The function of proteins, peptides, proteases and inhibitors of proteases in modulations of regulation mechanisms of gonadotrophins during the development of ovarian folicles has not been fully explained up to now. We can see difference reactions of ewes to superovulation stimulations in oestrous and anoestrous periods as shown by the variation of the antiproteolytic activity of blood plasma and cervical mucus. Trypsin is used as a model for serine protease, and trypsin inhibitory activity (TIA) was measured from the reduced rate of trypsin hydrolysis of the chromogenic substrate N-alpha-tosyl-L-arginine-4-nitroanilide (TAPA, Bartík et al., 1974). Full hydrolytic activity was determined as a change in absorbency at 405 nm = 1.0 after ten-minute incubation at 25 degrees C and pH = 8.1, and inhibition was expressed in percentage of full activity. Statistical analyses were performed by Student's t-test. Twenty-three ewes in anoestrus and twenty-eight ewes in oestrus were included in this experiment. They were of the Slovak Merino breed, two to three years old, with the mean live weight of thirty to forty kg. The ewes were treated with Ageline vaginal sponges (20 mg chlorsuperlutin/sponge) to provide for synchronization of ovarian activity, in the interval of eleven to twelve days. After removal of sponges, the ewes were stimulated for superovulation as shown in Tab. I. Heparinized blood plasma samples were stored at -25 degrees C. Low molecular TIA activities were determined in HClO4--treated blood plasma. Samples of cervical mucus were taken on cotton-wool tampons which were evaluated in 0.2 M Tris-HCl buffer, pH = 8.1. Figs. 1-6 shows TIA changes in blood plasma and cervical mucus on particular days. Fig. 7 shows the mean values of controls (I-initial) after synchronization (A) and after stimulation (S). TIA changes were different in anoestrous and oestrous periods. Differences in TIA changes in blood plasma and cervical mucus in oestrous period suggested certain local regulation mechanisms of synthesis and/or secretion of this activity in the cervix. Some values of TIA were maximum at the time of expected heat or ovulation and may be some of the factor(s) which influenced fertility of females after superovulation stimulations.
- MeSH
- cervikální hlen metabolismus MeSH
- chlormadinon analogy a deriváty farmakologie MeSH
- inhibitory trypsinu krev metabolismus MeSH
- ovce metabolismus MeSH
- superovulace účinky léků MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- chlormadinon MeSH
- chlorsuperlutin MeSH Prohlížeč
- inhibitory trypsinu MeSH
The antiproteolytic activities of the blood plasma (BP) and cervical mucus (CM) determined as trypsin inhibiting activities (TIA) where trypsin served as a model serine protease, were variable after superovulatory stimulations of ewes and after their gamma irradiation (2.45 Gy). TIA's were determined from the reduction in the bovine trypsin hydrolysis of the low-molecular chromogenic substrate N-alpha-tosyl-L-arginyl-p-nitro anilide (TAPA) (Bartík et al., 1974). The inhibition was expressed in per cent when delta A405 = 1.0 for 10-minute incubation at 25 degrees C, pH = 8.05, 0.2 mol/l tris-HCl buffer, was taken as 100%. Student's t-test was used for the statistical evaluation. A hundred ewes of the Merino breed were applied Agelin vaginal tampons (20 mg chlor-superlutin per head) on day 1 for 10 days in the anoestric period (May). A part of these ewes were subjected to whole-body gamma irradiation on days 6 to 11. When the irradiation was completed and the tampons were taken out, the ewes (three to four years old lambing ewes, yearling ewes) were stimulated to superovulations by an administration of 1500 IU serum gonadotropin (SG) or 450 IU follicle stimulating hormone (FSH). These parameters were followed in the subsequent 5-6 days: BP TIA (Figs. 1 and 4), fraction of low-molecular BP (n) TIA (Figs. 2 and 5), CM TIA (Figs. 3 and 6). Fig. 7 shows the average values of the results. The lambing ewes and yearling ewes had various responses to irradiation and superovulatory stimulation. Gamma irradiation eliminated the increase in BP TIA (P < 0.001) in the ewes after stimulations (Fig. 1a, b). The yearling ewes showed nonsignificant changes (ns) as their responses to SG stimulations and gamma irradiation, but the responses on the particular days of the trial were different (Fig. 1c, d--FSH stimulation). Similar changes were observed in the BP nTIA fraction. The changes in the average values of CM TIA were nonsignificant in the lambing ewes, the gamma irradiation reduced CM TIA only after FSH stimulation (P < 0.002). The amount of cervical mucus decreased after stimulations and irradiation, the thickness of ovarian epithelium and muscles was reduced in the particular parts, differently in lambing ewes and in yearling ewes.(ABSTRACT TRUNCATED AT 400 WORDS)
- MeSH
- anestrus * MeSH
- cervikální hlen metabolismus účinky záření MeSH
- estradiol farmakologie MeSH
- inhibitory proteas krev metabolismus účinky záření MeSH
- ovarium účinky léků účinky záření MeSH
- ovce krev MeSH
- progesteron farmakologie MeSH
- superovulace MeSH
- záření gama MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- estradiol MeSH
- inhibitory proteas MeSH
- progesteron MeSH
Serine proteases help to regulate the ovarian cycle at different levels and they are subjected to the control of gonadotropic hormones and protease inhibitors. Superovulation stimulations influence the activities of trypsin inhibitors (model serine protease) in blood plasma (BP) and in follicular fluid (FF), and also in dependence on the breed. Trypsin inhibiting activities were determined from the reduced rate of trypsin hydrolysis of a chromogenic substrate (TAPA) and they were determined in percent. A change in absorbancy at 405 nm = 1.0 after 10-minute incubation at 25 degrees C and pH = 8.05 was taken as 100%. The incubation mixture as a sample contained 100 microliters blood plasma or 10 microliters follicular fluid, diluted with gammaglobulin at 1:10. The differences in the trypsin inhibiting activities (TIA) of BP in ewes of the Merino, Tsigai and Wallachian breeds were insignificant, but Agelin synchronization (20 mg chlorsuperlutin per vaginal swab) induced statistically significant differences. The lowest TIA BP was recorded in the Tsigai breed (T), P less than 0.001 in comparison with the Wallachian (W) and Merino (M) breeds. Following the administration of 1,500 IU PMSG, the TIA BP within 120 hours decreased in W (P less than 0.001), it increased in T (P less than 0.1) and in M the changes in the TIA BP were insignificant. The average numbers of ovulations increased from 2.25 +/- 2.5 to 3.0 +/- 1.2 in W; from 0.25 +/- 0.43 to 2.5 +/- 1.6 in T and from 0.00 +/- 0.0 to 2.5 +/- 2.3 in M. Following the single administration of 2,000 IU PMSG after Agelin synchronization, the changes in M ewes were also insignificant, and there were no different responses in pregnant (1st to 2nd month) and nonpregnant ewes. In pregnant T ewes the TIA BP increased after Agelin synchronization and stimulation (P less than 0.01), in nonpregnant ewes these changes were not significant. In W lambing ewes the TIA BP increased (P less than 0.001), the effects of Agelin were greater than those of PMSG. The TIA of follicular fluid (FF) of antral follicles were on average tenfold if compared with BP. After hormonal treatment of ovaries, the TIA FF mostly increased at different levels of statistical significance. The TIA FF of follicles less than 10 mm were lower than in follicles greater than 10 mm (P less than 0.001 for M and W, P less than 0.1 for T).(ABSTRACT TRUNCATED AT 400 WORDS)
- MeSH
- folikulární tekutina chemie MeSH
- inhibitory trypsinu analýza krev MeSH
- ovce fyziologie MeSH
- superovulace účinky léků MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- inhibitory trypsinu MeSH
The synthesis and secretion of trypsin (trypsin model serine protease) inhibitors are regulated in ovarian follicles by gonadotropins. The superovulation stimulations with 400 IU FSH, 1000 IU PMSG, 1000 IU HCG, 750 IIU PMSG + 750 IU HCG influence in a different way the trypsin inhibiting activities (TIA) of blood plasma (BP) (Figs 1 and 2) and follicular fluid (fig. 3); this points to a possibility of local effects. An increase in the average values of TIA in BP was statistically significant during the whole experiment: P less than 0.05 to P less than 0.001 (following the administration of PMSG+HCG, or PMSG, and HCG); Antisergon administered in 68 hours after PMSG reduced this increase. The changes in the fraction of low-molecular TIA in BP (after BP treatment with perchloric acid) were of converse nature; a decrease in the average values ranged from P less than 0.02 to P less than 0.001 (following PMSG or other stimulations). Antisergon did not influence this decrease. The changes observed on particular days of the trial (Figs. 1 and 2) also indicate different effects of the preparations, mainly of the component LH, which resulted in the occurrence of large nonovulating follicles (greater than 10 mm--"cystic" ones). No such follicles were observed in nonstimulated ewes and after FSH stimulation. The administration of antisergon (goat's antiserum against PMSG) 68 hours after PMSG administration did not prevent their creation. The TIA of follicular fluid (FF) of antral follicles was on average tenfold in comparison with that of blood plasma; and the TIA FF of follicles greater than 10 mm was higher (up to P less than 0.001) than the TIA FF of follicles less than 10 mm. The administration of Antisergon in shorter intervals following PMSG administration (12, 24, 48 and 58 hours) influenced the average values of TIA BP in 120 hours (since PMSG administration) in dependence on time (Tab. I). The effects of Antisergon administered in 12 and 24 hours after PMSG administration on the TIA BP were insignificant if it was administered in 48 and 58 hours the TIA BP increased (P less than 0.02; P less than 0.001) in comparison with the interval of 12 hours. The TIA FF of follicles less than 5 mm, 5-10 mm and greater than 10 mm varied in dependence on the time intervals of Antisergon administration (Fig. 4). The statistical significance of these changes in shown in Tab. II.(ABSTRACT TRUNCATED AT 400 WORDS)
- MeSH
- folikulární tekutina chemie MeSH
- gonadotropiny koňské farmakologie MeSH
- inhibitory trypsinu analýza krev MeSH
- ovce metabolismus MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- gonadotropiny koňské MeSH
- inhibitory trypsinu MeSH
Superovulation treatment leaves alternations in the controlling regions of the hypothalamus and in the adjacent ependyme after ovulation. The test ewes were synchronized with Agelin (20 mg chlorsuperlutin in one vaginal sponge) and stimulated (after the removal of the sponges) with 750 IU PMSG + 750 IU HCG and with 1000 IU HCG and 750 IU PMSG + 5 ml Antisergon (goat antiserum against PMSG), administered 68 hours after PMSG (i.e. 40 hours after HCG). The control ewes were in different stages of the ovarial cycle. The experimental ewes were killed 120 to 130 hours after the start of stimulation. Routine histological techniques were used to treat the brain samples; this treatment was followed by assessment under light microscope. The ependyme epithelium of the third cerebral chamber was studied under scanning microscope. Preparations with different FSH:LH ratios had different effects on the nucleus ventromedialis. Antisergon administration influenced the secretion of NPV (prevented persistent stimulation), which was observed after administration of PMSG + HCG. On the surface of the lower part of the third cerebral chamber the administration of Antisergon slowed the formation of the miniblebs. Supraependyme cells disappeared after stimulation for superovulation.
- MeSH
- choriogonadotropin farmakologie MeSH
- ependym účinky léků ultrastruktura MeSH
- gonadotropiny koňské farmakologie MeSH
- hypothalamus účinky léků ultrastruktura MeSH
- indukce ovulace MeSH
- ovce anatomie a histologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- choriogonadotropin MeSH
- gonadotropiny koňské MeSH
The quantitative micromorphological changes of tertiary follicles and corpora lutea (CL) were studied in ewes in the autumn mating season after oestrus synchronization, induced by administration of PGF2 alpha (Oestrophan Spofa) at a rate of 125 micrograms, and after superovulation, induced by administration of PMSG (Antex Leo, Denmark) at a rate of 1000 I. U., or PMSG at rates of 750 and 1000 I. U. together with 50,000 I. U. vitamin A (Axerophthol Spofa). The highest number of ovulations was obtained in ewes treated with 1000 I. U. together with vitamin A (3.4 +/- 3.0) and after administration of 1000 I. U. PMSG alone (2.6 +/- 2.74). The highest number of tertiary follicles was recorded in ewes after administration of PGF2 alpha. The proportion of tertiary atretic follicles was the highest in ewes after administration of PMSG (64.6%). The occurrence of the luteinizing form of atresia was recorded only in ewes treated with PMSG (4% of the total number of atretic follicles). Using the caryometric analysis of the luteal cells of corpora lutea, the ewes of the experimental groups had two-peak variation curves; this corresponds to the theory of the presence of two luteal types in the tissue of the corpus luteum in ewes. As determined morphometrically, the smallest proportion of connective tissue out of the total volume of ewes' ovaries was found after administration of 1000 I. U. PMSG together with vitamin A. Administration of vitamin A together with PMSG had a favourable influence on the over-all follicular response, on the average number of ovulations, and on the proportion of non-atretic follicles.
- MeSH
- dinoprost farmakologie MeSH
- gonadotropiny koňské farmakologie MeSH
- ovariální folikul účinky léků MeSH
- ovarium cytologie účinky léků MeSH
- ovce anatomie a histologie MeSH
- superovulace účinky léků MeSH
- synchronizace říje * MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- dinoprost MeSH
- gonadotropiny koňské MeSH
In twenty ewes of the Slovak Merino breed coming from a demonstration farm at Zemplínska Teplica, to the age of two-three years biometrical variations of the sex organs and overall follicular response to PMSG and PGF2 alpha administration were investigated in the autumn period (October-November). In the ewes of all groups the heat was synchronized by i. m. administration of PGF2 alpha (Oestrophan Spofa) at a dose of 125 micrograms in the interval of 11 days. On the ninth day the ewes of the second and fourth group were given 1000 i. u. of PMSG (Antex Leo, Denmark), the ewes of the third group 750 i. u. of PMSG. The ewes of the third and fourth group were administered at the same time 50,000 i. u. vitamin A (Axerophtol Spofa) (each group). The weight and dimensions of the sex organs were investigated. The results demonstrate that the administration of PMSG in the mating period increases significantly the weight of oviducts and horns of uterus in the ewes while there are no variations of the weight of the other sex organs and of their length, or it is lower than in the ewes of the control group. The overall follicular response was not influenced by the higher weight of ovaries.
- MeSH
- biometrie MeSH
- chov MeSH
- dinoprost aplikace a dávkování MeSH
- gonadotropiny koňské aplikace a dávkování MeSH
- ovce anatomie a histologie MeSH
- ovulace * MeSH
- superovulace * MeSH
- synchronizace říje * MeSH
- ženské pohlavní orgány anatomie a histologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- dinoprost MeSH
- gonadotropiny koňské MeSH
The total quantitative changes of ovaries, proportion of atretic and non atretic follicles and changes of tertiary follicles in sheep after administration of increasing doses of PMSG during the anoestrous period were observed. In experimental groups the statistically significant increase of average weight, volume and dimensions of ovaries in comparison with control group were determined biometrically. The average number of tertiary follicles was greater in experimental groups but at the same time we observed a higher proportion of atretic follicles (64% of the total number in the control group; 71-77% in the experimental groups). In the group of sheep administered a dose of 1500 m.u. PMSG we determined a high proportion of luteinized follicles (as much as 21% of the total number of atretic follicles). The total number of small follicles in the so called transient phase in the comparison of experimental and control groups was not changed significantly. In the experimental group an increased incidence of preovulatory follicles and a reduction of tertiary follicle dimensions in the period of follicle cavity formation was determined.
- MeSH
- anestrus účinky léků fyziologie MeSH
- estrus účinky léků MeSH
- gonadotropiny koňské farmakologie MeSH
- ovariální folikul účinky léků MeSH
- ovce fyziologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- gonadotropiny koňské MeSH
The effect of the exposure of the whole body to continuous radiation and of the administration of serum gonadotropin (SG) was studied as exerted on the concentration of catecholamines (epinephrine and norepinephrine) in the hypothalamus, epiphysis and adrenal glands of ewes during the anoestric period with synchronized oestrus. The first group (young barren ewes) and second group (older ewes) were exposed to continuous radiation of 60Co for five days. The radiation was provided at the rate of 0.020 Gy per hour. After the termination of irradiation the ewes were subjected to hormonal stimulation by fractionated administration of 1500 I. U. SG. The third and fourth experimental groups of ewes were stimulated with 1500 I. U. SG without irradiation. Catecholamines were separated from the tissue supernatants by the adsorption chromatographic method and the catecholamine contents in the eluates were determined spectrofluorometrically. Protracted exposure to gamma radiation and hormonal stimulation with SG reduces the concentration of norepinephrine in the whole hypothalamus of the sheep. A statistically significant decrease (P less than 0.001) was recorded in the medial and caudal hypothalamus of the adult ewes and in the rostral and caudal hypothalamus regions of the young ewes. A decrease of norepinephrine concentration, statistically significant in the caudal (P less than 0.01) and medial hypothalamus, was recorded in the group of adult ewes after hormonal stimulation with SG without irradiation. The experimental group of young ewes responds to hormonal stimulation by a greater reduction of norepinephrine contents, as compared with combined exposure to radiation and hormonal stimulation. It is assumed that the decrease in catecholamine concentration after hormonal stimulation with SG is associated with the increase in the content of oestrogens, which act on the adrenergic receptors of the hypothalamus.
- MeSH
- epifýza mozková účinky léků metabolismus účinky záření MeSH
- gonadotropiny farmakologie MeSH
- hypothalamus účinky léků metabolismus účinky záření MeSH
- katecholaminy metabolismus účinky záření MeSH
- nadledviny účinky léků metabolismus účinky záření MeSH
- ovce metabolismus MeSH
- záření gama MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- gonadotropiny MeSH
- katecholaminy MeSH
The effect of exogenous hormonal preparations (PMSG, SG, PMSG + PGF2 alpha and FSH) was studied as exerted on neurosecretion and on changes in the nuclear volume of nucleus paraventricularis, nucleus ventromedialis, nucleus infundibularis and nucleus tuberomamillaris, and on histological changes in the third cerebral ventricle. Changes in catecholamine concentrations were determined in the rostral, medial, and caudal hypothalamus of the ewes. The study was performed with 28 ewes of the Slovak Merino breed during the oestrus period. The oestrus was synchronized by the Agelín vaginal tampons (Spofa). The ewes of four experimental groups were stimulated by the administration of 1000 I.U. PMSG (Antex-Leo), 1000 I. U. Serum Gonadotropin (Bioveta Ivanovice na Hané), 1000 I. U. PMSG with 250 micrograms Oestrophan inj. (Léciva Praha), and 450 I. U. Folistiman (VEB Arzneimittelwerk Dresden, GDR). The brain samples were processed by common histological methods. A caryometric analysis was performed at 3000-fold magnification and by measurement of 200 cells of one sample. The data were subjected to mathematical processing after Fischer and Inke (1956). The amount of neurosecretory material was evaluated by light microscopy (Nakahara, 1963). The multiplication of neurosecretion in the hypothalamic nuclei testifies to the fact that the administration of hormonal preparations stimulates the function of hypothalamic structures, which corresponds with the results of the caryometric analysis. It also ensues from the results that the administration of serum gonadotropins (PMSG and SG) at the dose of 1000 I. U. results in a statistically significant decline (P less than 0.001) of the concentrations of norepinephrine in the rostral, medial and caudal hypothalami of the ewes, although the PMSG preparation has a more pronounced effect when compared with SG. The hormonal stimulation with PGF2 alpha and with FSH causes no significant changes in the concentration of norepinephrine in the hypothalamus of the ewes. As norepinephrine is considered as a neurosecretion inhibitor, the multiplication of neurosecretion in the hypothalamus nuclei after stimulation with serum gonadotropins correlates with the decline of hypothalamic norepinephrine concentrations.
- MeSH
- chlormadinon analogy a deriváty farmakologie MeSH
- dinoprost farmakologie MeSH
- gonadotropiny hypofyzární farmakologie MeSH
- gonadotropiny koňské farmakologie MeSH
- hypothalamus cytologie účinky léků metabolismus MeSH
- katecholaminy metabolismus MeSH
- ovce anatomie a histologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- chlormadinon MeSH
- chlorsuperlutin MeSH Prohlížeč
- dinoprost MeSH
- gonadotropiny hypofyzární MeSH
- gonadotropiny koňské MeSH
- katecholaminy MeSH
