Mouse Urinary Bladder Research Articles

Differential activity of aspirin, ketoprofen and sulindac as cancer chemopreventive agents in the mouse urinary bladder.

In vivo studies were conducted to compare the activity of three non-steroidal anti-inflammatory drugs as inhibitors of urinary bladder carcinogenesis induced in B6D2F1 (BDF) mice by N-butyl-N-(4-hydroxybutyl)nitrosamine (OH-BBN). Mice received continuous dietary exposure to non-toxic doses of aspirin, sulindac or ketoprofen beginning 1 week prior to the first of eight weekly doses of 7.5 mg OH-BBN; studies were terminated at 24 weeks after the first carcinogen dose. Both dose levels of sulindac (200 and 400 mg/kg diet) and both dose levels of ketoprofen (40 and 80 mg/kg diet) reduced the incidence of transitional cell carcinoma of the urinary bladder by >70% from that seen in dietary controls. The high dose of sulindac conferred the greatest protection against bladder cancer induction. In contrast, when administered at 400 and 800 mg/kg diet aspirin was inactive as a chemopreventive agent in the OH-BBN/BDF bladder cancer model. The significant potency of sulindac and ketoprofen as inhibitors of urinary bladder carcinogenesis, when considered with their history of safe human use, suggests that these agents merit further study as drugs for cancer chemoprevention in this target tissue.

A comparative study of the desquamation of urothelial cells during gestation and in adults mice following moderate stress or endotoxin treatment

The present paper deals with the comparative EM study of the detachment of mouse urinary bladder epithelial cells under various physiological conditions, namely, during gestation and in adult mice following induction by endotoxin or by exposure to moderate stress. It has been shown that desquamation during gestation involves two distinct modes, shedding of single cells and formation of apoptotic bodies. Moderate stress in adult female mice induced by constant illumination for 72 or 96 hours, results in desquamation of superficial and intermediate cells. Application of LPS is followed by desquamation of single cells and whole sheets of cells. Cell detachment involves interruption of tight junctions between neighbouring cells and formation of numerous cup shaped vesicles, multivesicular bodies and vacuoles at the base of desquamating cells. LPS induces desquamation entails delivery of lysosomal enzymes extracellulary into the intercellular space. In the areas of sloughing the bladder epithelium lack permeability barrier. Desquamated cells in the bladder lumen are mostly alive. These results clearly demonstrate the existence of specific adhesion mechanisms of detachment and desquamation of uroepithelial cells.

Endocytosis during postnatal differentiation in superficial cells of the Mouse Urinary Bladder Epithelium.

This electron microscopical study was performed in order to follow the endocytic pathway of horseradish peroxidase and colloidal gold tracers and to determine the involvement of endocytosis in postnatal differentiation in superficial cells of the mouse urinary bladder epithelium. Morphometric analyses of late endosomes/multivesicular bodies from day of birth to day 25 were performed. The internalisation and intracellular transport of luminal plasmalemma to multivesicular bodies via endocytic vesicles, early endosomes and pleomorphic compartments was established. Dynamic changes in endocytic activity took place within the first few days of postnatal differentiation. During this period the number of multivesicular bodies changed in an inverse ratio to their size. After the third day endocytic activity gradually approached the low rate of adult urothelium.

Selective Staining of the Superficial Cells of Mouse Urinary Bladder Epithelium by Horseradish Peroxidase (HRP)

A simple method for staining the superficial cells of mouse urinary bladder epithelium with horseradish peroxidase (HRP) is presented. HRP solution was instilled into the bladder lumen in vivo. After fixation with glutaraldehyde, the bladder was treated with Triton X-100 and then reacted with diaminobenzidine (DAB). Intense DAB reactions were detected throughout the whole of the cytoplasm of superficial cells by both light and electron microscopy. No reaction was observed in intermediate or basal epithelial cells. This simple method will be useful for the three-dimensional reconstruction of cell shape and identification of the stratification of transitional epithelial cell layers.

Chemoprevention of mouse urinary bladder carcinogenesis by the naturally occurring carotenoid astaxanthin

The chemopreventive effects of two xanthophylls, astaxanthin (AX) and canthaxanthin (CX), on urinary bladder carcinogenesis induced by N-butyl-N(4-hydroxybutyl)nitrosamine (OH-BBN) was investigated in male ICR mice. Mice were given 250 p.p.m. OH-BBN in drinking water for 20 weeks and after a 1 week interval with tap water, water containing AX or CX at a concentration of 50 p.p.m. was administered during subsequent 20 weeks. Other groups of mice were treated with AX or CX alone or untreated. At the end of the study (week 41), the incidences of preneoplastic lesions and neoplasms in the bladder of mice treated with OH-BBN and AX or CX were smaller than those of mice given OH-BBN. In particular, AX administration after OH-BBN exposure significantly reduced the incidence of bladder cancer (transitional cell carcinoma) (P < 0.003). However, the inhibition of the frequencies of such lesions in mice treated with OH-BBN and CX was not significant. Treatment with AX or CX also decreased the number/nucleus of silver-stained nucleolar organizer region proteins (AgNORs), a new index of cell proliferation, in the transitional epithelium exposed to OH-BBN. Preneoplasms and neoplasms induced by OH-BBN, and the antiproliferative potential, was greater for AX than CX. These results indicate that AX is a possible chemopreventive agent for bladder carcinogenesis and such an effect of AX may be partly due to suppression of cell proliferation.

Endocytic compartments in superficial cells of the mouse urinary bladder epithelium: R. Romih and K. Jezernik (Institute of Cell Biology, Medical Faculty, Lipicˇeva 2, Ljubljana, Slovenia)

Endocytic compartments in superficial cells of the mouse urinary bladder epithelium: R. Romih and K. Jezernik (Institute of Cell Biology, Medical Faculty, Lipicˇeva 2, Ljubljana, Slovenia)

Paraneoplastic pemphigus in a child with a T-cell lymphoblastic lymphoma

We report a child with a T-cell lymphoblastic lymphoma, and a widespread erosive mucocutaneous eruption, whose clinical and immunopathological features were consistent with a diagnosis of paraneoplastic pemphigus. Direct immunofluorescence showed deposition of C3 in the epidermal intercellular spaces, and in a linear pattern along the dermo-epidermal junction. Circulating autoantibodies binding in a pemphigus-like pattern to mouse urinary bladder epithelium were found. By Western immunoblotting, the patient's serum recognized two epidermal polypeptides with molecular weights of 210 and 190 kDa. No reactivity with the 230-kDa bullous pemphigoid antigen and the 250-kDa desmoplakin I antigen was found.

Blood-urine barrier formation in mouse urinary bladder development.

Formation of the blood-urine permeability barrier in differentiating mouse transitional urothelium was studied. It was established that the development of superficial cell barrier is a two-phase process: beginning with formation of the tight junctions, followed by formation of fusiform vesicles and asymmetric apical plasma membranes. Fusiform vesicles differentiate during days 15 and 17 of gestation and fuse with the apical plasmalemma. Thus a thick membrane is formed before the excretion of hypertonic urine into the embryonic bladder. Through some degenerative superficial cells slough between fetal day 17 and the day of birth, the bladder epithelium in mice does not lack an effective permeability barrier.

ATP induced‐relaxation in the mouse bladder smooth muscle

1. The effect of adenosine 5'-triphosphate (ATP) on the free cytosolic Ca2+ concentration ([Ca2+]i) as measured with the fluorescent Ca(2+)-indicator fura-2, and on force was investigated in the intact smooth muscle strips of the mouse urinary bladder. 2. ATP elicited, when exogenously applied, a large increase of [Ca2+]i with limited force development resulting in a marked Ca(2+)-force dissociation. 3. Release of endogenous neurotransmitters by transmural electrical stimulation (TES) for 30 s induced a steady increase of [Ca2+]i and a peak contraction, followed within 15 s by a relaxation. 4. In carbachol-prestimulated preparations, ATP elicited an initial rise of [Ca2+]i followed by a return to the initial precontraction Ca(2+)-level. Force in contrast presented a biphasic pattern, i.e. an initial contraction was followed by a sustained relaxation. 5. In the K(+)-depolarized precontracted preparation, ATP elicited a slight initial rise of [Ca2+]i. The partial relaxation of the force during depolarization was not preceded by a transient contraction. 6. The ATP-induced relaxation of the K(+)-prestimulated preparations was not inhibited by 8-phenyltheophylline, a potent P1-purinoceptor antagonist. 7. The order of potency for relaxation of the ATP analogues was 2-MeSATP > ATP > beta gamma Me-ATP, which is characteristic for P2y-purinoceptors. 8. These results indicate that, besides its activating effect, ATP also relaxes the mouse urinary bladder. It is suggested that the relaxant effect, mediated through P2y-purinoceptors, is mainly responsible for the low contractile potency of ATP in the bladder.

Ustilaginoidea virens産生トキシンによるマウス肝, 腎および膀胱上皮のApoptosis誘発

Ustilaginoidea virens産生トキシンによるマウス肝, 腎および膀胱上皮のApoptosis誘発

Chemoprevention of OH-BBN-induced bladder cancer in mice by piroxicam.

Piroxicam inhibited induction of transitional cell carcinoma in mouse urinary bladder by N-butyl-N-(4-hydroxybutyl)-nitrosamine. At 15 mg piroxicam/kg diet, tumor incidence was reduced 82% (P < 0.0001) compared with carcinogen controls. At 30 mg piroxicam/kg diet, tumor incidence was reduced 70% (P < 0.001). Results at the higher dose level suggested that piroxicam also may have inhibited invasion slightly. Combination treatment with 2-difluoromethyl-ornithine (DFMO) or all-trans-N-(4-hydroxyphenyl)retinamide (4-HPR) or both agents did not improve the chemopreventive potential of piroxicam. However, the three-agent combination of 30 mg piroxicam/kg, 1200 mg DFMO/kg and 313 mg 4-HPR/kg diet was highly effective. Tumor incidence was reduced 91% (P < 0.0001) compared with carcinogen controls. Unfortunately, the high efficacy was somewhat compromised by a significant decrease in survival and body weight gain in mice receiving the combination of agents compared with the carcinogen control.

Photodynamic effect on mouse urinary bladder cancer with the combination of pheophorbide a and Nd:YAG laser

Photodynamic effect on mouse urinary bladder cancer with the combination of pheophorbide a and Nd:YAG laser

Fractionation sensitivity and latency of late radiation injury to the mouse urinary bladder

Transurethral bladder filling is a functional, non-invasive, in vivo assay of early and late radiation injury to the mouse bladder. Fractionated irradiations using single doses or 2, 3, 5, or 10 dose fractions in an overall time of 4 or 4.5 days, with a range of total doses, were given to the bladder of 12–14 week-old C 3D 2F 1/Bom mice. In 372 mice, bladder volume at an intravesical pressure of 20 mmHg was measured before irradiation and at regular intervals thereafter. The endpoint for late bladder injury was a volume of less than 50% of the median pretreatment volume in all animals, occurring more than 30 days after irradiation. This endpoint was reached after a latent period ranging between 35 and 401 days. Fractionation and latency parameters were estimated using a mixture model. There was a highly statistically significant dose-dependency of the latent period ( p < 10 −8). The α β ratio was estimated at 5.8 Gy [95% confidence limits (3.6; 8.8) Gy] for 250 kVp X-rays. Thus late radiation injury in the mouse urinary bladder is one of the least sensitive late endpoints with respect to change in dose per fraction. Introducing early bladder injury as a variable in the model improved the fit significantly ( p = 0.03), but the α β ratio remained unchanged. Thus the hypothesis that late bladder injury may be, at least in part, consequent upon early injury did not explain the relatively high α β ratio for this late endpoint.

Pharmacological characterization of adenosine A1 and A2 receptors in the bladder: evidence for a modulatory adenosine tone regulating non-adrenergic non-cholinergic neurotransmission.

1. The nerve-evoked contractions elicited by transmural electrical stimulation of mouse urinary bladders superfused in modified Krebs Ringer buffer containing 1 microM atropine plus 3.4 microM guanethidine were inhibited by adenosine (ADO) and related nucleoside analogues with the following rank order of potency: R-phenylisopropyladenosine (R-PIA) greater than cyclohexyladenosine (CHA) greater than 5'N-ethylcarboxamido adenosine (NECA) greater than ADO greater than S-phenylisopropyladenosine (S-PIA). Tissue preincubation with 8-phenyltheophylline (8-PT) displaced to the right, in a parallel fashion, the NECA concentration-response curve. 2. The contractions elicited by application of exogenous adenosine 5'-triphosphate (ATP) were also inhibited by ADO and related structural analogues. The rank order of potency to reduce the motor response to ATP was: NECA greater than 2-chloroadenosine (CADO) greater than R-PIA greater than ADO greater than CHA greater than S-PIA. 3. The ADO-induced ATP antagonism was of a non-competitive nature and was not specific. Tissue incubation with 10 microM NECA not only reduced the motor responses elicited by ATP, but also 5-hydroxytryptamine, acetylcholine and prostaglandin F2 alpha. The action of NECA was antagonized following tissue preincubation with 8-PT. The inhibitory action of NECA was not mimicked by 10 microM CHA. 4. The maximal bladder ATP contractile response was significantly increased by tissue preincubation with 5-30 microM 8-PT. 5. The 0.15 Hz evoked muscular twitch was significantly increased by 8-PT while dipyridamole consistently reduced the magnitude of the twitch response. These results are consonant with the hypothesis that an endogenous ADO tone modulates the bladder neurotransmission. 6. A working model is proposed suggesting the presence of ADO-Al and A2 receptors in the mouse urinary bladder. The A1 receptor subpopulation is probably of presynaptic origin whereas the smooth muscle membranes contain a population of the A2 receptor subtype.

Early Changes in Mouse Urinary Bladder Function Following Fractionated X Irradiation

Functional changes in the mouse urinary bladder following single-dose or fractionated irradiation were assessed by cystometry, i.e., by measuring the intravesical volume-pressure relationship during transurethral filling. The early response presented as a dose-dependent and transitory decrease in the reservoir function of the organ as defined by the intravesical volume at a filling pressure of 10 or 20 mm Hg, V10 or V20. The quantal dose response used in the present study was a reduction in the individual bladder volume (V10 and V20) by at least 50%. After single doses greater than or equal to 10 Gy, the reaction occurred between Days 7 and 25 with maximum prevalence between Days 7 and 14. The individual duration of the response was 3-9 days. Treatment with single doses and 2, 3, 5, and 10 fractions demonstrated a significant sparing effect with ED50 values of 18.3, 24.9, 26.8, 29.8, and 38.0 Gy, respectively. The linear-quadratic model fitted the data reasonably well when tested according to Tucker (Int. J. Radiat. Oncol. Biol. Phys. 10, 1933-1939, 1984). The alpha/beta ratios estimated with different two-step techniques ranged from 11.1 to 12.4 Gy. Analysis of the data as proposed by Thames et al. (Int. J. Radiat. Biol. 49, 999-1009, 1986) yielded an alpha/beta value of 13.9 Gy (95% confidence limits 8.4 and 24.6 Gy), illustrating a fractionation effect typical for acutely responding tissues, although no clear cell depletion occurred in the urothelium.

A Pharmacological in Vitro Study of the Mouse Urinary Bladder at the Time of Acute Change in Bladder Reservoir Function after Irradiation

Mouse urinary bladder strips were investigated as to whether the acute change in bladder reservoir function seen after irradiation might be due to major changes in basic nerve and smooth muscle functions. The release mechanism of acetylcholine, cholinergic and non-cholinergic nerve activation explored by indomethacin and potassium channel activation were investigated. It was concluded that the normal mouse bladder is partly cholinergically and partly non-cholinergically innervated. The role of acetylcholine is of the same importance as in other rodents. However, it was not possible to distinguish any difference between normal and irradiated mouse bladders in respect to nerve and smooth muscle function.

2-Aminofluorene-DNA adducts in mouse urinary bladder: effect of age, sex and acetylator phenotype

Formation of urinary bladder DNA-2-aminofluorene adducts in inbred and acetylator congenic mice was measured 3 h after a 60 mg/kg dose of the arylamine carcinogen. The sensitivity of 32P-postlabeling with HPLC analysis permitted quantitation of adducts in individual mouse bladders. Acetylator phenotype was a significant determinant of DNA damage in female mice as slow acetylators had higher levels of bladder DNA adducts than rapids. This correlation is the reverse of that seen with hepatic DNA. Age was also a significant determinant of DNA damage as older mice (20-23 weeks) formed more bladder DNA adducts than young (7 week) mice. The age-related increase in bladder adduct formation was seen in both sexes of all mouse lines. Male B6 mice exposed to 2-aminofluorene at 20-23 weeks of age showed a 26-fold higher level of bladder DNA adducts than males exposed at 7 weeks. In addition to the large increase in total adduct level, the older male B6 mice produced significant amounts of an unidentified, early-eluting adduct peak that had chromatographic properties similar to an aminofluorene-DNA adduct produced through peroxidative activation. These results indicate that age, sex and acetylator phenotype are all important determinants of aromatic amine-bladder DNA adduct formation in mice.

Multivesicular bodies in the transitional epithelium of the neonatal mouse urinary border

The origin of late endosomes — multivesicular bodies (MVBs) in the superficial cells of 16 and 17 embryonic old transitional epithelium of mouse urinary bladder was studied by electron microscopy, lectin labelling and HRP tracing. Analysis of hexagonally structured membrane particles, WGA, and RCA I binding sites revealed structural similarity between plasmalemma, fusiform vesicles and multivesicular bodies. Early endosomes are lined by symmetric unit membrane as well as by asymmetric thickened membrane regions. Multivesicular bodies and fusiform vesicles have asymmetric unit membranes. MVBs may be derived from primary endosomes as well as from fusiform vesicles in the cytoplasm.

Effects of various chemicals on the development of mouse urinary bladder tumors induced by N-butyl-N-(4-hydroxybutyl) nitrosamine.

Effects of various chemicals on the development of urinary bladder tumors in mouse treated with N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) were examined, Male C3H/He mice received 0.05% BBN in drinking water for 8 weeks, and were subsequently treated with basal diet containing trisodium nitrilotriacetate monohydrate (Na3NTA·H2O), nitrilotriacetic acid (H3NTA), sodium saccharin, and DL-tryptophan for 12 weeks. DL-tryptophan tended to induce a more significant increase in neoplastic and dysplastic lesions of the urinary bladder as compared to Na3NTA·H2O, H3NTA and sodium saccharin. These data showed that DL-tryptophan was more effective than Na3NTA·H2O, H3NTA and sodium saccharin with regard to promoting potential on the development of urinary bladder tumors in mouse treated with BBN.

Pharmacological analysis of agonist‐antagonist interactions at acetylcholine muscarinic receptors in a new urinary bladder assay

1. Agonist-antagonist interactions at acetylcholine (ACh) muscarinic receptors have been analysed by use of an improved urinary bladder assay, isolated and intact, from the mouse. With 5-methylfurmethide as agonist, validated cumulative concentration-effect curves were obtained in less than 7 min by re-dosing before the response plateaux began to fade. 2. The pKB value estimated for pirenzepine was 6.76. The pKB values estimated for atropine and N-methylatropine from data obtained at concentrations which produced dose-ratios greater than 20 and 60 were 8.90 and 9.58, respectively. 3. The deviation from simple competitive behaviour at low dose-ratios with atropine and N-methylatropine was consistent with the operation of saturable antagonist removal processes. The deviation observed with atropine was corrected by pre-incubation with methylbutyrate, an alternative substrate for 'atropine esterase'. 4. The simple competitive behaviour of N-methylatropine was restored following pre-incubation with the neuronal choline uptake blocker hemicholinium-3 (HC-3). However, the pKB estimated for N-methylatropine under these conditions was low. This latter result could be accounted for by the observed behaviour of HC-3 as a competitive antagonist of ACh muscarinic receptors (pKB = 4.01). 5. We conclude that the modified mouse urinary bladder assay is suitable for the quantitative analysis of muscarinic receptor interactions. In addition, we postulate the existence of a previously undescribed uptake mechanism for quaternary muscarinic receptor antagonists.