Rubem Carlos Araújo Guedes

After-effects of transcranial direct current stimulation (tDCS) on cortical spreading depression

Abnormal cortical excitability influences susceptibility to cortical spreading depression (CSD) in migraine. Because transcranial direct current stimulation (tDCS) is capable of inducing lasting changes of cortical excitability, we investigated the after-effects of tDCS on the propagation velocity of CSD in the rat. Twenty-five anesthetised rats received either anodal, cathodal or sham tDCS. The stimulation was applied for 20 min at a current strength of 200 microA after the recording of three baseline CSD measurements. Starting 5 min after tDCS, a further three CSDs were elicited and CSD velocity recorded at intervals of 20 min. tDCS and CSD recording was performed under anaesthesia with chloralose and urethane. As compared to the baseline velocity of 3.14 mm/min, anodal tDCS induced a significant increase of propagation velocity during the first post-tDCS recording (3.49 mm/min). In contrast to anodal tDCS, neither cathodal tDCS nor sham tDCS, which consisted of an initial ramped DC stimulation lasting only 20 s, showed a significant effect on CSD propagation velocity. As anodal tDCS is known to induce a lasting increase of cortical excitability in the clinical setting, our results support the notion that CSD propagation velocity reflects cortical excitability. Since cortical excitability and susceptibility to CSD is elevated in migraine patients, anodal tDCS - by increasing cortical excitability - might increase the probability of migraine attack in these patients, even beyond the end of its application.

Effects of transcranial direct current stimulation coupled with repetitive electrical stimulation on cortical spreading depression

We have recently shown that two techniques of brain stimulation - repetitive electrical stimulation (ES) (that mimics transcranial magnetic stimulation) and transcranial direct current stimulation (tDCS) - modify the velocity of cortical spreading depression (CSD) significantly. Herein we aimed to study the effects of these two techniques combined on CSD. Thirty-two Wistar rats were divided into four groups according to the treatment: sham tDCS/sham ES, sham tDCS/1 Hz ES, anodal tDCS/1 Hz ES, cathodal tDCS/1 Hz ES. Our findings show that 1 Hz ES reduced CSD velocity, and this effect was modified by either anodal or cathodal tDCS. Anodal tDCS induced larger effects than cathodal tDCS. Hereby CSD velocity was actually increased significantly after anodal tDCS/1 Hz ES. Our results show that combining two techniques of brain stimulation can modify significantly the effects of ES alone on cortical excitability as measured by the neurophysiological parameter of cortical spreading depression and therefore provide important insights into the effects of this new approach of brain stimulation on cortical activity.

Propagation of spreading depression inversely correlates with cortical myelin content

Cortical myelin can be severely affected in patients with demyelinating disorders of the central nervous system. However, the functional implication of cortical demyelination remains elusive. In this study, we investigated whether cortical myelin influences cortical spreading depression (CSD).

Reduction of intraspecific aggression in adult rats by neonatal treatment with a selective serotonin reuptake inhibitor

Most studies suggest that serotonin exerts an inhibitory control on the aggression process. According to experimental evidence, this amine also influences growth and development of the nervous tissue including serotoninergic neurons. Thus, the possibility exists that increased serotonin availability in young animals facilitates a long-lasting effect on aggressive responses. The present study aimed to investigate the aggressive behavior of adult rats (90-120 days) treated from the 1st to the 19th postnatal day with citalopram (CIT), a selective serotonin reuptake inhibitor (20 mg/kg, s.c., every 3 days). Aggressive behavior was induced by placing a pair of rats (matched by weight) in a box (20 x 20 x 20 cm), and submitting them to a 20-min session of electric footshocks (five 1.6-mA - 2-s current pulses, separated by a 4-min intershock interval). When compared to the control group (rats treated for the same period with equivalent volumes of saline solution), the CIT group presented a 41.4% reduction in the duration of aggressive response. The results indicate that the repeated administration of CIT early in life reduces the aggressive behavior in adulthood and suggest that the increased brain serotoninergic activity could play a role in this effect.

Manipulation of rat litter size during suckling influences cortical spreading depression after weaning and at adulthood.

Abstract Nutritional conditions early in life constitute one environmental factor that can influence brain electrophysiological features. Cortical spreading depression (SD) is a brain electrophysiological phenomenon that can be altered by the early nutritional status of organism. SD-velocity changes were presently studied in young (30–40 days old) and adult (90–120 days) rats suckled in litters formed by 3, 6, or 12 pups (called respectively small (S), medium (M) and large (L) litters). Body weights and SD propagation velocities in the 3 groups varied, respectively in an inverse and direct way, in relation to the litter sizes. The present investigation provides the first systematic description of the effectiveness of favorable and unfavorable lactation conditions (respectively suckling in Sand L litters) in altering cortical SD-propagation. The results confirm previous evidence in favor of permanent or at least long-lasting SD-changes associated to the prevailing nutritional status during the period of fast brain development.

Citalopram has an Antagonistic Action on Cortical Spreading Depression in Well-nourished and Early-malnourished Adult Rats

Adult, well-nourished (W) and early-malnourished (M) male Wistar rats were injected intraperitoneally for 7 days with 20 mg/kg CIT and cortical spreading depression (CSD) was recorded for 4h on the day following the treatment M-animals presented lower body weights, as well as higher CSD velocities of propagation, than the W ones, as previously reported. Compared to saline-injected controls, rats treated with CIT for 7 days presented comparable body weights and lower mean CSD velocities, per hour of recording, the differences being significant at the second hour (3.29 ± 0.31 versus 3.56 ± 0.40 mm/min; P < 0.05). Topical, cortical application of CIT (1- and 5 mg/ml solutions over the intact dura-mater) reduced dose-dependently the CSD velocity (maximal reductions of 16.3 and 55.8% for the 1 and 5 mg/ml solutions, respectively; P < 0.05), as well as the amplitude of the CSD-slow potential change (58.2 and 88.3%). In three out of seven W-rats and in one out of seven M-rats, topical CIT (5 mg/ml) blocked CSD propagation. The effects were reverted by flushing the treated region with saline. In the M-groups, CIT affected CSD in the same manner as in the W ones. The results reinforce previous evidence for an antagonistic influence of the serotoninergic activity on CSD.

Permanent and transitory morphometric changes of NADPH-diaphorase-containing neurons in the rat visual cortex after early malnutrition

We investigated the histochemical positivity to NADPH-diaphorase, which reveals nitric oxide synthase activity, in area 17 of rats malnourished early in life, both in the post-weaning period (group M1), and in adulthood after nutritional recovering (group M2). Control pups (C1 and C2 groups) received ad libitum after weaning the same diets as their mothers. Rats of group M2 were nutritionally recovered by receiving the control diet from post-natal day 42 until adulthood. Aldehyde-fixed sections (200-microm thick) through area 17 were processed for NADPH-diaphorase histochemistry following the malic enzyme indirect method. The features of NADPH-diaphorase-containing neurons of area 17 of malnourished young (M1) and adult (M2) rats were analyzed quantitatively in comparison to the matched groups C1 and C2. Permanent changes, represented by increase in the density and dendritic field areas of NADPH-diaphorase-positive cells, and transitory ones, represented by decreased values of soma areas, were observed in area 17 of the M1 and M2 cases. However, some other features, such as dendritic branch angle and number of dendrites per cell in the gray matter, remained unchanged after malnutrition. Thus, the findings indicate a possible relationship between early malnutrition and alterations in nitric oxide synthase-containing cells in the visual cortex. Physiological implications of these data may be related to synaptic plasticity and refinement of developmental brain circuits.

Dose‐Dependent Effects of Astaxanthin on Cortical Spreading Depression in Chronically Ethanol‐Treated Adult Rats

BACKGROUND The consumption of alcoholic drinks is a frequent drug-abuse situation, which is associated to a wide variety of pathological disturbances affecting several organs, including the brain. We have previously shown in the developing rat brain that ethanol intake facilitates the propagation of cortical spreading depression (CSD), an excitability-related neural phenomenon present in several animal species. This electrophysiological effect was attenuated by a shrimp (Litopenaeus vannamei) carotenoids extract. Here we investigated the effects of pure astaxanthin, the main carotenoid found in shrimp, on CSD. METHODS Adult Wistar rats were treated per gavage, during 18 days, with 2.5, 10 or 90 microg/kg/d astaxanthin dissolved in ethanol (3 g/kg) and CSD was recorded on the cortical surface 1 to 3 days thereafter. Four groups, treated respectively with ethanol, distilled water and soybean oil with- and without astaxanthin were also studied for comparison with the ethanol + astaxanthin groups. RESULTS Ethanol-treated rats displayed higher CSD-velocities (mean values, in mm/min, per hour of recording ranging from 4.08 +/- 0.09 to 4.12 +/- 0.16), compared to the distilled water-group (from 3.19 +/- 0.13 to 3.27 +/- 0.06). Addition of astaxanthin to ethanol lead to lower CSD-velocities in a dose-dependent manner, ranging from 3.68 +/- 0.09 to 3.97 +/- 0.22 for the 2.5 microg/kg/d-dose, from 3.29 +/- 0.09 to 3.32 +/- 0.07 for the 10 microg/kg/d-dose, and from 2.89 +/- 0.13 to 2.92 +/- 0.11 for the 90 microg/kg/d-dose. The velocities of the soybean oil groups (with and without astaxanthin) were not statistically different from the 10 microg/kg/d astaxanthin + ethanol and distilled water groups. CONCLUSION The results demonstrate the antagonistic effect of astaxanthin against the ethanol-induced facilitation of CSD propagation. Probably carotenoid antioxidant properties are involved in such effects.

Opioid mechanisms involved in the slow potential change and neuronal refractoriness during cortical spreading depression.

SummaryThe slow potential change (spc) accompanying spreading depression (SD) was studied in rats and in a seizure-sensitive strain of Mongolian gerbil under three different experimental paradigms, each involving the use of naloxone. Gerbils undergoing electroconvulsive shock treatment displayed SD during the post-ictal phase, which was blocked by the intraperitoneal (i.p.) administration of naloxone (20–50 mg kg-1). Topical application of naloxone to the exposed cortex of the anaesthetized gerbil and rat blocked the spc of SD evoked by KCl. Microiontophoretic ejection of naloxone during extracellular recordings reversed cell refractoriness following the spc, demonstrated by the observation of a maintained sensitivity to iontophoretic pulses of glutamate. The results suggest a possible involvement of naloxone-sensitive processes in the mechanism responsible for cortical SD.

Sexual differentiation of cortical spreading depression propagation after acute and kindled audiogenic seizures in the Wistar audiogenic rat (WAR)

SUMMARY Brain excitability diseases like epilepsy constitute one factor that influences brain electrophysiological features. Cortical spreading depression (CSD) is a phenomenon that can be altered by changes in brain excitability. CSD propagation was presently characterized in adult male and female rats from a normal Wistar strain and from a genetically audiogenic seizure-prone strain, the Wistar audiogenic rat (WAR), both previously submitted (RAS(+)), or not (RAS(-)), to repetitive acoustic stimulation, to provoke audiogenic kindling in the WAR-strain. A gender-specific change in CSD-propagation was found. Compared to seizure-resistant animals, in the RAS(-) condition, male and female WARs, respectively, presented CSD-propagation impairment and facilitation, characterized, respectively, by lower and higher propagation velocities (P<0.05). In contraposition, in the RAS(+) condition, male and female WARs displayed, respectively, higher and lower CSD-propagation rates, as compared to the corresponding controls. In some Wistar and WAR females, we determined estrous cycle status on the day of the CSD-recording as being either estrous or diestrous; no cycle-phase-related differences in CSD-propagation velocities were detected. In contrast to other epilepsy models, such as Status Epilepticus induced by pilocarpine, despite the CSD-velocity reduction, in no case was CSD propagation blocked in WARs. The results suggest a gender-related, estrous cycle-phase-independent modification in the CSD-susceptibility of WAR rats, both in the RAS(+) and RAS(-) situation.