Cytoskeletal and cytocontractile protein composition of smooth muscle cells in developing and obstructed rabbit bladder.
The differentiation patterns of smooth muscle cells (SMC) in rabbit bladder during development and in the hypertrophic response to partial outflow obstruction induced in adult animals were evaluated by biochemical and immunochemical techniques and by using a panel of monoclonal antibodies specific for desmin, vimentin, alpha-actin of smooth muscle (SM) type, SM myosin, and nonmuscle (NM) myosin isoforms. Desmin and SM alpha-actin were homogeneously distributed in SMC of developing, adult, and obstructed bladders. Conversely, marked changes in the ratio and antigenicity of SM myosin isoforms were observed by SDS electrophoresis and Western blotting, respectively. In particular, the 205 K (SM1) isoform was down-regulated with development whereas the 200 K (SM2) isoform was up-regulated around 7 days after birth and down-regulated in the obstructed bladder. Vimentin was expressed in SMC of the fetal bladder and declined markedly during postnatal, physiological hypertrophy of SMC, which occurs concomitantly with diminution of DNA synthesis. This polypeptide became detectable, however, in SMC of obstructed bladders. The 196 K (NM) myosin isoform recognized by NM-A9 antibody, present only in endothelium of blood vessels and in mucosa of normal fetal and adult bladders, became expressed in detrusor muscle, when SMC underwent a process of pathological hypertrophy. The reexpression of vimentin and the de novo appearance of NM myosin isoform in hypertrophic bladders can be reversed when the tissue mass is reduced, such as in bladders after 1-month recovery from partial obstruction. Thus, a specific NM myosin isoform can be used as a marker of SMC hypertrophy in obstructed bladder. In addition, the combined use of anti-vimentin and NM-A9 antibodies can distinguish between SMC which are in the physiological or in the pathological condition of adaptive bladder hypertrophy.
Chiavegato
A
A
Department of Biomedical Sciences, University of Padua, Italy.
Scatena
M
M
Roelofs
M
M
Ferrarese
P
P
Pauletto
P
P
Passerini-Glazel
G
G
Pagano
F
F
Sartore
S
S
D000818:Animals; D015153:Blotting, Western; D003285:Contractile Proteins / Q000032:analysis* / Q000737:chemistry / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000032:analysis* / Q000737:chemistry / Q000648:ultrastructure; D003893:Desmin / Q000032:analysis / Q000737:chemistry / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006984:Hypertrophy / Q000473:pathology; D007150:Immunohistochemistry; D007536:Isomerism; D008297:Male; D008854:Microscopy, Electron; D009130:Muscle, Smooth / Q000737:chemistry* / Q000166:cytology* / Q000648:ultrastructure; D009218:Myosins / Q000032:analysis / Q000737:chemistry / Q000648:ultrastructure; D011817:Rabbits; D001743:Urinary Bladder / Q000166:cytology / Q000196:embryology / Q000473:pathology; D001748:Urinary Bladder Neck Obstruction / Q000473:pathology*; D014746:Vimentin / Q000032:analysis / Q000737:chemistry / Q000648:ultrastructure
D003285:Contractile Proteins; D003598:Cytoskeletal Proteins; D003893:Desmin; D014746:Vimentin; D009218:Myosins