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K Semba

Publications and source records attributed to K Semba.

At least 91 records · Page 5Linked to original sources
PubMed1988

Ontogeny of histidine-decarboxylase-immunoreactive neurons in the tuberomammillary nucleus of the rat hypothalamus: time of origin and development of transmitter phenotype.

The ontogeny of the histidine decarboxylase (HDC)-immunoreactive neurons of the tuberomammillary (TM) nucleus was studied in the rat brain. The time of origin of TM neurons was studied by counting the percentage of HDC-immunopositive neurons double labelled by autoradiography in adult progeny of dams injected with [3H]-thymidine at various times during gestation. Neurogenesis began on embryonic day (E) 13, peaked on E16, and was complete by E18. HDC immunoreactivity was first detected in the fetal rat brain on E16. Experiments utilizing short-survival [3H]-thymidine autoradiography combined with HDC immunohistochemistry demonstrated that TM neurons undergo their final mitotic division prior to expression of their transmitter phenotype.

Animals↗
PubMed1988

Time of origin of cholinergic neurons in the rat basal forebrain.

The timing of the final mitotic division of basal forebrain cholinergic neurons was studied by injecting [3H]thymidine into timed pregnant rats and processing the brains of their progeny as young adults for immunohistochemistry with a monoclonal antibody to choline acetyltransferase (ChAT) followed by autoradiography. ChAT-positive neurons located caudally in the basal forebrain were found to become postmitotic mostly on embryonic (E) days 12 and 13, whereas the peak final mitosis of more rostrally located ChAT-positive neurons occurred increasingly later, with the most rostral ChAT-immunoreactive neurons leaving their final mitotic cycles on E15 and E16. In all basal forebrain regions, cholinergic neurogenesis was complete by E17. These results indicate that the cholinergic neurons in the basal forebrain become postmitotic in a caudal-to-rostral gradient over about 5 days. The continuity of the gradient suggests that these cholinergic neurons may derive from the same germinal source.

Animals↗
PubMed1988

Brainstem afferents to the magnocellular basal forebrain studied by axonal transport, immunohistochemistry, and electrophysiology in the rat.

Brainstem afferents to the magnocellular basal forebrain were studied by using tract tracing, immunohistochemistry and extracellular recordings in the rat. WGA-HRP injections into the horizontal limb of the diagonal band (HDB) and the magnocellular preoptic area (MgPA) retrogradely labelled many neurons in the pedunculopontine and laterodorsal tegmental nuclei, dorsal raphe nucleus, and ventral tegmental area. Areas with moderate numbers of retrogradely labelled neurons included the median raphe nucleus, and area lateral to the medial longitudinal fasciculus in the pons, the locus ceruleus, and the medial parabrachial nucleus. A few labelled neurons were seen in the substantia nigra pars compacta, mesencephalic and pontine reticular formation, a midline area in the pontine central gray, lateral parabrachial nucleus, raphe magnus, prepositus hypoglossal nucleus, nucleus of the solitary tract, and ventrolateral medulla. A similar but not identical distribution of labelled neurons was seen following WGA-HRP injections into the nucleus basalis magnocellularis. The possible neurotransmitter content of some of these afferents to the HDB/MgPA was examined by combining retrograde Fluoro-Gold labelling and immunofluorescence. In the mesopontine tegmentum, many retrogradely labelled neurons were immunoreactive for choline acetyltransferase. In the dorsal raphe nucleus, some retrogradely labelled neurons were positive for serotonin and some for tyrosine hydroxylase (TH); however, the majority of retrogradely labelled neurons in this region were not immunoreactive for either marker. The ventral tegmental area, substantia nigra pars compacta, and locus ceruleus contained retrogradely labelled neurons which were also immunoreactive for TH. Of the retrogradely labelled neurons occasionally observed in the nucleus of the solitary tract, prepositus hypoglossal nucleus, and ventrolateral medulla, some were immunoreactive for either TH or phenylethanolamine-N-methyltransferase. To characterize functionally some of these brainstem afferents, extracellular recordings were made from antidromically identified cortically projecting neurons, mostly located in the HDB and MgPA. In agreement with most previous studies, about half (48%) of these neurons were spontaneously active. Electrical stimulation in the vicinity of the pedunculopontine tegmental and dorsal raphe nuclei elicited either excitatory or inhibitory responses in 21% (13/62) of the cortically projecting neurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1988

Non-cholinergic basal forebrain neurons project to the contralateral basal forebrain in the rat.

Following injections of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) or the fluorescent tracer fluoro-gold into the magnocellular preoptic area and the horizontal limb of the diagonal band, retrogradely labelled neurons were found in the homotopic region of the contralateral basal forebrain. Labelled fibers apparently arising from these neurons travelled in the stria medullaris and the habenular commissure to terminate in the contralateral basal forebrain. Although the neurons retrogradely labelled with fluoro-gold in the contralateral basal forebrain were similar in size to choline acetyltransferase (ChAT)-immunoreactive neurons, and were intermingled with them, none was ChAT-positive. WGA-HRP injections into the nucleus basalis magnocellularis did not result in retrograde labelling in the contralateral basal forebrain. These findings suggest that non-cholinergic neurons may serve as a direct link between the two sides of selective magnocellular basal forebrain regions.

Animals↗
PubMed1988

Nucleotide sequence and chromosomal mapping of the human c-yes-2 gene.

We molecularly characterized the second gene, c-yes-2, of two copies of yes-related genes which we previously found to contain in the human genome. First, nucleotide sequence analysis revealed that the c-yes-2 gene is a pseudogene of the c-yes-1 gene. Second, by using two independent methods, hybridization of both DNAs from sorted chromosomes and metaphase spreads with c-yes-2 DNA, we assigned the c-yes-2 gene to chromosome 22q11.2. This chromosomal localization is consistent with that given in our previous report. The failure of proper mapping in our experiment might have been caused by instability of hybrid cell clones.

Base Sequence↗
PubMed1988

Different times of origin of choline acetyltransferase- and somatostatin-immunoreactive neurons in the rat striatum.

Two populations of aspiny interneurons have been identified in the mammalian striatum, one cholinergic and the other using the neuropeptide somatostatin as a neurotransmitter. The times at which these 2 cell populations undergo their final mitosis were studied by injecting tritiated thymidine into timed pregnant rats and then processing the brains of the progeny as young adults for immunohistochemistry with monoclonal antibodies to choline acetyltransferase and somatostatin followed by autoradiography. Choline acetyltransferase-immunoreactive neurons became postmitotic in a caudal-to-rostral gradient; the occurrence of final mitosis was maximal on embryonic day (E) 12 at the most caudal level and on E15 at the most rostral. A more subtle lateral-to-medial gradient was also observed in the precommissural striatum. In contrast, no obvious gradients were seen with somatostatin-immunoreactive neurons; regardless of their location within the striatum, these neurons underwent their final mitosis on days E15-16, towards the end of cholinergic neurogenesis. These results indicate that although both cholinergic and somatostatin-containing cells represent interneuronal populations in the striatum, they display distinctly different spatiotemporal patterns of neurogenesis.

Animals↗
PubMed1987

En bloc immunohistochemistry reveals extensive distribution of histidine decarboxylase-immunoreactive neurons on the ventral surface of the rat hypothalamus.

En bloc immunohistochemistry was used to examine the distribution of L-histidine decarboxylase (HDC)-immunoreactive neurons on the ventral surface of the rat hypothalamus. Following standard paraformaldehyde fixation, the ventral hypothalamus containing the tuber cinereum was carefully dissected free and incubated en bloc with antisera to HDC followed by standard avidin-biotin complex immunohistochemistry. Microscopic examination of these en block preparations revealed the existence of an extensive plexus of HDC-immunoreactive neurons located directly upon the ventral hypothalamic surface. HDC-immunoreactive neurons were largely restricted to the caudal half of the hypothalamic surface, and were multipolar, with 2-5 dendrites radiating in all directions from the soma. The proximity of these neurons to the ventral surface of the brain suggests that histaminergic neurons might be capable of responding to cerebrospinal fluid borne substances. Thus, one form of input to hypothalamic histaminergic neurons may be humoral rather than synaptic.

Animals↗
PubMed1987

Physiological evidence for subpopulations of cortically projecting basal forebrain neurons in the anesthetized rat.

Sixty-three cortically projecting basal forebrain neurons were identified in chloral hydrate anesthetized rats by antidromic activation from the cerebral cortex. Two subpopulations were noted: type I neurons exhibited two antidromic action potentials of constant latency and identical waveform in response to double pulse cortical stimulation. In contrast, type II neurons exhibited two antidromic action potentials of constant latency but differing waveforms in response to the double pulse paradigm. The phenomenon exhibited by type II cortically projecting basal forebrain neurons is interpreted as evidence for loss of the somatodendritic portion of the antidromic action potential with high frequency stimulation. The median latency to antidromic activation of type II neurons (13.5 ms) was significantly longer than that of type I neurons (3.9 ms). Spontaneous firing rates varied over a wide range (0-49 Hz), and there was no significant difference between the rates of type I and type II neurons. These data underscore the physiological heterogeneity of this presumptive cholinergic cortical afferent system. Anatomical studies have shown that most, but possibly not all cortically projecting basal forebrain neurons are cholinergic. The relative proportions of type I (87%) and type II (13%) neurons encountered in this study suggest that type I neurons might be cholinergic and type II neurons non-cholinergic. If substantiated, this hypothesis would permit cholinergic and non-cholinergic cortically projecting basal forebrain neurons to be distinguished using a simple test of antidromicity.

Afferent Pathways↗
PubMed1987

Morphology of cortically projecting basal forebrain neurons in the rat as revealed by intracellular iontophoresis of horseradish peroxidase.

The intracellular horseradish peroxidase technique was employed to study the morphology of basal forebrain neurons that were identified as cortically projecting by antidromic invasion from the cerebral cortex. Four neurons were examined in detail; they were located at different rostrocaudal levels within the basal forebrain. Their somata were large, 30-50 microns in longest dimension, and gave rise to three to eight primary dendrites, which ramified into third- to fifth-order dendrites. The longest observed dendrite in each neuron terminated at a distance of 600-900 microns from the soma. The sizes of soma and dendritic field of the two most rostrally located cells were smaller than those of the other two cells located more caudally. Dendritic spines were seen in all four cortically projecting basal forebrain neurons. Spines had shafts of variable lengths, and usually had spherical or elongated heads. The density of spines varied among the four neurons; one neuron, a type II cortically projecting basal forebrain neurons as defined physiologically by Reiner et al., had a much greater number of dendritic spines than the other three neurons, which were type I neurons. No somatic spines were observed. Presumptive axons were identified in three of the four cortically projecting basal forebrain neurons. These axons originated from either the soma or a primary dendrite, and two of them gave off local collaterals, which displayed occasional bouton-like swellings. The above observations confirm and extend previous findings that cortically projecting neurons in the basal forebrain are large multipolar cells, and provide evidence to support the conclusion that these cells, although somewhat variable in size, generally have extensive dendrites which display frequent spines.

Afferent Pathways↗
PubMed1987

Neurotransmitters in the mammalian striatum: neuronal circuits and heterogeneity.

The major input and output pathways of the mammalian striatum have been well established. Recent studies have identified a number of neurotransmitters used by these pathways as well as by striatal interneurons, and have begun to unravel their synaptic connections. The major output neurons have been identified as medium spiny neurons which contain gamma-aminobutyric acid (GABA), endogeneous opioids, and substance P. These neurons project to the pallidum and substantia nigra in a topographic and probably chemically organized manner. The major striatal afferents from the cerebral cortex, thalamus, and substantia nigra terminate, at least in part, on these striatal projection neurons. Striatal interneurons contain acetylcholine, GABA, and somatostatin plus neuropeptide Y, and appear to synapse on striatal projection neurons. In recent years, much activity has been directed to the neurochemical and hodological heterogeneities which occur at a macroscopic level in the striatum. This has led to the concept of a patch-matrix organization in the striatum.

Acetylcholine↗
PubMed1987

The yes-related cellular gene lyn encodes a possible tyrosine kinase similar to p56lck.

With v-yes DNA as the probe, a human cDNA library made from placental RNA was screened under relaxed conditions, and DNA clones derived from a novel genetic locus, termed lyn, were obtained. Nucleotide sequencing revealed that lyn could encode a novel tyrosine kinase that was very similar to mouse T-lymphocyte-specific tyrosine kinase p56lck and the v-yes protein as well as to the gene products of v-fgr and v-src. Northern hybridization analysis revealed that a 3.2-kilobase lyn mRNA was expressed in a variety of tissues of the human fetus. The pattern of lyn mRNA expression was different from those of related genes, such as yes and syn. Hybridization analysis of DNA from sorted chromosomes showed that the lyn gene is located on human chromosome 8 q13-qter.

Amino Acid Sequence↗
PubMed1987

Characterization of cDNA clones for the human c-yes gene.

Three c-yes cDNA clones were obtained from poly(A)+ RNA of human embryo fibroblasts. Sequence analysis of the clones showed that they contained inserts corresponding to nearly full-length human c-yes mRNA, which could encode a polypeptide of 543 amino acids with a relative molecular weight (Mr) of 60,801. The predicted amino acid sequence of the protein has no apparent membrane-spanning region or suspected ligand binding domain and closely resembles pp60c-src. Comparison of the sequences of c-yes and v-yes revealed that the v-yes gene contains most of the c-yes coding sequence except the region encoding its extreme carboxyl terminus. The region missing from the v-yes protein is the part that is highly conserved in cellular gene products of the protein-tyrosine kinase family.

Amino Acid Sequence↗
PubMed1987

Isolation and sequencing of cDNA clones homologous to the v-fgr oncogene from a human B lymphocyte cell line, IM-9.

Two c-fgr cDNA clones were isolated from a cDNA library derived from a human B lymphocyte cell line, IM-9. Sequence analysis of the clones showed that they contained inserts corresponding to nearly full-length human c-fgr mRNA, which could encode a polypeptide of 529 amino acids with a calculated molecular weight of 59,478. Although the amino acid sequence between Gly-78 and the carboxy-terminus of the c-fgr is highly homologous to the corresponding sequence of the c-yes protein, the homology between the two proteins is low in the amino-terminal proximal region. Northern blot hybridization analysis using the c-fgr specific sequence showed that the c-fgr mRNA was expressed at higher level in the liver than in the brain, lung, or kidney of a human fetus.

Amino Acid Sequence↗
PubMed1986

Dorsal horn cells in the cat responding to stimulation of the plantar cushion.

Dorsal horn cells (DHC) in the spinal cord of cats responding to low threshold mechanical stimulation of the plantar cushion (PC), labelled with intracellular horseradish peroxidase, displayed a wide variety of anatomical and physiological characteristics, some of which were quite consistent within a given lamina. These DHC had cell bodies located medially in spinal segment L7, in laminae III, IV and VI. Dendritic fields varied greatly, for the most part conforming to patterns corresponding to their locations in the dorsal horn. Axon collaterals of most DHC ramified near their cell bodies, chiefly projecting ventral ward within the same lamina as the cell body or into more ventral laminae. Cells with the shortest central delays had cell bodies in laminae III and IV. Response latencies of DHC to PC stimulation were only slightly greater than the latencies of primary afferent fibers. Lamina IV cells had mechanical thresholds comparable to those of the primary afferents, significantly lower than the thresholds of cells of laminae III or VI. The receptive fields (RF) of the lamina IV cells were significantly larger than those of the primary afferent fibers or of the lamina VI cells. Relationships among RF area, mediolateral locations of cell bodies and dendritic spread were investigated.

Animals↗
PubMed1986

The facial "motor" nerve of the rat: control of vibrissal movement and examination of motor and sensory components.

Rhythmical whisking of the mystacial vibrissae at about 7 Hz during exploration is one of the most conspicuous behavioral patterns in the rat. To identify the final common pathway for vibrissal movement, individual motor branches of the facial nerve, including the posterior auricular, temporal, zygomatic, buccal, marginal mandibular, cervical, stylohyoid, and posterior digastric branches, were cut, either singly or in various combinations. We found that vibrissal movement could be abolished only by transection involving the buccal branch and the upper division of the marginal mandibular branch. To trace back the central origins of the buccal and marginal mandibular, as well as the other branches of the facial nerve, all distal to the stylomastoid foramen, horseradish peroxidase (HRP) was applied to the cut proximal ends of these individual branches. The retrograde HRP labelling in the facial motor nucleus revealed topographical representation of these branches in which the buccal and marginal mandibular branches were represented laterally. The stylohyoid and posterior digastric branches originated from cells in the suprafacial nucleus. Consistent with earlier observations with intramuscular HRP injections, the motoneuronal population devoted to vibrissal movement did not seem to be substantially larger than that for other facial movements. An additional examination was made of the labelled afferent component of the facial motor nerve. We confirmed and extended previous findings that none of the above facial motor nerve branches, except the posterior auricular branch, contained a significant number of afferent fibers originating from the geniculate ganglion, the sensory ganglion of the seventh nerve. In addition, no labelling was seen in the mesencephalic trigeminal nucleus or trigeminal ganglion. These findings, in combination, suggest that, with the exception of the posterior auricular branch, all the facial motor nerve branches, including those involved in vibrissal movement, are almost entirely efferent.

Animals↗
PubMed1986

yes-related protooncogene, syn, belongs to the protein-tyrosine kinase family.

An src/yes-related novel gene named syn (SYN in human gene nomenclature) has been identified in the human genome on chromosome 6 and characterized by molecular cloning. Nucleotide sequence analysis of cDNA clones showed that the c-syn gene could encode a protein-tyrosine kinase that is very similar in primary structure to the v-yes and human c-src proteins. A 2.8-kilobase transcript of the c-syn gene, which differs in size from those of the c-yes, c-src, and c-fgr genes, was observed in various cell types. These results show that syn is a new member of the tyrosine kinase oncogene family.

Amino Acid Sequence↗
PubMed1986

Structure, expression, and chromosomal location of the human c-fgr gene.

The nucleotide sequence of seven exons of the human c-fgr gene, a cellular homolog of the oncogene of Gardner-Rasheed feline sarcoma virus, was determined. Twenty-six independent genomic clones were obtained from a human gene library with a DNA clone of Y73 avian sarcoma virus oncogene, v-yes, as a probe under relaxed hybridization conditions. Restriction mapping and partial sequence analyses revealed that two of these clones were derived from the c-fgr gene, distinct from the c-yes gene. Interestingly, the splicing points of the c-fgr gene were identical with those of the c-src gene throughout the seven exons, suggesting that the two proto-oncogenes were generated by gene duplication of an ancestral gene containing intervening sequences. On RNA blot hybridization the major transcript was found to be 2.6 kilobase long. Two additional transcripts of 3.5 and 4.7 kilobases were also detected. Furthermore, karyotype analysis of several human-mouse hybrid cells and Southern blot analyses of DNAs of the hybrids with a human c-fgr locus-specific probe showed that this gene is located on chromosome 1.

Animals↗