scholarly journals Serotonergic regulation of distention-induced ATP release from the urothelium

2016 ◽  
Vol 310 (7) ◽  
pp. F646-F655 ◽  
Author(s):  
Kazumasa Matsumoto-Miyai ◽  
Erika Yamada ◽  
Eriko Shinzawa ◽  
Yoshihisa Koyama ◽  
Shoichi Shimada ◽  
...  
Keyword(s):  
Urinary Bladder ◽  
Tryptophan Hydroxylase ◽  
Atp Release ◽  
Pcr Amplification ◽  
Inhibitory Effect ◽  
Visceral Sensation ◽  
Rt Pcr ◽  
Specific Expression ◽  
Bladder Distention ◽  
Gr 127935

Serotonin [5-hydroxytryptamine (5-HT)] is involved in both motor and sensory functions in hollow organs, especially in the gastrointestinal tract. However, the involvement of 5-HT in visceral sensation of the urinary bladder remains unknown. Because distention-induced ATP release from the urothelium plays an essential role in visceral sensation of the urinary bladder, we investigated the regulation of urothelial ATP release by the 5-HT signaling system. RT-PCR and immunohistochemical analyses of the urothelium revealed specific expression of 5-HT1D and 5-HT4 receptors. The addition of 5-HT did not affect urothelial ATP release without bladder distention, but it significantly reduced distention-induced ATP release by physiological pressure during urine storage (5 cmH2O). The inhibitory effect of 5-HT on distention-elicited ATP release was blocked by preincubation with the 5-HT1B/1D antagonist GR-127935 but not by the 5-HT4 antagonist SB-204070. mRNA encoding tryptophan hydroxylase 1 was detected in the urinary bladder by nested RT-PCR amplification, and l-tryptophan or the selective serotonin reuptake inhibitor citalopram also inhibited ATP release, indicating that 5-HT is endogenously synthesized and released in the urinary bladder. The addition of GR-127935 significantly enhanced the distention-elicited ATP release 40 min after distention, whereas SB-204070 reduced the amount of ATP release 20 min after distention. These data suggest that 5-HT4 facilitates the distention-induced ATP release at an earlier stage, whereas 5-HT1D inhibits ATP release at a later stage. The net inhibitory effect of 5-HT indicates that the action of 5-HT on the urothelium is mediated predominantly by 5-HT1D.

Expression of the ammonia transporter proteins Rh B glycoprotein and Rh C glycoprotein in the intestinal tract

2005 ◽  
Vol 288 (5) ◽  
pp. G1036-G1047 ◽  
Author(s):  
Mary E. Handlogten ◽  
Seong-Pyo Hong ◽  
Li Zhang ◽  
Allen W. Vander ◽  
Marshall L. Steinbaum ◽  
...  
Keyword(s):  
Gastrointestinal Tract ◽  
Epithelial Cells ◽  
Intestinal Tract ◽  
Pcr Amplification ◽  
Immunoblot Analysis ◽  
Rt Pcr ◽  
Specific Expression ◽  
Ammonia Metabolism ◽  
Transporter Family ◽  
Ammonia Transport

Ammonia metabolism is important in multiple aspects of gastrointestinal physiology, but the mechanisms of ammonia transport in the gastrointestinal tract remain incompletely defined. The present study examines expression of the ammonia transporter family members Rh B glycoprotein (RhBG) and Rh C glycoprotein (RhCG) in the mouse gastrointestinal tract. Real-time RT-PCR amplification and immunoblot analysis identified mRNA and protein for both RhBG and RhCG were expressed in stomach, duodenum, jejunum, ileum, and colon. Immunohistochemistry showed organ and cell-specific expression of both RhBG and RhCG. In the stomach, both RhBG and RhCG were expressed in the fundus and forestomach, but not in the antrum. In the forestomach, RhBG was expressed by all nucleated squamous epithelial cells, whereas RhCG was expressed only in the stratum germinativum. In the fundus, RhBG and RhCG immunoreactivity was present in zymogenic cells but not in parietal or mucous cells. Furthermore, zymogenic cell RhBG and RhCG expression was polarized, with apical RhCG and basolateral RhBG immunoreactivity. In the duodenum, jejunum, ileum, and colon, RhBG and RhCG immunoreactivity was present in villous, but not in mucous or crypt cells. Similar to the fundic zymogenic cell, RhBG and RhCG expression in villous epithelial cells was polarized when apical RhCG and basolateral RhBG immunoreactivity was present. Thus the ammonia transporting proteins RhBG and RhCG exhibit cell-specific, axially heterogeneous, and polarized expression in the intestinal tract suggesting they function cooperatively to mediate gastrointestinal tract ammonia transport.

scholarly journals Urothelium-Specific Deletion of Connexin43 in the Mouse Urinary Bladder Alters Distension-Induced ATP Release and Voiding Behavior

2021 ◽  
Vol 22 (4) ◽  
pp. 1594
Author(s):  
Jin Kono ◽  
Masakatsu Ueda ◽  
Atsushi Sengiku ◽  
Sylvia O. Suadicani ◽  
Osamu Ogawa ◽  
...  
Keyword(s):  
Urinary Bladder ◽  
Atp Release ◽  
Active Phase ◽  
Bladder Capacity ◽  
Wild Type ◽  
Sleep Phase ◽  
Cx43 Expression ◽  
Bladder Distention ◽  
Atp Signaling ◽  
Specific Deletion

Connexin43 (Cx43), the main gap junction and hemichannel forming protein in the urinary bladder, participates in the regulation of bladder motor and sensory functions and has been reported as an important modulator of day–night variations in functional bladder capacity. However, because Cx43 is expressed throughout the bladder, the actual role played by the detrusor and the urothelial Cx43 is still unknown. For this purpose, we generated urothelium-specific Cx43 knockout (uCx43KO) mice using Cre-LoxP system. We evaluated the day–night micturition pattern and the urothelial Cx43 hemichannel function of the uCx43KO mice by measuring luminal ATP release after bladder distention. In wild-type (WT) mice, distention-induced ATP release was elevated, and functional bladder capacity was decreased in the animals’ active phase (nighttime) when Cx43 expression was also high compared to levels measured in the sleep phase (daytime). These day–night differences in urothelial ATP release and functional bladder capacity were attenuated in uCx43KO mice that, in the active phase, displayed lower ATP release and higher functional bladder capacity than WT mice. These findings indicate that urothelial Cx43 mediated ATP signaling and coordination of urothelial activity are essential for proper perception and regulation of responses to bladder distension in the animals’ awake, active phase.

111 CLONING AND CHARACTERIZATION OF PIG VASA HOMOLOG GENE AND ITS SPECIFIC EXPRESSION IN GERM CELL LINEAGE

2005 ◽  
Vol 17 (2) ◽  
pp. 206
Author(s):  
G.S. Lee ◽  
S.H. Lee ◽  
H.S. Kim ◽  
E.B. Jeung ◽  
S.K. Kang ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Germ Line ◽  
Pcr Amplification ◽  
Molecular Probes ◽  
Cloning And Expression ◽  
Soil Nematode ◽  
Rt Pcr ◽  
Specific Expression ◽  
Cell Commitment

All of the vasa homologue genes in C. elegans (Caenorhabditis elegens, a free-living soil nematode), xenopus, zebrafish, mouse, human, chicken, trout, and rat exhibited a germ line-specific expression and are used as specific molecular probes to distinguish the developmental profile of germ cells. In order to determine a useful marker for the research of germ cell commitment and development in pigs, we investigated the cloning and expression profile of porcine vasa homolog gene (Pvh). A Pvh cDNA gene of size 2172 bps (submitted to NCBI gene Bank No. AY626785) was cloned from pig ovary by reverse transcription-polymerase chain reaction (RT-PCR) amplification. The amplification was repeated three times and each RT-PCR product was sequenced. The isolated cDNA had 724 deduced amino acids with significant homology to mouse (85%) or human (91%) vasa. The Pvh sequence presents five copies of the RGG motifs and the DEAD box. By RT-PCR amplification, the expression of Pvh mRNA was restricted to the ovary and testis and was undetectable in somatic tissues including brain, whole blood, heart, lung, kidney, spleen, intestine, and liver. When analyzed by RT-PCR amplification, during pre-implantation embryo development, Pvh was transcribed in oocytes and fertilized 2-cell embryos (no difference in the expression levels between oocytes and fertilized 2-cell embryos), but not in 4-cell, 8-cell, morula and blastocyst stages. Using mouse vasa antibody (kindly donated from Dr. Noce, Japan; tested in porcine cells with porcine oocytes and mouse oocytes as positive control and with porcine brain cells as negative control), immunohistochemical analysis of fetal (Day 100) and adult gonad sections revealed that Pvh protein was specifically expressed in proliferating primordial germ cells (PGC), oocytes and spermatocytes. Interestingly, Pvh protein was not expressed in embryonic germ cells, but it was strongly expressed in freshly isolated PGC. Our results indicate that Pvh gene is specifically transcribed in pig germ cells. This study was supported by grants from the Korean Ministry of Science and Technology (Biodiscovery) and the Biogreen 21-1000520030100000.

In vitro screen of inhibitors of rat brain serotonin synthesis

1969 ◽  
Vol 47 (5) ◽  
pp. 501-506 ◽  
Author(s):  
E. G. McGeer ◽  
D. A. V. Peters
Keyword(s):  
Rat Brain ◽  
Tryptophan Hydroxylase ◽  
Inhibitory Effect ◽  
Complexing Agents ◽  
Serotonin Synthesis ◽  
Structural Requirements ◽  
Class A ◽  
Numerous Data ◽  
Comparison Of The Results

Over 700 compounds were screened at 10−4 M concentration as inhibitors of the conversion of L-tryptophan-14C to serotonin-14C in crude rat brain homogenates. Most of the compounds had little or no inhibitory effect. Those with strong inhibitory properties were tested as inhibitors of 5-hydroxytryptophan decarboxylase and, if active on the decarboxylase, were assayed as tryptophan hydroxylase inhibitors. Except for a few oxidizing and complexing agents and for some substituted p-phenylenediamines, the compounds found to inhibit tryptophan hydroxylase by >50% belonged to the three types of inhibitors already known, i.e. catechols, phenylalanine and ring-substituted phenylalanines, and 6-substituted tryptophans. The numerous data in this screen make possible some comments as to the structural requirements for activity within each class. A comparison of the results on tryptophan hydroxylase with data on tyrosine hydroxylase inhibition in similar homogenates makes it clear that two separate, if somewhat similar, enzymes are involved.

Pharmacologic responses of the mouse urinary bladder

Open Medicine ◽  
2009 ◽  
Vol 4 (2) ◽  
pp. 192-197 ◽  
Author(s):  
A. Canda ◽  
Christopher Chapple ◽  
Russ Chess-Williams
Keyword(s):  
Nitric Oxide ◽  
Urinary Bladder ◽  
Electrical Field Stimulation ◽  
Inhibitory Effect ◽  
Minor Role ◽  
Detrusor Muscle ◽  
Field Stimulation ◽  
Muscarinic Agonists ◽  
Electrical Field ◽  
M3 Receptor

AbstractThe aim of the study was to determine pathways involved in contraction and relaxation of the mouse urinary bladder. Mouse bladder strips were set up in gassed Krebs-bicarbonate solution and responses to various drugs and electrical field stimulation were obtained. Isoprenaline (b-receptor agonist) caused a 63% inhibition of carbachol precontracted detrusor (EC50=2nM). Carbachol caused contraction (EC50=0.3µM), responses were antagonised more potently by 4-DAMP (M3-antagonist) than methoctramine (M2-antagonist). Electrical field stimulation caused contraction, which was inhibited by atropine (60%) and less by guanethidine and α,β-methylene-ATP. The neurogenic responses were not potentiated by inhibition of nitric oxide synthase. Presence of an intact urothelium significantly depressed responses to carbachol (p=0.02) and addition of indomethacin and L-NNA to remove prostaglandin and nitric oxide production respectively did not prevent the inhibitory effect of the urothelium. In conclusion, b-receptor agonists cause relaxation and muscarinic agonists cause contraction via the M3-receptor. Acetylcholine is the main neurotransmitter causing contraction while nitric oxide has a minor role. The mouse and human urothelium are similar in releasing a factor that inhibits contraction of the detrusor muscle which is unidentified but is not nitric oxide or a prostaglandin. Therefore, the mouse may be used as a model to study the lower urinary tract.

scholarly journals Identification and Differentiation of the Twenty Six Bluetongue Virus Serotypes by RT–PCR Amplification of the Serotype-Specific Genome Segment 2

PLoS ONE ◽  
2012 ◽  
Vol 7 (2) ◽  
pp. e32601 ◽  
Author(s):  
Narender S. Maan ◽  
Sushila Maan ◽  
Manjunatha N. Belaganahalli ◽  
Eileen N. Ostlund ◽  
Donna J. Johnson ◽  
...  
Keyword(s):  
Bluetongue Virus ◽  
Pcr Amplification ◽  
Genome Segment ◽  
Rt Pcr
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