THE POSSIBLE INVOLVEMENT OF TISSUE-TYPE PLASMINOGEN-ACTIVATOR IN LUTEOLYSIS OF RHESUS-MONKEY

Changes of plasminogen activators (PA) during different stages of development of the corpus luteum, and their possible physiological role in luteolysis were studied in rhesus monkeys. It was demonstrated for the first time that monkey corpus luteal cells not only produce PA, but that the function of the corpus luteum is also closely related to the activity of this enzyme system. Generally, the life span for a corpus luteum in monkey is approximately 14-16 days, its demise beginning thereafter. In the present study, we found that urokinase in the corpus luteum is higher on day 5 and day 10 after human chorionic gonadotrophin injection, while the tissue type (t) PA is mainly produced on day 13 when luteolysis may take place. Progesterone production remained high on day 5 and day 10 and decreased dramatically from day 13, indicating the important role of tPA but not urokinase (u) PA in suppressing luteal function. When purified tPA (but not uPA) monoclonal antibody was added to luteal cell culture to neutralize endogenously produced tPA activity, progesterone production in the cells was increased significantly. Interestingly, prolactin alone was capable of increasing PA production by luteal cells; prolactin together with luteinizing hormone, however, had a synergistic luteotrophic effect.

Ovulation in the tree shrew (Tupaia belangeri) induced by gonadotrophins

The induction of ovulation by exogenous gonadotrophins is an important approach for recovering oocytes used for studies on the reproductive biology of some mammals. In the present study, pregnant mare serum gonadotrophin (PMSG) and human chorionic gonadot

HORMONAL-REGULATION OF TISSUE-TYPE PLASMINOGEN-ACTIVATOR AND PLASMINOGEN-ACTIVATOR INHIBITOR TYPE-1 IN CULTURED MONKEY SERTOLI CELLS

Sertoli cells play a central role in the control and maintenance of spermatogenesis. Isolated Sertoli cells of mouse and rat testes have been shown to secrete plasminogen activator (PA) and a plasminogen activator inhibitor type-1 (PAI-1) in culture. In this study, we have investigated the hormonal regulation of PA and PAI-1 activities in cultured monkey Sertoli cells. Sertoli cells (5x10(5) cells/well) isolated from infant rhesus monkey testes were preincubated at 35 degrees C for 16 h in 24-well plates precoated with poly(D-lysine) (5 mu g/cm(2)) in 0.5 mi McCoy's 5a medium containing 5% of fetal calf serum and further incubated for 48 h in 0.5 mi serum-free medium with or without various hormones or other compounds, PA as well as PAI-1 activities in the conditioned media were assayed by fibrin overlay and reverse fibrin autography techniques respectively. The Sertoli cells in vitro secreted only tissue-type PA (tPA), no detectable amount of urokinase-type PA (uPA) could be observed, Monkey Sertoli cells were also capable of secreting PAI-1, Immunocytochemical studies indicated that both tPA and PAI-1 positive staining localized in the Sertoli cells, spermatids and residual bodies of the seminiferous epithelium; Northern blot analysis further confirmed the presence of both tPA and PAI-1 mRNA in monkey Sertoli cells. Addition of follicle-stimulating hormone (FSH) or cyclic adenosine monophosphate (cAMP) derivatives or cAMP-generating agents and gonadotrophin-releasing hormone (GnRH) agonist or phorbol ester (PMA) to the cell culture significantly increased tPA activity. PAI-1 activity in the culture was also enhanced by these reagents except 8-bromo-dibutyryl-cAMP, forskolin and 3-isobutyl-1-methylxanthin (MIX) which greatly stimulated tPA activity, whereas decreased PAI-1 activity, implying that neutralization of PAI-1 activity by tile high level of tPA in the conditioned media may occur. These data suggest that increased intracellular signals which activate protein kinase A (PKA), or protein kinase C (PKC) can modulate Sertoli cell tPA and PAI-1 activities, The concomitant induction of PA and PAI-1 by the same reagents in the Sertoli cells may reflect a finely tuned regulatory mechanism in which PAI-1 could limit the excession of the proteolysis.

Observations on the spawning behavior of artificially matured Japanese eels Anguilla japonica in captivity

Spawning behavior of artificially matured Japanese eels Anguillo japonica in captivity was investigated using a DVD Video image system. Following a routine hormone treatment technique for this fish, female eels were artificially matured by weekly intramuscular injections of salmon pituitary extracts (SPE) at a dosage of 40 mg kg(-1) BW for a total of 7-11 doses to induce ovarian maturation, while male eels received weekly intramuscular injections of human chorionic gonadotropin (HCG) at a dosage of 1000 IU kg(-1) BW for a total of 6-11 doses at 18 degrees C to induce testicular maturation in a separate aquarium. In this experiment, three pairs of such hormone-treated matured eels were acclimatized in seawater in 1.5 m(3) experimental aquaria with or without shelters at 20 degrees C for 24 h. Twenty four hours after the acclimatization terminated, the females received SPE injections to boost maturation and ovulation. Twenty four hours following these injections, the females received injections of HCG (1000 IU per fish, HCG injection) and 17 alpha-hydroxyprogesterone (2 mg per fish) to induce ovulation, while males were given HCG injections (1000 IU per fish, HCG injection) to induce spermiation. Video taping started after the 24 h acclimatization terminated and last for a total of 96 h. Before the HCG injections, both sexes were inactive, staying on the bottom or in shelters if available. Following these HCG injections, they became active and frequently left the bottom swimming in the water column. During the 24 h following HCG injections, activity accounted for 67% and 45% of the total activity in no shelter treatment for females and males, respectively, in comparison with 77% and 78% in shelter treatment. Activity was significantly more pronounced during this phase than during other phases for each sex in either shelter treatment. Egg release and sperm ejection occurred in the water column around the time eels' activity reached peaks. Eels either returned into the shelters or stayed motionlessly on the bottom of the aquaria after egg release and sperm ejection. Eight out of nine (89%) females in no shelter treatment spontaneously released eggs with a total of 11 batches 14-18 h following HCG injections, in contrast with four out of nine (44%) females releasing eggs for 4 batches 16-20 h in shelter treatment. Males arrived at activity peaks 11-13 h following HCG injections in no shelter treatment, 2-4 h ahead of the females (14-16 h), in comparison with 8-11 h in shelter treatment with 5-6 h ahead of the females (14-17 h). Courtship behavior indicative of spawning such as pairing, chasing and touching bodies was not observed in the eels in this study. However, on many occasions, eels of both sexes (male-female or female-female) were found to "cruise together" in water column for a short time period or frequently come together prior to releasing eggs and ejecting sperm, suggesting the possibility of group mating in artificially matured Japanese eels. (c) 2007 Elsevier B.V. All rights reserved.