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M Dym

Publications and source records attributed to M Dym.

At least 73 records · Page 4Linked to original sources
PubMed1985

Structural features of rat epididymal epithelial cells in vitro.

Epididymal epithelial fragments, free of stromal elements were isolated from mature rats using two sequential collagenase digestions. Within 24 h these attached efficiently to a variety of substrates including glass, plastic, placental collagen, type IV collagen and epididymal extracellular matrix material. Cells spreading away from the fragments rapidly assumed a flattened, overlapping, monolayer appearance typical of epithelial cells in culture. Cells still associated with the fragments or adjacent to them remained more polarized and more closely resembled epididymal principal cells in vivo than did cells that had migrated to the periphery of the monolayer. Apical microvilli characteristic of these cells in vivo were common during the first 4 days in culture but diminished in number and size thereafter. Cultured cells maintained many of the structural features characteristic of principal cells in vivo, including a well developed Golgi apparatus, coated pits and vesicles, and many multivesicular bodies. An extensive filamentous network, shown immunocytochemically to consist of keratin, was present in the cytoplasm of all cells but was more obvious in flattened cells at the periphery of the monolayer. Rhodamine phalloidin labelling of filamentous actin showed that concentrations of actin occurred corresponding to microvilli on the apical surface, in a continuous ring just below the apical surface, and also in stress fibres at the base of the cells. Cells isolated and cultured from the distal caput epididymidis possessed lobulated nuclei, in contrast to the round or oval nuclei found in cells cultured from the proximal caput epididymidis. Cells from the distal caput epididymidis were also characterized by the presence of many lipid droplets in their cytoplasm. Autofluorescent granules were observed in epithelial cells from both regions but were larger and more numerous in cells isolated from the distal caput epididymidis. Tritiated thymidine incorporation by the cells after 4 days in culture showed that cells adjacent to the parent epithelial fragment were dividing at a greater rate than cells that had migrated to the periphery of the monolayer.

Actins↗
PubMed1984

Immunocytochemical localization of alpha-lactalbumin in the male reproductive tract.

The immunocytochemical localization of the milk protein alpha-lactalbumin in the male reproductive tract is described. Using a primary antiserum raised against highly purified rat milk alpha-lactalbumin, specific staining was consistently shown in the supranuclear Golgi region of the principal cells of the proximal caput epididymidis but only occasionally in epithelial cells from other regions of the duct. Staining was also found in the epididymal lumen and associated with spermatozoa. This luminal staining persisted throughout the distal caput, corpus and cauda epididymidis. Staining was rarely associated with spermatozoa in the efferent ducts and initial segment. Alpha-lactalbumin immunoreactivity was also detected in the seminiferous epithelium. Staining was confined to the Golgi-acrosome region of spermatids. These results indicate that an alpha-lactalbumin-like molecule, or molecules, is present in the male reproductive tract and that it is localized specifically in principal cells from the proximal caput epididymidis and germ cells from the seminiferous epithelium.

Animals↗
PubMed1984

Receptor-mediated endocytosis of alpha 2-macroglobulin and transferrin in rat caput epididymal epithelial cells in vitro.

The endocytic activity of epithelial cells from the rat epididymis in vitro has been examined by following the uptake of tracer compounds conjugated to proteins. Transferrin-gold and alpha 2-macroglobulin-gold were taken up initially in coated pits, internalized and sequestered into tubular-vesicular structures, multivesicular bodies and, in the case of alpha 2-macroglobulin, into lysosomes. Uptake could be prevented by an excess of unlabeled protein. Studies using 125I-alpha 2-macroglobulin and 125I-transferrin also showed that the uptake of these proteins was specific and could be displaced with increasing amounts of unlabeled protein. In addition, binding of 125I-transferrin to cells was saturable at 4 degrees C. These studies indicate that transferrin and alpha 2-macroglobulin are taken up by receptor-mediated endocytosis. In contrast, a fluid phase marker, bovine serum albumin-gold (BSA-gold), was initially taken up predominantly in uncoated caveolae rather than coated pits, and could not be displaced with excess BSA. By virtue of their charge, polycationized ferritin and unlabeled colloidal gold were taken up and internalized by adsorptive endocytosis, a pathway which is similar to fluid phase endocytosis. The uptake and internalization of alpha 2-macroglobulin and transferrin differed in a number of respects. Uptake and internalization of alpha 2-macroglobulin but not of transferrin was dependent on extracellular calcium. Only alpha 2-macroglobulin was transferred into lysosomes, whereas transferrin was recycled to the cell surface. Although the proton ionophore, monensin, and the transglutaminase inhibitor, dansylcadaverine, did not stop uptake and internalization of either alpha 2-macroglobulin or transferrin, they did prevent the transfer of alpha 2-macroglobulin to lysosomes.

Adsorption↗
PubMed1983

Sertoli cells of the golden-mantled ground squirrel (Spermophilus lateralis): a model system for the study of shape change.

Sertoli cells of the ground squirrel (Spermophilus lateralis), a seasonal breeder, were examined by light and electron microscopy and their structure, particularly the organization of the cytoskeleton, was related to events that occur in the seminiferous epithelium during spermatogenesis. Among the events considered and described are the apical movement of elongate spermatids, withdrawal of residual cytoplasm from germ cells, transport of smooth endoplasmic reticulum (SER) between the base and apex of the Sertoli cells, and sperm release. These events are dramatically evident in this species because the seminiferous epithelium is thin, i.e., there are few germ cells, and both the germ cells and Sertoli cells are large. Sertoli cells of the ground squirrel have a remarkably well developed cytoskeleton. Microfilaments occur throughout the cell but are most evident in ectoplasmic specializations associated with junctions. Intermediate filaments occur around the nucleus, as a layer at the base of the cell, and adjacent to desmosome-like junctions with germ cells. Intermediate filaments, together with microtubules, are also abundant in regions of the cell involved with the transport of SER, in cytoplasm associated with elongate spermatids, and in processes that extend into the residual cytoplasm of germ cells. Our observations of ultrastructure are consistent with the hypothesis that Sertoli cell microtubules are involved with the movement of germ cells within the seminiferous epithelium, and further implicate these structures as possibly playing a role in the retraction of residual cytoplasm from germ cells and the intracellular transport of SER. The abundance and organization of intermediate filaments suggest that these cytoskeletal elements may also be involved with events that occur during spermatogenesis.

Animals↗
PubMed1983

Colchicine-induced changes in the cytoskeleton of the golden-mantled ground squirrel (Spermophilus lateralis) Sertoli cells.

Study of Sertoli cells of the ground squirrel provides a unique opportunity to examine cell structure and function. The cells are large, have an elaborate cytoskeleton, and undergo dramatic changes in organization during spermatogenesis. Microtubules (MTs) are prominent elements of the cytoskeleton and appear to be associated structurally with many of the events that occur during sperm production. To investigate the function of MTs, animals were injected subcutaneously with colchicine, and their seminiferous epithelia examined by light and electron microscopy. Some animals were injected with 30--80 mg of the drug per kg body weight and sacrificed 3 to 5 hr later. Others were given 0.3 mg/kg/day for 6 days and processed on day 7. Virtually no MTs were seen in Sertoli cells after short-term treatments, and their numbers were greatly reduced after the long-term injections. Intermediate filaments were very evident throughout the cytoplasm of treated cells, particularly in the short-term studies. Moreover, a close association of some of these filaments with centrioles was observed. In all cases, elongate spermatids which normally move apically did not do so. Indeed, some spermatids appear to have been pulled to a basal position after having moved apically prior to treatment. Also, smooth endoplasmic reticulum (SER) accumulated basally in the Sertoli cell, unlike controls, and the acrosomes of late spermatids developed abnormally or did not complete their shape changes. Cell junctions appeared normal and sperm release was observed. In conclusion, our data suggest that Sertoli cell MTs are necessary for the normal development and translocation of spermatids in the seminiferous epithelium and are involved with positional changes in Sertoli cell SER. They do not appear essential for the maintenance of cell junctions.

Animals↗
PubMed1983

Spermatogenesis in the vasectomized monkey: quantitative analysis.

The seminiferous epithelium in mature vasectomized Macaca fascicularis was examined quantitatively to assess spermatogenesis. Monkeys were bilaterally vasectomized and controls were bilaterally sham operated. At postoperative periods of 10 and 18 months, groups of monkeys were castrated and their testes prepared for morphologic analysis. Diameters were measured in 100 cross sections of seminiferous tubules from each animal. Numbers of spermatogonia (Ad and Ap), preleptotene spermatocytes, pachytene spermatocytes, and step 7 spermatids, relative 10 Sertoli cell nucleoli, were counted in stage VII tubules. Tubule diameter and germ cell numbers per Sertoli cell nucleoli were not altered by vasectomy. Our study demonstrates quantitatively that spermatogenesis in the monkey is not inhibited up to 18 months following vasectomy.

Animals↗
PubMed1982

Effects of age and 1,4,6-androstatriene-3,17-dione on activities of hCG-induced 19-hydroxylase and aromatase of rat testes.

The testicular aromatase activity was significantly increased by administration of hCG to 18-day old rats, but not increased in 29-day old rats. 1,4,6-Androstatriene-3,17-dione did not inhibit the in vitro conversion of testosterone to 19-hydroxytestosterone by the testicular cell-free homogenates of the hCG-treated 18-day old rats, but strongly suppressed production of estrogen from 19-hydroxyandrostenedione. On the other hand, the 19-hydroxylase activity of 18-day old rats stimulated by hCG was reduced to about 50% of the control value by SKF-525A, SU-4,885, SU-8,000 and SU-9,055 at their concentration of 10(-4) M. From our results, it is postulated that there are two distinct steps in the process of aromatization of testosterone, the one, the primary 19-hydroxylation which is inhibited by SKF-525A and SU-compounds, but not by 1,4,6-androstatriene-3,17-dione, and the other, aromatization of 19-hydroxylated androgen which is inhibited by both 1,4,6-androstatriene-3,17-dione and non-steroidal inhibitors.

Aging↗
PubMed1981

Immunocytochemical localization of androgen-binding protein in the male rat reproductive tract.

The localization of androgen-binding protein (ABP) in the reproductive tract of young adult male rats was studied with the peroxidase-antiperoxidase technique using frozen sections and light microscopy. Within the seminiferous tubules, a positive reaction was noted in the apical portion of the epithelium, apparently in spermatids and/or Sertoli cells. ABP was localized in granules in the apical cytoplasm of the principal epithelial cells of the proximal part of the caput epididymis and in the epithelial cells of the ductuli efferentes. The cells in the distal part of the caput as well as the corpus and cauda of the epididymis did not contain ABP. Numerous coated vesicles and multivesicular bodies were present in the supranuclear cytoplasm of the epididymal epithelium where ABP was taken up. The results indicate that ABP is taken up from the lumen by epithelial cells of the ductuli efferentes and proximal part of the caput epididymis.

Androgen-Binding Protein↗
PubMed1979

Testicular testosterone concentration following testicular vein ligation.

UNLABELLED: A rat model to study the local effects of testicular vein ligation is described. One hour after unilateral testicular vein ligation, the testicular concentration of testosterone was significantly greater (P less than 0.01) on the side that was ligated than in the contralateral testis (177.1 +/- 19.7 [SEM] ng/gm of tissue as compared with 108.8 +/- 11.8 ng/gm of tissue, respectively). This effect was not seen 1 week after the testicular vein ligation. The testosterone concentration in the ligated testis was also higher than that in sham-operated animals. These differences in testicular testosterone concentration were not associated with changes in peripheral serum testosterone levels. CONCLUSION: ligation of the testicular vein causes an acute rise in the testicular concentration of testosterone and may thus mediate changes in testicular function.

Animals↗
PubMed1979

Protein carboxyl-methylation in rat testes: a study of inherited and X-ray-induced seminiferous tubule failure.

Protein carboxyl-methylase (PCM), the enzyme that transfers methyl groups from S-adenosyl-methionine to free carboxyl groups on proteins, is highly localized in testes. The cellular distribution of PCM and its substrates, the methyl acceptor proteins, was investigated. Separation of testicular cells on an albumin gravity gradient revealed the preferential localization of both enzyme and substrates in spermatids. In young rats, PCM activity increases with age coincidently with germ cell maturation. Rats which are heterozygous for the Hre gene (Hre/+) are infertile as a result of germ cell depletion. In these animals, testicular PCM specific activity and total activity were, respectively, 4--6 and 40--50 times lower than in normal testes. Enzyme activity in testes from animals with x-ray-induced germ cell depletion was also very low. These observations suggest that PCM is located in germ cells.

Animals↗
PubMed1979

Is FSH required for maintenance of spermatogenesis in adult rats?

Administration of FSH antiserum to adult rats for 14 or 30 days had no or little effect on body, testis or accessory sex gland weights, androgen-binding protein, testosterone levels, germ cell numbers or fertility, thus indicating a relative insensitivity of the testis to withdrawal of FSH. Unlike immature rats, therefore, which do require FSH to initiate spermatogenesis, adult rats do not need this hormone to maintain spermatogenesis.

Age Factors↗