of object memory consolidation involves hippocampal extracellular signal-regulated kinase activation and membrane-bound estrogen receptors. J Neurosci 28:8660-8667]. However, it is unclear if estradiol modulates memory or ERK activation similarly in the presence of progesterone. Therefore, the present study investigated effects of combined estradiol and progesterone treatment on object memory consolidation and dorsal hippocampal ERK activation in young ovariectomized C57BL/6 mice. Object memory was tested in a novel object recognition task. Immediately after training, mice received intraperiotoneal (i.p.) injections of vehicle, 17beta-estradiol (E(2); 0.2 mg/kg), or E(2) plus 5, 10, or 20 mg/kg progesterone (P). Forty-eight hours later, mice receiving E(2) alone or E(2) plus 10 or 20 mg/kg P exhibited significantly enhanced memory for the novel object relative to chance, whereas those receiving vehicle or E(2) plus 5 mg/kg P spent no more time than chance with the novel object. Two weeks later, ERK phosphorylation was measured in the dorsal hippocampus 1 h after i.p. injection of vehicle, E(2), or E(2) plus P. Consistent with our previous work [Fernandez SM, Lewis MC, Pechenino AS, Harburger LL, Orr PT, Gresack JE, Schafe GE, Frick KM (2008) Estradiol-induced enhancement of object memory consolidation involves hippocampal extracellular signal-regulated kinase activation and membrane-bound estrogen receptors. J Neurosci 28:8660-8667], E(2) alone significantly increased phospho-p42 ERK protein levels in the dorsal hippocampus relative to vehicle controls. In contrast, no combination of E(2) and P affected dorsal hippocampal phospho-ERK levels. These data indicate that, unlike E(2) alone, the beneficial effects of combined E(2) plus P treatment on memory are not associated with ERK activation in the dorsal hippocampus 1 h after treatment, and suggest that E(2) alone and combined E(2) plus P may influence ERK activation in different time frames or enhance memory through different mechanisms.

Sex differences in contextual fear conditioning are associated with differential ventral hippocampal extracellular signal-regulated kinase activation.

Although sex differences have been reported in hippocampal-dependent learning and memory, including contextual fear memories, the underlying molecular mechanisms contributing to such differences are not well understood. The present study examined the extent to which sex differences in contextual fear conditioning are related to differential activation of the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK), a protein kinase critically involved in memory formation. We first show that male rats exhibit more long-term retention of contextual fear conditioning than female rats. During a tone test, females spent more time freezing than males, although both sexes exhibited robust retention of auditory fear learning. Using Western blot analysis, we then show that phosphorylated ERK levels in ventral, but not dorsal, hippocampus are higher in males than females, relative to same-sex controls, 60 minutes after fear conditioning. Post-conditioning increases in ERK activation were observed in the amygdala in both males and females, suggesting a selective effect of sex on hippocampal ERK activation. Together, these findings suggest that differential activation of the ERK signal transduction pathway in male and female rats, particularly in the ventral hippocampus, is associated with sex differences in contextual fear.

Environmental enrichment reduces the mnemonic and neural benefits of estrogen.

The degree to which memory is enhanced by estrogen replacement in postmenopausal women may depend on environmental factors such as education. The present study utilized an animal model of environmental enrichment to determine whether environmental factors influence the mnemonic and neural response to estrogen. Female mice were raised in standard (SC) or enriched (EC) conditions from weaning until adulthood (7 months). All mice were ovariectomized at 10 weeks, and tested in object recognition and water-escape motivated radial arm maze (WRAM) tasks at 6 months. Each day at the completion of training, mice received injections of 0.1 mg/kg cyclodextrin-encapsulated 17-beta-estradiol (E2), 0.2 mg/kg E2, or cyclodextrin vehicle (VEH). At the completion of behavioral testing, hippocampal levels of the presynaptic protein synaptophysin and of brain-derived neurotrophic factor (BDNF) were measured. Enrichment effects were evident in VEH-treated mice; relative to SC-VEH females, EC-VEH females committed fewer working memory errors in the WRAM and exhibited increased hippocampal synaptophysin levels. Estrogen effects depended on environmental conditions. E2 (0.2 mg/kg) improved object memory only in SC females. The same dose improved working memory in SC females, but somewhat impaired working memory in EC females. Furthermore, both doses reduced hippocampal synaptophysin levels in EC, but not SC, females. In contrast, E2 reduced hippocampal BDNF levels in SC, but not EC, females. This study is the first to compare the effects of estrogen on memory and hippocampal function in enriched and non-enriched female mice. The results suggest that: (1) estrogen benefits object and working memory more in mice raised in non-enriched environments than in those raised in enriched environments, and (2) the changes induced by estrogen and/or enrichment may be associated with alterations in hippocampal synaptic plasticity.

Estrogen replacement improves spatial reference memory and increases hippocampal synaptophysin in aged female mice.

Estrogen deficiency during menopause is often associated with memory dysfunction. However, inconsistencies regarding the ability of estrogen to improve memory in menopausal women highlight the need to evaluate, in a controlled animal model, the potential for estrogen to alleviate age-related mnemonic decline. The current study tested whether estrogen could ameliorate spatial reference memory decline in aged female mice. At the conclusion of testing, levels of the presynaptic protein synaptophysin, and activities of the synthetic enzymes for acetylcholine and GABA, were measured in the hippocampus and neocortex. Aged (27-28-month-old) female C57BL/6 mice were given daily subcutaneous injections of 1 microg or 5 microg of beta-estradiol-3-benzoate dissolved in sesame oil. Control mice received daily injections of sesame oil or no injections. Estradiol treatment began 5 days prior to behavioral testing and continued throughout testing. Spatial and non-spatial memory were assessed in the Morris water maze. The 5 microg dose of estradiol significantly improved spatial learning and memory in aged females. The performance of 5 microg females improved significantly more rapidly than that of control females; estradiol-treated females performed at asymptotic levels by session 2. Furthermore, 5 microg females exhibited a more robust spatial bias than controls during probe trials. In contrast, 1 microg of estradiol did not improve spatial task performance. Neither dose affected performance of the non-spatial task. In the hippocampus, synaptophysin was increased in 5 microg females relative to controls. Estrogen did not affect enzyme activities in either brain region. This study is the first to examine the effects of estrogen replacement on spatial reference memory and synaptophysin expression in aged post-estropausal female rodents. The results suggest that: (1) estrogen can profoundly improve spatial reference memory in aged females, and (2) this improvement may be related to increased hippocampal synaptic plasticity, but not modulation of the synthetic enzymes for acetylcholine and GABA.

Sex differences in neurochemical markers that correlate with behavior in aging mice.

Sex differences in neurochemical markers that correlate with behavior in aging mice NEUROBIOL AGING. We examined whether the enzymatic activities of choline acetyltransferase (ChAT) and glutamic acid decarboxylase (GAD) were altered similarly with age in male and female mice, and whether these changes were correlated with age-related alterations in memory and anxiety. ChAT and GAD activities were measured in neocortex, hippocampus, and striatum of behaviorally characterized male and female C57BL/6 mice (5, 17, and 25 months). Generally, ChAT activity was increased, and GAD activity decreased, with age. However, disparate changes were revealed between the sexes; activities of both enzymes were decreased in 17-month males, whereas alterations in females were not observed until 25-months. Furthermore, enzyme-behavior correlations differed between the sexes; in males, ChAT activity was related to one behavioral task, whereas in females, activities of both enzymes were correlated with multiple tasks. Significant enzyme-behavior correlations were most evident at 17 months of age, likely the result of behavioral and enzymatic sex differences at this age. These data represent the first comprehensive report illustrating differential alterations of ChAT and GAD activities in aging male and female mice.

Spatial reference memory and neocortical neurochemistry vary with the estrous cycle in C57BL/6 mice.

Estrous cycle-related variations of spatial reference memory and neurochemistry in intact female mice were examined. Spatial reference memory was tested in cycling females, ovariectomized (OVX) females, and males by using a 1-day water maze protocol. Choline acetyltransferase (ChAT) and glutamic acid decarboxylase (GAD) activities were measured in the hippocampus and neocortex. Estrus females exhibited worse spatial acquisition and 30-min retention than did proestrus and metestrus females, higher neocortical ChAT activity than proestrus females, and higher neocortical GAD activity than OVX females and males. Neocortical, rather than hippocampal, neurochemistry was more sensitive to hormonal modulation, suggesting that hormonal mediation of neocortical function may play a critical role in regulating spatial reference memory in female mice.

Mice are not little rats: species differences in a one-day water maze task.

The present study examined species differences in spatial and non-spatial memory in the Morris water maze. Male Wistar rats and C57BL/6 mice were tested in a one-day water maze task in which spatial learning, retention, and non-spatial learning were assessed within 3 h. Rats and mice appeared to use different strategies for locating the hidden escape platform. Whereas rats evinced a clear spatial strategy, mice appeared to rely less on spatial cues and more on alternative non-spatial strategies. The sensitivity of this behavioral protocol to subtle species differences highlights the potential use of this one-day water maze task as a tool for evaluating rapidly learning and memory in rodents.

Reference memory, anxiety and estrous cyclicity in C57BL/6NIA mice are affected by age and sex.

Age-related changes in learning and memory are common in rodents. However, direct comparisons of the effects of aging on learning and memory in both males and females are lacking. The present study examined whether memory deteriorates with increasing age in C57BL/6NIA mice, and whether age-related changes in learning and memory are similar in both sexes. Male and female mice (five, 17 and 25 months of age) were tested in a battery of behavioral tasks including the Morris water maze (spatial and non-spatial reference memory), simple odor discrimination (olfactory reference memory), plus maze (anxiety/exploration), locomotor activity, and basic reflexes. Five-month-old mice learned the water maze and odor discrimination tasks rapidly. Relative to five-month-old mice, 25-month-old mice exhibited impaired spatial and olfactory reference memory, but intact non-spatial reference memory. The spatial reference memory of 17-month-old mice was also impaired, but less so than 25-month mice. Seventeen-month-old mice exhibited intact non-spatial (visual and olfactory) reference memory. Five and 25-month-old mice had similar levels of plus maze exploration and locomotor activity, whereas 17-month-old mice were more active than both groups and were slightly less exploratory than five-month-old mice. Although sex differences were not observed in the five- and 25-month groups, 17-month-old females exhibited more impaired spatial reference memory and increased anxiety relative to 17-month-old males. Estrous cycling in females deteriorated significantly with increased age; all 25-month-old females had ceased cycling and 80% of 17-month-old females displayed either irregular or absent estrous cycling. This study is the first to directly compare age-related mnemonic decline in male and female mice. The results suggest that: (i) aged mice exhibit significant deficits in spatial and olfactory reference memory relative to young mice, whereas middle-aged mice exhibit only a moderate spatial memory deficit and; (ii) spatial reference memory decline begins at an earlier age in females than in males, a finding that may be related to the cessation of estrous cycling.

Cholinergic basal forebrain is critical for social transmission of food preferences.

Studies using selective lesions of basal forebrain cholinergic neurons suggest that these neurons play a role in attentional processing, but not learning and memory. However, the tests of learning and memory used thus far have been restricted largely to spatial tasks. In the present study, we examined whether the cholinergic basal forebrain plays a role in a form of nonspatial associative memory, the social transmission of food preferences. Sham-operated control rats were compared to rats with 192 IgG-saporin lesions of the medial septum/diagonal band cholinergic projections to hippocampus or nucleus basalis magnocellularis/substantia innominata cholinergic projections to neocortex. Both lesions impaired 24-h retention of a learned social food preference relative to controls, despite performance on an immediate retention trial that was indistinguishable from controls. Moreover, 24-h retention of the socially learned food preference correlated strongly with cholinergic enzymatic activity in the neocortex, but not in the hippocampus. Immunohistochemical data confirmed significant and selective lesion-induced cholinergic depletions in the intended brain regions. These data provide evidence that the cholinergic basal forebrain, particularly the cholinergic projection to neocortex, is involved in the formation and/or retrieval of social memories related to food preference, and suggest a role for cortical acetylcholine in consolidation of associative memory processes.

Presynaptic markers of cholinergic function in the rat brain: relationship with age and cognitive status.

The nature of age-related changes in cholinergic function and their relationship to age-related behavioral decline were examined in the present study. Male Fischer-344 rats of four ages (four, 11, 17 and 23 months) were tested in a battery of cognitive tasks. Discrete microdissections of brain areas involved in cognitive function were performed, and activity of choline acetyltransferase and levels of hemicholinium-3 binding were determined to assess the integrity of cholinergic innervation. Age-related changes in cholinergic markers occurred predominantly in the medial septal area and its target areas (hippocampus and cingulate cortex), and were also present in the posterior caudate. However, most of the age-related changes in cholinergic markers were already present at ages at which behavioral impairment was not yet maximal. There were some consistent correlations between behavioral and neurochemical measures, independent of age, but these accounted for relatively small proportions of variance in behavioral performance. For most of these correlations, lower levels of presynaptic cholinergic markers were related to better behavioral performance. In brain areas in which correlations changed with age, lower levels of presynaptic cholinergic markers were associated with better performance in young rats, whereas higher levels were associated with better performance in aged rats. Recent lesion studies using a toxin selective for basal forebrain cholinergic neurons have suggested that these neurons do not play as central a role in learning and memory in young and aged animals as was previously thought. When considered in this context, the present results suggest that preserved cholinergic function in old age might act indirectly to sustain cognitive ability. Changes in cholinergic function may represent one of a number of age-related neurobiological events that underlie behavioral impairments, or may be a permissive factor for other age-related processes that are more directly responsible for cognitive impairments.

The effects of nerve growth factor on spatial recent memory in aged rats persist after discontinuation of treatment.

Nerve growth factor (NGF) infusion significantly reduces spatial recent memory deficits in aged rats, an effect that has great relevance to the treatment of memory impairments characteristic of patients with Alzheimer's disease. The present study was designed to examine whether this NGF-induced improvement in spatial recent memory persists after the discontinuation of NGF treatment, an issue of crucial importance for the potential clinical use of this compound. Spatial recent memory was tested in a Morris water maze delayed nonmatch-to-position task. In addition to memory, sensorimotor skills were also examined. Four- and 22-month-old rats were tested preoperatively, infused intraventricularly with recombinant human NGF or vehicle, and tested both during the 4 week infusion period and during the 4 weeks after discontinuation of the infusion. NGF significantly improved spatial recent memory in 22-month-old rats only, during the 4th week of infusion and for up to 4 weeks after discontinuation of the infusion. Although NGF did not affect overall sensorimotor skills during infusion in either age group, sensorimotor skills were significantly improved both 2 and 4 weeks after discontinuation of infusion in 22-month-old rats. These findings demonstrate that the beneficial effects of NGF on spatial recent memory can persist for up to 1 month after discontinuation of infusion and suggest that NGF can be used intermittently for the treatment of age-associated memory dysfunction and Alzheimer's disease.

Oxotremorine infusions into the medial septal area of middle-aged rats affect spatial reference memory and ChAT activity.

Age-related spatial memory deficits are correlated with septohippocampal cholinergic system degeneration. The present study examined the effect of intraseptal infusions of the cholinergic agonist, oxotremorine, on spatial reference memory in middle-aged rats using place discrimination in the water maze, and on cholinergic activity using choline acetyltransferase (ChAT) activity. Oxotremorine mildly improved the rate of place discrimination acquisition of middle-aged rats during initial sessions only, but did not affect asymptotic levels of performance achieved. Of the brain regions assayed, ChAT activity increased with age in the temporal cortex and dorsal CA2/3 region of the hippocampus. Oxotremorine significantly decreased ChAT activity in the dorsal hippocampus. In contrast to our previous results in aged rats indicating a more robust effect of oxotremorine on spatial working memory, the present results suggest a modest effect of intraseptal oxotremorine on the acquisition of a spatial reference memory task.

Age-related spatial reference and working memory deficits assessed in the water maze.

Aged rats have spatial memory deficits relative to young rats. The extent of these deficits in intermediate-aged rats is not well established. The present study examined the pattern of age-related changes in spatial reference and working memory in four ages of Fischer-344 rats. Place discrimination (PD) in the Morris water maze measured spatial reference memory. Repeated acquisition (RA), a discrimination in which the escape platform location varied from session to session, measured spatial working memory. Fischer-344 rats, 4 months, 11 months, 17 months, and 24 months of age, were tested. Compared to 4-month-olds, 24-month-olds were significantly impaired on all six PD measures of performance, 17 months were significantly impaired on five PD measures, and 11 months were significantly impaired on only one PD measure. Only 24-month-olds had a significant working memory impairment in RA relative to 4 months. Reference and working memory measures were distinct as assessed by a principal components analysis. The results indicate a nonlinear age-related spatial memory decline in Fischer-344 rats from 4 to 24 months of age.

D-cycloserine, a novel cognitive enhancer, improves spatial memory in aged rats.

D-cycloserine, a partial agonist of the NMDA receptor-associated glycine site, can enhance cognition. The present experiment examines the behavioral effects of D-cycloserine on cognitive deficits in male Fischer-344 rats, 24 months old. Rats 24 months old (n = 42) received either vehicle or one of 3 doses of D-cycloserine prior to testing. Young rats, 4 months old (n = 13), received vehicle prior to testing. Place discrimination and repeated acquisition were tested in the water maze and a variety of sensorimotor tasks were given. Aging impaired performance in all tasks. D-cycloserine improved performance in place discrimination and repeated acquisition. No doses affected sensorimotor function. These results support the hypothesis that D-cycloserine has cognition enhancing properties and that it may be useful in treating disorders involving cognitive impairment.

Acetylcholine release in the hippocampus: effects of cholinergic and GABAergic compounds in the medial septal area.

The medial septal area (MSA) contains cholinergic and GABAergic neurons that send projections to the hippocampus. These neurons have both cholinergic and GABAergic receptors. This study was designed to determine the effects of intraseptal infusions of cholinergic and GABAergic drugs, which alter mnemonic processes, on hippocampal acetylcholine (ACh) release. Hippocampal ACh release was assessed using in vivo microdialysis and HPLC-EC. Oxotremorine and scopolamine produced a dose-dependent decrease in hippocampal ACh release. Muscimol decreased hippocampal ACh release at both high and low doses, although not in a dose-dependent manner. The effects of scopolamine and muscimol are consistent with a role of ACh in mnemonic processing.