Rapid communication: PstI, HindIII, and TaqI restriction fragment length polymorphisms in the bovine Uroplakin IB gene.
Explore the source record for details and available documents.
Alleles↗
Biomedical subjects
X R Wu
Publications and source records attributed to X R Wu.
[N-methyl-D-aspartate-sensitive glutamate receptors and seizure].
Explore the source record for details and available documents.
Animals↗
TaqI and BglII RFLPs for the bovine uroplakin III loci UPK3A and UPK3B.
Explore the source record for details and available documents.
Animals↗
[Roles of glutamate receptor in c-fos gene expression and epilepsy susceptibility in rats].
We use NMDA to induce expression of c-fos mRNA as a marker to observe the activity of NMDA receptor in neurons during development, and compare the activity of NMDA receptor between audiogenic epilepsy -prone (P77PMC) and audiogenic epilepsy resistant rats brain. In primary culture of rats cerebral cortical neurons NMDA induced c-fos mRNA expression exhibits dose and time-dependent changes, which can be prevented by antagonists. During the development of neurons, the NMDA -induced c-fos mRNA expression reaches a maximum at day 24. NMDA-induced c-fos mRNA expression of P77PMC rats is higher than that of controls during 6 to 24 days in vitro with significant difference (P < 0.05) at day 18. To present changes in c-fos mRNA expression induced by NMDA in cultured P77PMC rat cortical neurons may be one of the factors related to susceptibility of epilepsy in P77PMC rats.
Animals↗
A product of phosphatidylinositol-3 kinase is elevated in dividing HT29 colonic epithelial cells.
Phosphatidylinositol 3 kinase (PI-3 kinase) activity has been linked to cell proliferation and growth regulation. Therefore, we studied changes in phosphoinositide metabolism during the cell cycle of HT-29 cells, a colonic epithelial cell line. HT29 cells were treated with the microtubule disrupter, nocodozole, separated into mitotic and quiescent populations and their phospholipid composition was analyzed. Radiolabelled phospholipids from cells labelled with 32PO4 or [3H]myoinositol were analyzed by TLC and/or deacylated and analyzed by HPLC. In all cases, levels of phosphatidylinositol 3 phosphate from mitotic phase cells were double that in resting populations. Therefore, levels of a product of PI-3 kinase are elevated and may play a role in cell division.
Cell Adhesion↗
Excitatory amino acid regulation of the enkephalin phenotype in mouse embryonic spinal cord cultures.
Expression of the preproenkephalin gene in developing spinal cord-dorsal root ganglia (SC-DRG) cultures was determined by Northern analysis following treatments with different agonists and antagonists of the glutamate receptor. Cultures (10-12 days old) were treated with various concentrations (10(-7)-10(-3) M) of N-methyl-D-aspartate (NMDA), quisqualate, kainic acid (KA), 2-amino-5-phosphonovaleric acid (APV) and 5-methyl-10,11-dihydro-5H-dibenzo[a, d]cyclohepten-5,10-imine maleate (MK801) either with or without blocking spontaneous electrical activity with 1 microM tetrodotoxin (TTX). In electrically active cultures, treatments with NMDA and KA increased preproenkephalin transcripts (mRNAppENK), showing maximum effects at 1 microM (4-fold and 2-fold, respectively), while treatments with quisqualate and MK801 caused concentration-dependent down-regulation in mRNAppENK. The most effective concentrations of NMDA (1 microM) and quisqualate (10 microM) altered mRNAppENK levels within 4 h of treatment and peaked after 24 h for NMDA and 48 h for quisqualate treatment. Co-treatment with APV completely blocked the NMDA-induced rise of mRNAppENK. During electrical blockade, none of the concentrations of NMDA tested showed any effect on enkephalin expression, neither could NMDA pre-treatment prevent the TTX-induced down-regulation of mRNAppENK. Our results indicate that the activity-dependent establishment of the enkephalin phenotype is modulated through the selective activation of the NMDA-glutamate receptor.
Amino Acids↗
Chromosomal localization of uroplakin genes of cattle and mice.
The asymmetric unit membrane (AUM) of the apical surface of mammalian urinary bladder epithelium contains several major integral membrane proteins, including uroplakins IA and IB (both 27 kDa), II (15 kDa), and III (47 kDa). These proteins are synthesized only in terminally differentiated bladder epithelial cells. They are encoded by separate genes and, except for uroplakins IA and IB, appear to be unrelated in their amino acid sequences. The genes encoding these uroplakins were mapped to chromosomes of cattle through their segregation in a panel of bovine x rodent somatic cell hybrids. Genes for uroplakins IA, IB, and II were mapped to bovine (BTA) Chromosomes (Chrs) 18 (UPK1A), 1 (UPK1B), and 15 (UPK2), respectively. Two bovine genomic DNA sequences reactive with a uroplakin III cDNA probe were identified and mapped to BTA 6 (UPK3A) and 5 (UPK3B). We have also mapped genes for uroplakins IA and II in mice, to the proximal regions of mouse Chr 7 (Upk1a) and 9 (Upk2), respectively, by analyzing the inheritance of restriction fragment length variants in recombinant inbred mouse strains. These assignments are consistent with linkage relationships known to be conserved between cattle and mice. The mouse genes for uroplakins IB and III were not mapped because the mouse genomic DNA fragments reactive with each probe were invariant among the inbred strains tested. Although the stoichiometry of AUM proteins is nearly constant, the fact that the uroplakin genes are unlinked indicates that their expression must be independently regulated. Our results also suggest likely positions for two human uroplakin genes and should facilitate further analysis of their possible involvement in disease.
Animals↗
Levonorgestrel levels in serum and uterine flushings in women bearing LNG-Cu-IUD.
Radioimmunoassay was employed to measure levonorgestrel levels in serum and uterine flushings in women using intrauterine device releasing both copper ions and levonorgestrel (LNG-Cu-IUD) after one-year insertion. In the meanwhile physical characteristics of cervical mucus have been observed in women using LNG-Cu-IUD and compared with those using Tcu-220c-IUD. The mean concentrations of levonorgestrel in serum and uterine flushings (5 ml) were 84.53 pg/ml and 266.93 pg/ml respectively. In LNG-Cu-IUD group, the cervical mucus lacking in ferny crystals had a viscosity significantly higher than that in Tcu-220c-IUD group, suggesting that the local high concentration of levonorgestrel changed the environment of the uterine cavity and the cervical mucus status.
Adult↗
Molecular cloning of a 47 kDa tissue-specific and differentiation-dependent urothelial cell surface glycoprotein.
Despite the fact that bladder epithelium has many interesting biological features and is a frequent site of carcinoma formation, relatively little is known about its biochemical differentiation. We have shown recently that a 47 kDa glycoprotein, uroplakin III (UPIII), in conjunction with uroplakins I (27 kDa) and II (15 kDa), forms the asymmetric unit membrane (AUM)--a highly specialized biomembrane characteristic of the apical surface of bladder epithelium. Deglycosylation and cDNA sequencing revealed that UPIII contains up to 20 kDa of N-linked sugars attached to a core protein of 28.9 kDa. The presence of an N-terminal signal peptide sequence and a single transmembrane domain located near the C terminus, plus the N-terminal location of all the potential N-glycosylation sites, points to a type I (N-exo/C-cyto) configuration. Thus the mass of the extracellular domain (20 kDa plus up to 20 kDa of sugar) of UPIII greatly exceeds that of its intracellular domain (5 kDa). Such an asymmetrical mass distribution, a feature shared by the other two major uroplakins, provides a molecular explanation as to why the luminal leaflet of AUM is almost twice as thick as the cytoplasmic one. The fact that of the three major proteins of AUM only UPIII has a significant cytoplasmic domain suggests that this molecule may play an important role in AUM-cytoskeleton interaction in terminally differentiated urothelial cells.
Amino Acid Sequence↗
[Uroplakin III, a specific membrane protein of urothelial umbrella cells, as a histological markers for metastatic transitional cell carcinomas].
We have investigated, by immunohistochemical staining of various paraffin-embedded carcinoma sections, the tissue-specific expression of uroplakin III--a recently characterized constituent glycoprotein (47 kD) of the asymmetrical unit membrane which forms plaques on apical surface of urothelial umbrella cells. The apical membrane pattern of normal urothelial umbrella cells was in part maintained in papillary transitional cell carcinomas (TCCs). In addition, in both papillary and invasive TCCs, variously sized lumina exhibited positive membrane staining of uroplakin III. In some cases, basal cell membrane staining was seen. Positive reactions (which sometimes were very focal) were noted in 16/18 papillary non-invasive TCCs (89%), 21/37 invasive TCCs (57%) and 12/15 TCC metastases (80%). Non-TCC carcinomas of different origin (n = 63) were consistently negative. These results show that uroplakin III may serve as a useful marker for TCCs, revealing specific urothelial differentiation features to be expressed in such tumors even after metastasis. This marker, while of only intermediate sensitivity, is highly specific, thus opening interesting histodiagnostic possibilities in the case of unclear carcinoma metastases.
Biomarkers, Tumor↗
[Proto-oncogene c-fos and convulsion].
Explore the source record for details and available documents.
Animals↗
Effects of cholecystokinin octapeptide on genetically determined seizure susceptibility.
Utilizing audiogenic seizure-prone P77PMC rats, the effects of cholecystokinin octapeptide (CCK-8) on genetic seizure susceptibility were studied in vivo, in cerebral cortical synaptosomes, and in cortical neuronal cell cultures. The results showed that CCK-8 could decrease seizure susceptibility, and that the K(+)-stimulated release of GABA in cerebral cortex synaptosomes from seizure-prone animals was depressed. The presence of exogenous CCK-8 (10(-7) M) together with elevated K+ (25 mM) causes a higher increased magnitude in GABA release from synaptosomes (enhanced by 100%) and cell cultures (17 days in vitro, increased by 177%) derived from seizure-prone rats than the controls (increased by 42%, in synaptosomes; and 107% in cell cultures). These preliminary results raise the possibility that the developmental abnormalities in modulation effect of CCK-8 on GABA release in central nervous system may play a role causing greater seizure susceptibility in genetic seizure-prone rats. The analysis of the brain tissue level and gene-expression of CCK-8 will be the important step of further investigation.
Animals↗
Dynamic changes of myometrial activity, levels of PGF2 alpha and E2 in rabbits after insertion of four types of IUDs.
Myometrial activity, levels of PGF2 alpha and E2 in uterine flushings were measured in rabbits before and after insertion of four types of IUDs. The results showed that IUDs could increase the uterine contraction during the early stages of insertion. Thereafter, the uterine activity tended to be stable. The sequences of uterine contractility generally were: Cu-IUD greater than SS-IUD greater than LNG-Cu-IUD greater than LNG-IUD. The measurement of prostaglandins, determined by RIA, indicated that the concentrations of PGF2 alpha and E2 in Cu-IUD and SS-IUD groups were higher than those of controls for the early phase of insertion, which was not found after four months of use. Our results suggested that hyperactivity of the uterus in the early stage after insertion of IUDs might be relevant to side-effects like expulsion and pain. Copper released by Cu-IUD stimulates the uterine activity partially through increasing PGF2 alpha and E2 contents. In rabbits wearing LNG-IUD or LNG-Cu-IUD, both PGF2 alpha and E2 levels were low. The relatively low uterine mechanical and electrical activities were also observed in these two groups. Based on these data, it is concluded that development of LNG-Cu-IUD is feasible for reducing expulsion of IUDs.
Animals↗
Large scale purification and immunolocalization of bovine uroplakins I, II, and III. Molecular markers of urothelial differentiation.
The differentiation of mammalian urothelium culminates in the formation of asymmetrical unit membrane (AUM). Using gradient centrifugation and detergent wash, we purified milligram quantities of AUMs which, interestingly, contained three major proteins (15, 27, and 47 kDa) that appeared to be identical to the three immunoaffinity purified, putatively AUM-associated proteins that we described earlier (Yu, J., Manabe, M., Wu, X.-R., Xu, C., Surya, B., and Sun, T.-T. (1990) J. Cell Biol., 111, 1207-1216). Peptide mapping and immunoblotting established that these three proteins were distinct molecules. Using monospecific antibodies to these three proteins, we showed that they were all restricted to the superficial urothelial cells and were AUM-associated. The 27- and 15-kDa proteins were detected exclusively on the luminal side of mature, apical AUMs. In contrast, epitopes of the 47-kDa protein were detected on both sides of apical AUMs suggesting a transmembranous configuration. These results (i) provide the strongest evidence thus far that AUM contains three major proteins (the 27-kDa uroplakin I, 15-kDa uroplakin II, and 47-kDa uroplakin III) which form an extremely insoluble complex, (ii) suggest that uroplakin II, like uroplakin I (Yu, J., Manabe, M., Wu, X.-R., Xu, C., Surya, B., and Sun, T.-T. (1990) J. Cell. Biol. 111, 1207-1216), translocates from one side of the membrane to another during AUM maturation, (iii) indicate that uroplakin III may play a different structural role than uroplakins I and II in AUM formation, and (iv) establish the three uroplakins as markers for an advanced stage of urothelial differentiation.
Animals↗
New prostaglandin synthetase inhibitors--di-and triphenylacrylonitriles.
16 triphenylacrylonitriles (TPE) or diphenylacrylonitriles (DPE) were synthesized by condensation of various benzophenones or benzaldehydes with various phenylacetonitriles. The pharmacological potency of these compounds was studied by incubation of bovine seminal vesicle microsomes and PG-RIA. The results showed that the potency of inhibition of PG biosynthetase of DPE was stronger than that of TPE. Compounds with electron-releasing functional groups were proved to be more effective than those with electron-attracting groups. Compound DPE-9 was the most active one, the potency of which was 40 times stronger than that of naproxen. The structure of some compounds has been analysed by X-ray diffraction. The relationship between structure and activity was investigated by means of X-ray diffraction, UV and NMR.
Acrylonitrile↗
The elastic modular changes of the rat uterine cervix under different physiological conditions.
In this paper, the elastic moduli of the rat uterine cervix were measured under different physiological conditions. It was found that the moduli significantly changed with the sex cycle, being higher in diestrus than in estrus, and highest in early pregnancy. The results suggest that active changes occur in the function of the uterine cervix under the modulation of hormones.
Animals↗
Uroplakin I: a 27-kD protein associated with the asymmetric unit membrane of mammalian urothelium.
The luminal surface of mammalian urothelium is covered with numerous plaques (also known as the asymmetric unit membrane or AUM) composed of semi-crystalline, hexagonal arrays of 12-nm protein particles. Despite the presumed importance of these plaques in stabilizing the urothelial surface during bladder distention, relatively little is known about their protein composition. Using a mouse mAb, AE31, we have identified a 27-kD protein that is urothelium-specific and is differentially expressed in superficial umbrella cells. This protein (pI approximately 5.8) partitions into the detergent phase during Triton X-114 phase separation. Pulse-chase experiments using cultured bovine urothelial cells showed that this protein is synthesized as a 32-kD precursor that is processed through a 30-kD intermediate, to the mature 27-kD form. In cytoplasmic vesicles containing immature AUM, the AE31 epitope is detected in patches on the cytoplasmic side, but in mature, apical AUM it is detected exclusively on the luminal side. This suggests an unusual translocation of the AE31 epitope during AUM maturation; more data are required, however, to substantiate this interpretation. Immunoaffinity purification of the 27-kD protein results in the copurification in approximately molar ratio of a 15-kD protein, as well as a small and variable amount of a 47-kD protein. Immunoblotting data indicate that these three proteins are immunologically distinguishable. This copurified 15-kD protein is relative basic (pI approximately 8.0). Like the 27-kD protein, it is urothelium-specific and is present mainly in the umbrella cells. Together, our data indicate that a 27-kD protein is urothelial plaque-associated (uroplakin I). Based on complex formation data, we provisionally name the 15-kD protein uroplakin II; additional data will be required to determine whether this and the 47-kD protein are integral parts of AUM. The identification of these AUM-associated and -related proteins, plus the availability of a culture system capable of synthesizing and processing some of these molecules, offer new opportunities for studying the detailed structure, assembly, and function of asymmetrical unit membrane.
Animals↗