Proteins of the membrane skeleton in rat Sertoli cells

1986 ◽  
Vol 86 (1) ◽  
pp. 145-154
Author(s):  
E. Ziparo ◽  
B.M. Zani ◽  
A. Filippini ◽  
M. Stefanini ◽  
V.T. Marchesi
Keyword(s):  
Plasma Membrane ◽  
Sertoli Cell ◽  
Sertoli Cells ◽  
Primary Cultures ◽  
Membrane Skeleton ◽  
Permeabilized Cells ◽  
Protein 4.1 ◽  
Molecular Weights ◽  
Gamma Subunits ◽  
Alpha Beta

Analogues of the alpha, beta and gamma subunits of human spectrin and erythroid protein 4.1 have been detected in rat Sertoli cell primary cultures. Immunofluorescence of permeabilized cells showed that erythroid type spectrin, protein 4.1 and actin co-distribute within the cells as filamentous structures. Fodrin-like molecules were distributed in a diffuse manner, mostly associated with the plasma membrane. Immunoprecipitation and immunoblotting experiments indicated that the polypeptides present in rat Sertoli cells are immunologically related and display molecular weights similar to their analogues in the human erythroid and non-erythroid membrane skeleton.

scholarly journals Agorins: major structural proteins of the plasma membrane skeleton of P815 tumor cells.

1986 ◽  
Vol 103 (2) ◽  
pp. 351-360 ◽  
Author(s):  
J R Apgar ◽  
M F Mescher
Keyword(s):  
Plasma Membrane ◽  
Plasma Membranes ◽  
Matrix Protein ◽  
Membrane Skeleton ◽  
Insoluble Fraction ◽  
Molecular Weights ◽  
Cell Plasma ◽  
The Matrix ◽  
Section Electron ◽  
Mastocytoma Cells

Plasma membranes of P815 mastocytoma cells contain a set of proteins that remain selectively insoluble upon extraction of the membranes with Triton X-100, and appear to form a membrane skeletal matrix independent of the filamentous cytoskeletal systems. EGTA treatment of the matrix was found to release approximately 25% of the protein as polypeptides of 70, 69, 38, and 36 kD, all of which appear to be peripheral components associated with the cytoplasmic face of the plasma membrane via divalent cation-dependent interactions. About 75% of the total matrix protein was recovered in the EGTA-insoluble fraction. Actin accounted for approximately 5% of the total protein in the EGTA-insoluble fraction. The rest was accounted for by two novel proteins of 20 and 40 kD which, despite their relatively low molecular weights, do not enter SDS PAGE gels. Together these proteins account for approximately 15% of the total plasma membrane protein, and are thus present in much higher amounts than any other characterized protein of nucleated cell plasma membranes. Based on the extensive associations of these proteins to form very large detergent-insoluble structures, we propose that they may be named agorin I, the 20-kD protein, and agorin II, the 40-kD protein, from the Greek agora meaning assembly. The amount and properties of these proteins and the appearance of the EGTA-insoluble material in thin-section electron micrographs indicate that the agorins are the major structural elements of the membrane matrix, and thus of the putative membrane skeleton.

scholarly journals Establishment of plasma membrane domains in hepatocytes. I. Characterization and localization to the bile canaliculus of three antigens externally oriented in the plasma membrane.

1983 ◽  
Vol 97 (6) ◽  
pp. 1823-1833 ◽  
Author(s):  
J Cook ◽  
E Hou ◽  
Y Hou ◽  
A Cairo ◽  
D Doyle
Keyword(s):  
Plasma Membrane ◽  
Membrane Fraction ◽  
Gradient Centrifugation ◽  
Primary Cultures ◽  
Limited Proteolysis ◽  
Bile Canaliculus ◽  
Molecular Weights ◽  
Crossed Immunoelectrophoresis ◽  
Membrane Antigens ◽  
Sinusoidal Surface

A membrane fraction denoted N2 upper was isolated from homogenates of rat liver by sucrose gradient centrifugation. This fraction, which was enriched 65-fold over the homogenate in 5'-nucleotidase activity, was used as an immunogen in goats. The antisera obtained contained antibodies to three predominant polypeptides in the N2 upper membrane fraction, as shown by crossed immunoelectrophoresis. These polypeptides had molecular weights of 105,000, 110,000, and 160,000 after recovery from the crossed immunoelectrophoretic gels and are denoted PM105, PM110, and PM160. Each was a distinct polypeptide, as shown by the distinct peptide patterns resulting from limited proteolysis in the presence of detergents. The three polypeptides were synthesized by primary cultures of hepatocytes and were externally oriented at the surface of these cells, as shown by their accessibility in situ to iodination catalyzed by lactoperoxidase. They were not detectable in the serum by crossed immunoelectrophoresis. The three antigens were present at very low (PM110) or nondetectable (PM105, PM160) concentrations in intracellular membrane fractions derived from the Golgi and smooth and rough endoplasmic reticulum of liver. The antigens also were reduced in concentration in a plasma membrane fraction most likely derived from the sinusoidal surface of the hepatocyte. The three membrane antigens bind to concanavalin A; hence, they are probably glycoprotein constituents of a discrete domain of the hepatocyte plasma membrane. Immune complexes were isolated after crossed immunoelectrophoresis and injected into rabbits. Each of the antisera obtained was reactive to one of the membrane polypeptides. Sections of fixed rat livers were reacted with each of the antibodies and then the primary antibody was localized by indirect immunocytochemical methods using horseradish peroxidase or colloidal gold as labels. Each of the three antigens was localized by this method to the bile canalicular domain of the hepatocyte plasma membrane.

scholarly journals Multiple Promoter Elements Contribute to Activity of the Follicle-Stimulating Hormone Receptor (FSHR) Gene in Testicular Sertoli Cells

1998 ◽  
Vol 12 (10) ◽  
pp. 1499-1512 ◽  
Author(s):  
Leslie L. Heckert ◽  
Melissa A. F. Daggett ◽  
Jiangkai Chen
Keyword(s):  
Sertoli Cell ◽  
Cell Lines ◽  
Sertoli Cells ◽  
Primary Cultures ◽  
Cell Types ◽  
Mobility Shift ◽  
Promoter Elements ◽  
Fshr Gene ◽  
E Box ◽  
Sequence Requirements

Abstract The FSH receptor (FSHR) is expressed only in granulosa cells of the ovary and Sertoli cells of the testis. This highly specific pattern of gene expression asserts that transcriptional events unique to these two cell types are responsible for activation of the FSHR gene. We have characterized the promoter elements required for activity of the rat FSHR gene in a Sertoli cell line MSC-1, primary cultures of rat Sertoli cells, and two non-Sertoli cell lines. Transient transfection analysis of deletion and block replacement mutants identified several elements, both 5′ and 3′ to the transcriptional start sites, that are essential for full promoter activity in Sertoli cells. These studies confirmed the use of an important E box element (CACGTG), which had the single greatest impact on promoter function. Bases within the core CACGTG of the E box, as well as flanking sequences, were shown to be essential for its function. Electrophoretic mobility shift assays identified both upstream stimulatory factor 1 (USF1) and USF2 as primary components of the complexes binding the E box. Sequence requirements for USF binding in vitro modestly diverged from the sequence requirements for in vivo function of the element. Comparison of the E box binding proteins in different cell types revealed that similar proteins bind the E box in Sertoli and non-Sertoli cell lines. Extracts from primary cultures of rat and mouse Sertoli cells have a second E box-binding complex that cross-reacts with USF antibodies that is not present in the cell lines.

Ultrastructual studies on spematids and sertoli cells during early spermogenesis in the bandicoot Peramelea nasuta geoffroy (Marsupialia)

1967 ◽  
Vol 15 (5) ◽  
pp. 881 ◽  
Author(s):  
CS Sapsford ◽  
CA Rae ◽  
KW Cleland
Keyword(s):  
Plasma Membrane ◽  
Nuclear Envelope ◽  
Sertoli Cell ◽  
Sertoli Cells ◽  
Close Contact ◽  
Cell Cytoplasm ◽  
Axial Filament ◽  
Stage Of Development ◽  
The Future ◽  
Nuclear Protrusion

The present paper deals with spermiogenesis up to and including the attachment of spermatids to Sertoli cells. The first observed step in spermatid differentiation was the development of the anlage of the middle piece and principal piece. This anlage, called the axial filament complex, has the structure of a cilium and arises from the future longitudinal centriole, while the latter, together with the future transverse centriole, lies in the vicinity of the Golgi complex. The definitive acrosomal vacuole, which ultimately becomes attached to and invaginates the nuclear envelope, is formed by the enlargement and coalescence of Golgi vacuoles. While this definitive vacuole is developing, the centrioles and attached axial filament complex migrate to the opposite pole of the nucleus. Before and during migration a number of accessory structures are developed in association with the centrioles, and one of these structures, the proximal junctional body, invaginates the nuclear envelope when the centrioles reach their definitive abacrosomal position. During this period, a cytoplasmic canal forms around the intraspermatid part of the axial filament complex. The definitive acrosomal vacuole ultimately extends out to make close contact with the plasma membrane of the spermatid. This stage of development is followed by a process of nuclear protrusion, initiated by the migration of the nucleus towards the region of contact between acrosomal vacuole and spermatid plasma membrane. During the migratory phase, that part of the nuclear envelope previously invaginated by the acrosomal vacuole becomes everted and the latter collapses, finally becoming sandwiched in between the nucleus and the plasma membrane of the spermatid. The nucleus subsequently projects from the surface of the spermatid, its acrosome-covered apex becoming coneshaped. During these phases of development the accessory structures elaborated in association with the centrioles, and which now lie in the neck region of the spermatid, have become more highly organized. The manchette begins to develop in spermatids at the stage at which the acrosome has become sandwiched in between the nucleus and the plasma membrane of the spermatid. Concurrently the spermatids become surrounded on all sides by Sertoli cell cytoplasm. In the later stages of nuclear protrusion, the manchette elongates and its walls become thicker. The protruding nuclei become orientated with their acrosome-covered apices facing towards the basement membrane of the tubules. Aggregations of finely granular material appear in Sertoli cell cytoplasm in the region of contact with the acrosomal vacuole. The possible role of the manchette and of Sertoli cell cytoplasm in the phenomenon of nuclear protrusion and orientation is discussed.

Cytoskeleton and Extracellular Matrix Interactions During Sertoli Cell-Cell Adhesion in Vitro

1982 ◽  
Vol 40 ◽  
pp. 290-291
Author(s):  
Glenn L. Decker ◽  
Daniel Carson ◽  
William J. Lennarz
Keyword(s):  
Extracellular Matrix ◽  
Sertoli Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Primary Cultures ◽  
Ultrastructural Studies ◽  
Matrix Interactions ◽  
Extracellular Matrix Interactions ◽  
Sertoli Cell Line

The cytoskeletal components of Sertoli cells appear to play an important role both in the maintenance of elaborate basolateral septate junctions (presumptive blood-testes barrier) and in the development and movement of germ cells into the semeniferous tubule lumen. Additionally, Sertoli cells contain numerous endocytotic vesicles, subjacent to the basal lamina, which may be involved in delivery of serum components to germ cells during maturation. Until now ultrastructural studies have been confined to semeniferous tubules or primary cultures of Sertoli cells. We have initiated studies to visualize the cytoskeleton and extracellular matrix of an established Sertoli cell line.

Freeze-etch observations on the tight junctions of sertoli cells grown in organ culture with FSH

1978 ◽  
Vol 36 (2) ◽  
pp. 340-341
Author(s):  
Rita Meyer ◽  
Zoltan Posalaky ◽  
Dennis Mcginley
Keyword(s):  
Organ Culture ◽  
Tight Junctions ◽  
Sertoli Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Barrier Function ◽  
Cell Junction ◽  
Intramembranous Particles ◽  
Junctional Complexes ◽  
Oriented Parallel

The Sertoli cell tight junctional complexes have been shown to be the most important structural counterpart of the physiological blood-testis barrier. In freeze etch replicas they consist of extensive rows of intramembranous particles which are not only oriented parallel to one another, but to the myoid layer as well. Thus the occluding complex has both an internal and an overall orientation. However, this overall orientation to the myoid layer does not seem to be necessary to its barrier function. The 20 day old rat has extensive parallel tight junctions which are not oriented with respect to the myoid layer, and yet they are inpenetrable by lanthanum. The mechanism(s) for the control of Sertoli cell junction development and orientation has not been established, although such factors as the presence or absence of germ cells, and/or hormones, especially FSH have been implicated.

Effect of nitric oxide on Sertoli cell microtubule of piglets

2009 ◽  
Vol 6 (3) ◽  
pp. 257-263 ◽  
Author(s):  
Yang Li ◽  
Wang Xian-zhong ◽  
Yang Meng-bo ◽  
Zhang Jia-hua
Keyword(s):  
Nitric Oxide ◽  
Enzyme Activity ◽  
Protein Kinase ◽  
Cell Viability ◽  
Sodium Nitroprusside ◽  
Sertoli Cell ◽  
Sertoli Cells ◽  
High Concentration ◽  
Treatment Results ◽  
Anti Oxidant

AbstractTo illustrate the effect of nitric oxide (NO) on the microtubules of Sertoli cells (SC), SCs of piglets were treated with sodium nitroprusside (SNP). Changes in cell viability, anti-oxidant activity, enzyme activity and p38 mutagen-activated protein kinase (p38MAPK) activation were detected. The results were as follows. A low concentration of NO can keep SC microtubule and cell viability normal, and a high concentration of NO could increase p38MAPK activation, decrease anti-oxidant activity and transferrin secretion, and destroy the structure and distribution of the microtubules. The results suggest that SNP treatment results in an increase in NO in SCs and decreased cell anti-oxidant activity. The high concentration of NO destroys cell microtubules by activating p38MAPK.

scholarly journals Role of G protein beta gamma subunits in the regulation of the plasma membrane Ca2+ pump.

1992 ◽  
Vol 267 (4) ◽  
pp. 2375-2379 ◽  
Author(s):  
S Lotersztajn ◽  
C Pavoine ◽  
P Deterre ◽  
J Capeau ◽  
A Mallat ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
G Protein ◽  
Gamma Subunits

Sertoli cell expression of the cystic fibrosis transmembrane conductance regulator

1998 ◽  
Vol 274 (4) ◽  
pp. C922-C930 ◽  
Author(s):  
Fredric R. Boockfor ◽  
Rebecca A. Morris ◽  
Dennis C. DeSimone ◽  
D. Margaret Hunt ◽  
Kenneth B. Walsh
Keyword(s):  
Cystic Fibrosis ◽  
Sertoli Cell ◽  
Sertoli Cells ◽  
Transmembrane Conductance Regulator ◽  
Transmembrane Conductance ◽  
Patch Clamp Technique ◽  
Immunoreactive Material ◽  
Normal Spermatogenesis ◽  
Cell Expression ◽  
Cystic Fibrosis Transmembrane

Mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene have been associated with a number of male reproductive problems, including testis abnormalities and a reduction in germ cell quality and number. To establish at least one site of functional CFTR expression in the testis, we subjected cultured Sertoli cells to analysis of message, protein, and channel activity for CFTR. With reverse transcription-polymerase chain reaction, we obtained evidence for the presence of CFTR RNA when CFTR primers were used with RNA from cultured Sertoli cells. Western analysis performed with both anti-R and anti-C domain CFTR antibodies revealed immunoreactive material in extracts from primary Sertoli cell cultures that seemed consistent with CFTR previously identified in other cells and tissues. This led us to perform more detailed studies using the whole cell arrangement of the patch-clamp technique. Application of the membrane-soluble cAMP analog, 8-chlorophenylthio-cAMP, resulted in the activation of a Cl− current that displayed a permeability sequence of Br− > I− ≥ Cl− and was blocked by diphenylamine-2-carboxylate and glibenclamide. In addition, a 13-pS conductance Cl− channel was measured in excised membrane patches exposed to the catalytic subunit of protein kinase A. When taken together, our findings of evidence of CFTR message, immunoreactive material that appeared consistent with CFTR, and Cl− channels with properties similar to those reported for CFTR provide strong evidence that Sertoli cells express a functional CFTR-like protein. The presence of CFTR in these cells may be needed to maintain the specific nutritional and fluid balance in the seminiferous tubule that is vital for normal spermatogenesis.

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