Muscarinic Receptor Transcript and Protein Density in Hypertrophied and Atrophied Rat Urinary Bladder

Overview

Journal Neurourol Urodyn

Date 2005 Nov 24

PMID 16304675

Citations 4

Authors

Alan S Braverman

Michael R Ruggieri Sr

Affiliations

Soon will be listed here.

Abstract

Aims: Our previous studies showed that bladder hypertrophy shifts the muscarinic receptor subtype mediating contraction from M(3) towards M(2) along with increased M(2) and decreased M(3) protein concentration. We quantified mRNA for M(1) through M(5) receptors to determine whether the changes in M(2) and M(3) protein levels was due to changes in transcription.

Methods: Bladder hypertrophy was induced by bladder outlet obstruction (BOO), major pelvic ganglion electrocautery (DEN), and major pelvic ganglion decentralization (DEC). Bladder atrophy was induced by ureteral diversion (DIV). Additional groups included denervated and diverted (DEN-DIV), sham operated (SHAM), and normal (NOR) controls. Transcripts were quantified using a multiplex ribonuclease protection assay (RPA) and receptor protein density was determined by immunoprecipitation. Receptor transcripts were expressed per unit total RNA.

Results: Although all five receptor subtype transcripts were detected in all experimental groups, the densities of M(1), M(4), and M(5) were much lower than for the M(2) and M(3) subtype. There were more M(2) receptor transcripts than all the others, consistent with M(2) protein determinations. M(2) transcripts were significantly increased in DEN and BOO bladders. Surprisingly, M(3) transcripts were also significantly increased in BOO. There was a significant correlation (r=0.98, P<0.001) between protein density and transcript density for the M(2) but not the M(3) receptor among the different experimental groups.

Conclusions: Changes in mRNA concentration are reflected by changes in protein density for the M(2) receptor but not for the M(3) receptor. Extrapolation of functional effects from transcript density data is invalid for M(3) mediated bladder contractions.

Citing Articles

Muscarinic receptor subtype mRNA expression in the human prostate: association with age, pathological diagnosis, prostate size, or potentially interfering medications?.

Witte L, Teitsma C, de la Rosette J, Michel M Naunyn Schmiedebergs Arch Pharmacol. 2013; 387(2):207-14.

PMID: 24193354 DOI: 10.1007/s00210-013-0934-4.

Blebbistain, a myosin II inhibitor, as a novel strategy to regulate detrusor contractility in a rat model of partial bladder outlet obstruction.

Zhang X, Seftel A, DiSanto M PLoS One. 2011; 6(10):e25958.

PMID: 22016792 PMC: 3189232. DOI: 10.1371/journal.pone.0025958.

Temporal morphological and functional impact of complete urinary diversion on the bladder: a model of bladder disuse in rats.

Liu G, Lin Y, Li M, Xiao N, Daneshgari F J Urol. 2010; 184(5):2179-85.

PMID: 20850838 PMC: 4113019. DOI: 10.1016/j.juro.2010.06.090.

Effects of bladder outlet obstruction on properties of Ca2+-activated K+ channels in rat bladder.

Kita M, Yunoki T, Takimoto K, Miyazato M, Kita K, de Groat W Am J Physiol Regul Integr Comp Physiol. 2010; 298(5):R1310-9.

PMID: 20200132 PMC: 2867513. DOI: 10.1152/ajpregu.00523.2009.

References

Caulfield M . Muscarinic receptors--characterization, coupling and function. Pharmacol Ther. 1993; 58(3):319-79. DOI: 10.1016/0163-7258(93)90027-b. View

Thellin O, Zorzi W, Lakaye B, de Borman B, Coumans B, Hennen G . Housekeeping genes as internal standards: use and limits. J Biotechnol. 2000; 75(2-3):291-5. DOI: 10.1016/s0168-1656(99)00163-7. View

Revillion F, Pawlowski V, Hornez L, Peyrat J . Glyceraldehyde-3-phosphate dehydrogenase gene expression in human breast cancer. Eur J Cancer. 2000; 36(8):1038-42. DOI: 10.1016/s0959-8049(00)00051-4. View

Sienna N, Larson D, SELLS B . Dexamethasone stimulates ribosomal protein L32 gene transcription in rat myoblasts. Mol Cell Endocrinol. 2000; 167(1-2):127-37. DOI: 10.1016/s0303-7207(00)00282-3. View

Levin R, HAUGAARD N, OConnor L, Buttyan R, Das A, Dixon J . Obstructive response of human bladder to BPH vs. rabbit bladder response to partial outlet obstruction: a direct comparison. Neurourol Urodyn. 2000; 19(5):609-29. DOI: 10.1002/1520-6777(2000)19:5<609::aid-nau7>3.0.co;2-h. View

Braverman A, Tallarida R, Ruggieri Sr M . Interaction between muscarinic receptor subtype signal transduction pathways mediating bladder contraction. Am J Physiol Regul Integr Comp Physiol. 2002; 283(3):R663-8. PMC: 3277793. DOI: 10.1152/ajpregu.00116.2002. View

Glare E, Divjak M, Bailey M, Walters E . beta-Actin and GAPDH housekeeping gene expression in asthmatic airways is variable and not suitable for normalising mRNA levels. Thorax. 2002; 57(9):765-70. PMC: 1746418. DOI: 10.1136/thorax.57.9.765. View

Braverman A, Ruggieri Sr M . Hypertrophy changes the muscarinic receptor subtype mediating bladder contraction from M3 toward M2. Am J Physiol Regul Integr Comp Physiol. 2003; 285(3):R701-8. PMC: 3277086. DOI: 10.1152/ajpregu.00009.2003. View

Chess-Williams R . Muscarinic receptors of the urinary bladder: detrusor, urothelial and prejunctional. Auton Autacoid Pharmacol. 2002; 22(3):133-45. DOI: 10.1046/j.1474-8673.2002.00258.x. View

10.

Sigala S, Mirabella G, Peroni A, Pezzotti G, Simeone C, Spano P . Differential gene expression of cholinergic muscarinic receptor subtypes in male and female normal human urinary bladder. Urology. 2002; 60(4):719-25. DOI: 10.1016/s0090-4295(02)01819-8. View

11.

Pontari M, Braverman A, Ruggieri Sr M . The M2 muscarinic receptor mediates in vitro bladder contractions from patients with neurogenic bladder dysfunction. Am J Physiol Regul Integr Comp Physiol. 2004; 286(5):R874-80. PMC: 3274770. DOI: 10.1152/ajpregu.00391.2003. View

12.

Bas A, Forsberg G, Hammarstrom S, Hammarstrom M . Utility of the housekeeping genes 18S rRNA, beta-actin and glyceraldehyde-3-phosphate-dehydrogenase for normalization in real-time quantitative reverse transcriptase-polymerase chain reaction analysis of gene expression in human T lymphocytes. Scand J Immunol. 2004; 59(6):566-73. DOI: 10.1111/j.0300-9475.2004.01440.x. View

13.

Malkowicz S, Wein A, Elbadawi A, Van Arsdalen K, Ruggieri M, Levin R . Acute biochemical and functional alterations in the partially obstructed rabbit urinary bladder. J Urol. 1986; 136(6):1324-9. DOI: 10.1016/s0022-5347(17)45331-6. View

14.

Sirover M . Role of the glycolytic protein, glyceraldehyde-3-phosphate dehydrogenase, in normal cell function and in cell pathology. J Cell Biochem. 1997; 66(2):133-40. View

15.

Sohn U, Harnett K, Cao W, Rich H, Kim N, Behar J . Acute experimental esophagitis activates a second signal transduction pathway in cat smooth muscle from the lower esophageal sphincter. J Pharmacol Exp Ther. 1998; 283(3):1293-304. View

16.

Shinohara M, Loros J, Dunlap J . Glyceraldehyde-3-phosphate dehydrogenase is regulated on a daily basis by the circadian clock. J Biol Chem. 1998; 273(1):446-52. DOI: 10.1074/jbc.273.1.446. View

17.

Braverman A, Kohn I, Luthin G, Ruggieri M . Prejunctional M1 facilitory and M2 inhibitory muscarinic receptors mediate rat bladder contractility. Am J Physiol. 1998; 274(2):R517-23. PMC: 3275803. DOI: 10.1152/ajpregu.1998.274.2.R517. View

18.

Zou K, Ing N . Oestradiol up-regulates oestrogen receptor, cyclophilin, and glyceraldehyde phosphate dehydrogenase mRNA concentrations in endometrium, but down-regulates them in liver. J Steroid Biochem Mol Biol. 1998; 64(5-6):231-7. DOI: 10.1016/s0960-0760(97)00194-5. View

19.

Braverman A, Luthin G, Ruggieri M . M2 muscarinic receptor contributes to contraction of the denervated rat urinary bladder. Am J Physiol. 1998; 275(5):R1654-60. PMC: 3275805. DOI: 10.1152/ajpregu.1998.275.5.R1654. View

20.

Kim J, Kim S, Han S, Paik S, Hur S, Kim Y . Increased glyceraldehyde-3-phosphate dehydrogenase gene expression in human cervical cancers. Gynecol Oncol. 1998; 71(2):266-9. DOI: 10.1006/gyno.1998.5195. View