Hippocampal Neurotransmission Research Articles

Magnetic resonance spectroscopy and tissue protein concentrations together suggest lower glutamate signaling in dentate gyrus in schizophrenia.

Hippocampal dysfunction in schizophrenia is widely acknowledged, yet the mechanism of such dysfunction remains debated. In this study we investigate the excitatory and inhibitory hippocampal neurotransmission using two complementary methodologies, proton magnetic resonance spectroscopy (MRS) and tissue biochemistry, sampling individuals with schizophrenia in vivo and postmortem hippocampal tissue in vitro. The results show significantly lower glutamate concentrations in hippocampus in schizophrenia, an in vivo finding mirrored by lower GluN1 protein levels selectively in the dentate gyrus (DG) in vitro. In a mouse model with a DG knockout of the GRIN1 gene, we further confirmed that a selective decrease in DG GluN1 is sufficient to decrease the glutamate concentrations in the whole hippocampus. Gamma-aminobutyric acid (GABA) concentrations and GAD67 protein were not significantly different in hippocampus in schizophrenia. Similarly, GABA concentrations in the hippocampi of mice with a DG knockout of the GRIN1 gene were not significantly different from wild type. These findings provide strong evidence implicating the excitatory system within hippocampus in the pathophysiology of schizophrenia, particularly indicating the DG as a site of pathology.

Simultaneous quantification of seven hippocampal neurotransmitters in depression mice by LC-MS/MS.

There is no method available to simultaneously detect GABA, Glu, Epi, NE, DA, 5-HT and 5-HIAA in mouse hippocampus. A rapid and sensitive LC-MS/MS method has been developed for simultaneously measuring seven neurotransmitters in mouse hippocampus. The analytes were detected in positive mode with multiple reaction monitoring (MRM) and the procedure was completed in less than 9min. This method exhibited excellent linearity for all of the analytes with regression coefficients higher than 0.99, and showed good intra- and inter-day precisions (RSD<15%) with good accuracy (80-120%). Moreover, the method was successfully applied for the quantitative determination of neurotransmitters in a mouse depression model induced by successive methylprednisolone injections. The results indicated that this depression model was closely associated with the decreased level of Epi (p=0.002) and elevated ratio of 5-HIAA/5-HT (p=0.01), which has never been reported elsewhere. Compared with previous methods, current approach is more convenient without any pre-column derivatization of the analytes but enhances detectability with incremental neurotransmitter profile and shortens detection time. This work represents the first accurate simultaneous determination of seven neurotransmitters in the mouse depression model induced by methylprednisolone. The reliable method will benefit the research of neurological diseases with the altered neurotransmitter profile in brain.

Effect of realgar on extracellular amino acid neurotransmitters in hippocampal CA1 region determined by online microdialysis–dansyl chloride derivatization–high-performance liquid chromatography and fluorescence detection.

An online microdialysis (MD)–dansyl chloride (Dns) derivatization–high-performance liquid chromatography (HPLC) and fluorescence detection (FD) system was developed for simultaneous determination of eight extracellular amino acid neurotransmitters in hippocampus. The MD probe was implanted in hippocampal CA1 region. Dialysate and Dns were online mixed and derivatized. The derivatives were separated on an ODS column and detected by FD. The developed online system showed good linearity, precision, accuracy and recovery. This online MD-HPLC system was applied to monitor amino acid neurotransmitters levels in rats exposed to realgar (0.3, 0.9 and 2.7 g/kg body weight). The result shows that glutamate concentrations were significantly increased (p<0.05) in hippocampal CA1 region of rats exposed to three doses of realgar. A decrease in γ-aminobutyric acid concentrations was found in rats exposed to medium and high doses of realgar (p<0.05). Elevation of excitotoxic index (EI) values in hippocampal CA1 region of realgar-exposed rats was observed (p<0.05). Positive correlation was found between EI values and arsenic contents in hippocampus of realgar-exposed rats, which indicates that the change in extracellular EI values is associated with arsenic accumulation in hippocampus. The developed online MD–Dns derivatization–HPLC–FD system provides a new experimental method for studying the effect of toxic Chinese medicines on amino acid neurotransmitters.

The effects of chronic ethanol self-administration on hippocampal 5-HT1A receptors in monkeys

BackgroundChronic alcohol consumption reduces brain serotonin and alters the synaptic mechanisms involved in memory formation. Hippocampal 5-HT1A receptors modulate these mechanisms, but the neuroadaptive response of 5HT1A receptors to chronic alcohol self-administration is not well understood. MethodsHippocampal tissue from monkeys that voluntarily self-administered ethanol for 12 months (n=9) and accompanying controls (n=8) were prepared for in vitro receptor autoradiography and laser capture microdissection. The 5-HT1A receptor antagonist, [3H]MPPF, and the agonist, [3H]8-OH-DPAT, were used to measure total and G-protein coupled 5-HT1A receptors respectively. The expression of the genes encoding the 5-HT1A receptor and its trafficking protein Yif1B was measured in microdissected dentate gyrus (DG) granule cells and CA1 pyramidal neurons. ResultsAn increase in G-protein coupled, but not total, receptors was observed in the posterior pyramidal cell layer of CA1 in ethanol drinkers compared to controls. Chronic ethanol self-administration was also associated with an up-regulation of total and G-protein coupled 5-HT1A receptors in the posterior DG polymorphic layer. Changes in receptor binding were not associated with concomitant changes in 5-HT1A receptor mRNA expression. Chronic ethanol self-administration was associated with a significant increase in Yif1B gene expression in posterior CA1 pyramidal neurons. ConclusionsChronic, ethanol self-administration up-regulates hippocampal 5-HT1A receptor density in a region-specific manner that does not appear to be due to alterations at the level of transcription but instead may be due to increased receptor trafficking. Further exploration of the mechanisms mediating chronic ethanol-induced 5-HT1A receptor up-regulation and how hippocampal neurotransmission is altered is warranted.

Diabetes Alters KIF1A and KIF5B Motor Proteins in the Hippocampus

Diabetes mellitus is the most common metabolic disorder in humans. Diabetic encephalopathy is characterized by cognitive and memory impairments, which have been associated with changes in the hippocampus, but the mechanisms underlying those impairments triggered by diabetes, are far from being elucidated. The disruption of axonal transport is associated with several neurodegenerative diseases and might also play a role in diabetes-associated disorders affecting nervous system. We investigated the effect of diabetes (2 and 8 weeks duration) on KIF1A, KIF5B and dynein motor proteins, which are important for axonal transport, in the hippocampus. The mRNA expression of motor proteins was assessed by qRT-PCR, and also their protein levels by immunohistochemistry in hippocampal slices and immunoblotting in total extracts of hippocampus from streptozotocin-induced diabetic and age-matched control animals. Diabetes increased the expression and immunoreactivity of KIF1A and KIF5B in the hippocampus, but no alterations in dynein were detected. Since hyperglycemia is considered a major player in diabetic complications, the effect of a prolonged exposure to high glucose on motor proteins, mitochondria and synaptic proteins in hippocampal neurons was also studied, giving particular attention to changes in axons. Hippocampal cell cultures were exposed to high glucose (50 mM) or mannitol (osmotic control; 25 mM plus 25 mM glucose) for 7 days. In hippocampal cultures incubated with high glucose no changes were detected in the fluorescence intensity or number of accumulations related with mitochondria in the axons of hippocampal neurons. Nevertheless, high glucose increased the number of fluorescent accumulations of KIF1A and synaptotagmin-1 and decreased KIF5B, SNAP-25 and synaptophysin immunoreactivity specifically in axons of hippocampal neurons. These changes suggest that anterograde axonal transport mediated by these kinesins may be impaired in hippocampal neurons, which may lead to changes in synaptic proteins, thus contributing to changes in hippocampal neurotransmission and to cognitive and memory impairments.

Vesicular storage of glycine in glutamatergic terminals in mouse hippocampus

Glycine acts as a neuromodulator to regions rich in glutamatergic synapses, such as the forebrain. However, recent evidences for synaptic release of glycine in hippocampal cultured neurons and synaptosomes argue for the existence of functional glycinergic synapses in the hippocampus. It is well established that GABA and glycine act in concert at inhibitory synapses, while the existence of synapses which utilize both glutamate and glycine is less common. The purpose of the present study was to investigate the distribution of glycine and its role in hippocampal neurotransmission. Using immunohistochemistry, we demonstrate that vesicular glycine is preferentially stored in glutamatergic, rather than GABAergic presynaptic terminals. Using the sniffer patch technique, we found that glycine could be released upon presynaptic activity. Furthermore, using whole-cell patch-clamp recordings, we show for the first time the presence of a postsynaptic strychnine-sensitive chloride current in response to presynaptic stimulation. The small amplitude of this current is likely due to the paucity of postsynaptic glycine receptors rather than a low level of glycine release. Taken together, our results suggest that glycine is stored in glutamatergic presynaptic terminals. It is likely that the major role of glycine that is released from presynaptic terminals is to modulate N-methyl-d-aspartate receptor function but may also play a role in decreasing neuronal excitability by opposing glutamatergic neurotransmission in pathological states such as epilepsy or ischemia.

Evidências neuroquímicas e comportamentais do efeito ansiogênico da deltametrina em ratos

Pyrethroid insecticides are extensively used for pest control around the house, flea prevention for pets, and plant sprays for the home and in agriculture. Deltamethrin (DTM) is a Type II pyrethroid insecticide used to control a variety of insects in agriculture and domestic environments. The present study investigated the possible anxiogenic effects of DTM (1, 3, and 10 mg/kg) in rats using behavioral and neurochemical methods. We assessed general locomotor activity and behavior in the elevated plus maze and open field test. Striatal and hippocampal neurotransmitter and metabolite levels were also measured. DTM (i) reduced locomotion and rearing frequency, (ii) slightly increased the duration of immobility, (iii) reduced the time engaged in social interaction, (iv) reduced the percentage of entries into and time spent on the open arms of the elevated plus maze, (v) reduced the number of center crossings in the elevated plus maze, (vi) Striatal and hippocampal neurotransmitter and metabolite levels were also measured. DTM (i) reduced locomotion and rearing frequency, (ii) slightly increased the duration of immobility, (iii) reduced the time engaged in social interaction, (iv) reduced the percentage of entries into and time spent on the open arms of the elevated plus maze, (v) reduced the number of center crossings in the elevated plus maze, (vi) increased striatal serotonin neurotransmitter and its metabolite, and (vii) did not alter motor coordination on the rotarod, grooming duration in the open field test, rectal temperature, or hippocampal neurotransmitter levels. These data suggest that DTM at the present doses and under these experimental conditions presented a similar profile to that of anxiogenic drugs, unrelated with the increased serotonin neurotransmission.

Globular and Protofibrillar Aβ Aggregates Impair Neurotransmission by Different Mechanisms

In Alzheimer's disease, substantial evidence indicates the causative role of soluble amyloid β (Aβ) aggregates. Although a variety of Aβ assemblies have been described, the debate about their individual relevance is still ongoing. One critical issue hampering this debate is the use of different methods for the characterization of endogenous and synthetic peptide and their intrinsic limitations for distinguishing Aβ aggregates. Here, we used different protocols for the establishment of prefibrillar Aβ assemblies with varying morphologies and sizes and compared them in a head-to-head fashion. Aggregation was characterized via the monomeric peptide over time until spheroidal, protofibrillar, or fibrillar Aβ aggregates were predominant. It could be shown that a change in the ionic environment induced a structural rearrangement, which consequently confounds the delineation of a measured neurotoxicity toward a distinct Aβ assembly. Here, neuronal binding and hippocampal neurotransmission were found to be suitable to account for the synaptotoxicity to different Aβ assemblies, based on the stability of the applied Aβ aggregates in these settings. In contrast to monomeric or fibrillar Aβ, different prefibrillar Aβ aggregates targeted neurons and impaired hippocampal neurotransmission with nanomolar potency, albeit by different modalities. Spheroidal Aβ aggregates inhibited NMDAR-dependent long-term potentiation, as opposed to protofibrillar Aβ aggregates, which inhibited AMPAR-dominated basal neurotransmission. In addition, a provoked structural conversion of spheroidal to protofibrillar Aβ assemblies resulted in a time-dependent suppression of basal neurotransmission, indicative of a mechanistic switch in synaptic impairment. Thus, we emphasize the importance of addressing the metastability of prefacto characterized Aβ aggregates in assigning a biological effect.

Alcohol administration blocks stress-induced impairments in memory and anxiety, and alters hippocampal neurotransmitter receptor expression in male rats

Chronic exposure to stress has many deleterious effects on behavior, which can often lead to self-medication with anxiolytics, antidepressants, or alcohol. We determined the effects of alcohol administration following a stressor on established behavioral, physiological, and neural responses to stress. Male Sprague–Dawley rats received: No alcohol/No stress (CON), Alcohol alone (ALC), Stress alone (STR), or Stress plus Alcohol (STR+ALC). For seven consecutive days, two cohorts received an oral dose of 2.0g/kg of either 20% ethanol or saline. In Cohort 1, behavioral testing began after the final treatment (day-8). Memory was tested using the object recognition (OR) and Y-maze, anxiety on the plus maze, and depression on the forced swim task. Memory on OR and Y-maze tasks was impaired in the ALC and STR groups. This deficit was reversed in the STR+ALC group, which performed not differently from the CON group. Stress alone was associated with increased anxiety, which was alleviated with alcohol treatment. No treatment effects were found in the forced swim task. In Cohort 2, hippocampal GABAα4 was upregulated in the STR+ALC group and GluN2B was upregulated in the ALC and STR+ALC groups. The STR+ALC group in Cohort 1 showed enhanced corticosterone levels after forced swim. The STR+ALC group in Cohort 2 showed increased corticosterone levels on day-1 of treatment and a habituation by day-7. In conclusion, this study found a reversal of stress-induced deficits in cognition and anxiety when alcohol was given post-stress, and changes in neurotransmitter receptor expression may contribute to these behavioral effects.

Changes in the hippocampal extracellular amino acid neurotransmitter of kindled rats by kainic acid

Objective To establish and evaluate the animal model for temporal lobe epilepsy kindled by kainic acid (KA)in rats,to observe the changes in glutamic amino acid (Glu),taurine amino acid (Tau),and γ-aminobatyric amino acid (GABA) of the extracellular fluid in the hippocampus of kindled rats,and to analyze the role of neurotransmitters in epileptogenesis.Methods In order to establish an animal model for temporal lobe epilepsy in rats,the characteristics of praxiological changes,electrophysiological changes,and ultrastructural examinations were observed.The contents of Glu,Tau,and GABA were detected in the hypppocampus by microdialysis combined with high performance liquid chromatography (HPLC).Results Typical seizures were observed in the models.Epileptic discharges,such as spike,slow,spike-slow,and sharp waves appeared in the electroencephalogram (EEG).Electron microscopic analysis of the hippocampal tissues revealed an increase of excitatory amino acid levels.The levels of Glu,Tau,and GABA were higher than groups that were injected with normal sodium (P ＜ 0.05).Conclusion Excitatory amino acids and inhibitory amino acids in the presence of epilepsy play important roles and is one of the main causes of seizure. Key words: Epilepsy; Kainic acid; Glutamic amino acid; γ-aminobutyric amino acid ; Taurine amino acid

Direct Alteration of a Specific Inhibitory Circuit of the Hippocampus by Antidepressants

Correct brain functioning relies on the precise activity of a myriad of synapses assembling neurons in complex networks. In the hippocampus, highly diverse inhibitory circuits differently govern several physiologically relevant network activities. Particularly, perisomatic inhibition provided by specific interneurons was proposed to control emotional states, and could therefore be affected by mood disorders and their therapy. We found that both chronic and acute administration of two major antidepressants, imipramine and fluoxetine, strongly and directly altered GABA-mediated (GABAergic) hippocampal neurotransmission in mice and rats, independently of their effects on amine reuptake systems. These drugs affected GABA release from synapses formed by fast-spiking cells, but not interneurons expressing cannabinoid receptor type 1, resulting in the disruption of γ oscillations. This differential effect, shared by two types of antidepressants, suggests a new mechanism of action of these medications, and a possible role of perisomatic inhibition in depressive disorders.

Key Physiological Parameters Dictate Triggering of Activity-Dependent Bulk Endocytosis in Hippocampal Synapses

To maintain neurotransmission in central neurons, several mechanisms are employed to retrieve synaptically exocytosed membrane. The two major modes of synaptic vesicle (SV) retrieval are clathrin-mediated endocytosis and activity-dependent bulk endocytosis (ADBE). ADBE is the dominant SV retrieval mode during intense stimulation, however the precise physiological conditions that trigger this mode are not resolved. To determine these parameters we manipulated rat hippocampal neurons using a wide spectrum of stimuli by varying both the pattern and duration of stimulation. Using live-cell fluorescence imaging and electron microscopy approaches, we established that stimulation frequency, rather than the stimulation load, was critical in the triggering of ADBE. Thus two hundred action potentials, when delivered at high frequency, were sufficient to induce near maximal bulk formation. Furthermore we observed a strong correlation between SV pool size and ability to perform ADBE. We also identified that inhibitory nerve terminals were more likely to utilize ADBE and had a larger SV recycling pool. Thus ADBE in hippocampal synaptic terminals is tightly coupled to stimulation frequency and is more likely to occur in terminals with large SV pools. These results implicate ADBE as a key modulator of both hippocampal neurotransmission and plasticity.

Stratum oriens stimulation‐evoked modulation of hippocampal long‐term potentiation involves the activation of muscarinic acetylcholine receptors and the inhibition of Kv7/M potassium ion channels

Acetylcholine is considered to be an endogenous modulator of hippocampal neurotransmission and synaptic plasticity. The activation of muscarinic acetylcholine receptors (mAChRs) reportedly enhances hippocampal synaptic plasticity, which plays an important role in memory function; however, the mechanism by which it enhances synaptic plasticity remains unclear. Here, we examined the involvement of the inhibition of Kv7/M K(+) channels, which are targets of mAChR modulation, during mAChR activation-induced enhancement of long-term potentiation (LTP) at rat hippocampal Schaffer collateral (SC)-CA1 synapses. When an electrical stimulus was applied to the stratum oriens before tetanic stimulation of the SCs, the magnitude of the induced SC-CA1 synapse LTP was enhanced as compared with that induced without stratum oriens stimulation. In the presence of the mAChR antagonist atropine, tetanic stimulation induced stable LTP, but the stratum oriens stimulation-evoked enhancement of LTP was abolished. The additional application of XE991, a selective blocker of Kv7/M K(+) channels, rescued the atropine-induced inhibition of LTP enhancement. The phospholipase C (PLC) inhibitor U-73122 inhibited the stratum oriens stimulation-evoked enhancement of LTP. Application of the T/R-type voltage-dependent Ca(2+) channel (VDCC) blocker Ni(2+) abolished the stratum oriens stimulation-evoked enhancement of LTP. In addition, tetanic stimulation with preceding stratum oriens stimulation was able to induce LTP during N-methyl-d-aspartate receptor blockade. We therefore propose that stratum oriens stimulation inhibits Kv7/M K(+) channels through mAChR activation-induced PLC activation, which leads to VDCC activation, and hence causes sufficient Ca(2+) influx to enhance LTP.

Thiamine deficiency degrades the link between spatial behavior and hippocampal synapsin I and phosphorylated synapsin I protein levels

The links between spatial behavior and hippocampal levels of synapsin I and phosphosynapsin I were assessed in normal rats and in the pyrithiamine-induced thiamine deficiency (PTD) rat model of Wernicke–Korsakoff's syndrome. Synapsin I tethers small synaptic vesicles to the actin cytoskeleton in a phosphorylation-dependent manner, is involved in neurotransmitter release and has been implicated in hippocampal-dependent learning. Positive correlations between spontaneous alternation behavior and hippocampal levels of both synapsin I and phosphorylated synapsin I were found in control rats. However, spontaneous alternation performance was impaired in PTD rats and was accompanied by a significant reduction (30%) in phosphorylated synapsin I. Furthermore, no correlations were observed between either form of synapsin I and behavior in PTD rats. These data suggest that successful spontaneous alternation performance is related to high levels of hippocampal synapsin I and phosphorylated synapsin I. These results not only support the previous findings that implicate impaired hippocampal neurotransmission in the spatial learning and memory deficits associated with thiamine deficiency, but also suggest a presynaptic mechanism.

The sodium‐driven chloride/bicarbonate exchanger in presynaptic terminals

The sodium-driven chloride/bicarbonate exchanger (NDCBE), a member of the SLC4 family of bicarbonate transporters, was recently found to modulate excitatory neurotransmission in hippocampus. By using light and electron microscopic immunohistochemistry, we demonstrate here that NDCBE is expressed throughout the adult rat brain, and selectively concentrates in presynaptic terminals, where it is closely associated with synaptic vesicles. NDCBE is in most glutamatergic axon terminals, and is also present in the terminals of parvalbumin-positive γ-aminobutyric acid (GABA)ergic cells. These findings suggest that NDCBE can regulate glutamatergic transmission throughout the brain, and point to a role for NDCBE as a possible regulator of GABAergic neurotransmission.

Axonal Gradient of Arachidonic Acid-containing Phosphatidylcholine and Its Dependence on Actin Dynamics

Phosphatidylcholine (PC) is the most abundant component of lipid bilayers and exists in various molecular forms, through combinations of two acylated fatty acids. Arachidonic acid (AA)-containing PC (AA-PC) can be a source of AA, which is a crucial mediator of synaptic transmission and intracellular signaling. However, the distribution of AA-PC within neurons has not been indicated. In the present study, we used imaging mass spectrometry to characterize the distribution of PC species in cultured neurons of superior cervical ganglia. Intriguingly, PC species exhibited a unique distribution that was dependent on the acyl chains at the sn-2 position. In particular, we found that AA-PC is enriched within the axon and is distributed across a proximal-to-distal gradient. Inhibitors of actin dynamics (cytochalasin D and phallacidin) disrupted this gradient. This is the first report of the gradual distribution of AA-PC along the axon and its association with actin dynamics.

Hippocampal neurotransmitter efflux during one-trial novel object recognition in rats

Several lines of evidence point to a role for the hippocampal formation and contiguous temporal lobe structures in a variety of learning and memory paradigms. Presumably, these cognitive phenomena are mediated (and accompanied) by dynamic changes in neurochemical transmission that may differ between learning and recall phases. However, the neurotransmitter correlates of most memory-related tasks have not been thoroughly investigated. Here we used a one-trial object recognition paradigm paired with in vivo microdialysis to assess hippocampal acetylcholine (ACh), glutamate and GABA efflux when rats were exposed to familiar objects, and when given the option to explore familiar and novel objects. Rats preferentially explored the novel object over the familiar one when presented with the option. Regardless of object familiarity, object exploration was accompanied by an increase in hippocampal ACh efflux, while GABA efflux was unaffected. However, glutamate efflux was not increased above baseline levels by presentation of familiar objects, but was significantly enhanced in the presence of the novel object. These data suggest that the hippocampus, and in particular, hippocampal glutamate, may be involved in memory processes during novelty recognition paradigms.

The Effects of Chronic Ethanol Self-Administration on Hippocampal Serotonin Transporter Density in Monkeys

Evidence for an interaction between alcohol consumption and the serotonin system has been observed repeatedly in both humans and animal models yet the specific relationship between the two remains unclear. Research has focused primarily on the serotonin transporter (SERT) due in part to its role in regulating extracellular levels of serotonin. The hippocampal formation is heavily innervated by ascending serotonin fibers and is a major component of the neurocircuitry involved in mediating the reinforcing effects of alcohol. The current study investigated the effects of chronic ethanol self-administration on hippocampal SERT in a layer and field specific manner using a monkey model of human alcohol consumption. [3H]Citalopram was used to measure hippocampal SERT density in male cynomolgus macaques that voluntarily self-administered ethanol for 18 months. Hippocampal [3H]citalopram binding was less dense in ethanol drinkers than in controls, with the greatest effect observed in the molecular layer of the dentate gyrus. SERT density was not correlated with measures of ethanol consumption or blood ethanol concentrations, suggesting the possibility that a threshold level of consumption had been met. The lower hippocampal SERT density observed suggests that chronic ethanol consumption is associated with altered serotonergic modulation of hippocampal neurotransmission.

Neuronal localization of 5-HT2A receptor immunoreactivity in the rat hippocampal region

The 5-HT2A receptor subtype (5-HT2Ar) plays an important role in the modulation of the hippocampal region activity and it has been associated with learning and memory processes. In the present study, the 5-HT2Ar was immunohistochemically localized in the rat hippocampal region, which includes the hippocampal formation and the parahippocampal region. In the hippocampal formation (dentate gyrus, hippocampus proper and subiculum) and entorhinal cortex, the colocalization of the 5-HT2Ar with the inhibitory transmitter γ-aminobutyric acid (GABA) was studied using double immunofluorescence confocal microscopy. The patterns of immunostaining were very different in non-injected and colchicine-injected rats. In untreated rats, the immunoreactivity could be attributed especially to neuropil. Interestingly, in non-injected rats, the 5-HT2Ar immunoreactivity was located in the mossy fibers, suggesting that serotonin acts presynaptically via this receptor subtype directly on glutamate axons. Pretreatment with colchicine increased the number of 5-HT2Ar-immunoreactive somata. Morphological and double immunofluorescence analyses indicated that the 5-HT2Ar was located on both the excitatory and the inhibitory neurons of the rat hippocampal region. The results of the present study suggest that the 5-HT2Ar could participate in the hippocampal neurotransmission by acting on different neuronal populations.

Stress during development alters anxiety-like behavior and hippocampal neurotransmission in male and female rats

Epidemiological data indicate that early stress increases vulnerability to psychiatric disorders, including anxiety and depression. In the present study we sought to investigate the long-term behavioral and neurochemical consequences of increased and sustained corticosterone levels induced by a 24 h bout of maternal deprivation (DEP) imposed on postnatal day 11 (DEP11). As adults, animals were exposed to the elevated plus maze for assessment of anxiety-like behavior and corticosterone response to this challenge, or decapitated for determination of monoamines and amino acid neurotransmitters content in the hippocampus by HPLC method. The results showed that DEP11 male and female rats displayed increased time in the central hub of the maze and more risk assessment behavior, reflecting increased anxiety-like behavior; in addition, these animals continuously secreted corticosterone in response to the behavioral test until the latest time-point, e.g., 60 min post-stress. In males, maternal deprivation increased aspartate and glutamate levels and reduced taurine levels compared to non-deprived (NDEP) rats. DEP11 females displayed reduced noradrenaline, aspartate and GABA levels compared to NDEP counterparts. These results indicate that maternal deprivation at 11 days of age produced changes in hippocampal neurotransmission that may mediate the increased anxiety-like behavior observed in male and female deprived rats.This article is part of a Special Issue entitled ‘Anxiety and Depression’.