Manipulation of the somatosensory cortex modulates stimulus-induced repetitive ear movements in a seizure-sensitive strain of gerbil.

Some Mongolian gerbils (Meriones unguiculatus) respond to stimulation by seizures, the pattern of which changes progressively during development. We previously established a seizure-sensitive strain, MGS/Idr, in which all animals exhibit such stimulus-induced seizures. We have now noted that all adults of this strain also show repetitive backward movements of the ears at the ears at the beginning of stimulus-induced seizures, although the incidence varies with the individual. We examined whether the cerebral cortex was involved in these movements and found that electrical stimulation of an area of the somatosensory cortex elicited strong backward movement of the ear on the contralateral side, and that unilateral application of bicuculline, a GABAA receptor antagonist, induced spontaneous repetitive backward movements of the same ear. In this area, sharp waves appeared in the electrocortigram during the repetitive ear movements induced by seizure-inducing stimuli. Unilateral ablation of this area abolished stimulus-induced repetitive movements of the contralateral ear, but had no effects on those of the ipsilateral ear. These results suggest that, in certain types of seizure-susceptible subjects, it may be possible to modify stimulus-induced repetitive movements by manipulating a certain area of the somatosensory cortex which is related to these movements and that this gerbil strain may be useful in research on this subject.

Cortical somatosensory evoked potentials of seizure-sensitive and seizure-resistant gerbils.

The Mongolian gerbil is known as an animal model that often exhibits spontaneous seizures that are characteristic of human epilepsy. Whereas there is much more information available relating this phenomenon to anatomical and electrophysiological characteristics of the hippocampal formation, the somatosensory cortex has rarely been the focus of attention. Given the existence of the fine grain cortical barrels developed in an orderly matrix, the vibrissa ascending system was thought the best sensory channel in which the gerbil neocortical excitability was to be tested. In the present study, we compared cortical evoked potentials to electric stimulation of vibrissa (whisker) follicle between the seizure-sensitive (SS) and seizure-resistant (SR) gerbils. Whereas our standard stimulation of the whisker follicle elicited a positive-negative biphasic somatosensory cortical potential in SR animals, it evoked only a positive monophasic potential in SS animals. Although the amplitude of positivity was generally larger in SR animals, the latency to reach this peak was significantly smaller than in the SS animals. Apparently there was no group difference in the laminar profile or the size of barrels that represent, in one-to-one fashion, the array of whisker follicles. It was suggested that an altered level of excitability in the absence of gross anatomy differentiates the seizure-sensitive gerbils from the seizure-resistant counterparts. Possible functional differences between the cortices of the two groups were discussed in light of the known synaptic electrophysiology.

Two types of aggregate in the cerebral cortex of a seizure-sensitive strain of the Mongolian gerbil.

A 70-kDa protein, P70, found mostly in the pyramidal cells of the cerebral cortex of cobalt-induced epileptogenic rats, has been implicated in epileptogenesis. The presence of a P70-like substance was searched for immunohistochemically in the cerebral cortex of MGS/ldr, a seizure-sensitive strain of the Mongolian gerbil (Meriones unguiculatus) that we previously established. Immunoreactive aggregates were observed in the pyramidal neurons of the motor cortex and the primary somatosensory cortex. Analysis using confocal laser scanning microscopy revealed that the aggregates were often colocalized with a second type of aggregate with red autofluorescence at the marginal zone of the cell somata. Both aggregates appeared and increased before the appearance of generalized tonic-clonic convulsion. These may be involved in some change of physiological function of the cerebral cortex but their presence itself is not enough to determine the occurrence of epileptic seizure because the gerbils that showed no such seizure had both aggregates.

Site of cortical utricular representation with special reference to the somatosensory barrel field in the gerbil.

In mammals, the osseous semicircular canals of the vestibular labyrinths are usually embedded in the pyramis of the temporal bone. Thus, the osseous semicircular canals are a cavity system that can only be visualized by injection molding. Exceptionally, the walls of the osseous semicircular canals of the Mongolian gerbil are not embedded, but exposed in the hollow space in the temporal bone. Under urethane anesthesia, a window was made in the periotic capsule of the gerbil to reach this hollow space (semicircular canal triangle), and a pair of wire electrodes were inserted through an opening made in the horizontal semicircular canal toward the utricular nerve endings. Repetitive electric stimuli at 80 Hz were applied, and the direction of eye movements was noted. Subsequently, the stimulation frequency was reduced to 0.3 Hz, and evoked potentials were recorded from the neocortex. Positive surface potentials and negative deep potentials were recorded in the somatosensory area and, more specifically, in the vibrissa "barrel field," as judged by later histochemical staining of the cortical specimens. This unique anatomic feature of the gerbil labyrinth offers an opportunity whereby the vestibular organ can be reached without any heavy surgical insult, and the presence of fine-grain vibrissa barrels in this species (seven rows instead of five rows in most rodents) will help elucidate functional interactions between vestibular and somatosensory sensations.

Age-related appearance of dystrophic axon terminals in cerebellar and vestibular nuclei of Mongolian gerbils.

In the brains of 360-day-old Mongolian gerbils, numerous swellings immunoreactive to anti-neurofilament antibody were observed in cerebellar and vestibular nuclei. The number of these swellings was the same in two gerbil strains with different susceptibility to spontaneous motor seizures by various stimuli, but much more numerous in gerbils as compared with the 360-day-old Slc:Wistar rats. Such swellings were only occasionally found before 60 days of age in gerbils, but they increased in number about fivefold from 60 to 180 days of age and about quadruple from 180 to 360 days of age. Electron microscopic observation showed that these swellings were dystrophic axon terminals (DATs) whose cytoplasms were occupied with large bundles of neurofilaments, numerous vesicular structures containing membranous and/or granular materials, and many rod-shaped mitochondria. Additionally, other types of DATs displaying degenerative changes of cytoplasmic organelles were observed. ACPase cytochemistry showed that the vesicular structures in the DATs contained ACPase and released it into the cytoplasm.

Sequencing of cDNA encoding serum albumin and its extrahepatic synthesis in the Mongolian gerbil, Meriones unguiculatus.

We have sequenced serum albumin cDNA from liver of the Mongolian gerbil, Meriones unguiculatus. The deduced amino acid sequence showed 82.6% and 73.6% identity with the corresponding proteins from rats and humans, respectively. Identical cDNA was detected in pancreas by reverse transcription followed by polymerase chain reaction (RT-PCR). Further amplification of cDNA by nested PCR revealed the presence of the same cDNA in the brain and kidney. These results indicate that serum albumin is expressed in some extrahepatic tissues. In rats, an albumin-related 70-kDa protein (P70) has been proposed to be associated with cobalt-induced epilepsy (Onozuka et al. (1995) Neurochem. Res., 20, 901-905). We intensively searched for a P70-like protein in the brain of an epilepsy-prone gerbil strain, MGS/Idr, by the RT-PCR and nested PCR using several pairs of primers based on the albumin cDNA sequence. However, we found only mRNA for albumin itself.

Altered axon terminals containing concentric lamellar bodies of cerebellar Purkinje cells in Mongolian gerbil.

Altered axon terminals containing concentric lamellar bodies were observed in cerebellar and vestibular nuclei of the Mongolian gerbil. The terminals increased in number from 30 days of age onward, and reached about tenfold at 360 days. The numbers were the same in two gerbil strains with different susceptibility to spontaneous motor seizures by various stimuli, but about threefold those in Slc:Wistar rat.

The surface evoked potential and parvalbumin-immunoreactivity in the somatosensory cortex of the developing rat.

The development of the rat somatosensory system was followed electrophysiologically and immunohistochemically. In the surface evoked potential elicited in the primary somatosensory cortex by electrical stimulation of the whisker C3 follicle, a short-latency positive wave was first recorded on postnatal Day 2. A long-latency positive wave was recorded in some pups on postnatal Day 7 and in most pups on postnatal Day 8. On postnatal Day 10, a P/N complex appeared between the short- and long-latency positive waves. Parvalbumin, believed to appear with functional maturation, appeared mainly after postnatal Day 7 in Layer V in the underlying area, although a few weakly stained cells appeared on postnatal Day 5. On postnatal Day 10, weakly stained cells appeared in the area containing barrels; their staining increased with time. In this system, electrophysiological and immunohistochemical parameters changed by the 3rd postnatal week with the most marked changes occurring within 2 postnatal weeks.

Intrinsic and drug-induced seizures of adult and developing gerbils.

Seizures elicited by posture change and intraperitoneal administration of convulsants were studied ontogenetically in the Mongolian gerbil (Meriones unguiculatus). In posture change, the first signs of seizure appeared after age 6 weeks with maximal frequency at 8-9 weeks. Adults developed complex, but stereotyped, seizures. Facial twitch was followed by the generalized convulsion, further progressing to trembling of the limbs and then kicking of the hindlimb (full seizure) after 55 days of age. Pentylenetetrazole induced a seizure similar to the full event in gerbils as young as 37 days of age. The seizure pattern elicited by strychnine or glutamate was different from that of pentylenetetrazole.

Calmodulin and calbindin localization in retina from six vertebrate species.

Calmodulin is abundant in the central nervous system, including the retina. However, the localization of calmodulin in the retina has not been described in detail. We therefore decided to investigate calmodulin localization in retinae from six vertebrate species, by using immunohistochemical labeling with four different rabbit polyclonal antibodies against calmodulin. The localization of calbindin-D28k, another calcium-binding protein already well described in retina, was compared. We found that calmodulin distribution is more highly conserved among species, contrasting with calbindin variability. The most striking result emerging is that calmodulin could not be detected in photoreceptors although other layers are intensely calmodulin-immunoreactive, casting doubt about a direct role of calmodulin in phototransduction. Horizontal cells are weakly calmodulin-immunoreactive, bipolar cells are calmodulin-immunoreactive except in turtle retina, numerous amacrine and ganglion cells are labeled in all species, and the fiber layer is always labeled. These data demonstrate that, while the calmodulin distribution in retina is similar among vertebrate species, selective differences in localization can be detected not only among the same cell types in different species but also among different cell types in the same species. The results showing differences in calmodulin immunoreactivity among cell types also provide further evidence that calmodulin expression in eukaryotes is not constitutive, in the sense that not every cell expresses similar levels of calmodulin.

Parvalbumin immunoreactivity in the central auditory system of the gerbil: a developmental study.

Changes in parvalbumin-like immunoreactivity were studied during ontogenetic development of the central auditory system of the Mongolian gerbil (Meriones unguiculatus). The nucleus of the trapezoid body contained cells that could be stained on the day of birth while in the superior olivary complex, prominent staining of cell somata was only found at P15 (postnatal day 15). Neurons located in the ventral nucleus, in the dorsal nucleus of the lateral lemniscus and in the inferior colliculus developed parvalbumin immunoreactivity mostly between P11-P15 (ventral nucleus), P15-P19 (dorsal nucleus) and P15-P19 (inferior colliculus), respectively. The accumulation of PV correlates reasonably well with the functional maturation of the neurons in these areas.

Coexistence of parvalbumin and glycine in the rat brainstem.

The coexistence of glycine- and PV-immunoreactivities was studied immunocytochemically in the nuclei of the superior olive, trapezoid body, cochlea and lateral lemniscus. All of the PV-immunoreactive neurons in the nuclei of the superior olive and trapezoid body were immunoreactive to glycine but not to GABA. In the dorsal cochlear nucleus, PV-positive neurons were sometimes immunoreactive to glycine. In the ventral nucleus of the lateral lemniscus, PV-positive cells were immunoreactive neither to glycine nor to GABA. Consequently, it was concluded that PV-immunoreactivity was distributed not only in the GABAergic neurons, but also in the glycinergic neurons and possibly in wider neuronal populations.

Calbindin D28K as a marker for the degeneration of the striatonigral pathway in Huntington's disease.

An antibody raised against the chick calcium-binding protein calbindin D28K was used in immunohistochemical studies of normal post mortem human brain and the brains of individuals with Huntington's disease. Calbindin D28K immunoreactivity in the caudate nucleus and putamen coincided with the distribution of areas of acetylcholinesterase staining termed the matrix. In the matrix, calbindin D28K immunoreactivity was present in medium-sized neurons, the major neuronal population, however a further minor population of more strongly stained large neurons was detected. In Huntington's disease there was a dramatic loss of the majority of matrix calbindin D28K immunoreactive neurons and a parallel loss of calbindin D28K immunoreactivity from the substantia nigra. In contrast to the medium-sized calbindin D28K immunoreactive neurons which degenerate in Huntington's disease, the larger immunoreactive neurons were relatively preserved.

Appearance of parvalbumin-specific immunoreactivity in the cerebral cortex and hippocampus of the developing rat and gerbil brain.

Developmental changes in the distribution of parvalbumin-specific immunoreactivity in the brain, in particular in the cerebral cortex and hippocampus, were followed immunohistochemically in two different species, the rat and the Mongolian gerbil (Meriones unguiculatus) using an antibody raised against for rat parvalbumin. The gerbil is known to develop its auditory and visual capacity later than rat. In both the rat and gerbil, parvalbumin-specific immunoreactivity appeared after birth in both the cerebral cortex and hippocampus. The timing of the development of expression of parvalbumin varied among different parts of the cerebral cortex. The parietal cortex showed evidence of the earliest expression of parvalbumin whilst the occipital and temporal cortices expressed parvalbumin at a later stage of a development. This feature was common to both the rat and gerbil but occurred at a relatively later stage in the gerbil. The profile of the distribution of parvalbumin in the brain of the developing and adult gerbil was similar to that of the rat, but there were some differences. The frequency of bead-like structures on the dendrites of the parvalbumin-positive cells in the CA1 region of the hippocampus was markedly lower in the gerbil; instead, straight non-beaded fibers which ran vertically into the pyramidal layer were stained. Parvalbumin-positive fibers were also found in the cerebral cortex of the gerbil.

Localization of parvalbumin mRNA in rat brain by in situ hybridization histochemistry.

Parvalbumin mRNA was localized in rat brain by in situ hybridization using a 35S labelled rat parvalbumin cDNA and a synthetic oligodeoxyribonucleotide (corresponding to base sequences 140 to 183 of rat parvalbumin cDNA). Strongest hybridization signals were detected in the Purkinje cells of the cerebellum and in neurones of the reticular nucleus of the thalamus. Signal was also detected in the cerebral cortex, hippocampus, basal ganglia and brain stem in agreement with the distribution of parvalbumin immunoreactivity.

[Calmodulin and neuron: immunohistochemical studies].

Calmodulin is present in higher concentrations in brain tissues. The content rapidly increased during the 2nd postnatal week in rat brain. Although the protein is ubiquitous in all eukaryotic cells, immunohistochemical studies have revealed that calmodulin is mainly localized in the neurons, exhibiting a similar distribution to that of gamma-type neuron-specific enolase. In the mouse retina, both calmodulin and gamma-enolase were found to be localized in optic nerves, ganglion cells, and inner and outer plexiform layers. The development study showed that gamma-enolase increased in the 2nd postnatal week and that the levels of calmodulin did not significantly change in that stage. In the mouse retina with an inherited retinal dysplasia (C3H), in which all the photoreceptor cells degenerate during the 2nd and 3rd postnatal weeks, calmodulin-specific staining decreased in the residual layers. Calmodulin is also enriched in mammalian testes. In the mouse testis, levels of calmodulin were high in the spermatocytes and in the spermatids, as compared to the level in spermatozoa. This suggests that the large amount of calmodulin in the testis may be associated with miotic divisions and/or spermatogenesis. Immunocytochemical staining of calmodulin in C6 glioma cells and PC12 pheochromocytoma cells showed a high level of calmodulin to be localized on the half spindles between poles and chromosomes in mitotic cells. The protein was also shown to be localized on fibrous structures in the interphase of those cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Loss of matrix calcium-binding protein-containing neurons in Huntington's disease.

In post-mortem brain specimens from patients dying with a clinical diagnosis of Huntington's disease (HD) immunohistochemistry showed a substantial loss from the neostriatum of neurons containing the calcium-binding protein calbindin 28K. These calbindin neurons, and the straital compartment in which they are sited, are particularly damaged in HD, suggesting that a failure of calcium buffering or homeostasis may contribute to cell death in HD.

Lateral motion of fluorescent molecules embedded into cell membranes of clonal myogenic cells, L6, changes upon cell maturation.

The lateral motion of fluorescent molecules embedded into cell membranes of myogenic cell line, L6, was measured. The motion of S-F-ConA became faster at cell fusion stage, and became slower after fusion. On the other hand, the motion of lipid analog, F18, was not changed at cell fusion stage. However, after fusion when myotubes were formed, the motion of F18 became slower. At cell fusion stage, there was a large variation in the motion of S-F-ConA. This means that at this stage the properties of myoblasts change drastically and rapidly.

The effect of axotomy and denervation on calmodulin content in the superior cervical sympathetic ganglion of the rat.

Calmodulin (CaM) in the superior cervical sympathetic ganglion (SCG) of the rat and its changes after transection of the pre- or postganglionic nerves were studied biochemically and immunohistochemically. The concentration of CaM in the SCG was assayed using the extent of activation of CaM-dependent adenylate cyclase from bovine neural retina. In the SCG, CaM concentration was 4.5 micrograms/mg protein, a level similar to that in another peripheral ganglion, the nodose ganglion (4.9 micrograms/mg protein). Following denervation of the SCG, the total amount of ganglionic CaM did not change significantly within a week. On the other hand, CaM content per ganglion increased gradually 3 days after axotomy. Immunohistochemical examination revealed that CaM in unoperated SCG is present both in the neuronal fibers and in the cell soma of sympathetic neurons. But the intensity of the staining with CaM-antibody in the cytoplasmic soma varied among cells. The gross profile of the CaM immunostaining seemed to be unchanged one week after either denervation or axotomy, but the immunoelectron microscopic study showed heavy staining of the innercellular membranes of irregular shape in the axotomized SCG.