Yukio Hattori

Increase in the calcium level following anodal polarization in the rat brain

The accumulation of calcium ions (Ca) was examined in the rat brain by means of 45Ca autoradiography following the application of a weak anodal direct current to the surface of the sensorimotor cortex. Repetition of the anodal polarization with 3.0 microA for 30 min caused more Ca to accumulate in the cerebral cortex. The degree and extent of accumulation was greater in the hemisphere ipsilateral to the polarization than in the other. Accumulation was also noted in the hippocampus and thalamus. Ca accumulation was detected after 24 h and it remained virtually constant up to 72 h after the last polarization. The results suggest that a long-lasting disturbance of Ca homeostasis is involved in the cortical plastic changes seen following anodal polarization.

Biphasic effects of polarizing current on adenosine-sensitive generation of cyclic AMP in rat cerebral cortex

Cyclic AMP accumulation elicited by adenosine was investigated in cortical slices of rats following the application of a weak anodal direct current (anodal polarization) to the unilateral sensorimotor cortex. Anodal polarization with 3.0 microA for 30 min caused an increase in the adenosine-elicited accumulation of cyclic AMP in the polarized cortex, in which case the increase in the polarized cortical region was highlighted by repeated applications of the currents. Polarization with 0.3 microA for 30 min decreased the cyclic AMP accumulation, and polarization with 30.0 microA for 30 min had no effect. When applied for 3 h, the polarizing currents at all of the intensities tested decreased the cyclic AMP accumulation. The results indicate that anodal polarization has biphasic effects on adenosine-elicited accumulation of cyclic AMP in the cortex. Alterations in the cyclic AMP generation are suggested to form the neurochemical basis of central and behavioral activity induced by anodal polarization.

Atrial natriuretic polypeptide depresses angiotensin II induced excitation of neurons in the rat subfornical organ in vitro

The effects of atrial natriuretic polypeptide (ANP) on the extracellularly recorded activity of neurons in the subfornical organ (SFO) were investigated in rat brain slice preparations by adding the peptide to the perfusion medium. Eight (14%) of 56 SFO neurons were inhibited and none of the cells were excited by ANP at 10(-7) M. Of 42 SFO neurons tested with both ANP and angiotensin II (AII) at 10(-7) M, 4 (10%) cells were inhibited by ANP and excited by AII, the remainder responded to either one or other of the peptides but not both or were unresponsive. In 13 (87%) of 15 SFO neurons, ANP at 10(-7) M depressed by more than 40% the excitation induced by AII at 10(-7) M, while ANP did not always depress the excitation induced by raising the extracellular potassium concentrations in 6 SFO cells tested. We conclude that ANP strongly depresses AII-induced excitation in all SFO neurons, although it has very weak inhibitory effects on spontaneous activity, thus ANP may act as a neuromodulator in the SFO.

Increases in brain polyamine concentrations in chemical kindling and single convulsion induced by pentylenetetrazol in rats

Concentrations of the polyamines, putrescine, spermidine and spermine were investigated in rat brains, in which chemical kindling or single convulsion had been induced by intraperitoneal injection of pentylenetetrazol (PTZ). A single injection of 60 mg/kg of PTZ produced tonic-clonic convulsion and increased the putrescine concentration 8 h after the injection. At lower doses of PTZ (10 and 30 mg/kg), neither marked behavioral seizure nor significant change in any polyamine concentration was observed. On the other hand, repeated injections of 30 mg/kg of PTZ eventually resulted in intense motor seizures (PTZ kindling) and increased the concentrations of all three polyamines. The most marked increase was detected in putrescine 1-48 h after the intense seizures. The increase in putrescine was clearly higher in PTZ kindling than in single convulsion. These results suggest that increases in polyamine concentrations are involved in neuronal excitability in the epileptic brain.

c-Fos Expression Mediated by N-Methyl-d-aspartate Receptors Following Anodal Polarization in the Rat Brain

c-Fos protein-like immunoreactivity (IR) was investigated in the rat brain following an application of weak anodal direct current to the surface of the unilateral sensorimotor cortex in an attempt to elucidate the cellular and molecular bases of central plasticity. Anodal polarization resulted in a massive increase in c-Fos protein-like IR in neurons of the cingulate, piriform, frontoparietal cortices, and hippocampus ipsilateral to the polarization. The effects were dependent upon the duration and intensity of currents applied. The time-dependent induction of c-Fos protein-like IR was maximal at 1 h, became weaker by 6 h, and almost returned to the baseline within 24 h following polarization. When MK-801 [(+)-5-methyl-10,11-di-hydro-5H- dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate], a noncompetitive antagonist for N-methyl-D-aspartate (NMDA) receptors, was injected intraperitoneally, the induction of this nuclear protein was reduced or completely blocked in both hemispheres, except around the polarized point itself, as a function of the time and dosage. These results suggest that the proto-oncogene c-fos is rapidly and transiently activated in the brain following anodal polarization and this activation is mediated by NMDA receptors.

Involvement of Putrescine in the Development of Kindled Seizure in Rats

Abstract: During the development of kindling by daily electrical stimulations applied to the left amygdala of rats, concentrations of the polyamines putrescine, spermidine, and spermine were measured in the left amygdala and the remainder of the cerebrum. A significant increase of putrescine concentration appeared first at the left amygdala in prekindled rats and then propagated to the remainder of the cerebrum with the development of kindling. This increase in putrescine concentration in the left amygdala was higher in prekindled rats than in fully kindled rats and lasted for at least 24 h after the final kindling stimulation. The concentrations of spermidine and spermine were slightly increased in a fully kindled state. To clarify the role of putrescine in kindling, the development of amygdaloid kindling was examined in rats after microinjections of α‐difluoromethylornithine, a specific inhibitor of polyamine synthesis, and putrescine into the ipsilateral amygdala. Pretreatment with α‐difluoromethylornithine (50 nmol) for 10 days accelerated both the development of behavioral kindling and the propagation of the afterdischarge from the left amygdala to the frontal cortex. In contrast, pretreatment with putrescine (200 nmol) for 10 days retarded the development of kindling. These results suggest that the increase in putrescine concentration in the kindled brain has an inhibitory effect on the development of kindling.

Electrocorticographic characterization of chronic iron-induced epilepsy in rats.

Electrocorticograms were recorded from rats which were unilaterally injected with ferrous chloride solution into the sensorimotor cortex to induce chronic epileptic activity. All of the iron-injected rats showed isolated spikes near the injection site and in the contralateral cortex immediately after the injection. The injection produced 3 kinds of responses in the rats according to the frequency of the isolated spikes. Spike and wave complexes appeared bilaterally approximately 30 days or more after the injection in the rats in which the frequency of the isolated spikes was dominant on the side ipsilateral to the injection site or nearly equal on the two sides. These results suggest that there are at least 2 stages in the development of chronic iron-induced epilepsy.

Changes in polyamine concentrations in amygdaloid-kindled rats

Concentrations of the polyamines putrescine, spermidine, and spermine were investigated in the left and right amygdala and in the remaining cerebrum, in which kindling was induced by repeated application of electrical stimulation of the left amygdala of rats. In kindled rats, the concentrations of spermidine and spermine increased slightly, but elevations did not reach significant levels in any brain regions. The most profound increase was detected in the putrescine concentration in all parts of the cerebrum 1–8 h after the final stimulation. These results suggest that the increases in concentrations of polyamines, particularly of putrescine, are involved in the pathogenesis of amygdaloid kindling.

Development of epileptic activity induced by iron injection into rat cerebral cortex: electrographic and behavioral characteristics.

Unilateral injection of ferrous chloride solution into the rat sensorimotor cortex produced epileptic discharges in the electrocorticograms (ECoGs). The discharges were isolated spikes and spike and wave complexes, and the epileptic activity lasted for more than 12 months after the injection. Isolated spike activity often appeared on the left or right side of the cortex, whereas spike and wave complex activity appeared bilaterally. In rats showing dominant isolated spike activity in the secondary epileptic cortex, there was a deviation in somatosensory evoked potentials (SEPs). Rats showing isolated spikes and spike and wave complexes exhibited vibrissa tremors and head nodding. Rats showing only isolated spikes exhibited no abnormal behavior, but their convulsion thresholds to pentylenetetrazol were lowered. The results including ECoGs, SEPs, behavior and convulsion threshold were characterized with reference to the development of iron-induced epilepsy. The profiles of ECoG discharge activity and SEP configuration suggest that the process of iron-induced epilepsy consists of 3 stages.

Regional difference in responsiveness of norepinephrine-sensitive cyclic AMP-generating systems of rat cerebral cortex with iron-induced epileptic activity

Responsiveness of norepinephrine (NE)–sensitive cyclic AMP (cAMP)‐generating systems was determined in slices from different areas of the rat cerebral cortex in which FeCl2 solution was injected unilaterally into the sensorimotor cortex to induce epileptic activity. In anterior cortical areas of rats in which the appearance of electrographic isolated spikes was dominant either ipsilaterally or contralaterally to the injection site 8–10 days after the injection, the cAMP accumulations elicited by NE and an NE‐phentolamine combination were greater on the side of dominant spike activity than on the other. In anterior cortical areas of rats showing dominant spike activity on either side of the cortex 31–60 days after the injection, the cAMP accumulation elicited by NE was smaller on the dominant side than on the other. In anterior cortical areas of rats showing nearly equal spike activity on the two sides 31–60 days after the injection, the cAMP accumulations elicited by NE and an NE‐phentolamine combination were greater on the side ipsilateral to the injection site than on the other. In anterior and posterior cortical areas of rats in which the appearance of spike and wave complexes, as well as isolated spikes, was detected 31–60 days after the injection, the cAMP accumulations elicited by NE and combinations of NE and phentolamine or propranolol were greater on the side ipsilateral to the injection site than on the other. The elicitation by an NE‐propranolol combination, but not by an NE‐phentolamine combination, of cAMP accumulation was almost completely inhibited by 8‐phenyltheophylline. These results suggest that alterations in NE‐sensitive cAMP‐generating systems of the cortex involving α‐ and β‐adrenoceptors and adenosine receptors are related to the neurochemical process of iron‐induced epilepsy.