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1.
Biol Reprod ; 43(1): 15-24, 1990 Jul.
Article in English | MEDLINE | ID: mdl-2393687

ABSTRACT

We analyze by immunocytochemistry the in vivo distribution in rat Sertoli cells of Cyclic Protein-2 (CP-2), which is maximally synthesized and secreted in vitro at stages VI and VII of the cycle of the seminiferous epithelium. This analysis demonstrates that CP-2 staining is strongest in Sertoli cells in stage VI and VII tubules. Additionally, we demonstrate that the staining for CP-2 within a stage VII tubule differs from the staining of another Sertoli cell secretory product, androgen-binding protein. CP-2 is not detected by immunocytochemistry in any other tissues of the reproductive tract, though immunoblot analysis demonstrates the presence of CP-2 in rete testis and epididymal fluids. CP-2 was immunocytochemically detected in only three other organs: the kidney, the brain (with greatest concentration in the supraoptic and paraventricular nuclei), and the posterior pituitary. The presence of CP-2 in the kidney was confirmed by metabolic radiolabeling, immunoprecipitation, and peptide analysis. The presence of CP-2 in the brain was confirmed by immunoblot analysis of radioinert protein immunoprecipitated from the anterior hypothalamus.


Subject(s)
Brain/metabolism , Kidney Tubules, Proximal/analysis , Proteins/analysis , Sertoli Cells/analysis , Androgen-Binding Protein/analysis , Animals , Blotting, Western , Brain/cytology , Epididymis/analysis , Kidney/analysis , Kidney/cytology , Male , Pituitary Gland, Posterior/analysis , Precipitin Tests , Prostate/analysis , Proteins/ultrastructure , Rats , Rats, Inbred Strains , Rete Testis/analysis , Seminal Vesicles/analysis , Sertoli Cells/cytology , Vas Deferens/analysis
2.
Cancer ; 64(3): 770-5, 1989 Aug 01.
Article in English | MEDLINE | ID: mdl-2663132

ABSTRACT

A case of adenocarcinoma of the rete testis was encountered in a 36-year-old white man. The tumor fulfilled established criteria for determining origin in the rete and showed an unusual biphasic morphology with papillary adenocarcinoma mixed with a prominent component of cytologically malignant spindle cells. Immunohistochemical study demonstrated a positive reaction in the epithelium for cytokeratin and epithelial membrane antigen, and the cytoplasm of a few of the spindle cells also reacted with these antibodies. Electron microscopic study confirmed the biphasic pattern, showing epithelial gland formation and mesenchymal cells. The results indicate that this tumor is a metaplastic carcinoma of the rete testis. Recognition of this pattern of rete carcinoma may further enhance our knowledge of primary tumors at this unusual site.


Subject(s)
Adenocarcinoma/pathology , Testicular Neoplasms/pathology , Adenocarcinoma/analysis , Adenocarcinoma/secondary , Adenocarcinoma/ultrastructure , Adult , Humans , Immunoenzyme Techniques , Male , Rete Testis/analysis , Rete Testis/pathology , Rete Testis/ultrastructure , Testicular Neoplasms/analysis , Testicular Neoplasms/ultrastructure
3.
J Androl ; 9(6): 397-402, 1988.
Article in English | MEDLINE | ID: mdl-3215825

ABSTRACT

Concentrations of gossypol in blood plasma, rete testis fluid, and fluid from the caudia epididymidis were measured simultaneously by high performance liquid chromatography in rats treated with gossypol (15 mg/kg daily for 3 weeks). Antispermatogenic effects were demonstrated by loss of sperm motility in the cauda epididymidis and structural changes in the testis. It was found in these treated rats that concentrations of gossypol were lower in rete testis fluid compared with blood plasma but increased significantly in fluid from the cauda epididymidis. The results indicate a restriction of the blood-testis barrier to gossypol and its local concentration in the epididymis after fluid resorption.


Subject(s)
Epididymis/analysis , Gossypol/analysis , Rete Testis/analysis , Testis/analysis , Animals , Epididymis/drug effects , Gossypol/analogs & derivatives , Gossypol/blood , Gossypol/pharmacology , Male , Rats , Rats, Inbred Strains , Rete Testis/drug effects , Sperm Motility/drug effects , Testis/cytology
4.
Pathol Res Pract ; 183(3): 326-8, 1988 Jun.
Article in English | MEDLINE | ID: mdl-2458580

ABSTRACT

A systematic search for endocrine cells in the excurrent duct system of the testis was carried out by means of histochemical and immunohistochemical techniques. A panel of antibodies against amine and polypeptide hormones was used. 80 specimens comprising representative areas of rete testis, ductuli efferentes, ductus epididymis and 30 examples of ductus deferens were investigated. Cells immunoreactive for serotonin were detected in four out of 110 specimens. They were invariably in normal-appearing ductuli efferentes. A salient feature was their rarity and focal distribution. We failed to detect any endocrine cells in other segments of the excurrent duct system and notably not among epididymal epithelium. It seems of interest that serotonin cells are specifically distributed throughout remnants of excretory mesonephric tubules in both males and females.


Subject(s)
Endocrine Glands/cytology , Serotonin/analysis , Testis/cytology , Endocrine Glands/analysis , Epididymis/analysis , Epididymis/cytology , Humans , Male , Rete Testis/analysis , Rete Testis/cytology , Seminiferous Tubules/analysis , Seminiferous Tubules/cytology , Staining and Labeling , Testis/analysis , Vas Deferens/analysis , Vas Deferens/cytology
5.
Biochemistry ; 27(11): 4079-88, 1988 May 31.
Article in English | MEDLINE | ID: mdl-3415974

ABSTRACT

Clusterin is a protein present in the rete testis fluid of the ram that elicits aggregation of erythrocytes and Sertoli cells in vitro. In view of its possible biologic function in relation to cell-cell interaction in the testis, we isolated this protein from ram rete testis fluid using sequential high-performance liquid chromatography columns and performed a detailed physicochemical characterization. This protein consists of two molecular variants designated form I and form II clusterin. Each form of clusterin consists of two subunits with an apparent molecular weight of 40,000. It is of note that the two subunits have no homology in their N-terminal amino acid sequences. However, the N-terminal amino acid pairs of the two subunits derived for the two forms of clusterin are identical. Using o-phthalaldehyde to block the Lys residue at the fourth amino acid pair from the N-terminus which leaves the Pro residue free for subsequent Edman degradation, we have deduced the N-terminal sequence of each of the two subunits for form I clusterin. Comparison of the NH2-terminal sequences of the two subunits of clusterin with the release 10.0 of the protein sequence data base of the Protein Identification Resource indicated no homology between either of the subunits of clusterin and any of the known proteins in the data base. A highly specific radioimmunoassay developed for clusterin was used to measure its concentrations in the fluids of the rete testis and cauda epididymis. Since a significant amount of immunoreactive clusterin was found in serum, the protein was partially purified from this source by immunoaffinity chromatography. Immunoreactive serum clusterin was smaller than the testicular clusterin (Mr 37,000 vs 40,000), but both proteins share common epitopes as demonstrated by radioimmunoassay and immunoblots. However, serum clusterin does not possess the biologic activity of the testicular clusterin in that it does not elicit cell aggregation in vitro. It is of note that deglycosylation of testicular clusterin can also eliminate this in vitro biologic activity, suggesting that the serum clusterin might be a deglycosylated form of the testicular protein and the carbohydrate core plays an important role in determining the cell aggregation activity. Studies on the distribution of this protein in the reproductive compartment indicate that it is highly concentrated in the rete testis and the cauda epididymal fluids. This suggests that this protein might have some important functions in the reproductive tract.


Subject(s)
Glycoproteins/analysis , Molecular Chaperones , Rete Testis/analysis , Testis/analysis , Amino Acid Sequence , Amino Acids/analysis , Animals , Body Fluids/analysis , Chromatography, Affinity , Chromatography, High Pressure Liquid , Clusterin , Electrophoresis, Polyacrylamide Gel , Glycoproteins/isolation & purification , Immunochemistry , Male , Molecular Weight , Radioimmunoassay , Sheep
6.
Biol Reprod ; 37(1): 135-46, 1987 Aug.
Article in English | MEDLINE | ID: mdl-3651540

ABSTRACT

A major 68-kDa protein in ram rete testis fluid (RTF) is shown to be chemically and immunologically indistinguishable from albumin in ovine serum. Data obtained with two-dimensional gel electrophoresis of RTF demonstrate the presence of additional proteins with a molecular mass of 68 kDa that do not react with antisera against sheep serum albumin. Biochemical characteristics of albumin preparations isolated by immunoaffinity chromatography from ovine serum and from RTF were compared. Albumin from both sources had the same apparent molecular mass of 68 kDa, the same isoelectric point of approximately 4.2, and neither bound specifically to Concanavalin A. Analysis of tryptic peptide maps, obtained with reverse-phase high-pressure liquid chromatography, indicated no significant differences between digests of the two purified albumin preparations. Results indicate that RTF albumin and serum albumin are the same protein, which implies that RTF albumin may originate from serum. Albumin levels in RTF, collected from different rams and measured by radioimmunoassay, varied between 46 and 164 micrograms/ml, constituting between 11 and 17% of total RTF protein, while albumin levels in sheep plasma were 40,000 micrograms/ml. The protein composition of RTF is discussed in relation to the relative amounts of various components contributed by testis cells and the amounts derived from serum.


Subject(s)
Albumins/isolation & purification , Rete Testis/analysis , Testis/analysis , Animals , Body Fluids/analysis , Chromatography, High Pressure Liquid , Electrophoresis, Polyacrylamide Gel , Isoelectric Focusing , Male , Molecular Weight , Sheep
7.
Biol Reprod ; 36(3): 753-68, 1987 Apr.
Article in English | MEDLINE | ID: mdl-3593846

ABSTRACT

Spermatozoa, fluids, and principal cells from different regions of the epididymis were characterized by two-dimensional electrophoresis. Rete testis fluid was collected after 36-h efferent duct ligation, and cauda epididymal fluid was collected by retrograde perfusion through the vas deferens. Spermatozoa were collected after their exudation from minced caput and corpus epididymal tissue. Principal cells were recovered after enzymatic disaggregation and centrifugal elutriation of epididymides. Two-dimensional polyacrylamide gel electrophoresis was used to prepare protein profiles of all samples. Comparison of the proteins found in rete testis fluid versus those found in cauda epididymal fluid revealed a dramatic change in composition, including the loss, addition, or retention of specific proteins as well as changes in the relative concentrations of certain proteins. Prominent cauda epididymal fluid proteins, possibly contributed by the epididymal epithelium, were detected at 16, 23, and 34 kDa. After epididymal transit, a considerable decrease was observed in the number of aqueous-soluble sperm proteins. Differences in the protein composition of epididymal epithelial principal cells from the caput versus corpus epididymidis were also noted, suggesting that functional differences exist for these epididymal regions. Of particular interest was the occurrence of a prominent protein of approximately 20-23 kDa found in all sperm samples, in fluids, and in caput and corpus principal cells. However, this protein was absent in cauda epididymal sperm after 36-h efferent duct ligation. The rapid loss of this protein from sperm after efferent duct ligation suggests that this surgical intervention may affect spermatozoa residing within the epididymis.


Subject(s)
Epididymis/analysis , Proteins/isolation & purification , Spermatozoa/analysis , Animals , Body Fluids/analysis , Electrophoresis, Polyacrylamide Gel , Epididymis/cytology , Male , Rats , Rats, Inbred Strains , Rete Testis/analysis
8.
Biol Reprod ; 33(1): 177-86, 1985 Aug.
Article in English | MEDLINE | ID: mdl-3904851

ABSTRACT

Clusterin, a glycoprotein that elicits cell aggregation, has previously been isolated from ram rete testis fluid, and has been partially characterized. In experiments reported, we have used monoclonal antibodies against clusterin in combination with indirect immunofluorescence microscopy to investigate the distribution of clusterin in the adult ram testis, rete testis, and excurrent ducts. Tissue blocks (5 mm3) were fixed in periodate/lysine/paraformaldehyde containing 0.1% glutaraldehyde and, after embedding, 5-microM sections were prepared for immunolocalization. In the testis, 2 basic patterns were observed: 1) strong to moderate staining for clusterin in the adluminal region with little staining in the basal region of the seminiferous epithelium and germinal cells; and 2) moderate staining throughout the seminiferous epithelium between germinal cells. In the rete testis, strong clusterin staining was localized intracellularly in the rete epithelial cells, most often associated with the luminal surface. In the epididymis, intracellular clusterin was localized in some principal cells of the caput epididymidis. The luminal surfaces and spermatozoa within the lumen were strongly positive. In the vas deferens, clusterin staining was associated with the luminal surface only. The presence of clusterin was clearly detected in unwashed isolated epididymal spermatozoa, but not in spermatozoa washed with phosphate-buffered saline containing 0.05% Tween 20.


Subject(s)
Testis/analysis , Animals , Antibodies, Monoclonal , Fluorescent Antibody Technique , Immunologic Techniques , Male , Rete Testis/analysis , Sheep
9.
J Androl ; 6(1): 20-34, 1985.
Article in English | MEDLINE | ID: mdl-3972717

ABSTRACT

Luminal fluid samples were collected by micropuncture of the seminiferous tubule, rete testis, and defined levels of the epididymal tubule. After removal of spermatozoa by centrifugation, the supernatant fluids were analyzed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and an ultrasensitive silver staining procedure to define the sequential change in protein composition along the excurrent duct system. Fluid from each segment displayed a characteristic 2-D PAGE map composed of numerous polypeptides. Seminiferous tubule fluid contained a wide array of polypeptides, with most concentrated in the 45 Kd to 90 Kd range, but, in contrast, rete testis fluid lacked most of these polypeptides. The major complex of rete testis fluid comigrated with serum albumin and was present in all distal segments. Other major rete testis components were not noted distally. Fluid from the caput was characterized by new major components of 30 to 37 Kd, 28 to 30 Kd, 24 Kd, and 23 Kd, each of which consisted of multiple spots of apparent isoelectric variants; all except the 30 to 37 Kd complex were present in the fluid from more distal segments. Proceeding distally, there was a temporal appearance of new polypeptides, especially in the molecular weight range below 30 Kd. Two-dimensional PAGE analysis of detergent extracts of washed spermatozoa indicate that a specific subset of these fluid polypeptides are sperm associated.


Subject(s)
Epididymis/analysis , Peptides/analysis , Testis/analysis , Animals , Body Fluids/analysis , Detergents , Electrophoresis, Polyacrylamide Gel/methods , Male , Rats , Rats, Inbred Strains , Rete Testis/analysis , Seminiferous Tubules/analysis , Solubility , Spermatozoa/analysis
10.
Arch Int Physiol Biochim ; 91(2): 69-80, 1983 Jul.
Article in French | MEDLINE | ID: mdl-6195989

ABSTRACT

We describe a method to estimate binding of human 125I-FSH to a preparation of bovine testicular receptors. Various experimental conditions are tested and the validity of the method is demonstrated. Using this method, the presence of biological substances modifying the FSH binding is measured in various fractions of ram retetestis fluid submitted to several steps of purification by chromatography. FSH receptor binding inhibitor (FSHRBI) activity is obtained in a low molecular weight fraction and FSH receptor binding stimulator activity in a larger one. These cybernin activities are isolated in fractions different from the ones observed with inhibin and GnRH like activities.


Subject(s)
Follicle Stimulating Hormone/antagonists & inhibitors , Follicle Stimulating Hormone/metabolism , Hormone Antagonists/isolation & purification , Inhibins , Receptors, Cell Surface/metabolism , Testis/metabolism , Animals , Body Fluids/analysis , Cattle , Follicle Stimulating Hormone/isolation & purification , Iodine Radioisotopes , Male , Methods , Receptors, FSH , Rete Testis/analysis , Rete Testis/physiology , Sheep
15.
J Reprod Fertil ; 57(2): 397-400, 1979 Nov.
Article in English | MEDLINE | ID: mdl-513029

ABSTRACT

Boar rete testis fluid was tested for its capacity to induce Müllerian regression in 14.5-day-old rat Müllerian ducts. Weak activity was present in crude RTF, but after gel filtration 5-fold concentration, greater activity was detected in 1 our of 7 pools of the eluted fractions. The biologically active fraction (mol. wt 160 000-310 000) coincided with the elution of authentic labelled anti-Müllerian hormone, obtained from bovine fetal testes. These results indicate that a small amount of anti-Müllerian hormone is still synthesized in post-natal life.


Subject(s)
Mullerian Ducts/drug effects , Rete Testis/analysis , Testicular Hormones/pharmacology , Testis/analysis , Animals , Male , Rats , Swine , Testicular Hormones/analysis
16.
J Reprod Fertil ; 56(1): 105-11, 1979 May.
Article in English | MEDLINE | ID: mdl-469829

ABSTRACT

Luminal fluid was collected by micropuncture techniques from the testis and epididymis of the rat, hamster, rabbit, boar and ram and the concentration of free L-carnitine in the fluid was estimated using enzymic methods. Carnitine was present in the testicular fluid of the rat in concentrations less than 1 mM but increased down the epididymis to reach 53 mM in luminal fluid from the cauda epididymidis, approximately 2000 times higher than in blood plasma. A high concentration was first found in the luminal fluid from the distal caput epididymidis, at about the point where the spermatozoa become motile. Carnitine was also present in the epididymal luminal fluid of the other species studied; the amounts were not as high as those in the rat but were still higher than those in blood plasma.


Subject(s)
Carnitine/analysis , Epididymis/analysis , Testis/analysis , Animals , Body Fluids/analysis , Carnitine/blood , Cricetinae , Male , Mesocricetus , Rabbits , Rats , Rete Testis/analysis , Seminiferous Tubules/analysis , Sheep , Swine
19.
Endocrinology ; 99(6): 1618-30, 1976 Dec.
Article in English | MEDLINE | ID: mdl-137112

ABSTRACT

Ten steroids which may have a role in the process of sperm maturation within the epididymis were quantified by competitive protein binding or radioimmunoassay. Rete testis fluid (RTF) carrying testicular sperm into the epididymis was rich in dehydroepiandrosterone and testosterone (21 +/- 2 and 33 +/- 3 ng/ml) while cauda eipididymal plasma (CEP) around sperm which have completed maturation had high levels of progesterone, dihydrotestosterone, 3beta-androstanediol, dehydroepiandrosterone and testosterone (7.4 +/- 0.8, 20.3 +/- 1.1, 6.5 +/- 0.4, 8.0 +/- 0.7 and 11.5 +/- 0.7 ng/ml). About 4 mug of steroids enter the epidymis daily in RTF, but less than 1% was found in CEP; the balance presumably was absorbed by the epithelium in the proximal caput epididymidis. Nevertheless, tissue levels of total 17beta-OH androgens were lower in the proximal caput than in the distal caput or corpus epididymidis. In all zones of the epididymis, dihydrotestosterone accounted fro about 70% of the total 17beta-OH androgens found in the nuclear fraction. In the cytoplasmic fraction, however, dihydrotestosterone predominated only in the distal caput and corpus epididymidis. In the cauda epididymidis, CEP and sperm probably accounted for less than 35% of the total 17beta-OH androgens and less than 25% of the dihydrotesterone. The progesterone concentration of the cauda than in the caput epidymidis. Twice washed testicular sperm contained more testosterone than cauda epididymal or ejaculated sperm (16.6 +/- 1.9, 1.6 +/- 0.2 and 1.5 +/- 0.3 ng/10(9) sperm, respectively), but less progesterone (0.5 +/- 0.1, 1.3 +/- 0.2 and 1.0 +/- 0.4 ng/10(9) sperm, respectively). As a consequence of mixture with estrogen-rich prostatic fluid (150 +/- 9 pg/ml), ejaculated sperm contained a relatively high amount of estrogens (112 +/- 15 pg/10(9) sperm). These studies revealed marked differences in steroid profiles of fluids entering and leaving the epididymis and of infertile testicular and fertile cauda epididymal sperm.


Subject(s)
Epididymis/analysis , Semen/analysis , Steroids/analysis , Androstane-3,17-diol/analysis , Androstenediols/analysis , Androstenedione/analysis , Androsterone/analysis , Animals , Cattle , Cell Nucleus/analysis , Cytoplasm/analysis , Dehydroepiandrosterone/analysis , Dihydrotestosterone/analysis , Estradiol/analysis , Estrone/analysis , Male , Progesterone/analysis , Prostate/analysis , Rete Testis/analysis , Seminal Vesicles/analysis , Steroids/blood , Testosterone/analysis
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