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  • 1
    ISSN: 1432-0711
    Keywords: Ovary ; FSH ; Inhibin ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The effects of a highly purified preparation of human follicular fluid inhibin (hFF-Inhibin) on the ovary of immature rats have been studied. The highly purified preparation was obtained by gel and ion-exchange chromatography and ultrafiltration of the acetone dried extract of human follicular fluid. Administration of 5, 10, or 20 μg of inhibin daily for 10 days starting from day 11 postnatum resulted in (1) a significant reduction of basal FSH levels by the 20 μg dose but not by lower doses, (2) atresia of preantral and antral follicles in an apparently dose-related manner, (3) hyperplasia of the theca interna, and (4) degeneration of the granulosa cells, maximum damage being in the group treated with 20 μg. In inhibin-induced atretic follicles there were no signs of (a) granulosa cell-luteinisation, (b) connective tissue invasion, (c) hyalinisation of the basement membrane, or (d) vascularisation. Simultaneous administration of human menopausal gonadotrophin (hMG) led to luteinisation of the atretic follicles. However, the other structural alterations mentioned above (b-d), and those of the theca cells could not be overcome by hMG. In addition, there was no significant difference in the ovarian weight of animals treated with hMG when compared to those treated with inhibin and hMG, but the serum levels of the oestradiol in the latter were significantly lower. Our results suggest that hFF-inhibin acts not only at the pituitary, but also at the ovarian level.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1434-0879
    Keywords: Key words Reverse transcription-polymerase chain reaction (RT-PCR) ; Micrometastases ; Prostatic cancer ; Prostate membrane-specific antigen (PSM)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Detection of prostate-specific membrane antigen (PSM)-mRNA expression in blood samples using reverse transcription polymerase chain reaction (RT-PCR) is discussed as a new diagnostic marker of circulating micrometastases in prostate cancer patients. We applied the RT-PCR technique to different human tissues and obtained positive signals for PSM transcripts in human genital and multiple extra-genital tissue sites. The cDNAs were prepared from different human tissues and prostatic cell lines. RT-PCR and nested RT-PCR for PSM was performed with primers derived from the published PSM cDNA. The RT-PCR fragments obtained were cloned and showed 100% sequence homology to PSM. Southern blot hybridization with labeled probes was used to confirm the specificity of the amplicons. In addition to the known PSM expression in the human brain, PSM-mRNA was detected in cDNA isolated from human testis, epididymis and seminal vesicles and in the PC-3 prostatic cancer cell line. Furthermore, we found PSM-mRNA in heart, liver, lung, kidney, spleen, and thyroid gland. The results indicate that PSM expression is not restricted to the prostate gland, but represents a more general component of genital and extra-genital human tissues. This must be considered when RT-PCR and nested RT-PCR screening for PSM expression is performed as a diagnostic measure in blood from prostate cancer patients.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 144 (1974), S. 1-18 
    ISSN: 1432-0568
    Keywords: Mammals ; Mole ; Ovary ; Interstitial cells ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Das Ovar des Maulwurfs wurde licht- und elektronenmikroskopisch untersucht. Der Feinbau der verschiedenen Follikel und der Keimzellen entspricht dem anderer Species. Die Granulosa wird vermutlich erst mit der Ovulation luteinisiert. Sie enthält reichlich rauhes endoplasmatisches Reticulum und Sprossen von glattem endoplasmatischem Reticulum. Die Zellen der Theka interna leiten sich von Fibrocyten ab und enthalten neben reichlich glattem ER auch granuläres ER sowie Lipidtropfen und Mitochondrien mit Cristae und Tubuli. Diese Zellen ähneln in ihrer Feinstruktur stark den Zwischenzellen des ovarialen Marks. Dieses enthält in der Marksträngen Epithelien, die einer kräftigen Basalmembran aufsitzen. Sie werden teils als embryonal persistierende Vorstufen von Granulosa- oder Sertoli-Zellen, teils als Granulosasprossen aus der Rinde gedeutet. Für die Zwischenzellen des Marks erscheint eine Analogie mit den Hiluszellen des menschlichen Ovarialmarks bzw. eine Homologie mit den Leydig-Zellen des Hodens zweifelhaft. Als mögliche Quelle für diesen Zelltyp werden neben embryonal liegengebliebenen Anteilen auch Thekakeile und Epoophoronzellen sowie Fibroblasten diskutiert. Neben einer Steroid-synthese dürfte ihnen eine Reserve- und Speicherfunktion zukommen.
    Notes: Summary In order to make possible comparison between relatively primitive and relatively specialized gonads, the ovary of the mole was studied by light and electron microscopy. The ultrastructure of primary, secondary and tertiary follicles and of the germ cells is similar to that of other species. The granulosa cells of secondary and early tertiary follicles contain abundant rough endoplasmic reticulum and a small number profiles of smooth endoplasmic reticulum. The cells of the theca interna, which develop from simple fibrocytes are rich in smooth and rough endoplasmic reticulum and contain many lipid droplets and mitochondria, which possess both cristae and tubuli mitochondriales. At the time of ovulation, the granulosa cells are luteinized and their ultrastructure changes correspondingly. The medulla of the ovary is composed of the medullary cords and the interstitial cells. The medullary cords are solid epithelial cords, which are surrounded by a prominent basement membrane. They may be derived from embryonic precursors of granulosa—or Sertoli-cells or bud from the cortical zonae granulosae. There is a striking morphological similarity between the theca and interstitial cells. The interstitial cells of the ovarian medulla differ from the hilus cells of the human ovary and the Leydig-cells of the testis. They may develop either from embryonic rudiments of Leydig—or hilus-cell precursors, or bud from the theca or the epoophoron, or they may develop from fibrocytes. In addition to their suggested activity in steroid biosynthesis, the interstitial cells may have a trophic or storage function.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0568
    Keywords: Human testis ; Sertoli cells ; Spermatogenesis ; Spermatogenetic disturbances ; Intermediate filaments ; 3D reconstruction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The presence, distribution and spatial arrangement of vimentin-type intermediate filaments in Sertoli cells from human testis biopsies, were studied in semithin and ultrathin sections using a polyclonal rabbit antiserum. At the ultrastructural level, vimentin immunoreactivity was seen concentrated around the nuclei, along fibrillary material within the cytoplasm and at the ectoplasmic specializations of the Sertoli cell junctions, as well as throughout the periphery of the Sertoli cell processes. It is therefore well suited as a marker for Sertoli cell configuration. In computer-aided 3D reconstructions of 20 serial sections, Sertoli cells displayed particular configurations of intermediate filaments in the different stages of spermatogenesis. Two basic configurations, named AS (before spermiation, stages V, VI, I and II), and PS (after spermiation, stages III and IV) respectively, could be differentiated. In addition to the reconstruction and morphological analysis of vimentin filaments in Sertoli cells from patients with unaltered spermatogenesis (obstructive azoospermia), pathological specimens (spermatogenetic arrest, Sertoli cells only-syndrome) were studied with respect to vimentin immunohistochemistry. The results indicate that vimentin filaments play an important role in the adaptation of Sertoli cells to the varying configurations of neighbouring cells during spermatogenesis as well as under pathological conditions.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  In the present study we examined the expression and release of the extracellular matrix glycoprotein fibronectin (FN) in a prostate cancer cell line (LNCaP) and in primary prostatic stromal cells using the reverse transcription–polymerase chain reaction (RT-PCR) and by an enzyme-linked immunosorbent assay. Perturbation experiments in vitro using antibodies directed against FN and the FN receptor were also performed. Immunohistochemistry was used to show the in vivo distribution of FN and the FN receptor in tissue sections of normal human prostate, benign prostatic hyperplasia, and prostate carcinoma. The expression of the oncofetal FN ED-B segment in benign prostatic hyperplasia and prostate carcinoma tissue was investigated by RT-PCR. The FN mRNA was expressed by LNCaP and primary prostatic stromal cells, respectively. Both cell types released FN into the medium in a time-dependent manner, whereby FN secretion was about 2.5-fold higher in cultures of stromal cells relative to LNCaP cells. Blocking FN with anti-FN antibodies resulted in a significant decrease in cell adhesion for LNCaP cells and a change in morphology for the primary stromal cells. FN was located mainly in the stromal compartment of the prostate, showing a distinct distribution pattern in prostate carcinoma, whereas the FN receptor was detectable only in the prostate epithelia. RT-PCR experiments showed the expression of the oncofetal FN ED-B segment in benign prostatic hyperplasia and prostate carcinoma tissue, with a 3.5-fold higher expression in the prostate carcinoma probes. Our data point to an important role for FN in cell adhesion of prostatic cells and show that an alternatively spliced FN mRNA is upregulated in the pathologically altered human prostate.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-1335
    Keywords: Small cell lung cancer ; Cell lines ; Serum free medium ; Growth properties ; Morphological characteristics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cell lines from SCLC were established with a success rate of 43% from different metastatic sites of treated and untreated patients. All 6 SCLC cell lines grew as floating cell aggregates without substrate adherence. The degree of aggregation ranged from very tight spheroids to very loose sheets and chains. This gross morphological property showed a striking correlation to the PDT, with short PDTs in loose growing cell lines and long PDTs in tight growing cell lines. Cell size and nuclear features, i.e., chromatin pattern and nucleolar prominence, also seemed to correlate with the PDT and gross morphology. All SCLC cell lines had dense core granules by electron microscopical examination. Several different serum-free and serum-supplemented growth media were tested for their feasibility in estabilishing and permanently growing SCLC. Serum-free SIT medium and SIT 2.5 medium provided the best results in liquid culture. For semisolid SCLC cultivation, R10 medium was suprior to all other media tested. These cell lines are currently under intensive biochemical, molecular biological, and cytogenetical investigation in different laboratories and thus provide a tool for studying the biology of lung cancer.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-4935
    Keywords: seminal plasma protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We isolated the major protein of apparent Mr of 15,000–16,000 from seminal plasma as well as from seminal veiscle secretion of bull and proved by amino acid analysis and tryptic peptide mapping that the two proteins were identical. An antiserum against this major protein was employed to quantitate and identify the major protein in seminal plasma as well as seminal vesicle secretion. The antiserum did not cross-react with proteins from bovine or human plasma or follicular fluid respectively. Cell-free translation of poly(A)RNA from seminal vesicle tissue and immunoprecipitation yielded one major species with apparent Mr of 18,000. Using the anti-major protein antiserum, this major species was specifically immuno absorbed. Cloning and sequencing of a major protein-specific cDNA led to the identification of clone pMP17, encoding a precursor of the major protein of 128 amino acid residues. We proved that the major protein is identical to protein PDC 109 (Eschet al., Biochem. Biophys. Res. Comm. 113:861–867, 1983). The seminal vesicles synthesize major protein in an androgen-dependent fashion. In addition to intraluminal secretion of the vas deferens, ampullary spermatozoa revealed an intense immunoreaction which was restricted to the neck region of the sperm head and the middle piece, while the principal piece of the tail as well as the sperm head were devoid of immunoreactive material. Epididymal epithelium (as well as calf seminal vesicle epithelium) showed no immunoreactivity with major protein antiserum. Immunoelectron microscopy demonstrated that only spermatozoa devoid of a plasma membrane around the middle piece were able to bind the antiserum against major protein. After removal of the plasma membrane from epididymal spermatozoa, binding of major protein to subplasmalemmal binding sites was visualised using gold-labeled MP. Transblotting with gold-labeled MP demonstrated a protein of about 66 kDa which appears to represent the major protein-receptor. Binding of major protein to the receptor (after loss of the plasma membrane in the mid-piece region of the spermatozoa after contact with secretions from seminal vesicles) is interpreted as a phyisological process presumably related to the onset of sperm motility.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-0878
    Keywords: Spermatozoa ; Seminal vesicles ; Seminal fluid ; Sperm motility ; Immunocytochemistry ; Bull
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The seminal vesicles synthesize in an androgen-dependent manner a neutral protein of 13.5 kDa molecular weight that makes up about 40% of their secretion (“major protein”). An antiserum against this protein raised in rabbits was used to localize the antigen within the seminal vesicles. In addition to intraluminal secretion of the seminal vesicles and the ampulla of the vas deferens, ejaculated and ampullary spermatozoa revealed an intense immunoreaction, which was restricted to the neck region of the sperm head and the middle piece, while the principal piece of the tail as well as the sperm head were devoid of immunoreactive material. Comparison of spermatozoa taken from the tail of the epididymis with ampullary spermatozoa showed that about 90% of the latter, but only 10–20% of the former presented this distributional pattern of immunoreactive sites. Epididymal epithelium as well as calf seminal vesicle epithelium showed no immunoreactivity with major protein antiserum. Using a pre-embedding staining technique with gold-labeled primary or secondary antibodies, respectively, no immunostaining could be achieved at the ultrastructural level. Incubation experiments of epididymal spermatozoa in EGTA-containing solutions in the absence of calcium resulted in a gradual labilization and eventual loss of the plasma membrane of the sperm middle piece. After removal of (at least part of) the plasma membrane, bound major protein could be visualized immunohistochemically close to the mitochondria of the middle piece using a gold-labeled primary or secondary antibody. The acceptor site for major protein therefore seems to reside inside the plasma membrane of the sperm middle piece. Incubation of epididymal spermatozoa in phospholipase-containing solutions removed the acceptor site from the spermatozoa. Separation by polyacrylamide treatment of proteins from epididymal sperm cells extracted by sodium hydroxide or phospholipase treatment, subsequently transblotted on nitrocellulose sheets and directly labeled with gold-tagged major protein, demonstrated a protein duplet with a molecular weight of 65 and 67 kDa, respectively, which appears to represent the specific binder of major protein underneath the sperm surface. Binding of major protein to this ∼66 kDa acceptor site is regarded as a physiological event that may be related to the onset of hyperactivated sperm motility.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 319-332 
    ISSN: 1059-910X
    Keywords: Rat ; Prostate ; Epithelium ; Stroma ; Cytodifferentiation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Instructive influences of fetal mesenchyme were examined in heterotypic tissue recombinants consisting of urogenital sinus mesenchyme (UGM) from male and female rats and distal ductal tips from adult rat prostate. Tissues were grown under the renal capsule of male hosts for periods up to 28 days. Resultant growths exhibited typical prostate histology. Expression of lobe-specific proteins for the ventral (prostatic steroid binding protein [PSBP]) lateral (seminal vesicle secretion II [SVS II]), and dorsal prostate (secretory transglutaminase [TGase]) were examined by immunocytochemistry. Male or female UGM combined with terminal segments of the ventral or dorsal prostate and immunolabeled with antibodies to lobe-specific proteins demonstrated expression of all three secretory products. The pattern of staining was consistent with a compound inductive response from the UGM. Unique to this study was our ability to use a defined mesenchymal tissue (female ventral mesenchymal pad [VMP]). This tissue is specifically associated with ductal branching morphogenesis and cytodifferentiation of the ventral prostate. Distal ductal tips from the dorsal lobe of the adult male prostate when recombined with female VMP and grown in vivo exhibited transformation of secretory phenotype, and the epithelium expressed mRNAs for PSBP. Immunocytochemistry of serial sections did not demonstrate labeling for TGase in the new epithelial growth. Ultrastructural analysis of the heterotypic recombinants indicated that the epithelium had similar characteristics to those of normal ventral prostate. Early stages of the mesenchymal-epithelial interactions resulted in dedifferentiation of the adult epithelium to solid cords of stratified cells. These findings illustrate the potent instructive capacity of a defined fetal UGM to influence development and cytodifferentiation of adult prostate epithelium. © 1995 Wiley-Liss, Inc.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
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