Ketamine and fluoxetine exert similar actions on prelimbic and infralimbic responsivity to lateral septal nucleus stimulation in Wistar rats.

Ketamine is a dissociative anesthetic that has been proposed to be a useful alternative in cases of a poor response to other treatments in patients with depression. Remarkably, beneficial clinical actions of ketamine are detected once its psychotropic actions disappear. Therefore, clinical actions may occur independently of dose. Most current studies focus on actions of ketamine on neurotrophic factors, but few studies have investigated actions of ketamine on neural structures for which actions of antidepressants have been previously explored. Lateral septal nucleus (LSN) stimulation reduces neural activity in the prelimbic cortex (PL) and infralimbic cortex (IL) subregions of the medial prefrontal cortex (mPFC). Fluoxetine increases inhibitory responsivity of the LSN-IL connection. In the present study, actions of an anesthetic dose of ketamine were compared with a high dose of fluoxetine on behavior and neural responsivity 24 h after drug administration. Fluoxetine reduced immobility in the forced swim test without changing locomotor activity in the open field test. Ketamine strongly decreased locomotor activity and did not produce changes in immobility. In another set of Wistar rats that received similar drug treatment regimens, the results indicated that LSN stimulation in saline-treated animals produced a long-lasting inhibitory afterdischarge in these mPFC subregions. Actions of ketamine on the LSN-mPFC connection reproduced actions of fluoxetine, consisting of accentuated inhibition of the LSN action on the mPFC. These findings suggest that independent of different actions on neurotransmission, the common final pathway of antidepressants lies in their actions on forebrain structures that are related to emotional regulation.

Prelimbic and infralimbic responsivity to amygdala input is modified by gonadal hormones in parallel to low anxiety-like behavior in ovariectomized rats.

Gonadal hormones may influence sexual activity by reducing anxiety. The basolateral amygdala (BLA) and prelimbic (PL) and infralimbic (IL) cortical regions comprise a loop that is related to fear, anxiety, and social behavior. In female ovariectomized rats, actions of estradiol, progesterone, and sequential estradiol and progesterone administration were explored in the open field test (OFT) and plus maze test (PMT) to evaluate signs of anxiety-like behavior. The three hormonal treatments reduced indicators of anxiety in the PMT but did not influence behavior in the OFT. In the same behaviorally tested rats under urethane anesthesia, single-unit extracellular recordings were obtained from the PL and IL during electrical stimulation of the BLA. The analysis of 250 ms peristimulus histograms showed that BLA stimulation produced two kinds of response. A small group of neurons increased their firing rate after BLA stimulation. Most neurons exhibited a reduction of spiking. Neurons that increased their firing rate after BLA stimulation did not show any difference with the hormonal treatments. In neurons that were inhibited by BLA stimulation, estradiol reduced the neuronal firing rate in the PL and IL, and progesterone alone and the sequential administration of estradiol followed by progesterone administration 24 h later (priming) increased the firing rate during the 240 ms before BLA stimulation. Analyses of responsivity of the PL and IL during electrical stimulation of the BLA indicated that estradiol, progesterone, and estradiol followed by progesterone administration 24 h later (priming) reduced inhibitory actions of the BLA on the PL but not IL. In the BLA-IL connection, progesterone exacerbated the inhibitory response. These findings indicate that anxiolytic actions of estradiol, progesterone, and estradiol followed by progesterone administration 24 h later (priming) correspond to lower BLA-PL responsivity. Actions of progesterone on BLA-IL responsivity appear to contribute to sexual activity by interacting with other forebrain structures that are also related to sexual receptivity.

Anxiety Constitutes an Early Sign of Acute Hypoglycemia.

INTRODUCTION: Research in humans has identified a link between hypoglycemia and anxiety. The present study examined anxiety-like behaviors in rats that were subjected to hypoglycemia that was produced by an acute injection of insulin. Healthy female Wistar rats were subjected to a battery of tests to explore anxiety (elevated plus maze) and locomotion (open field test). METHODS: The control (CT) group received 0.9% saline (3 mL/kg, p.o.). Three other groups received 50% glucose (3 mL/kg, p.o.), insulin (0.1 UI, s.c.), or insulin + glucose (normalized glycemia [NG] group). RESULTS: Normal glycemic values were found in the CT and NG groups. Therefore, a single control (CT-NG) group was formed for statistical comparisons. The highest glycemic value was found in the glucose-induced hyperglycemia group. The lowest glycemic value was found in the insulin-induced hypoglycemia group. In the open field test, the most significant change was a higher number of rearings in the hypoglycemia group. In the elevated plus maze test, the CT-NG group and hyperglycemia groups exhibited similar behavior, whereas the hypoglycemia group spent a shorter time on the open arms and a longer time on the closed arms and had the highest Anxiety Index. Hyperglycemia is a typical characteristic of diabetes. Insulin normalizes glycemia. In the present study, insulin produced anxiety only when it produced hypoglycemia. CONCLUSION: The main effect of acute hypoglycemia is anxiety, which may be considered an early sign of hypoglycemia in an allostatic process.

Insulin and fluoxetine produce opposite actions on lateral septal nucleus-infralimbic region connection responsivity.

Some diabetes patients develop depression, the main treatment for which is antidepressants. Pharmacological interactions between insulin and antidepressants (e.g., fluoxetine) are controversial in the literature. Some authors reported hypoglycemic actions of fluoxetine, whereas others reported antidepressant-like actions. In healthy rats, insulin produces an antidespair-like action in rats through an increase in locomotor and exploratory activity, but differences in actions of insulin and fluoxetine on neuronal activity are unknown. The present study evaluated Wistar healthy rats that were treated with saline, insulin, fluoxetine, or fluoxetine + insulin for 3 days (short-term) or 21 days (long-term). The model consisted of electrical stimulation of the lateral septal nucleus (LSN) while we performed single-unit extracellular response recordings in the prelimbic cortex (PL) and infralimbic cortex (IL) subregions of the medial prefrontal cortex (mPFC). Stimulation of the LSN produced an initial brief excitatory paucisynaptic response and then a long-lasting inhibitory afterdischarge in the PL and IL. Treatment with saline and fluoxetine, but not insulin, minimally affected the paucisynaptic response. Differences were found in LSN-IL responsivity. The inhibitory afterdischarge was clearly enhanced in the long-term fluoxetine group but not by insulin alone or fluoxetine + insulin. These findings suggest that insulin produces some actions that are opposite to fluoxetine on LSN-mPFC connection responsivity, with no synergistic actions between the actions of insulin and fluoxetine.

2-Heptanone reduces inhibitory control of the amygdala over the prelimbic region in rats.

Basolateral amygdala (BLA) nuclei and their reciprocal connections with prelimbic (PL) and infralimbic (IL) regions of the medial prefrontal cortex (mPFC) are involved in the regulation of fear. 2-Heptanone is released in urine in stressed rats, and the olfactory detection of this odor produces immediate avoidance and alarm reactions and modifies neuronal activity in limbic connections in non-stressed rats. If 2-heptanone acts as a danger signal, then long-lasting actions would be expected. The aim of the present study was to investigate whether the forced inhalation of 2-heptanone modifies the response capacity of the BLA-mPFC circuit in the long term (48 h). Single-unit extracellular recordings were obtained from the PL and IL during electrical stimulation of the BLA (square-wave pulses; 1 ms, 20 µA, 0.3 Hz, 110 stimuli over a total duration of 360 s) in three groups of Wistar rats: control group (no sensory stimulation), unpredictable auditory stimulation group, and 2-heptanone stimulation group. A brief-latency (1 ms), short-duration (5 ms) paucisynaptic response followed BLA stimulation and was unaffected by any sensorial stimulation. The paucisynaptic response was followed by a mostly inhibitory and long-lasting (>750 ms) afterdischarge in the control and auditory stimulation groups. In the 2-heptanone group, the inhibitory afterdischarge shifted to an excitatory afterdischarge after ∼250 ms in the PL and after ∼500 ms in the IL. Importantly, the rats that were included in this study were born in local housing facilities. Thus, these animals were never in contact with predators and instead in contact with only conspecifics. These results indicate that the forced inhalation of 2-heptanone is able to modify BLA-mPFC responsivity in the long term. 2-Heptanone decreases inhibitory control of the amygdala over mPFC activity. Disinhibition of the mPFC may lead to the adaptive expression of defensive behaviors, even in animals that are not in the presence of predators.

Putative Anti-Immobility Action of Acute Insulin Is Attributable to an Increase in Locomotor Activity in Healthy Wistar Rats.

BACKGROUND/AIMS: Anti-immobility actions of insulin in diabetic rats that are subjected to the forced swim test (FST) have been reported. In this test, low doses of antidepressants exert actions after long-term treatment, without affecting locomotor activity in healthy rats. Few studies have compared acute and chronic actions of insulin with antidepressants in healthy rats. METHODS: We hypothesized that if insulin exerts a true anti-immobility action, then its effects must be comparable to fluoxetine in both a 1-day treatment regimen and a 21-day treatment regimen in healthy, gonadally intact female Wistar rats. RESULTS: The results showed that low levels of glycemia were produced by all treatments, including fluoxetine, and glycemia was lower in proestrus-estrus than in diestrus-metestrus. None of the treatments or regimens produced actions on indicators of anxiety in the elevated plus maze. Insulin in the 1-day regimen increased the number of crossings and rearings in the open field test and caused a low cumulative immobility time in the FST. These actions disappeared in the 21-day regimen. Compared with the other treatments, fluoxetine treatment alone or combined with insulin produced a longer latency to the first period of immobility and a shorter immobility time in the chronic regimen in the FST, without affecting locomotor activity, and more pronounced actions were observed in proestrus-estrus (i.e., a true anti-immobility effect). CONCLUSION: These results indicate that insulin does not produce a true antidepressant action in healthy rats. The purported antidepressant effects that were observed were instead attributable to an increase in locomotor activity only in the 1-day regimen.

¿Los protocolos experimentales son un símil real de la diabetes humana? / Are experimental protocols an authentic simile of human diabetes?

Resumen Para el estudio de la diabetes se dispone de diversas estrategias metodológicas en modelos animales, tales como, técnicas quirúrgicas, modificaciones dietéticas, incluso manipulación genética y la administración de fármacos específicos, por su toxicidad. En animales, la diabetes experimental se logra con el uso de fármacos, como la aloxana o la estreptozotocina, los cuales producen daño irreversible en las células β-pancreáticas, aunque causan una alta mortalidad, debido a la cetosis asociada al daño agudo de estas células pancreáticas. El objetivo de este trabajo fue analizar los protocolos farmacológicos y otras estrategias disponibles, para determinar si la diabetes experimental realmente emula la diabetes humana. La diabetes es un proceso progresivo y crónico, en el que la mayor parte de las alteraciones clínicas son consecuencia, en el largo plazo, de alteraciones micro y macrovasculares. Por ello, es conveniente diferenciar entre los efectos de una hiperglucemia aguda, con aquellos que se observan cuando la hiperglucemia se prolonga a lo largo del tiempo, a fin de establecer analogías, entre el modelo experimental animal, con el síndrome diabético humano, mediante datos de laboratorio y de tipo clínico, de uso habitual en el diagnóstico y manejo de la diabetes humana.

Abstract For the study of diabetes, several methodological strategies use animal models. Such methodologies involve surgical techniques, diet modifications, some genetic manipulations and specific toxic drugs. The experimental production of diabetes in animal models use the administration of alloxan or streptozotocin and these drugs produce irreversible damage to pancreatic β-cells. However, its use is associated to a ketosis high mortality rate due to the acute damage of pancreatic cells. The aim of this review consisted in the analysis of the pharmacological diabetes production protocols as well as other available strategies, in order to elucidate which is potentially the ideal protocol that emulates human diabetes. Diabetes is a progressive and chronic process, in which most of the clinical alterations are a long-term consequence of micro and macrovascular alterations. Therefore, it is convenient to establish a difference between the effects of acute hyperglycemia, with those effects observable when hyperglycemia is present over the long-term in order to reach enough analogies between the animal experimental model with the human diabetes syndrome, through the use of laboratory and clinical indicators commonly employed for the diagnoses and management of human diabetes.

Estrogen and progesterone priming induces lordosis in female rats by reversing the inhibitory influence of the infralimbic cortex on neuronal activity of the lateral septal nucleus.

The lateral septal nucleus (LSN) exerts inhibitory control over lordosis in female rats, but the influence of forebrain structures, such as prelimbic (PL) and infralimbic (IL) regions of the medial prefrontal cortex (mPFC), on LSN activity during sexual receptivity is unknown. We hypothesized that the neural responsivity of these connections may differ depending on sexual receptivity. Gonadally intact female Wistar rats received sequential priming injections of estradiol and progesterone (E2-P4). The presence of lordosis was then confirmed by exposing the female rats to a sexually experienced male rat. Intromission was not allowed. Vaginal smear analyses verified that the rats were in proestrus-estrus of the estrous cycle. The results were compared with a diestrus group, which was verified by vaginal smears and the absence of lordosis. Under ethyl-carbamate anesthesia, single-unit extracellular recordings of the LSN were performed during electrical stimulation of the PL and IL to evaluate possible changes in the responsivity of neural connections. Stimulation of the PL or IL produced a short-latency, brief-duration (paucisynaptic) excitatory response in the LSN, followed by a period of afterhyperpolarization. Responsivity of the PL-LSN pathway was unaffected by E2-P4 priming. The paucisynaptic response of the IL-LSN pathway was significantly greater in the E2-P4-primed group than in the diestrus group, and the afterhyperpolarization response decreased to nearly zero. These findings indicate that the IL exerts inhibitory control over the LSN during diestrus in rats, but this inhibitory control decreases under the action of gonadal steroids, seemingly favoring sexual receptivity.

Reciprocal interactions between the basolateral amygdala and infralimbic and prelimbic regions of the mPFC: Actions of diazepam.

The basolateral amygdala (BLA) and prelimbic (PL) and infralimbic (IL) cortices are interconnected structures that participate in the regulation of fear. Unknown are the reciprocal functional influence of these regions on neuronal responsivity and the action of an anxiolytic drug. We performed multiunit activity recordings from one neuronal pool while applying electrical stimulation to another neuronal pool. In the same experimental session, PL-BLA and IL-BLA sequences of stimulation-recording were applied, followed by the inverse sequence (BLA-PL and BLA-IL). Using this procedure, we obtained information from the same neuronal pool that was stimulated and recorded. Using peristimulus histograms, we analyzed neuronal responsivity. In the saline-treated group, the PL-BLA stimulation-recording sequence produced an inhibitory response. The inverse sequence, BLA-PL, produced an excitatory response. For BLA-IL and IL-BLA stimulation-recording sequences, a minimal response was observed in the saline control group. Diazepam minimally affected responsivity of the PL-BLA and BLA-IL connections. Diazepam blocked the initial excitatory response of the IL-BLA connection. Under control conditions, the PL and BLA appeared to regulate each other. Under the action of diazepam, the IL exerted an inhibitory influence on the BLA. Because of the well-known actions of this anxiolytic drug on the BLA, this combined action may result in the synergistic control of fear.

Responsivity of lateral septum-mPFC connections in alloxan-induced hyperglycemia.

The lateral septal nucleus (LSN) is related to the actions of antidepressants, and the prelimbic cortex (PL) and infralimbic cortex (IL) modulate responses to fear. However, unknown is whether experimental diabetes that is induced by alloxan alters the responsivity of these regions. We used a method in which one forebrain region (LSN) was electrically stimulated while single-unit extracellular recordings were performed in another mPFC region (PL and IL). Several experimental groups were tested: (a) animals that were subjected to long-term (42-day) alloxan-hyperglycemia and protected with insulin, (b) healthy animals that received a low dose of insulin that does not produce changes in glycemia, and (c) animals that received long-term treatment with fluoxetine. Additional healthy groups of animals received insulin or fluoxetine and underwent the forced swim test. Biological measurements indicated the induction of diabetes in alloxan-treated rats. In this group, a shift toward an inhibitory response to LSN stimulation was observed in the PL and IL compared with the control group. A low dose of insulin or fluoxetine produced similar changes in LSN-PL and LSN-IL responsivity. Long-term hyperglycemia increased inhibitory responsivity in the LSN-PL and LSN-IL, but this action was less pronounced than the action that was exerted by insulin and fluoxetine, which produced similar actions. Such similar actions were confirmed in the forced swim test, in which the antidepressant-like effects of insulin partially resembled the effects of fluoxetine. The changes that were observed in the alloxan group appeared to be related to neuronal damage, and a low dose of insulin exerted some antidepressant-like actions.

Fluoxetine and stress inversely modify lateral septal nucleus-mpfc neuronal responsivity.

Several clinically effective antidepressants increase the neuronal firing rate in the lateral septal nucleus (LSN), a forebrain structure that is anatomically related to medial prefrontal cortex (mPFC) regions. mPFC function is related to depression and the regulation of fear. However, unknown is whether antidepressant treatment or chronic stress modifies the responsivity of neuronal LSN-mPFC connections. We performed single-unit extracellular recordings in the anterior cingulate cortex (ACC) and prelimbic (PL) and infralimbic (IL) regions of the mPFC during stimulation of the LSN in anesthetized male Wistar rats that received fluoxetine (1 mg/kg, 21 days) or were subjected to chronic mild stress (5 weeks). The results were compared with a control group (saline treatment, devoid of behavioral manipulations). Stimulation of the LSN produced an initial excitatory paucisynaptic response, followed by an afterdischarge, characterized by an increase in the neuronal firing rate. Opposite changes were induced by fluoxetine treatment and chronic stress exposure. Peristimulus histograms and unit-activity ratio analyses indicated that LSN-mPFC responsivity differed between fluoxetine treatment and chronic stress exposure. Fluoxetine reduced neuronal responsivity in the LSN-PL and LSN-IL, and stress increased neuronal responsivity in the same regions. In both cases, the changes were more pronounced in the IL region. The lower responsivity of LSN-PL and LSN-IL connections that was produced by fluoxetine may reflect a higher threshold for fear, and lower responsivity of this connection may be related to states of fear. The LSN and mPFC comprise a portion of a limbic-cortical circuit where neuronal responses depend on specific conditions.

An alarm pheromone reduces ventral tegmental area-nucleus accumbens shell responsivity.

2-Heptanone (methyl n-amyl ketone) is a ketone that produces alarm reactions in insects (e.g., bees and ants). As an olfactory stimulus, 2-heptanone produces anxiety reactions in the short term and despair in the long term in rodent models. Among the anatomical connections of the olfactory system that integrate behavioral responses, connections between the amygdala and nucleus accumbens are important, which in turn form a circuit with the ventral tegmental area (VTA). 2-Heptanone increases the firing rate of amygdala neurons without participation of the vomeronasal organ. The olfactory amygdala-VTA-nucleus accumbens circuit may integrate defensive behaviors, but the possible actions of 2-heptanone on the responsivity of VTA-nucleus accumbens connections have not yet been explored. In the present study, multiunit activity recordings were obtained in adult Wistar rats from the core and shell subregions of the nucleus accumbens during electrical stimulation of the VTA under basal conditions and later during simultaneous stimulation of the VTA and olfactory exposure to 2-heptanone. 2-Heptanone reduced the responsivity of the VTA-nucleus accumbens shell but did not influence the responsivity of the VTA-nucleus accumbens core. The lower VTA-nucleus accumbens shell excitability may be related to a primary defensive warning when exposed to an alarm pheromone.

Sensitivity to diazepam after a single session of forced swim stress in weaning Wistar rats.

The present study investigated the sensitivity to stress and diazepam in weaning (21-day old) Wistar rats. A single 15-min session of forced swimming was used to induce anxiety-like behavior. The group that was forced to swim exhibited an increase in anxiety-like behavior in the elevated plus maze (EPM) and open field test (OFT) compared to the non-stressed group. Diazepam (1 h before the tests) reduced anxiety-like behavior in rats forced to swim compared to the vehicle stressed group. The dose-response curve for diazepam indicated that the 0.5 mg kg-1 dose (1 h before the EPM and OFT) was the minimum effective dose in reducing anxiety-like behavior without altering locomotor activity in weaning rats. These results indicate that weaning rats can develop anxiety-like behavior after a brief, single session of stress, and that rats at this age are seemingly more sensitive to diazepam than adult rats, which may be taken into account for clinical applications.

Myristic acid in amniotic fluid produces appetitive responses in human newborns.

BACKGROUND: A mixture of eight fatty acids (lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, elaidic acid, and linoleic acid) that are contained in human amniotic fluid, colostrum, and milk produces appetitive responses in newborns, suggesting the existence of a transition of sensorial cues that guide newborns to the maternal breast. OBJECTIVE: To explore the ability of each of these eight fatty acids individually to produce appetitive responses in newborns. METHODS: The study included 12 healthy human newborns<24h after birth. Using a longitudinal design, cotton swabs that were impregnated with each of the eight fatty acids and control substances (i.e., vehicle, saline, and vanilla) were placed approximately 1cm from the newborns' nostrils for 30s. Positive responses that were suggestive of acceptance included appetitive movements (i.e., suckling) and sniffing that were directed toward the cotton swab. Lateral movements of the head away from the swab were considered negative responses. Remaining stationary with no changes in facial expressions was considered indifference. RESULTS: Compared with controls (i.e., vehicle, saline, and vanilla) and the other fatty acids tested, myristic acid produced the longest duration of positive facial responses (suckling and sniffing). No significant differences in negative facial responses were observed in response to the odoriferous stimuli. No reactions that were suggestive of disgust were observed. CONCLUSION: A complex combination of stimuli, including the odor of myristic acid, may integrate sensory cues that guide newborns to the maternal breast.

Cognitive impairment in diabetes and poor glucose utilization in the intracellular neural milieu.

The main characteristic of diabetes is hyperglycemia. Depending on whether diabetes is type-1 or type-2, it is characterized by deficiencies in insulin secretion, insulin receptor sensitivity, hexokinase activity, and glucose transport. Current drug treatments are able to lower circulating glucose but do not address the problem of glucose utilization in the intracellular milieu, the consequence of which is tissue damage. In the long-term, such changes can produce structural damage in many cortical and subcortical brain areas that are related to cognitive function. Many epidemiological reports consider anxiety and depression as clinical entities that accompany diabetes. However, anxiety and depression in diabetes appear to occur in parallel and do not follow a causal relationship. From a behavioral perspective, anxiety may be considered adaptive, whereas depression can be considered reactive in response to changes in lifestyle and ailments that are caused by the disease. Therefore, the main alteration in diabetes seems to be cognitive function. We hypothesized that in type-2 diabetes, hypoglycemic medications do not restore the function of glucose in the intracellular compartment, which may have deleterious effects on neural tissue with behavioral consequences. In such a case, it is important to develop pharmacological interventions that directly restore plasma insulin levels, insulin receptor function, and hexokinase activity, thereby avoiding damage to neural tissue that is associated with cognitive deficits in diabetic patients, particularly patients with type-2 diabetes. The better management of such alterations in diabetes should be directed toward improving glucose utilization by neural tissue.

Progesterone modifies the responsivity of the amygdala-mPFC connection in male but not female Wistar rats.

Amygdala-medial prefrontal cortex (mPFC) connections partially regulate fear, anxiety, and the acquisition of conditioned fear. Progesterone exerts some effects on anxiety and fear. Currently unknown, however, are the actions of progesterone on the responsivity of amygdala-mPFC connections and possible sex differences. We performed single-unit extracellular recordings from the prelimbic (PL) and infralimbic (IL) cortices of the mPFC during stimulation of the basal amygdala (BA) in anesthetized male and diestrus female rats. Basal amygdala stimulation produced an initial excitatory paucisynaptic response that was similar between sexes and unaffected by progesterone. A long-lasting inhibitory response followed the initial brief excitatory response, which was more pronounced in the PL region in males. The unit activity ratio analysis indicated that progesterone negated the sex difference in the PL region response to BA stimulation. The results suggest that progesterone decreases the responsivity to amygdala stimulation, particularly in males compared with diestrus females, which may be related to sex differences in the strategies to cope with threatening situations.

A Fatty Acids Mixture Reduces Anxiety-Like Behaviors in Infant Rats Mediated by GABAA Receptors.

Fatty acids (C6-C18) found in human amniotic fluid, colostrum, and maternal milk reduce behavioral indicators of experimental anxiety in adult Wistar rats. Unknown, however, is whether the anxiolytic-like effects of fatty acids provide a natural mechanism against anxiety in young offspring. The present study assessed the anxiolytic-like effect of a mixture of lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, elaidic acid, and linoleic acid in Wistar rats on postnatal day 28. Infant rats were subjected to the elevated plus maze, defensive burying test, and locomotor activity test. Diazepam was used as a reference anxiolytic drug. A group that was pretreated with picrotoxin was used to explore the participation of γ-aminobutyric acid-A (GABAA) receptors in the anxiolytic-like effects. Similar to diazepam, the fatty acid mixture significantly increased the frequency of entries into and time spent on the open arms of the elevated plus maze and decreased burying behavior in the defensive burying test, without producing significant changes in spontaneous locomotor activity. These anxiolytic-like effects were blocked by picrotoxin. Results suggest that these fatty acids that are contained in maternal fluid may reduce anxiety-like behavior by modulating GABAergic neurotransmission in infant 28-day-old rats.

Myristic acid produces anxiolytic-like effects in Wistar rats in the elevated plus maze.

A mixture of eight fatty acids (linoleic, palmitic, stearic, myristic, elaidic, lauric, oleic, and palmitoleic acids) at similar concentrations identified in human amniotic fluid produces anxiolytic-like effects comparable to diazepam in Wistar rats. However, individual effects of each fatty acid remain unexplored. In Wistar rats, we evaluated the separate action of each fatty acid at the corresponding concentrations previously found in human amniotic fluid on anxiety-like behaviour. Individual effects were compared with vehicle, an artificial mixture of the same eight fatty acids, and a reference anxiolytic drug (diazepam, 2 mg/kg). Myristic acid, the fatty acid mixture, and diazepam increased the time spent in the open arms of the elevated plus maze and reduced the anxiety index compared with vehicle, without altering general locomotor activity. The other fatty acids had no effect on anxiety-like behaviour, but oleic acid reduced locomotor activity. Additionally, myristic acid produced anxiolytic-like effects only when the concentration corresponded to the one identified in human amniotic fluid (30 µg/mL) but did not alter locomotor activity. We conclude that of the eight fatty acids contained in the fatty acid mixture, only myristic acid produces anxiolytic-like effects when administered individually at a similar concentration detected in human amniotic fluid.

Defensive burying test in postweaning rats: use of a small round chamber.

The defensive burying test is an experimental model that is used to explore anxiety-like behavior in adult rats. Because the expression of anxiety-like behavior may differ between infant and adult rats, we tested the impact of chambers with different sizes and shapes on defensive burying in 28-day-old Wistar rats. The first two chambers had base areas of 560 cm, but one was rectangular and the other round. The base areas of the other two chambers were 282 cm, also with one rectangular and one round. We examined the effects of vehicle and 1 mg/kg diazepam on defensive burying in the various chambers. Locomotor activity was also measured to identify or exclude any sedative effects. Independent of the treatments used, the infant rats showed a shorter burying latency in the three modified chambers and a longer cumulative burying time compared with the original apparatus. The effects of diazepam (i.e. increased latency and decreased burying time) were only significant in the small round chamber, without significant effects on general motor activity. These results suggest that a small round chamber that is used to test burying behavior is sensitive to the anxiolytic actions of diazepam when the experimental subjects are very young rats.

Participation of GABAA chloride channels in the anxiolytic-like effects of a fatty acid mixture.

Human amniotic fluid and a mixture of eight fatty acids (FAT-M) identified in this maternal fluid (C12:0, lauric acid, 0.9 µ g%; C14:0, myristic acid, 6.9 µ g%; C16:0, palmitic acid, 35.3 µ g%; C16:1, palmitoleic acid, 16.4 µ g%; C18:0, stearic acid, 8.5 µ g%; C18:1 cis, oleic acid, 18.4 µ g%; C18:1 trans, elaidic acid, 3.5 µ g%; C18:2, linoleic acid, 10.1 µ g%) produce anxiolytic-like effects that are comparable to diazepam in Wistar rats, suggesting the involvement of γ -aminobutyric acid-A (GABA(A)) receptors, a possibility not yet explored. Wistar rats were subjected to the defensive burying test, elevated plus maze, and open field test. In different groups, three GABA(A) receptor antagonists were administered 30 min before FAT-M administration, including the competitive GABA binding antagonist bicuculline (1 mg/kg), GABA(A) benzodiazepine antagonist flumazenil (5 mg/kg), and noncompetitive GABA(A) chloride channel antagonist picrotoxin (1 mg/kg). The FAT-M exerted anxiolytic-like effects in the defensive burying test and elevated plus maze, without affecting locomotor activity in the open field test. The GABA(A) antagonists alone did not produce significant changes in the behavioral tests. Picrotoxin but not bicuculline or flumazenil blocked the anxiolytic-like effect of the FAT-M. Based on the specific blocking action of picrotoxin on the effects of the FAT-M, we conclude that the FAT-M exerted its anxiolytic-like effects through GABA(A) receptor chloride channels.