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K K Vihko

Publications and source records attributed to K K Vihko.

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Local synthesis of plasminogen by the seminiferous tubules of the testis.

Plasminogen activation occurs through conversion of plasminogen to plasmin by plasminogen activators. In adult mammals, liver has been the only known site for plasminogen synthesis. Seminiferous tubules secrete plasminogen activator, but are behind a barrier that excludes the entrance of many macromolecules to this tissue. Therefore, it became of interest to study the existence of plasminogen in this system. After metabolic labeling, a 90 kDa lysine-binding polypeptide was found. This protein could be immunoblotted by anti-plasminogen antibody, and was shown to possess urokinase-dependent proteolytic activity. The findings suggest that plasminogen is synthesized by seminiferous tubules. We propose that local plasminogen synthesis may occur in tissues which are separated by specific anatomical barriers.

Affinity Labels↗

Testicular plasminogen activators during postnatal development in the rat.

In the seminiferous epithelium, Sertoli cells secrete plasminogen activator (PA) under regulation of follicle stimulating hormone, cyclic AMP and neighbouring spermatogenic cells. Recent observations suggest that preleptotene spermatocytes upon their release from the basement membrane of the seminiferous tubule are important regulators of PA secretion. To study further the role of PA's in the seminiferous tubules, we have analyzed the endogenous levels and secretion rates of PA at various ages during postnatal development, and performed biochemical analyses of the types of PA in the testis and spent media from seminiferous tubular cultures. Cyclic secretion of PA started at the age of 28 days, and from 40 days onwards, the high secretion rates were localized in stages VII and VIII of the cycle of the seminiferous epithelium. The secreted PA is most obviously of the urokinase type; both urokinase-type and tissue-type PA-like activities were found in seminiferous tubular homogenates. The increase in testicular PA levels concomitant to the onset of meiosis in the epithelium was due to the urokinase-type PA-like activity.

Aging↗

Regulation of stages VI and VIII of the rat seminiferous epithelial cycle in vitro.

Plasminogen activator (PA) is secreted cyclically (in stages VII and VIII) by rat seminiferous tubules. To investigate whether this can be maintained and influenced in vitro, tubule segments from stages VI and VIII of the epithelial cycle were cultured for 3 days in chemically defined medium supplemented with testosterone, FSH, or a combination of testosterone, FSH, insulin and retinoic acid (4F). Morphological and flow cytometric analyses of stage VI tubules suggested a roughly normal differentiation to stage VIII. They developed an increased PA secretion on day 3 of culture. Stage VIII tubules, however, did not develop all the characteristics of stage XII. Step 8 spermatids did not elongate and step 19 spermatids failed to develop into spermatozoa. Secretion of PA on day 3 was not significantly different to that on day 1. The 4F combination very significantly stimulated PA secretion in both stages, but FSH alone was effective only in stage VIII. Most of the secreted PA had a molecular weight of 43,000 in both stages, suggesting that it is of urokinase type. The results suggest that stage VI is more able to differentiate in vitro for 3 days than stage VIII; the cyclic secretion pattern of PA was partially maintained in tubule segments from stage VI. Follicle-stimulating hormone had an effect on PA secretion only in stage VIII, whereas the 4F combination was stimulatory in both stages. The retinoic acid in this combination may be of importance in the regulation of PA secretion by seminiferous tubules.

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Cell interactions in the rat seminiferous epithelium with special reference to the cellular distribution of calmodulin.

Spermatogenesis is dependent on stimulation by pituitary gonadotropins, FSH and LH. Targets for these hormones are Sertoli and Leydig cells, respectively. The effect of LH on spermatogenesis is mediated by testosterone. In addition to hormones, interactions between neighbouring cells seem to regulate spermatogenesis. This is reflected by cyclic secretion of several proteins by the seminiferous epithelium, of which plasminogen activator is a good example. While it is controlled by FSH a factor in preleptotene spermatocytes may also influence its cyclic secretion pattern. Both testosterone and FSH have a cyclic action in the seminiferous epithelium. The androgens seem to predominate in stages where spermiation, onset of meiosis and the highest rate of RNA transcription occur (VII-XI). FSH is most active in stages that contain meiotic divisions and early spermiogenesis (XIII-V), greatly stimulating the production of cyclic AMP. To investigate further the "second messengers" of FSH action in the seminiferous epithelium, the cellular distribution of calmodulin was analyzed using an indirect immunocytochemical method. In addition to their clear cyclic distribution in primary spermatocytes and in spermatids, Sertoli cells also showed a bright calmodulin immunofluorescence that was apparently cyclic. These observations suggest a local calmodulin and calcium regulation of spermatogenesis.

Androgens↗

Cellular regulation of plasminogen activator secretion during spermatogenesis.

The secretion of plasminogen activator (PA) has been found to be highly stage-specific during rat spermatogenesis. It is maximal in Stages VII and VIII of the cycle. At these stages, seminiferous tubules contain primitive type A1 spermatogonia, preleptotene and midpachytene primary spermatocytes, round and maturation-phase spermatids and Sertoli cells. The last cell types are the most likely sources of PA. To investigate which cell type might be involved in the regulation of PA secretion, we have sequentially isolated 1-mm segments of rat seminiferous tubules from Stages VI-IX with transillumination-assisted microdissection and measured PA secretion using 125I-labeled fibrinogen as substrate. In another experiment, spermatogenia were killed by 300 rads of x-rays and PA secretion was analyzed during the absence of desired germ cell classes. The results support the idea that upon their detachment from the basal lamina, preleptotene-stage primary spermatocytes have the major stimulatory action on the PA secretion of the seminiferous tubules. Other phenomena such as spermiation, phagocytosis of residual bodies or opening up of the Sertoli cell junctions seem to influence PA secretion to a lesser extent.

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