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Prenatal maternal stress (PS) can adversely affect the development of the central nervous system in offspring. The effects of PS become evident laterin life and may be in‑ volved in the pathogenesis of neurological and mental dis‑ orders. The dorsal raphe nucleus (DRN), as a major source of serotonin (5-HT) in the mammalian forebrain, plays a key role in the stress response. The DRN is also involved in the development of stress-related disorders. GABA-ergic and glutamatergic transmission in the DRN are modulated by the 5-HT7 receptor, however, little is known about the effects of PS on the activity of the DRN neuronal network. The aim of this study was to determine the effects of PS by analysing excitatory and inhibitory synaptic transmis‑ sion, and its modulation by the 5-HT7 receptor, in the DRN of rat adolescent offspring of stressed rat dams. Pregnant Sprague-Dawley rats were subjected daily to three restraint stress sessions, from the 14th day of pregnancy until birth. During each stress session, rats were placed in plastic cyl‑ inders and exposed to bright light for 45 min. Control preg‑ nant females were left undisturbed in their home cages. The effects of PS were studied in slices of the DRN prepared from adolescent male offspring of control and stressed mothers. Whole-cell recordings were carried out from pu‑ tative 5-HT neurons. Spontaneous excitatory (sEPSCs) and inhibitory (sIPSCs) postsynaptic currents were recorded to assess glutamatergic and GABA-ergic transmission, respec‑ tively. 5-CT, in the presence of WAY 100635, was applied to the ACSF to selectively activate the 5-HT7 receptor. In pre‑ natally-stressed rats an increased frequency of sEPSCs and a decreased frequency of sIPSCs were evident, compared to control animals. In slices originating from control rats, ac‑ tivation of the 5-HT7 receptor resulted in a decrease in the mean frequency of sEPSCs and an increase in the mean fre‑ quency of sIPSCs. These effects were absent from slices ob‑ tained from prenatally-stressed rats. These results suggest that prenatal maternal stress in rats causes an enhance‑ ment of glutamatergic transmission and an attenuation of GABA-ergic transmission and affects the function of the 5-HT7 receptor in the DRN of their adolescent offspring. These effects may be related to prenatal stress-induced ab‑ normalities in the functioning of the serotonergic system. Support: This study was supported by grant 2015/17/N/ NZ4/02455, National Science Centre Poland.
Maternal separation (MS) procedure is an experimental paradigm for studying disturbances in brain functions that result from adverse events occurring during development. However, the effects of early life stress on synaptic plasticity in the amygdala of adolescent animals are poorly understood. This study investigated the effects of MS on LTP in the cortical (CoI) and the thalamic input (ThI) to the lateral amygdala (LA) of rats. Wistar dams with their offspring were housed under 12:12 L/D conditions. On each postnatal day (PND) 1–21 rats were subjected to MS (3h/day). For electrophysiological ex vivo experiments rats between PND 35 and PND 55 were used. The animals were anesthetized and brain slices containing LA (450 µm) were cut. Field potentials (FPs) were evoked by the stimulation of CoI or ThI to LA. LTP was induced using theta-burst stimulation protocol. In slices prepared from control, animal facility reared (AFR) rats, the amplitude of FPs after LTP induction in CoI amounted 185.9 ± 17.04. In slices obtained from MS-subjected rats LTP in the cortical input was weaker (117.7 ± 7.7%). Amplitude of FPs after LTP induction in ThI, amounted 128.99 ± 9.47%, while in MS-subjected rats amplitude of FPs recorded after LTP induction amounted 110.41 ± 8.91%. These results demonstrate that MS impairs LTP in the cortical and in the thalamic input to LA.
Prenatal maternal stress (PS) adversely influences the development of the central nervous system. Its effects become evident later in life and may lead to mental and neurological disorders. The dorsal raphe nucleus (DRN) is a major source of serotonin in the mammalian brain. DRN plays a key role in regulation of the stress response and is involved in the development of stress-related psychiatric disorders. Little is known of the effect of prenatal stress on the DRN. In particular, it is not known how PS influences excitatory and inhibitory synaptic transmission and the properties of neurons in the DRN. The aim of this study was to determine the effects of prenatal stress on glutamatergic and GABAergic inputs to DRN serotonergic neurons of the rat. Pregnant Sprague-Dawley rats were subjected daily to three restraint stress sessions, from 14th day of pregnancy until the delivery. The effects of this treatment were studied in slices of the DRN prepared from adolescent male offspring of control and stressed mothers. Whole-cell recordings were carried out from putative serotonergic neurons in DRN slices. Spontaneous excitatory (sEPSCs) and inhibitory (sIPSCs) postsynaptic currents were recorded to assess glutamatergic and GABA-ergic transmission, respectively. Prenatal stress caused an increase in the frequency of sEPSCs and a decrease in the frequency of sIPSCs. Basic electrophysiological properties of serotonergic neurons in rat dorsal raphe nucleus, such as resting membrane potential, input resistance and excitability were not changed after prenatal stress. These results suggest that prenatal maternal stress causes an enhancement of glutamatergic transmission and an attenuation of GABAergic transmission in the DRN of adolescent offspring rats. These effects are likely to affect the function of the serotonergic system. FINANCIAL SUPPORT: This study was supported by grant 2015/17/N/NZ4/02455, National Science Centre Poland. Joanna Sowa is a holder of scholarship from the KNOW sponsored by Ministry of Science and Higher Education, Republic of Poland.
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Increased level of corticoid hormons during stress results in activation of hippocampal mineralocorticosteroid receptors (MRs) and glucocorticoid receptors (GRs). Our earlier studies showed impairment of LTP in the dentate gyrus (DG) of mice after 3 daily sessions of brief (10 min) restraint stress and LTP augmentation after 14 and 21 daily sessions of immobilization. In contrast, 1 and 7 daily sessions of immobilization did not affect LTP level. We demonstrated that the GR antagonist RU38486 selectively blocks the effect of 3 sessions of restraint. In the present study we investigated whether the effects of repeated brief restraint stress are mediated by MRs. C57BL/6 male mice were exposed to the restraint, lasting 10 minutes, for 3 and 14 days. 1 hour before immobilization the animals were subcutaneously injected with the MR receptor antagonist spironolactone. Mice were sacrifi ced 24 h after the last exposure to restraint and hippocampal slices were prepared. Field excitatory postsynaptic potentials were evoked by the stimulation of the lateral perforant path and recorded from the molecular layer of the DG. LTP induction was attempted by applying 4 trains of high-frequency stimulation (HFS, 100 Hz, 1 s, repeated every 25 s). In both experimental groups immediately after HFS recorded responses were potentiated but 2 h later they were diminished relative to control. Thus, the present results show that spironolactone blocks LTP induction after 3 and 14 days of daily restraint.
Using extracellular recording we studied changes in the reactivity of rat frontal cortical slices to the 5-HT1A, 5-HT2 and 5-HT4 receptor agonists, (±)-2-dipropyloamino-8-hydroxy-1,2,3,4-tetrahydronaphtalene hydrobromide (8-OH-DPAT), (±)-2,5-dimethoxy-4-iodoamphetamine hydrochloride (DOI) and zacopride, respectively, induced by an earlier treatment of animals with corticosterone lasting 1 or 3 weeks. Spontaneous bursting activity was recorded in ex vivo slices incubated in a medium devoid of Mg2+ ions and containing picrotoxin (30 µM). Repetitive, but not single, corticosterone administration resulted in an attenuation of the effect of the activation of 5-HT1A receptors and in an enhancement of the effect related to 5-HT2 receptors. The effect of 5-HT4 receptor activation remained unchanged. In separate two sets of experiments rats were treated with corticosterone for 3 weeks and additionally with imipramine or citalopram, beginning on the eighth day of corticosterone administration. In the corticosterone plus imipramine as well as corticosterone plus citalopram groups the effects of 8-OH-DPAT and DOI were not different from control indicating that corticosterone-induced functional modifications in the reactivity of 5-HT1A and 5-HT2 receptors were reversed by antidepressant treatments.
Stressful experiences during the early stages of life can distort normal brain development. In humans, mother-infant interactions may represent a key factor for disease susceptibility which can manifest itself as cognitive and behavioral disorders later in life. The maternal separation (MS) procedure of rat pups represents a useful experimental paradigm to study disturbances in brain function that might occur in response to adverse events during development. MS-subjected animals, as adults, express behavioral and neuroendocrine signs of elevated stress reactivity and cognitive deficits. However, mechanisms by which early life stress exerts its impact on the development and maturation of the brain are poorly understood. This study was aimed at finding the efects of repeated MS on long-term potentiation (LTP) in the lateral amygdala (LA) of adolescent rats. Wistar dams with their offspring were housed under 12:12 h light/dark cycle with food and water available ad libitum. On each of postnatal days (PNDs) 1 - 21 the dams were removed from the maternity cages for 3 h and placed individually in holding cages, while the litter stayed in homecages. Then, the dams were returned to the maternity cages. MS animals, as well as control, animal facility reared rats, were weaned at PND 28 and then housed in groups (4 - 6 animals). For electrophysiological experiments rats between PND35 and PND 50 were used. They were anesthetized and coronal brain slices (450 µm thick) containing the lateral amygdala were cut. Field potentials (FPs) were evoked in LA by the stimulation of the external capsule. LTP was induced using repetitive theta burst stimulation (TBS) protocol. While in slices prepared from control rats FP amplitude, 90 min after TBS, amounted 185.9 ± 17.04 % of baseline, in slices obtained from MS animals LTP was significantly weaker (117.7 ± 7.7%; p=0.0002). These preliminary results indicate that MS stress impairs LTP in the lateral amygdala of adolescent rats.
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