Differential Effects of Pentoxifylline on Learning and Memory Impairment Induced by Hypoxic-ischemic Brain Injury in Rats.

OBJECTIVE: Hypoxic-ischemic (HI) brain injury in the human perinatal period often leads to significant long-term neurobehavioral dysfunction in the cognitive and sensory-motor domains. Using a neonatal HI injury model (unilateral carotid ligation followed by hypoxia) in postnatal day seven rats, the present study investigated the long-term effects of HI and potential behavioral protective effect of pentoxifylline. METHODS: Seven-day-old rats underwent right carotid ligation, followed by hypoxia (FiO2 = 0.08). Rats received pentoxifylline immediately after and again 2 hours after hypoxia (two doses, 60‒100 mg/kg/dose), or serum physiologic. Another set of seven-day-old rats was included to sham group exposed to surgical stress but not ligated. These rats were tested for spatial learning and memory on the simple place task in the Morris water maze from postnatal days 77 to 85. RESULTS: HI rats displayed significant tissue loss in the right hippocampus, as well as severe spatial memory deficits. Low-dose treatment with pentoxifylline resulted in significant protection against both HI-induced hippocampus tissue losses and spatial memory impairments. Beneficial effects are, however, negated if pentoxifylline is administered at high dose. CONCLUSION: These findings indicate that unilateral HI brain injury in a neonatal rodent model is associated with cognitive deficits, and that low dose pentoxifylline treatment is protective against spatial memory impairment.

The effects of intra-hippocampal L-thyroxine infusion on long-term potentiation and long-term depression: A possible role for the αvß3 integrin receptor.

Although the effects of long-term experimental dysthyroidism on long-term potentiation (LTP) and long-term depression (LTD) have been documented, the relationship between LTP/LTD and acute administration of L-thyroxine (T4) has not been described. Here, we investigated the effects of intra-hippocampal administration of T4 on synaptic plasticity in the dentate gyrus of the hippocampal formation. After a 15-minute baseline recording, LTP and LTD were induced by application of high- and low-frequency stimulation protocols, respectively. Infusions of saline or T4 and tetraiodothyroacetic acid (tetrac), a T4 analog that inhibits binding of iodothyronines to the integrin αvß3 receptor, either alone or together, were made during the stimulation protocols. The averages of the excitatory postsynaptic potential (EPSP) slopes and population spike (PS) amplitudes, between 55 to 60 minutes, were used as a measure of the LTP/LTD magnitude and were analyzed by two-way univariate ANOVA with T4 and tetrac as between-subjects factors. The input-output curves of the infusion groups were comparable to each other, as shown by the non significant interaction observed between stimulus intensity and infused drug. The magnitude of the LTP in T4-infused rats was significantly lower as compared to saline-infused rats. Both the PS amplitude and the EPSP slope were depressed more markedly with T4 infusion than with saline, tetrac, and T4 + tetrac infusion. Data of this study provide in vivo evidence that T4 can promote LTD over LTP via the integrin αvß3 receptor, and that the effect of endogenous T4 on this receptor can be suppressed by tetrac in the hippocampus. © 2016 Wiley Periodicals, Inc.

Adult-onset hyperthyroidism impairs spatial learning: possible involvement of mitogen-activated protein kinase signaling pathways.

Given evidence that mitogen-activated protein kinase (MAPK) activation is part of the nongenomic actions of thyroid hormones, we investigated the possible consequences of hyperthyroidism for the cognitive functioning of adult rats. Young adult rats were treated with L-thyroxine or saline. Twenty rats in each group were exposed to Morris water maze testing, measuring their performance in a hidden-platform spatial task. In a separate set of rats not exposed to Morris water maze testing (untrained rats), the expression and phosphorylated levels of p38-MAPK and of its two downstream effectors, Elk-1 and cAMP response element-binding protein, were evaluated using quantitative reverse transcriptase-PCR and western blotting. Rats with hyperthyroidism showed delayed acquisition of learning compared with their wild-type counterparts, as shown by increased escape latencies and distance moved on the last two trials of daily training in the water maze. The hyperthyroid rats, however, showed no difference during probe trials. Western blot analyses of the hippocampus showed that hyperthyroidism increased phosphorylated p38-MAPK levels in untrained rats. Although our study is correlative in nature and does not exclude the contribution of other molecular targets, our findings suggest that the observed impairments in acquisition during actual learning in rats with hyperthyroidism may result from the increased phosphorylation of p38-MAPK.

Low-frequency stimulation induces a durable long-term depression in young adult hyperthyroid rats: the role of p38 mitogen-activated protein kinase and protein phosphatase 1.

Long-term potentiation and long-term depression (LTD) are cellular mechanisms of learning and memory in the mammalian brain. We have previously shown that adult hyperthyroid rats showed a delay in the acquisition of a place learning task and attenuated long-term potentiation. However, changes in LTD in hyperthyroidism remain unclear. Rats were administered 0.2 mg/kg/day of L-thyroxine for 21 days starting at postnatal day 40 to induce hyperthyroidism. LTD was induced in the dentate gyrus using low-frequency stimulation (LFS) of the perforant pathway. The mRNA expressions of p38 mitogen-activated protein kinase (p38-MAPK) and protein phosphatase 1 (PP1) were evaluated using a quantitative reverse transcriptase PCR. In control rats, a standard LFS protocol induced a slight depression of the population spike (PS) amplitude during the induction phase of LTD (76±13% vs. baseline), but a small potentiation of the PS amplitude was observed in the early (107±18%) and late (111±20%) phases of LTD. Interestingly, in the hyperthyroid rats, the same LFS protocol induced a reliable LTD in the dentate gyrus of the hippocampus as evidenced by a marked depression in the PS amplitude during the induction (54±6% vs. baseline) and the early phases (56±8%) of LTD. Elevated mRNA levels of p38-MAPK and PP1 were observed in the hippocampus of the LFS-treated hyperthyroid rats compared with the hippocampus of the vehicle-treated hyperthyroid rats. No significant change in p38-MAPK or PP1 mRNA expression was observed in the euthyroid rats. The present study shows that a standard LFS protocol can induce a durable depression of synaptic strength and an upregulation of PP1 and p38-MAPK mRNA in hyperthyroid rats. We conclude that hyperthyroidism can induce molecular changes associated with degeneration of the hippocampus. The relationship between the levels of thyroid hormone and dementia requires further investigation.

Effects of selenium treatment on 6-n-propyl-2-thiouracil-induced impairment of long-term potentiation.

The goal of this study was to evaluate whether sodium selenite could afford protection against the effects of hypothyroidism on long-term potentiation (LTP), which is thought to be the cellular basis for learning and memory. Hypothyroidism was induced in young-adult rats by the administration of 6-n-propyl-2-thiouracil (PTU) in tap water for 21 days. Half of these hypothyroid and euthroid rats were given 10ppM selenium with their drinking water. Field potentials were recorded from the dentate gyrus in response to stimulation of the medial perforant pathway in vivo. PTU treatment resulted in a significant reduction in both free T3 and free T4 levels, whereas selenium administration to PTU-treated rats restored only the levels of free T3 to their control values. Thyroid hormone levels were not affected by selenium in euthyroid rats. PTU-treated rats exhibited an attenuation of population spike (PS) - LTP, but a comparable potentiation of excitatory postsynaptic potential (EPSP) was found among these rats. The administration of selenium to PTU-treated rats was partially able to attenuate impairment of LTP, but not of potentiation during the LTP induction protocol in hypothyroid rats. Interestingly, the hypothyroid rats that were supplemented with selenium had a lower EPSP potentiation during induction protocol than the control rats. The present study suggests a possible importance of T3 in Se-induced rescue of impaired PS-LTP in hypothyroidism.

Effects of cold exposure on behavioral and electrophysiological parameters related with hippocampal function in rats.

AIM: Behavioral and mental changes may occur in people exposed to cold stress by decreasing their work efficiency and their mental capacity while increasing the number of accidents on the job site. The goal of this study was to explore the effect of cold stress in spatial learning performance excitability and LTP. MATERIALS AND METHODS: Three to four month old rats were randomly divided into four groups to form a control group and a cold stress group for each sex. The groups of cold stressed animals were placed in a cold room ambient temperature of 4°C for 2 h day. Adrenal glands and body weight (g) were recorded in control and stressed rats during the cold exposure. Spatial learning (acquisition phase) and memory (probe trial) were tested in the Morris water maze (MWM) immediately after daily exposure. Latency to locate the hidden platform, distance moved (DM), mean distance to platform, swim speed (SS) and time spent in the platform quadrant were compared between genders and treatments. Field potential recordings were made, under urethane anesthesia, from the dentate gyrus (DG) granule-cell layer, with stimulation of the medial perforant pathway 2 h after the probe trial. This study examined spatial memory as measured by MWM performance and hippocampal long-term potentiation (LTP) in the DG after exposure to cold in a repeated stress condition for 2 h/day for 5 days. RESULTS: The cold-exposed female rats needed less time to find the hidden platform on day 1 (43.0 ± 13.9 s vs. 63.2 ± 13.2 s), day 2 (18.2 ± 8.4 s vs. 40.9 ± 12.2 s) and on day 4 (8.0 ± 2.1 s vs. 17.2 ± 7.0 s) while cold-exposed male rats showed a decreased escape latency (EL) on day 1 only (37.3 ± 12.5 s vs. 75.4 ± 13.1 s). Cold-exposed male rats spent less time in the target quadrant (30.08 ± 6.11%) than the control male rats (37.33 ± 8.89%). Two hour cold exposure decreased population spike (PS) potentiation during both induction (218.3 ± 21.6 vs. 304.5 ± 18.8%) and maintenance intervals (193.9 ± 24.5 vs. 276.6 ± 25.4%) in male rats. Meanwhile cold exposure did not affect the body weight (C: 221 ± 2.5 vs. S: 222 ± 1.7) but it impacts the adrenal gland relative weight (S: 27.1 ± 1.8 mg vs. C: 26.2 ± 1.4 mg). CONCLUSION: Overall, the results show that repeated cold exposure can selectively improve spatial learning in adult female rats, but impaired retention memory for platform location in male rats. It is possible that impaired LTP underlies some of the impaired retention memory caused by cold exposure in the male rats.

Maternal intake of Omega-3 essential fatty acids improves long term potentiation in the dentate gyrus and Morris water maze performance in rats.

Omega-3 fatty acid deprivation during development reduces performance in learning tasks, and dietary DHA supplementation improves learning ability and enhances long term memory in both young and old animals. However, little attention has been paid to the effect of maternal intake of Omega-3 fatty acids on hippocampal function in their pups. Randomly some of the pregnant dams were supplemented with Omega-3 essential fatty acid, others with tap-water, during pregnancy and breast-feeding by gavage daily. Spatial learning and memory was tested in Morris water maze. Field potentials from the dentate gyrus were recorded in response to medial perforant pathway in urethane-anesthetized pups. Omega-3-treated rats found the platform less traveled and closer to platform than control animals. However the pups from both groups show the same performance in retrieval task. No differences were found between corresponding animal groups in the input-output curves of the field potential slopes, suggesting no effect of Omega-3 supplementation on basal synaptic efficacy. Potentiation of population spike amplitude was much higher in pups of Omega-3 treated dams than control. Up to now Omega 3 fatty acid has been shown to be beneficial on the synaptic plasticity only under some pathological conditions. For the first time, we showed improved dentate gyrus-LTP and enhanced Morris water maze performance in healthy pups from healthy dams treated with Omega-3 fatty acids during pregnancy and breast-feeding period. Molecular studies are needed to explain Omega-3 effect on hippocampal synaptic plasticity.

Experimentally induced hyperthyroidism disrupts hippocampal long-term potentiation in adult rats.

BACKGROUND: Manipulating thyroid hormones has been shown to influence learning and memory. Although a large body of literature is available on the effects of thyroid hormone deficiency on learning and memory functions during developmental or adult-onset hypothyroidism, electrophysiological findings are limited. This limitation is especially notable with respect to thyroxine administration in adult, normothyroid animals. METHODS: Experiments were carried out on 12 adult male Wistar rats, each 9-10 months of age. Rats were randomly divided into hyperthyroid (0.2 mg/kg/day intraperitoneal thyroxine injection, for 21 days) and control groups (n = 6 animals in each group). Following spatial learning performance tests on hyperthyroid and control groups, rats were anesthetized with urethane and placed in a stereotaxic frame. A bipolar, tungsten electrode was used to stimulate the medial perforant path. A glass micropipette was inserted within the granule cell layer of the ipsilateral dentate gyrus to record field excitatory postsynaptic potentials (fEPSP). Following a 15-min baseline recording of fEPSPs, long-term potentiation (LTP) was induced by four sets of tetanic pulse trains. RESULTS: Thyroxine-treated rats showed significantly worse performance in the spatial memory task and attenuated input-output relationships in the electrophysiological analyses. Treated rats also showed a lower efficacy of LTP induction when compared with controls. CONCLUSION: The present study provides clear in vivo evidence for the action of L-thyroxine in the impairment of synaptic plasticity and in inducing spatial memory task deficits in adult rats. These findings may explain the complaints of cognitive function reductions in hyperthyroid patients.