Identification and Characterization of Myosin from Rat Testicular Peritubular Myoid Cells

被引:12
作者
Fernandez, Dario
Bertoldi, Maria V.
Gomez, Laura
Morales, Alfonsina
Callegari, Eduardo [2 ]
Lopez, Luis A. [1 ]
机构
[1] Univ Nacl Cuyo, Fac Ciencias Med, Inst Histol & Embriol, Lab Cytoskeleton & Cell Cycle,Ctr Univ, RA-5500 Mendoza, Argentina
[2] Univ S Dakota, Sanford Sch Med, Div Basic Biomed Sci, Vermillion, SD 57069 USA
基金
美国国家卫生研究院;
关键词
male reproductive tract; myosin; peritubular myoid cells; seminiferous tubule; testis;
D O I
10.1095/biolreprod.107.066472
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
In the mammalian testis, peritubular myoid cells (PMCs) surround seminiferous tubules. These cells are contractile, express the cytoskeletal markers of true smooth muscle alpha- isoactin and F-actin -and participate in the contraction of seminiferous tubules during the transport of spermatozoa and testicular fluid to the rete testis. Myosin from PMCs (PMCmyosin) was isolated from adult rat testis and purified by cycles of assembly-disassembly and sucrose gradient centrifugation. PMC-myosin was recognized by a monoclonal anti-smooth muscle myosin antibody, and the peptide sequence shared partial homology with rat smooth muscle myosin-II, MYH11 (also known as SMM-II). Most PMC-myosin (95%) was soluble in the PMC cytosol, and purified PMC-myosin did not assemble into filaments in the in vitro salt dialysis assay at 48C, but did at 208C. PMC-myosin filaments are stable to ionic strength to the same degree as gizzard MYH11 filaments, but PMC-myosin filaments were more unstable in the presence of ATP. When PMCs were induced to contract by endothelin 1, a fraction of the PMC-myosin was found to be involved in the contraction. From these results we infer that PMCs express an isoform of smooth muscle myosin-II that is characterized by solubility at physiological ionic strength, a requirement for high temperature to assemble into filaments in vitro, and instability at low ATP concentrations. PMC-myosin is part of the PMC contraction apparatus when PMCs are stimulated with endothelin 1.
引用
收藏
页码:1210 / 1218
页数:9
相关论文
共 42 条
[31]  
SHIRINSKY VP, 1993, J BIOL CHEM, V268, P16578
[32]   CLONING OF THE CDNA-ENCODING THE MYOSIN HEAVY-CHAIN OF A VERTEBRATE CELLULAR MYOSIN - (DNA-SEQUENCE COMPARATIVE AMINO-ACID SEQUENCES MESSENGER-RNA EXPRESSION) [J].
SHOHET, RV ;
CONTI, MA ;
KAWAMOTO, S ;
PRESTON, YA ;
BRILL, DA ;
ADELSTEIN, RS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (20) :7726-7730
[33]   Sites of interaction between kinase-related protein and smooth muscle myosin [J].
Silver, DL ;
Vorotnikov, AV ;
Watterson, DM ;
Shirinsky, VP ;
Sellers, JR .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (40) :25353-25359
[34]   MYOSIN-FILAMENTS HAVE NON-PHOSPHORYLATED LIGHT-CHAINS IN RELAXED SMOOTH-MUSCLE [J].
SOMLYO, AV ;
BUTLER, TM ;
BOND, M ;
SOMLYO, AP .
NATURE, 1981, 294 (5841) :567-569
[35]   STRUCTURE AND FUNCTION OF CHICKEN GIZZARD MYOSIN [J].
SUZUKI, H ;
ONISHI, H ;
TAKAHASHI, K ;
WATANABE, S .
JOURNAL OF BIOCHEMISTRY, 1978, 84 (06) :1529-1542
[36]   Direct visualization of rat peritubular myoid cell contraction in response to endothelin [J].
Tripiciano, A ;
Filippini, A ;
Giustiniani, Q ;
Palombi, F .
BIOLOGY OF REPRODUCTION, 1996, 55 (01) :25-31
[37]   Assembly of cytoplasmic and smooth muscle myosins [J].
Trybus, Kathleen M. .
CURRENT OPINION IN CELL BIOLOGY, 1991, 3 (01) :105-111
[38]  
TRYBUS KM, 1984, J BIOL CHEM, V259, P8564
[39]   CHARACTERIZATION OF RAT TESTICULAR PERITUBULAR MYOID CELLS IN CULTURE - ALPHA-SMOOTH MUSCLE ISOACTIN IS A SPECIFIC DIFFERENTIATION MARKER [J].
TUNG, PS ;
FRITZ, IB .
BIOLOGY OF REPRODUCTION, 1990, 42 (02) :351-365
[40]   MYOSIN-II FILAMENT ASSEMBLIES IN THE ACTIVE LAMELLA OF FIBROBLASTS - THEIR MORPHOGENESIS AND ROLE IN THE FORMATION OF ACTIN FILAMENT BUNDLES [J].
VERKHOVSKY, AB ;
SVITKINA, TM ;
BORISY, GG .
JOURNAL OF CELL BIOLOGY, 1995, 131 (04) :989-1002