[Analysis of the path of the movement of the neural crest cells of the mammalian head region].
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Animals↗
Biomedical subjects
Y Ninomiya
Publications and source records attributed to Y Ninomiya.
Isolation and sequencing of cDNAs and genomic DNAs encoding the alpha 4 chain of basement membrane collagen type IV and assignment of the gene to the distal long arm of human chromosome 2.
We cloned three overlapping cDNAs covering 2,452 base pairs encoding a new basement membrane collagen chain, alpha 4(IV), from rabbit corneal endothelial cell RNA. Nucleotide sequence analysis demonstrated that the clones encoded a triple-helical domain of 392 1/3 amino acid residues and a carboxyl non-triple-helical (NC1) domain of 231 residues. We also isolated a genomic DNA fragment for the human alpha 4(IV) chain, which contained two exons encoding from the carboxyl end of the triple-helical domain to the amino end of the NC1 domain. Identification of the clones was based on the amino acid sequence identity between the cDNA-deduced amino acid sequence and the reported amino acid sequence obtained from a fragment of the alpha 4(IV) collagen polypeptide M28+ (Butkowski, R. J., Shen, G.-Q., Wieslander, J., Michael, A. F., and Fish, A. J. (1990) J. Lab. Clin. Med. 115, 365-373). When compared with four other type IV collagen chains, the NC1 domain contained 12 cysteinyl residues in positions identical to those of the residues in those chains. The domain demonstrated 61, 70, 55, and 60% amino acid similarity with human alpha 1, human alpha 2, bovine alpha 3, and human alpha 5 chains, respectively. The human genomic DNA fragment allowed us to map the alpha 4(IV) gene (COL4A4) to the 2q35-2q37.1 region of the human genome.
Amino Acid Sequence↗
Genetically variable taste sensitivity to D-amino acids in mice.
Behavioral and neural responses to D-amino acids were compared between two inbred strains, C57BL and BALB mice. In both strains, an aversion conditioned to D-valine, D-leucine, D-methionine, D-histidine or D-tryptophan generalized to sucrose, whereas an aversion to D-alanine or D-serine did not generalize to sucrose. Generalization patterns across various test stimuli for each of these 7 D-amino acids were significantly correlated between two strains. However, an aversion conditioned to D-phenylalanine generalized to sucrose in C57BL mice, but not in BALB mice. Application of a proteolytic enzyme, Pronase E, to the tongue reduced chorda tympani responses to sucrose and D-amino acids to which a conditioned aversion generalized to sucrose. Again, only in C57BL mice, Pronase inhibited D-phenylalanine responses. These comparable results indicate that sweet taste response is genetically highly variable only to D-phenylalanine among 8 D-amino acids tested.
Amino Acids↗
Tissue-specific response of estrogen receptor gene expression to estrogen in chick.
We have developed a sensitive assay system by RT (reverse transcription)-PCR(polymerase chain reaction) to detect the low level of chicken estrogen receptor (cER) transcript. Using this system, the differential expression of cER gene in chick tissues was observed. Moreover, we found that the oviduct cER transcript levels in chick was not affected, albeit the remarkable growth of this tissue, by exogenous estrogen. In contrast, estrogen enhanced the hepatic cER transcript level several folds. Thus, the present study clearly showed a novel tissue-specific response of cER gene expression to estrogen in chick.
Animals↗
alpha 1(VIII)-collagen gene transcripts encode a short-chain collagen polypeptide and are expressed by various epithelial, endothelial and mesenchymal cells in newborn mouse tissues.
Type-VIII collagen is a major constituent of Descemet's membrane and contains two genetically distinct alpha chains, alpha 1(VIII) and alpha 2(VIII). We have previously cloned the human alpha 1(VIII) and alpha 2(VIII) genes and determined that they are located on chromosomes 3 and 1, respectively. Comparison of the alpha 1(VIII) and alpha 2(VIII) genes with the alpha 1(X)-collagen gene showed that the structure of the three genes and the sequence of their collagen polypeptides were strikingly similar. Therefore, we have grouped the three genes in a common subclass of collagens which we have named the short-chain collagens because of the relatively small size of their triple-helical domains. In the present study, we have isolated and characterized a mouse gene fragment encoding the entire mouse alpha 1(VIII)-collagen chain and determined the complete primary structure of the polypeptide chain. The size of mouse alpha 1(VIII)-collagen mRNA, as estimated by Northern-blot analysis, was 4.2 kb, compared with 2.8 kb previously reported for the corresponding rabbit mRNA. By cloning and sequencing of four overlapping cDNA, we demonstrate that this larger size of mouse alpha 1(VIII) mRNA is due to a larger 3' untranslated region in the mouse transcript. Using the gene fragment as a probe, we performed Northern-blot hybridization analysis of RNA prepared from newborn mice and demonstrated that alpha 1(VIII) collagen mRNA is expressed at high levels in the calvarium, eye and skin. In situ hybridization revealed that alpha 1(VIII) RNA is present in skin keratinocytes, corneal epithelial and endothelial cells, lens epithelial cells, as well as mesenchymal cells surrounding cartilage and calvarial bone and in the meninges surrounding the brain.
Amino Acid Sequence↗
Taste synergism between monosodium glutamate and 5'-ribonucleotide in mice.
1. Strain differences of mice were found in the taste synergism between monosodium L-glutamate (MSG) and disodium 5'-guanylate (GMP). 2. Magnitudes of chorda tympani responses to the mixture of MSG and GMP over the sum of responses to each component were greater in the order of C3H/HeSlc(C3H) greater than C57BL/6CrSlc(C57BL) greater than BALB/cCrSlc(BALB) mice. The greatest synergism was observed in response to the mixture of 0.03 M MSG and 0.1 mM GMP, to which responses were about 2.6, 1.8 and 1.4 times greater than the sum of each component in C3H, C57BL and BALB mice, respectively. 3. Magnitudes of inhibition of MSG and mixture responses by the lingual treatment of proteolytic enzyme, Pronase E, were greater in the same order of C3H greater than C57BL greater than BALB mice as that observed in magnitudes of the synergism. These results suggest that there exists quantitative differences in receptors responsible for taste synergism between MSG and GMP among three mouse strains.
Animals↗
Immunohistological analysis of P53 expression in human skin tumors.
The p53 expression in various skin tumors was immunohistologically evaluated using two mouse monoclonal anti-p53 antibodies, PAb421 and PAb1801. The p53 expression was not detected in the normal epidermal cells. Nuclear staining suggested that the p53 expression was observed in 10 of 26 squamous cell carcinomas (SCCs) from 24 patients, in one undifferentiated carcinoma, one proliferating trichilemmal cyst, one malignant proliferating trichilemmal tumor and in one metastatic carcinoma of breast cancer. None off four cases of Bowen's disease (SCC in situ) showed nuclear staining. In the SCCs, five of 20 primary lesions, three of four recurrent lesions and both of two metastatic lesions had positive nuclei. There was one case of SCC in which a primary lesion was negative but a recurrent lesion was positive. Thus, p53 expression was more frequently observed in SCCs at more clinically advanced stages. This may suggest that p53 has some relevance to progression of SCC. Nuclear staining was not detected in any of the following cases: two cases of seborrheic keratosis, one eccrine poroma, one keratoacanthoma, 11 basal cell epitheliomas, two mammary Paget's disease, three genital Paget's disease, one sebaceous carcinoma, four malignant melanomas, six lymphomas, two leukemia cutis and two angiosarcomas.
Antibodies, Monoclonal↗
Autonomic nervous system regulation of baseline heart rate in the fetal lamb.
OBJECTIVE: We examined 29 chronically instrumented fetal lambs from 125 to 143 days' gestation to investigate the effects of fetal behavioral states and autonomic nervous system maturation on baseline fetal heart rate. STUDY DESIGN: Behavioral states were defined from electrocorticographic analysis as low-voltage fast activity or high-voltage slow activity. Decrease and increase in baseline fetal heart rate subsequent to administration of propranolol and methylatropine represented beta-sympathetic and parasympathetic activity. RESULTS: Baseline fetal heart rate decreased with gestation in both states, with steeper regression in low-voltage fast activity (p less than 0.001). Positive correlation was noted between gestational age and percent decrease baseline fetal heart rate in both states with steeper regression in high-voltage slow activity (p less than 0.001), and between gestational age and percent increase baseline fetal heart rate with steeper regression in low-voltage fast activity (p less than 0.001). Fetal heart rate beta-sympathetic and parasympathetic tones increased with age in both states, with elevation of beta-sympathetic tone in high-voltage slow activity and parasympathetic tone in low-voltage fast activity. CONCLUSION: Sympathetic and parasympathetic systems influence baseline fetal heart rate in these behavioral states and with age.
Animals↗
Physiologic baroreceptor activity in the fetal lamb.
OBJECTIVE: We investigated the influences of gestational age and behavioral state on fetal baroreceptor activity. STUDY DESIGN: Two protocols were used to analyze low-voltage fast activity and high-voltage slow activity in eight chronically instrumented ovine fetuses at days 127 to 142. In protocol 1 correction of fetal heart rate to mean arterial blood pressure was assessed under physiologic conditions, and the linear regression slope of fetal heart rate on mean arterial pressure was calculated. In protocol 2 an injection of 7.5 micrograms of norepinephrine was given during consecutive behavioral cycles. The ratio of fetal heart rate decrease to mean arterial pressure increase was determined as representing baroreceptor activity. RESULTS: Under protocol 1 the slope was steeper in high-voltage slow activity (2.33 +/- 1.03) than in low-voltage fast activity (0.89 +/- 0.37) (n = 21, p less than 0.001), indicating that fetal heart rate changes are sensitive to blood pressure changes during high-voltage slow activity. Under protocol 2 inverse relationships were found between the ratio of fetal heart rate decrease to mean arterial pressure increase and gestational age in low-voltage fast activity and high-voltage slow activity. CONCLUSION: Fetal baroreceptor activity increased during high-voltage slow activity and decreased with gestational age.
Animals↗
The complete primary structure of the human alpha 1 (VIII) chain and assignment of its gene (COL8A1) to chromosome 3.
Type VIII collagen molecules, expressed by corneal and vascular endothelial cells, appear to be heterotrimers composed of two genetically distinct polypeptides, alpha 1 (VIII) and alpha 2(VIII), in the ratio of 2:1. Characterization of the rabbit alpha 1(VIII) gene has demonstrated that it consists of only four exons, one of which is large and encodes the entire triple-helical and carboxyl non-triple-helical domains. A similar exon organization has been found for the chicken alpha 1(X) and the mouse and human alpha 2(VIII) collagen genes. The genes encoding alpha 1(VIII), alpha 2(VIII), and alpha 1(X) collagen chains are therefore homologous members of a unique class of genes within the collagen superfamily. In the present paper we describe for the first time the primary structure of the human alpha 1(VIII) collagen chain, based on isolation and sequencing of a genomic DNA fragment. The results indicate that the human alpha 1(VIII) chain has the same domain structure as the rabbit protein. We also demonstrate that the gene is localized on the long arm of the human chromosome 3. The availability of genomic DNA encoding the human alpha 1(VIII) collagen chain and information about its chromosomal location should make it possible to examine whether hereditary diseases are linked to abnormalities in the structure or expression of the alpha 1(VIII) gene.
Amino Acid Sequence↗
The alpha 2(VIII) collagen gene. A novel member of the short chain collagen family located on the human chromosome 1.
Type VIII collagen is a major component of Descemet's membrane, the specialized basement membrane of corneal endothelial cells. Sequence analysis of a cDNA isolated from a library made with mRNA from rabbit corneal endothelial cells has indicated that type VIII molecules contain a polypeptide chain, alpha 1(VIII), consisting of a short triple-helical domain of 454 amino acid residues flanked by non-triple-helical domains of 117 and 173 amino acid residues at the amino and carboxyl ends, respectively (Yamaguchi, N., Benya, P. D., van der Rest, M., and Ninomiya, Y. (1989) J. Biol. Chem. 264, 16022-16029). The sequence of alpha 1(VIII) is strikingly similar to that of alpha 1(X) collagen, a product of hypertrophic chondrocytes. Also, characterization of the alpha 1(VIII) and alpha 1(X) collagen genes has shown that they are quite similar in their exon organization. It has been concluded, therefore, that they are homologous members of a distinct subclass of collagen genes (Yamaguchi, N., Mayne, R., and Ninomiya, Y. (1991) J. Biol. Chem. 266, 4508-4513). We have given this subclass the name short chain collagens because of the relatively small size of the triple-helical domain. In the present study, we report on the identification and characterization of a collagen gene encoding a polypeptide which is co-expressed with the alpha 1(VIII) chain in corneal endothelial cells. This collagen chain contains a triple-helical and a carboxyl non-triple-helical domain encoded by a single, large exon both in mice and humans. We conclude, therefore, that the genes encodes a novel member of the short chain collagen family, and we have given this chain the designation alpha 2(VIII) collagen. By in situ hybridization we demonstrate that the alpha 2(VIII) gene is located in the p32.3-p34.3 region of the short arm of chromosome 1.
Amino Acid Sequence↗
The alpha 1 (VIII) collagen gene is homologous to the alpha 1 (X) collagen gene and contains a large exon encoding the entire triple helical and carboxyl-terminal non-triple helical domains of the alpha 1 (VIII) polypeptide.
We recently cloned and sequenced alpha 1 (VIII) collagen cDNAs and demonstrated that type VIII collagen is a short-chain collagen that contains both triple helical and carboxyl-terminal non-triple helical domains similar to those of type X collagen (Yamaguchi, N., Benya, P., van der Rest, M., and Ninomiya, Y. (1989) J. Biol. Chem. 264, 16022-16029). We report here on the structural organization of the gene encoding the rabbit alpha 1 (VIII) collagen chain. The alpha 1 (VIII) gene contains four exons, whose sizes are 69, 120, 331, and 2278 base pairs. The first and second exons encode only 5'-untranslated sequences, whereas the third exon codes for a very short (3 nucleotides) stretch of 5'-untranslated sequence, the signal peptide, and almost the entire amino-terminal non-triple helical (NC2) domain (109 1/3 codons). Interestingly, the last exon encodes the rest of the translated region, including 7 2/3 codons of the NC2 domains, the complete triple helical domain (COL1, 454 amino acid residues), the entire carboxyl-terminal non-triple helical domain (NC1, 173 amino acid residues), and the 3'-untranslated region. This exon-intron structure is in stark contrast to the multi-exon structure of the fibrillar collagen (types I, II, III, V, and XI) genes, but it is remarkably similar to that of the type X collagen gene (LuValle, P., Ninomiya, Y., Rosenblum, N. D., and Olsen, B. R. (1988) J. Biol. Chem. 263, 18278-18385). The data suggest that the alpha 1 (VIII) and the alpha 1 (X) genes belong to the same subclass within the collagen family and that they arose from a common evolutionary precursor.
Amino Acid Sequence↗
Spatiotemporal pattern of type X collagen gene expression and collagen deposition in embryonic chick vertebrae undergoing endochondral ossification.
We examined the spatio-temporal pattern of type X collagen mRNA and its protein in the embryonic chick vertebrae undergoing ossification by in situ hybridization and immunohistochemistry. Hypertrophic chondrocytes, producing type X collagen, were developed as islands of cells in a few vertebral body segments of stage 36 embryos. These cells were increased in number at stages 37 and 38 and they expressed high levels of type X collagen mRNA and deposited its protein in the matrix. Blood vessels entered from the perichondrium at stage 37 and invaded deeply into hypertrophic cartilage at stage 38. As the vertebrae grew further at stage 40, the leading front of active hypertrophic chondrocytes with high levels of type X mRNA shifted from the midvertebral perivascular area towards intervertebral borders, while the perivascular area retained a number of inactive hypertrophic chondrocytes with low levels of type X mRNA. Type X collagen was found in large amounts throughout the matrix areas containing both active and inactive hypertrophic chondrocytes. Calcium was detected by von Kossa's technique in hypertrophic cartilage matrix in a small amount at stage 37, in parts of the matrix with type X collagen deposition in succeeding stages, and finally in almost the entire area of type X collagen deposition at stage 45. The vertebral segments of stage 45 embryos also showed a clearly reversed pattern of expression between type X collagen mRNA and types II and IX collagen mRNAs. The results demonstrate that the production of type X collagen by hypertrophic chondrocytes precedes both vascular invasion and mineralization of the matrix, suggesting that hypertrophic chondrocytes have an important role in regulating these events.
Animals↗
Gustatory neural responses in preweanling mice.
Taste sensitivity of preweanling mice was studied by examining responses of the chorda tympani (CT) and glossopharyngeal (GL) nerves to various taste stimuli, and was compared to that of adult mice. In mice of 7-10 days of age, comparing to that of the CT nerve, threshold of the GL nerve for monosodium l-glutamate (MSG) was low, but those for sucrose and NaCl were high. Sensitivities to HCl and quinine-HCl were similar between the CT and GL nerves, although that to quinine-HCl was larger in the GL nerve than in the CT nerve in adult mice. Enhancement of MSG responses by addition of GMP was observed in the CT nerve but not in the GL nerve in this age group. In mice of 8-16 weeks of age, threshold of the GL nerve for MSG became higher but that for NaCl became lower. Enhancement of MSG responses by addition of GMP appeared also in the GL nerve. Inhibition of NaCl responses by amiloride was observed in the CT nerve. These results suggest that, in mice, the GL nerve is important taste input for umami substances especially during the preweanling period, whereas the CT nerve is for sweet and salty substances. Properties of umami and salt receptor systems change during the postweanling period.
Animals↗
On the taste of umami in chimpanzee.
Whole and single fiber chorda tympani nerve recordings were obtained in 5 chimpanzees to stimulation with MSG (monosodium phosphate) and GMP (guanosine 5'-monophosphate, disodium salt) alone and in combination. The overall chorda tympani nerve activity was recorded to 5 concentrations of MSG, ranging from 1 to 100 mM with and without 0.3 mM GMP, and to 5 concentrations of GMP, ranging from 0.1 to 10 mM, with and without 30 mM MSG. A synergistic effect was recorded between MSG and GMP in 3 out of 4 animals. The effect of stimulation with MSG and GMP alone and mixed was studied in approximately 25 single fiber recordings against a background of the stimulating effects of 11 different sweeteners, 3 acids, 3 bitter compounds and 3 different salts. The fibers showed a high taste specificity and fell into groups which corroborated with the human concepts of the taste qualities. The umami compounds elicited moderate responses which were largest in the sweet fibers. In the 6 sweet fibers that responded to the umami compounds. 0.3 mM GMP was a more effective stimulus than 10 mM MSG. In 3 of these fibers a synergistic effect was recorded to the mixture of GMP and MSG. It is interesting that the response to GMP and MSG was unaffected by gymnemic acid, although it blocked the response to the sweet compounds. Three out of 10 salt fibers responded to MSG and GMP but no synergistic effect was recorded. No specific umami fibers were recorded. However, more data must be collected before the final conclusion on the presence or absence of specific umami fibers can be drawn.
Animals↗
Cloning of alpha 1(IV) and alpha 2(IV) collagen cDNAs from rabbit corneal endothelial cell RNA.
To understand the role of type IV collagen in embryogenesis, regeneration, and tissue repair of the cornea at the molecular level, the authors isolated clones coding for rabbit alpha 1(IV) and alpha 2(IV) chains from a cDNA library constructed with rabbit corneal endothelial cell RNA. The isolated alpha 2(IV) cDNA clones encode a part of a 5' untranslated region, the signal peptide, the 7S domain, part of the triple-helical domain, and the entire carboxyl-terminal nontriple-helical (NC1) domain. By cross hybridization, using one of the alpha 2(IV) cDNA inserts, a cDNA clone encoding the alpha 1(IV) chain was isolated from a lambda gt10 library primed with oligo(dT). The clone covered a short portion of the triple-helical domain, the entire NC1 domain, and a short 3' untranslated region. The nucleotide-sequence analysis of these clones provides, for the first time to the authors' knowledge, the primary structure of the carboxyl-terminal portion of both rabbit alpha 1(IV) and alpha 2(IV) collagen chains. Between the alpha 1(IV)NC1 and the alpha 2(IV)NC1, 61% and 66% sequence similarities were observed at the amino acid and nucleotide levels, respectively. The locations of all the 12 cysteinyl residues were conserved in the two NC1 sequences.
Amino Acid Sequence↗
[A study on interleukin 2 and soluble interleukin 2 receptor in serum and bronchoalveolar lavage fluid from patients with farmer's lung].
Interleukin 2 (IL-2) and soluble Interleukin 2 receptor (IL-2R) were assayed in serum and bronchoalveolar lavage fluid (BALF) from patients with farmer's lung. Patients with farmer's lung had significantly higher levels of soluble IL-2R in serum and BALF than normal controls. The level of soluble IL-2R was correlated with the disease activity, and fell to within the normal range after successful treatment with corticosteroids. On the other hand, asymptomatic dairy farmers with precipitating antibodies against Micropolyspora faeni antigen or Thermoactinomyces vulgaris antigen had the same levels of soluble IL-2R in their serum and BALF as normal controls. IL-2 levels in serum and BALF were significantly higher in patients with farmer's lung than in normal controls; however, asymptomatic patients had low levels comparable to those of normal controls. These results indicate that soluble IL-2R could be a sensitive parameter for evaluation of the activity of this disease. It is also suggested that the IL-2/IL-2R system is important in the pathogenesis of hypersensitivity pneumonitis.
Adult↗
[Primary chronic glomerulonephritis in diabetic patients].
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Adult↗