Step-by-step approach: Stereotaxic surgery for in vivo extracellular field potential recording at the rat Schaffer collateral-CA1 synapse using the eLab system.

In vivo extracellular field potential recording is a commonly used technique in modern neuroscience research. The success of long-term electrophysiological recordings often depends on the quality of the implantation surgery. However, there is limited use of visually guided stereotaxic neurosurgery and the application of the eLab/ePulse electrophysiology system in rodent models. This study presents a practical and functional manual guide for surgical electrode implantation in rodent models using the eLab/ePulse electrophysiology system for recording and stimulation purposes to assess neuronal functionality and synaptic plasticity. The evaluation parameters included the input/output function (IO), paired-pulse facilitation or depression (PPF/PPD), long-term potentiation (LTP), and long-term depression (LTD).•Provides a detailed picture-guided procedure for conducting in vivo stereotaxic neurosurgery.•Specifically covers the insertion of hippocampal electrodes and the recording of evoked extracellular field potentials.

Differential impact of a ghrelin receptor antagonist or inverse agonist in the electrical kindling model of epilepsy.

Ghrelin is a peptide, which has been shown to affect seizures. However, there is not a consensus about its real impact on the control of seizure severity. We assessed the influence of intra-amygdala injections of a ghrelin receptor (GHSR) antagonist, as well as a GHSR inverse agonist on the electrical kindling-induced seizures. Two unipolar electrodes and a tripolar electrode twisted with a guide cannula were implanted in the skull surface or the basolateral amygdala of adult male rats, respectively. A rapid electrical kindling protocol was applied for kindling epileptogenesis. The stimulations were applied until rats showed three consecutive stage five seizures. Each rat was considered as its control. D-Lys-3-GHRP-6 (1, 12.5, and 25 µg/rat) or [D-Arg, D-phe, D-Trp, heu] substance P (D-SP) (50, 500 and 5000 ng/rat) as the GHSR antagonist or inverse agonist were injected into the basolateral amygdala. Seizure parameters including after-discharge duration (ADD), stage five duration (S5D), and seizure stage (SS) were documented thirty minutes following administration of the drugs or saline. Antagonism of the GHSR in the amygdala, significantly increased seizure induction in the kindled rats, in a dose-dependent manner, and induced spontaneous seizures leading to status epilepticus. Conversely, D-SP had a dose-dependent anticonvulsant activity, indicated by decreased ADD and S5D. The results show that GHSR inverse agonism suppressed seizure severity in the rat amygdala kindling model, whereas GHSR antagonism made seizures more severe. Therefore, when considering the ghrelin system to modulate seizures, it is crucial to note the differential impact of various GHSR ligands.

Down regulation of the hippocampal ghrelin receptor type-1a during electrical kindling-induced epileptogenesis.

Numerous studies have shown that the ghrelin hormone is involved in epileptic conditions. However, the profile of ghrelin or its functional receptor mRNAs in seizure-susceptible brain areas was not assessed during epileptogenesis. Here, we measured the expression levels of the hippocampal ghrelin or its receptor mRNAs during electrical kindling-induced epileptogenesis. The study was conducted on twenty adult male Wistar rats. One tri-polar and two uni-polar electrodes were stereotaxically implanted in the baso-lateral amygdala or skull surface, respectively. Animals were divided into four groups, consisting of two sham groups (sham1 and sham2), and two other groups, which were partially or fully kindled. After the establishment of partial or full kindling, the hippocampi of the animals and that of the corresponding sham groups were removed. A quantitative real-time PCR technique was used to measure the expression levels of ghrelin or its functional receptor mRNAs. The results indicated that the expression levels of ghrelin did not alter in either of the partially or fully kindled rats compared to the corresponding sham groups. Ghrelin receptor (ghrelinR) down regulated, significantly in the fully-kindled rats, compared to sham2 group. Meanwhile, the mRNA expression levels of ghrelinR did not change in partially-kindled rats compared to sham1 group. The outcomes of the current study highlight the crucial, unknown impact of the hippocampal ghrelinR through the development of electrical kindling epileptogenesis, and points out the importance of ghrelinR as a goal to adjust epileptogenic progression.

Antagonism of the ghrelin receptor type 1a in the rat brain induces status epilepticus in an electrical kindling model of epilepsy.

Studies have shown the anti-seizure properties of the ghrelin hormone in different models of epilepsy. Nevertheless, the role of the endogenous ghrelin is unknown in the electrical kindling model of epilepsy. In this study, we evaluated the effect of the antagonism of the ghrelin receptors in the brain of fully kindled rats. Adult male Wistar rats weighing 300 g were used. Animals were stereotaxically implanted with two uni-polar electrodes in the skull surface and a tri-polar electrode in the basolateral amygdala, and a guide cannula in the left lateral ventricle. Animals underwent a rapid kindling protocol. After showing three consecutive stages of five seizures, the animals were considered fully kindled. D-Lys-3-GHRP-6 (1, 50, and 100 µg/rat) was injected intracerebroventricularly (i.c.v.) in the kindled animals. Each rat was considered as its control and received a single dose of D-Lys-3-GHRP-6. Seizure parameters including after discharge duration (ADD), seizure stage (SS), stage four latency (S4L), and stage five duration (S5D) were recorded. The paired t test indicated a significant increase in seizure induction. D-Lys-3-GHRP-6 (1 µg/rat; i.c.v.) prolonged ADD in the kindled rats, significantly. D-Lys-3-GHRP-6 (50 and 100 µg/rat; i.c.v.) induced spontaneous seizures, which led to status epilepticus in the kindled rats. The results indicate that the antagonism of the ghrelin functional receptors prolongs seizures and induces status epilepticus in the kindling model of epilepsy, and propose that the endogenous ghrelin signaling has crucial antiepileptic properties.

Alteration of intrapancreatic serotonin, homocysteine, TNF-α, and NGF levels as predisposing factors for diabetes following exposure to 900-MHz waves.

Exposure to mobile phone radiation causes deleterious health effects on biological systems. The objects of this study were to investigate the effect of 900-MHz radiofrequency waves (RFW) emitted from base transceiver station antenna on intrapancreatic homocysteine (Hcy), tumor necrosis factor-α (TNF-α), and nerve growth factor (NGF) as predisposing factors involved in pancreatic beta cell damage. Thirty male rats (Sprague-Dawley, 200 ± 10 g) were randomly divided into the control (without any exposure) and exposed groups: short time (2 h/day), long time (4 h/day), and exposed to 900-MHz RFW for 30 consecutive days. On the last days of the experiment, animals were killed and pancreas tissue was dissected out for evaluation of serotonin, Hcy, TNF-α, and NGF. There was a significant decrease in the serotonin and NGF levels in the pancreatic tissue of exposed groups compared to the control group (p < 0.05). Also, the levels of serotonin and NGF in the long-time exposure were significantly lower than the short-time exposure (p < 0.05). However, levels of Hcy and TNF-α were significantly increased in the pancreas of exposed groups compared to the control groups (p < 0.05). Exposure to 900-MHz RFW decreased pancreatic NGF and serotonin levels and increased the proinflammatory markers (Hcy and TNF-α), which can be a predisposing factor for type 2 diabetes.

Prenatal and early postnatal exposure to radiofrequency waves (900 MHz) adversely affects passive avoidance learning and memory.

Prenatal and early postnatal are the most sensitive and high-risk periods to expose to electromagnetic fields (EMFs). This study aimed to investigate the effect of prenatal and early postnatal exposure to 900 MHz radiofrequency waves (RFWs) emitted from a base transceiver station antenna on passive avoidance learning and memory (PALM) and hippocampus histomorphology. Female Sprague Dawley rats (190-230 g) were paired with males. The mated rats, confirmed by observing a vaginal plug, were divided into two groups; control and exposed. The control group (n = 7) was not exposed to RFW. The exposed group was divided into three subgroups (n = 8); exposed Ⅰ, exposed during the gestational period (fetal life), and exposed Ⅱ and Ⅲ (postnatal exposure), exposed to RFW during the first 21 days of life, for 2 h/d and 4 h/d, respectively. PALM was evaluated by a shuttle box in 45-day-old pups. Learning and memory of animals were demonstrated as the duration of remaining within the light area, which is called the lighting time. Histological sections were prepared from brain tissues and stained with hematoxylin and eosin. An impairment in the PALM performance was noticed in all exposed subgroups (Ⅰ, Ⅱ, and Ⅲ) (p < 0.05). Learning (short-term memory) and retention (long-term memory) behaviors were more affected in exposed subgroup Ⅰ (prenatal exposed) compared to other postnatal exposed subgroups (Ⅱ and Ⅲ). Also, a mild decrease in the density of pyramidal cells was observed in the hippocampus of exposed subgroups (Ⅰ and Ⅲ). Prenatal and early postnatal exposure to 900 MHz RFW adversely affected PALM performance and hippocampus tissue in rat pups with more impact for prenatal period exposure.

The protective effect of vitamin supplementation (E and E + C) on passive avoidance learning and memory during exposure to 900 MHz RFW emitted from BTS.

Deleterious effects of exposure to electromagnetic radiation on public health have been widely studied. This study was conducted to evaluate the protective effect of vitamin supplementation (E or E + C) on passive avoidance learning (PAL) and memory in rats subjected to 900 MHz radiofrequency waves (RFW). Thirty adult male Sprague-Dawley rats (190 ± 20 g) were randomly divided into six groups as: control I (vehicle), control II (vitamin E 250 mg/kg), control III (vitamin E 100 mg/kg + l-ascorbic acid 200 mg/kg), and three exposed groups to RFW as: sham-exposed, treatment I (vitamin E), and treatment II (vitamin E + C). The duration of exposure was 30 continuous days (4 h/day). The PAL was evaluated on the last day by the shuttle box. Learning and memory of animals demonstrated as the duration of remaining within the light area, which is called the light time (LT). The sham-exposed group showed a significant decrease in LT on the learning, consolidation, and retention days compared to other groups (p < 0.05). Pretreatment with vitamins (E and E + C) could protect PAL against adverse effects of RFW, and the administration of vitamin E + C improved PAL performance in control III compared to control I and treatment II groups (p < 0.05). Administration of vitamin E + C to exposed group (treatment II) caused a significant increase in LT on the learning (p = 0.013), consolidation, and retention (p = 0.009) sessions compared to the treatment group I (vitamin E). Long-term exposure to 900 MHz RFW impaired PAL and memory, and pretreatment of vitamin (E or E + C) prevented these effects, which may be a new potential mechanism against side effects of RFW.

Trace elements homeostasis in brain exposed to 900 MHz RFW emitted from a BTS-antenna model and the protective role of vitamin E.

Advances in telecommunication and their broad usage in the community have become a great concern from the health aspect. The object of the present study was to examine the effects of exposure to 900 MHz RFW on brain Iron (Fe), Copper (Cu), Zinc (Zn) and Manganese (Mn) concentration, and the protective role of pre-treatment of vitamin E on mentioned elements homoeostasis. Twenty adult male Sprague-Dawley rats (200 ± 20 g) randomly were divided into four groups. Control group (without any exposure, received distilled water), treatment control group (orally received 250 mg/kg BW/d vitamin E), treatment group (received 250 mg/kg BW/d vitamin E and exposed to 900 MHz RFW) and sham-exposed group (exposed to 900 MHz RFW). Animals (with freely moving in the cage) were exposed to RFW for 30 consecutive days (4 hr/day). The levels of the above mentioned elements in the brain tissue were determined on the last day using atomic absorption spectrophotometry. Exposure to 900 MHz RFW induced a significant increase in the Fe, Cu, Mn levels and Cu/Zn ratio accompanied by a significant decrease in Zn level in the sham-exposed group compare to control group. Vitamin E pre-treatment improved the level of Fe, Cu, Mn and Cu/Zn ratio, except in the Zn concentration. Exposure to 900 MHz RFW caused disrupted trace elements homoeostasis in the brain tissue and administration of vitamin E as an antioxidant and neuroprotective agent improved the situation.

Alteration of testicular regulatory and functional molecules following long-time exposure to 900 MHz RFW emitted from BTS.

The aim of this investigation was to evaluate changes in testosterone and some of the functional and regulatory molecules of testis such as P450scc, steroidogenic acute regulatory protein (StAR), tumour necrosis factor-α (TNF-α), interleukin-1α (IL-1α), interleukin-1ß (IL-1ß) and nerve growth factor (NGF) following exposure to 900 MHz radio frequency (RF). Thirty adult male Sprague Dawley rats (190 ± 20 g BW) were randomly classified in three equal groups, control (sham, without any exposure), short-time exposure (2 hr) (STE) and long-time exposure (4 hr) (LTE). The exposure was performed for 30 consecutive days. The testosterone level in both exposed groups was significantly less than control (p < .05). Level of TNF-α in both exposed groups was significantly greater than control (p < .05). IL-1α and NGF levels in LTE were significantly higher than the STE and control groups (p < .05). Level of IL-1ß in LTE was significantly higher than control (p < .05). Expression of both P450scc and StAR mRNA was significantly down-regulated in both exposed groups compared to control (p < .05). Our results showed that RFW can affect testis and reproductive function through changes in factors, which are important during steroidogenesis, and also through changes in inflammatory factors, which regulate Leydig cell functions.