Biomedical subjects
Publications and source records attributed to H Takagi.
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The ontogeny of the neurotensin (NT) neuron system in the forebrain and diencephalon of the rat was investigated by means of indirect immunofluorescence. NT-positive structures first appear in the primordium of the olfactory bulb, anterior amygdaloid area, piriform cortex, amygdaloid complex, nucleus tractus diagonalis, lateral preoptic area, and lateral hypothalamus, etc., of the fetus of gestational day 16. From this day, NT-positive structures appear gradually in various areas of the forebrain and diencephalon and reach their maximum content on postnatal day 7. In contrast, NT-positive structures decrease slightly in number as the rat grows. However, even in adult rats, numerous NT-positive structures are still identifiable in various areas of the forebrain and diencephalon.
The Met-enkephalin contents in the dorsal horn of the lumbar enlargement and the nucleus reticularis gigantocellularis of the medulla oblongata of the rat were measured, using a sensitive and specific radioimmunoassay for Met-enkephalin, and the effects of morphine and noxious stimuli on the Met-enkephalin contents in these regions were examined. In this radioimmunoassay, the IC50 and assayable limits for Met-enkephalin were 45 and 5 fmol/tube respectively, and the IC50 for Leuenkephalin was 0.56 nmol/tube (0.008% cross reactivity between Met-and Leu-enkephalins). The contents of Met-enkephalin-like immunoreactivity in the dorsal horn of the lumbar enlargement and the nucleus reticularis gigantocellularis were not altered by either subcutaneous injection of morphine or thermal (hot plate) and chemical (formalin injection) noxious stimuli applied to the hind-paws.
Substance P-like immunoreactivity (SPI) was investigated in the chicken retina by means of the indirect immunofluorescence method with flat-mounts. SPI cells were located mostly in the peripheral retinal regions, while in the central region, none or only a few cells were seen. Based upon their 3-dimensional profiles, SPI cells can be divided into two types: one type belongs to the stratified amacrine cells of the first sublayer and the other to those of the third to fifth sublayer.
The afferent source of enkephalin-like immunoreactive (ELI) fibers in the lateral septal area of the rat was elucidated experimentally by means of an indirect immunofluorescence technique. These ELI fibers had almost completely disappeared on the operated side after the destruction of the area just ventrolateral to the anterior hypothalamic nucleus, where a number of ELI cells was detected. This fact strongly suggests that these ELI cells project ipsilaterally to the lateral septal area.
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Ontogeny of the leucine-enkephalin (L-Enk) neuron system in the lower brainstem of the rat was investigated by means of indirect immunofluorescence. L-Enk-containing cells first appear in the primordium of the medullary reticular formation just medial to the n. tractus spinalis nerve trigemini at the level of the the rostral half of the inferior olivary nucleus, in the n. cuneiformis, and in the mesencephalic reticular formation of the fetus at gestational day 16 (14-15-mm embryos). From that time onward, L-Enk-containing cells appear in various areas of the lower brainstem one after another until birth. After birth, although L-Enk-containing cells decrease slightly in number as the rats grow, L-Enk-containing fibers continue to increase in number.
The ontogeny of the somatostatin (SRIF) neuron system in the forebrain and diencephalon the rat was investigated by means of the indirect immunofluorescence method. SRIF-positive cells first appear in the primordium of the hypothalamus surrounding the fasciculus mammillothalamicus of the fetus at gestational day 14 (10-12-mm embryo). At gestational days 16-17 (14-17-mm embryo, SRIF-positive cells appear in the developing piriform cortex and entopeduncular nucleus. The fetus at gestational days 18-19 (17-26-mm embryos) showed a remarkable increase in the number of SRIF-positive cells and numerous groups of such cells are detectable in various forebrain and diencephalic areas such as the hypothalamic periventricular zone, zona incerta, area lateral to the commissura posterior (ACP), area between the optic tract and capsula interna, pars retrolenticularis (AOR), n. caudatus putamen, hippocampus, somatic sensory cortex, and n. accumbens, etc. At gestational day 20, SRIF-positive cells newly appear in the septum, olfactory bulb, diagonal band of Broca, claustrum, lateral preoptic area, and lateral habenular nucleus. The majority of SRIF-positive structures found in the forebrain and diencephalon increase in number during the perinatal stage (between gestational day 21 and postnatal day 2) and more or less maintain their immunoreactivity even in adult rats. However, SRIF-positive cells located in the AOR, ACP, and lateral septal area, etc., begin to decrease in number during the perinatal stage and no or only a few SRIF-containing cells are identified in these areas of infant and adult rats.
To examine the physiological role of the analgesic dipeptide, kyotorphin (Tyr-Arg), which was isolated from the mammalian brain, its subcellular localization was studied. Kyotorphin was determined using high-performance liquid chromatography (HPLC) with an electrochemical detector. This dipeptide was found to be concentrated in the crude mitochondrial (P2) fraction. Further subfractionation of the P2 fraction revealed that kyotorphin was exclusively localized in the synaptosomal fraction.This finding suggests the possibility that kyotorphin has a neurotransmitter/neuromodulator role in the brain.
Two different antisera to leucine-enkephalin were used to study the localization of enkephalin-like immunoreactive material in the neostriatum and globus pallidus of the rat, by means of the unlabelled antibody-enzyme method. Thin immunoreactive varicose fibres are scattered throughout the neostriatum. In the ventral striatum, fibres come together and follow a relatively straight course for several micrometers, forming tube-like structures which can be traced to cell bodies; these cell bodies are completely surrounded by immunoreactive fibers. Occasional immunoreactive varicose fibres are also found close to another type of neuron throughout the whole neostriatum. Examination by electron microscopy of immunoreactive structures that had been identified first in the light microscope, showed that each of the nearly 200 varicosities examined was a vesicle-containing bouton that formed a synaptic contact. Rarely were asymmetrical synaptic contacts found between immunoreactive boutons and dendritic spines. All other synapses formed by enkephalin-immunoreactive boutons were symmetrical. Two types of postsynaptic neuron were identified; the first type was a medium-sized neuron with the ultrastructural features of a typical striatal spiny neuron. The second type had a larger perikaryon surrounded by numerous immunoreactive varicosities that were found to be boutons forming symmetrical synapses. The long dendrites of this second type of neuron likewise received a dense input of immunoreactive boutons forming symmetrical synapses; such ensheathed dendrites were found to be the tube-like structures seen in the light microscope. The ultrastructural features of these neurons, notably a highly indented nucleus, were those of a rare type of striatonigral neuron. In the globus pallidus, all the enkephalin-immunoreactive boutons studied formed symmetrical synapses with ensheathed dendrites and perikarya that were similar to the latter type of postsynaptic neuron in the neostriatum. Axo-axonic synapses involving immunoreactive boutons were not seen in our material. The results are consistent with the view that enkephalin-like substances may be synaptic transmitters in the neostriatum and that they may have different actions according to the nature of the postsynaptic target. The finding that one type of neostriatal neuron, and a very similar neuron in the globus pallidus, receives multiple enkephalin-immunoreactive boutons all over its perikaryon and along its dendrites indicates a potentially important role of enkephalin in the convergence of information within the neostriatum and pallidum on to output neurons.
Ten patients who developed neurological deficits associated with angiographically proven cerebral vasospasm caused by ruptured aneurysm were treated with hyperdynamic therapy induced by administration of a large amount of human serum albumin. No vaso-active drugs were administered. Cardiopulmonary function and intracranial pressure were monitored during the treatment. Marked improvement of neurological function was observed in all cases. Nine patients recovered completely without any neurological residual following treatment. The degree of the improvement observed during treatment closely correlated with the decrease in total peripheral resistance. Infusion of albumin did not cause elevation of intracranial pressure. It was concluded that the hyperdynamic therapy induced by administration of albumin has a dramatic effect on the ischaemic cerebral insult caused by vasospasm. It is postulated that the main effect of this treatment is produced by cerebrovascular dilatation.
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A pain-producing substance, bradykinin (0.63--1.25 ng in 0.5--1.0 microliter of distilled water), was applied onto the tooth pulp of the lower incisor of unrestrained rats through a cannula fixed on the surfaces of the incisors using a microsyringe. Such a microapplication of bradykinin induced biting response and other aversive behaviors. When the microapplications were repeated at intervals of 60 min, the biting response seemed most characteristic and reproducible. Therefore, the duration of biting response was used as a measure in studying the effects of drugs on trigeminal pain. If biting duration was inhibited 90% or more after drug administration the effect was considered analgesic. Carbamazepine and phenytoin, which are clinically employed for treating trigeminal neuralgia, and morphine, a narcotic analgesic, produced dose-dependent analgesic effects in this method and the corresponding ED50 values were 13.1 intraperitoneally (ip), 75.0 ip, and 3.00 subcutaneously (sc) mg/kg, respectively. On the other hand, pentobarbital (15 mg/kg, ip), diazepam (1.0 mg/kg, ip), and aspirin (150 mg/kg, ip) were not effective in suppressing the biting response. These results indicate that this newly devised method in the rat is reliable and feasible in evaluating pain related the trigeminal system.
The cochlearofugal somatostatin (SRIF) neuron system of very young rats (between newborn and 1 week old) was investigated by means of the indirect immunofluorescence technique of Coons. Cochlear nuclei (both dorsal and ventral cochlear nuclei) of this stage contain numerous SRIF-positive cells which are scarcely found in adult rats. Based upon the experimental findings obtained by this study, the majority of SRIF-positive fibers originating from these nuclei reach the contralateral inferior colliculus via well-established cochlearo-inferior collicular tracts such as dorsal acoustic striae, intermediate acoustic striae and trapezoid body, respectively. The possible significance of SRIF in the development of the auditory system is briefly discussed.
The buffered picric acid paraformaldehyde fixative originally recommended for electronmicroscopy and which has since been used occasionally for light-microscopic immunocytochemistry, has been supplemented with glutaraldehyde and used as primary fixative for the perfusion of rat brains. In the basal ganglia and preoptic area, substance P, somatostatin and leu-enkephalin immunoreactive material was localized with the unlabelled antibody enzyme method in thick sections cut from freeze-thaw treated blocks. Good penetration of the antibodies without the use of detergents and the light background of the osmium-treated sections allowed the selection for electron-microscopy of immunoreactive structures as small as individual boutons that had been identified at the light-microscopic level. It is suggested that the procedure may be useful for electron-microscopic sampling of immunoreactive structures occurring infrequently over a large area or for the electron-microscopic study of light-microscopically classified neurons.
With the indirect immunofluorescence technique, the localization (including the postnatal ontogeny) of substance P in the cerebellum, and the ways of entry of substance P-containing fibers into the cerebellum were explored. In the newborn rat cerebellum, dense fiber bands of axons with substance P-like immunoreactivity which can be traced to the lower brain stem are found. These fibers are also traceable to the developing granular cell layer. Two weeks after birth, however, substance P-containing structures seen in the cerebellum begin to decrease progressively and in the cerebellum of the adult rats, only a small amount of substance P-containing structures is observed. The present study established that substance P-containing fibers are mostly derived from extracerebellar substance P-containing cells and demonstrated the presence of three sites of entry of these substance P-containing fibers to the cerebellum, via (1) the inferior cerebellar peduncle, (2) the fasciculus uncinatus and (3) the middle cerebellar peduncle, respectively. Following deafferentation of the cerebellum, substance P-accumulating fibers are observed only ventral to the lesion (i.e. on the brain stem side), while in the cerebellum a remarkable decrease of substance P-containing fibers is seen and no substance P-accumulating fibers are found dorsal to the lesion (cerebellar side).
The ontogeny of substance P-containing neuron system in the forebrain and upper brain stem of the rat was investigated by means of the indirect immunofluorescence technique. Substance P-positive structures first appeared in the primordium of the epithalamus and the area which surrounded the commissura posterior of the rat fetus corresponding to gestational day 14 (10-12 mm embryos). On and after gestational day 14, substance P-positive structures gradually made their appearance in various areas of the forebrain and upper brain stem. Substance P-positive structures thus continued to increase in number and in density during the fetus and perinatal stage and showed histochemically maximum content at the stage between postnatal days 5 and 15. After then, substance P-positive neurons tended to decrease in number as the rats grew, while substance P-positive fibers maintained in general their strong immunoreactivity even in the adult rats. The present study demonstrates that substance P-positive structures appear at a very early ontogenetical stage. This suggests that substance P might play an important role in the development of the forebrain and upper brain stem in addition to its neurotransmitter or neuromodulator functions.