Bladder smooth muscle organ culture preparation maintains the contractile phenotype.
Smooth muscle cells, when subjected to culture, modulate from a contractile to a secretory phenotype. This has hampered the use of cell culture for molecular techniques to study the regulation of smooth muscle biology. The goal of this study was to develop a new organ culture model of bladder smooth muscle (BSM) that would maintain the contractile phenotype and aid in the study of BSM biology. Our results showed that strips of BSM subjected to up to 9 days of organ culture maintained their contractile phenotype, including the ability to achieve near-control levels of force with a temporal profile similar to that of noncultured tissues. The technical aspects of our organ culture preparation that were responsible, in part, for the maintenance of the contractile phenotype were a slight longitudinal stretch during culture and subjection of the strips to daily contraction-relaxation. The tissues contained viable cells throughout the cross section of the strips. There was an increase in extracellular collagenous matrix, resulting in a leftward shift in the passive length-tension relationship. There were no significant changes in the content of smooth muscle-specific α-actin, calponin, h-caldesmon, total myosin heavy chain, protein kinase G, Rho kinase-I, or the ratio of SM1 to SM2 myosin isoforms. Moreover the organ cultured tissues maintained functional voltage-gated calcium channels and large-conductance calcium-activated potassium channels. Therefore, we propose that this novel BSM organ culture model maintains the contractile phenotype and will be a valuable tool for the use in cellular/molecular biology studies of bladder myocytes.
Dept. of Pharmacology and Physiology, Drexel Univ. College of Medicine, 245 N 15th St., MS 488, Philadelphia, PA 19102, USA.
Tanchun
T
Wang
Derek M
DM
Kendig
Shaohua
S
Chang
Danielle M
DM
Trappanese
Samuel
S
Chacko
Robert S
RS
Moreland
D000199:Actins / Q000378:metabolism; D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D008297:Male; D008840:Microfilament Proteins / Q000378:metabolism; D023421:Models, Animal; D009119:Muscle Contraction / Q000502:physiology*; D009130:Muscle, Smooth / Q000502:physiology*; D018995:Myosin Heavy Chains / Q000378:metabolism; D009924:Organ Culture Techniques / Q000379:methods*; D010641:Phenotype; D011817:Rabbits; D001743:Urinary Bladder / Q000502:physiology*; D054460:rho-Associated Kinases / Q000378:metabolism; D000096985:Calponins
D000199:Actins; D002135:Calcium-Binding Proteins; D002148:Calmodulin-Binding Proteins; D017869:Cyclic GMP-Dependent Protein Kinases; D008840:Microfilament Proteins; D018995:Myosin Heavy Chains; D054460:rho-Associated Kinases; D000096985:Calponins