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K Semba

Publications and source records attributed to K Semba.

At least 109 records · Page 6Linked to original sources

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↗

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↗

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↗

Localization of a novel v-erbB-related gene, c-erbB-2, on human chromosome 17 and its amplification in a gastric cancer cell line.

The c-erbB-2 gene is a v-erbB-related proto-oncogene which is distinct from the gene encoding the epidermal growth factor receptor. By using two independent methods, hybridization of both sorted chromosomes and metaphase spreads with cloned c-erbB-2 DNA, we mapped the c-erbB-2 locus on human chromosome 17 at q21, a specific breakpoint observed in a translocation associated with acute promyelocytic leukemia. Furthermore, we observed amplification and elevated expression of the c-erbB-2 gene in the MKN-7 gastric cancer cell line. These data suggest possible involvement of the c-erbB-2 gene in human cancer.

Cell Line↗

Nakahara memorial lecture. Non-receptor type protein-tyrosine kinases closely related to src and yes compose a multigene family.

We have determined the total coding sequence of human c-yes, a non-receptor type protein-tyrosine kinase gene, and found that the c-yes gene closely resembles the c-src gene. Recently, two new genes, syn and lyn, were found to encode proteins closely related to the yes product. In addition, we also determined the partial sequence of fgr. These genes together with lck reported by two American groups have very closely related structures and are thought to compose a closely related group of non-receptor type protein-tyrosine kinases. Partial analysis of the structures of these genes indicated that they have identical splicing junctions at all sites so far examined. On the other hand, the erbB-1/EGF (epidermal growth factor) receptor gene and the erbB-2/neu gene have completely different splicing junctions from those of the above gene group even in the kinase domain, although these genes also have protein kinase activity specific for tyrosine residues and the erbB-1 and -2 genes share splicing sites. These results suggest that the genes of the group of six non-receptor type kinases and those of the erbB-1 and erbB-2 gene group are descendants evolved by duplication of two distinct ancestor genes and are members of two distinct multi-gene families. The genes coding for protein kinases may be members of a super-family including multiple distinct gene families.

Amino Acid Sequence↗

4-Aminopyridine induces expansion of cutaneous receptive fields of dorsal horn cells.

Systemic administration of 4-aminopyridine (4-AP) increased the size of the cutaneous receptive fields of 9 of the 15 dorsal horn cells tested. These receptive fields were on the feet and toes of the hind limbs of cats. Receptive field sizes increased with increasing doses of 4-AP. However, 4-AP administration did not change the responses of dorsal horn cells to graded mechanical stimuli administered near the centers of their receptive fields.

4-Aminopyridine↗

Location of the c-yes gene on the human chromosome and its expression in various tissues.

Analysis of DNA from human embryo fibroblasts showed that ten Eco RI fragments were hybridizable with the Yamaguchi sarcoma virus oncogene (v-yes). Four of the Eco RI fragments were assigned to chromosome 18 and one to chromosome 6. There was evidence for multiple copies of yes-related genes in the human genome; however, only a single RNA species, 4.8 kilobases in length, was related to yes in various cells.

Animals↗

An electron microscopic study of terminals of rapidly adapting mechanoreceptive afferent fibers in the cat spinal cord.

The intra-axonal horseradish peroxidase technique was used to examine the central terminals of 7 A beta primary afferent fibers from rapidly adapting (RA) mechanoreceptors in the glabrous skin of the cat's hindpaw. At the light microscopic level, labelled collaterals were seen to bear occasional boutonlike swellings, mostly (75-82%) of the en passant type. These swellings were distributed more or less uniformly from lamina III to a dorsal part of lamina VI in the dorsal horn, over a maximum longitudinal extent of about 4 mm. At the electron microscopic level, we observed that labelled boutons of RA afferent fibers were 1.0 to 3.3 micrometers in longest sectional dimension, and contained clear, round synaptic vesicles. They frequently formed asymmetric axospinous and axodendritic synapses and commonly appeared to receive contacts from unlabelled structures containing flattened or pleomorphic vesicles plus occasional large dense-cored vesicles. The examination of synaptic connectivity over the entire surface of individual boutons indicated that RA afferent boutons each made contacts with an average of one spine and one dendrite and, in addition, appeared to be postsynaptic to an average of two unlabelled vesicle-containing structures. This synaptic organization was, in general, more complex than that we had seen previously in Pacinian corpuscle (PC) and slowly adapting (SA) type I mechanoreceptive afferent fibers. Our findings indicate that RA, SA, and PC afferent terminals, while displaying some differential synaptic organizations, have many morphological and synaptological characteristics in common. These afferent terminals, in turn, seem to be generally distinguishable from the terminals of muscle spindle Ia afferents or unmyelinated primary afferents.

Animals↗

A v-erbB-related protooncogene, c-erbB-2, is distinct from the c-erbB-1/epidermal growth factor-receptor gene and is amplified in a human salivary gland adenocarcinoma.

From a human genomic library, we obtained six v-erbB-related DNA clones. A DNA probe prepared from one of the clones, lambda 107, hybridized to EcoRI fragments of 6.4 and 13 kilobase pairs of human DNA. Neither of these fragments was amplified in A431 vulva carcinoma cells, in which the gene encoding the epidermal growth factor receptor is amplified. In addition, the probe from lambda 107 hybridized with a single, 4.8-kilobase poly(A)+ RNA species and did not react with EGF receptor mRNA. Thus, we conclude that clone lambda 107 represents a v-erbB-related gene (c-erbB-2) that is distinct from the EGF receptor gene. In contrast, the other five clones were shown to represent the EGF receptor gene (c-erbB-1). Partial nucleotide sequence analysis of the lambda 107 insert showed that this clone contained at least seven putative exons and that six of them could encode the kinase domain characteristic of protein products of the src oncogene family. Southern blot analysis showed close similarity of the restriction patterns of the rat c-erbB-2 gene and the rat neu oncogene, suggesting possible involvement of c-erbB-2 in human cancer. In fact, approximately 30-fold amplification of c-erbB-2 was observed in a human adenocarcinoma of the salivary gland.

Adenocarcinoma↗

Human c-f gr gene does not contain coding sequence for actin-like protein.

f gr is a member of the protein kinase oncogene family and was found to have the highest homology to yes gene among the members of the family. Using a v-yes probe, a molecular clone of human c-f gr locus was isolated from a genomic library. Nucleotide sequence determination revealed that the actin gene-like sequence found in v-f gr gene was absent at the corresponding position of the c-f gr gene. Thus, the v-f gr gene product is considered to be a tri-chimeric gene product consisting of viral gag gene and two distinct cellular genes, actin gene and the proto-f gr gene.

Actins↗

Regional mapping of the human proto-oncogene c-yes-1 to chromosome 18 at band q21.3.

Human proto-oncogene c-yes-1 homologous to the viral oncogene (v-yes) of an avian Y73 sarcoma virus was mapped to region q21.3 of chromosome 18 by in situ hybridization. This finding confirmed and more rigidly substantiated the previous assignment which was based on analyses of human X mouse somatic cell hybrids and Southern blots.

Chromosome Mapping↗

Examination of geniculate ganglion cells contributing sensory fibers to the rat facial 'motor' nerve.

Using a method to visualize HRP-containing cells in the geniculate ganglion (GG) in situ after decalcifying surrounding bone, we found that about 30% of the total (about 1000) GG cells contributed sensory fibers to the posterior auricular branch of the facial motor nerve. These cells are relatively large for GG cells in general. The remaining facial motor nerve branches, including those involved in vibrissal movement, contained few sensory afferent fibers originating from GG cells.

Animals↗

Trigeminal sensorimotor mechanisms and eating in the rat.

Photographic, electrophysiological and neurobehavioral analyses were used to examine the contribution of trigeminal inputs to the behavioral organization of eating in the rat. During eating, jaw opening was always preceded by a period of perioral contact with the food source. Mechanical or electrical stimulation of oral and perioral areas in anesthetized animals elicited compound action potentials in the mylohyoid nerve (jaw-opener innervation) at short latencies and low stimulus intensities. Trigeminal orosensory deafferentation (sparing jaw muscle afferents and efferents) abolished or significantly reduced mouth opening during eating. We conclude that trigeminal orosensory inputs provide an essential link in the stimulus-response chain mediating eating in the rat.

Animals↗

Ultrastructure of pacinian corpuscle primary afferent terminals in the cat spinal cord.

The glabrous skin of the hindlimb of the cat contains 3 types of low-threshold mechanoreceptors: Pacinian corpuscles (PC), and slowly and rapidly adapting receptors. In the present study, 12 primary afferent fibers transmitting impulses from PC were injected intra-axonally with horseradish peroxidase (HRP) in the spinal cord to examine the morphology of their terminals in the dorsal horn. At the light microscopic level, terminal arborizations were observed in laminae II-VI of the dorsal horn, extending up to 7 mm rostrocaudally in and near the seventh lumbar segment. Bouton-like swellings, predominantly (67%) of the en passant type, were distributed in two discrete clusters, one concentrated rostrally in Rexed's laminae III-IV, and the other concentrated caudally in lamina V. At the electron microscopic level, a combination of morphometric and serial reconstructive analyses with 3 fibers revealed the following. Boutons labelled with HRP invariably contained clear round vesicles, approximately 40 nm in diameter. Labelled bouton sections had longest dimensions of 1.84 +/- 0.63 micron. Their shapes varied from rounded to elongated forms with occasional scalloped appearances. A majority (73%) of the contacts associated with HRP-filled boutons were made with dendritic spines and shafts. Thick postsynaptic densities were usually associated with these synapses, although thinner densities were also observed. 24% of the contacts made by labelled boutons were synapse-like contacts with unlabelled vesicle-containing structures. The vesicles in the unlabelled structures were usually pleomorphic, but sometimes round. These contacts were identified as 'synapse-like' because labelling obscured possible landmarks necessary for definitive identification of synapses. However, in most of these contacts, there was an accumulation of vesicles near the cleft on the unlabelled side, suggesting that the labelled boutons were postsynaptic. Only 3% of the contacts made by labelled boutons were axosomatic. The lengths of contacts with dendritic spines (0.49 +/- 0.23 micron) or with dendrites proper (0.45 +/- 0.20 micron) were significantly longer than those with vesicle-containing unlabelled structures (0.31 +/- 0.18 micron). The portions of cross-sectional bouton contours devoted to synaptic or synapse-like contacts accounted for 9-13% of the perimeters. The larger the bouton, the longer the summed lengths of contacts. Serial reconstruction of selected labelled boutons revealed both simple and quite complex synaptic organizations, including glomeruli with labelled boutons as the central component.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗

Amplification and enhanced expression of the epidermal growth factor receptor gene in A431 human carcinoma cells.

The sequence of the human epidermal growth factor (EGF) receptor shows great homology with the avian erythroblastosis virus v-erb B oncogene, raising the possibility that the receptor gene is identical to the c-erb B protooncogene. Human A431 epidermoid carcinoma cells, which have an unusually high number of EGF receptors, were examined to determine whether elevated EGF receptor levels correlate with gene amplification. Southern blots of genomic DNA's from A431 and other human cell lines were probed with either a v-erb B gene fragment or a human EGF receptor complementary DNA clone (pE7), previously isolated from an A431 complementary DNA library. When either probe was used to analyze Eco RI- or Hind III-generated DNA fragments, EGF receptor DNA sequences were amplified about 30-fold in A431. Differences in the banding pattern of A431 DNA fragments relative to normal fibroblast DNA indicate the occurrence of a rearrangement in the region of the receptor gene. Furthermore, A431 cells contain a characteristic, prominent 2.9-kilobase RNA. These results are consistent with the hypothesis that, in A431 cells, gene amplification, possibly associated with a translocation event, may result in the overproduction of EGF receptor protein or the appearance of the transformed phenotype (or both).

Alpharetrovirus↗