scholarly journals Determination of glycosidase activity in porcine oviductal fluid at the different phases of the estrous cycle

Reproduction ◽  
2008 ◽  
Vol 136 (6) ◽  
pp. 833-842 ◽  
Author(s):  
Luis César Carrasco ◽  
Raquel Romar ◽  
Manuel Avilés ◽  
Joaquín Gadea ◽  
Pilar Coy

Sperm–oocyte binding and gamete–oviductal epithelium interactions are carbohydrate-mediated events occurring in the oviductal fluid (OF). Thus, knowledge about the activities of glycosidases (enzymes catalyzing hydrolytic cleavage of terminal sugar residues) in this milieu would help us understand the molecular mechanisms involved in these events. This work was carried out to investigate the glycosidase activity, protein content, and volume of OF collected from gilts and sows. Oviducts were classified into four phases of the estrous cycle (early follicular, late follicular, early luteal, and late luteal) based on the appearance of the ovaries. OF was aspirated, centrifuged, measured for volume, and frozen until assay. Substrates conjugated to 4-methylumbelliferyl were used to screen the activities of seven different glycosidases at physiological pH (7.2). α-l-Fucosidase and β-N-acetyl-glucosaminidase activities increased at the late follicular phase to decrease after ovulation. β-d-Galactosidase, α-d-mannosidase, and β-N-acetyl-galactosaminidase showed higher activities at the early follicular phase, which decreased after ovulation. N-Acetyl-neuraminidase and α-d-galactosidase did not show activity at any phase of estrous cycle neither in sows nor in gilts at pH 7.2, although it did at acidic pH (4.4) in the follicular and luteal phase samples. Total protein also changed during the cycle showing the maximum secretion at the late follicular phase (2118.6±200.7 μg/oviduct). The highest volumes of OF were collected from the oviducts at the late follicular phase (50.7±1.3 μl/oviduct). These results indicate that OF from sows and gilts shows glycosidase activity varying throughout the estrous cycle suggesting a role of these enzymes in carbohydrate-mediated events.

1993 ◽  
Vol 6 (1) ◽  
pp. 44-46
Author(s):  
Michael W. Jann ◽  
Y.W. Francis Lam

The discipline of pharmacogenetics will continue to expand as scientific and clinical investigations increase our understanding of genetic variabilities in drug metabolism and response. These research efforts will include determination of molecular mechanisms for the different polymorphisms and evaluation of their clinical significance. The availability of molecular methodologies such as restriction fragment length polymorphisms analysis, polymerase chain reaction, and expression of cDNAs in cell cultures will further the investigative work in detection of normal and mutant alleles, identification of new substrates for different polymorphic metabolizing enzymes, and evaluation of mechanisms of individual susceptibility to biological disorders. Other areas such as the role of pharmacogenetics in drug development and regulatory control, in evaluation of potential drug-drug interactions, ethnic variation in polymorphic metabolism, and response, also need to be evaluated.


1992 ◽  
Vol 133 (3) ◽  
pp. 341-NP ◽  
Author(s):  
H. M. Fraser ◽  
K. B. Smith ◽  
S. F. Lunn ◽  
G. M. Cowen ◽  
K. Morris ◽  
...  

ABSTRACT The putative endocrine role of inhibin in the control of FSH secretion during the luteal phase in the primate was investigated by immunoneutralization. Antisera against the 1–23 amino acid sequence of the N-terminus of the human inhibin α subunit were raised in a ewe and three macaques. Antisera (10–20 ml) were administered to macaques on day 8/9 of the luteal phase and serum samples collected during the treatment cycle and post-treatment cycle for determination of FSH, oestradiol and progesterone. In addition, localization of inhibin within the macaque ovary at this stage of the luteal phase was investigated using the ovine antiserum. Intense immunostaining was localized within the granulosa-lutein cells of the corpus luteum with absence of staining in the thecalutein cells or other ovarian compartments. Administration of antisera was without significant effect on serum concentrations of FSH when compared with control animals, either during the first 24 h of detailed observation or for the following 10-day period of the late luteal phase and subsequent early follicular phase. These results provide further evidence that the corpus luteum is the major source of inhibin immunoreactivity during the primate menstrual cycle, but fail to support an endocrine role for inhibin in the suppression of FSH secretion. Journal of Endocrinology (1992) 133, 341–347


2018 ◽  
Vol 99 (6) ◽  
pp. 931-935
Author(s):  
E M Alieva ◽  
N E Akhundova

Aim. To evaluate the effectiveness of correction of the ovarian hyperandrogenism (OHA) in girls in puberty. Methods. 38 girls with OHA syndrome were examined. The average age of the girls was 14.39 ± 0.27 years. Ferriman-Gallwey hirsutism score was 19.7 ± 0.47, the hormonal score 18.14 ± 2.43, the indifferent score 2.34 ± 0.13. The comparison group included 20 girls matched by age (14.74 ± 0.15 years) with physiological course of puberty. The study included the evaluation of hirsutism severity by Ferriman-Gallway score, determination of the hormones of hypothalamus-pituitary-adrenal-ovarian system in early follicular phase on day 5-7 of menstrual cycle. Results. The causes of OHA syndrome in the pubertal period were found to be polycystic ovary syndrome in 68.42 % and tumor-like formations (follicular cysts) in 31.58 % of girls. Compared to healthy girls, the girls with OHA syndrome had statistically significantly higher values of luteinizing hormone (LH) - 9.19 ± 0.96 mME/ml, LH/FSH ratio - 1.8 ± 0.15, total testosteron (Ttotal) - 1.3 ± 0.11 ng/ml, estrone (E1) - 109.5 ± 4.88 ng/ml, androstenedione (An) - 4.01 ± 0.12 ng/ml as well as low values of estradiol (E2) - 49.1 ± 2.6 pg/ml and sex hormone-binding globulin (SHBG) - 45.7 ± 2.14 nmol/l. The study demonstrated that correction of ovarian hyperandrogenism, promotes an increase in the level of E2, SHBG and decrease of the level of LH, LH/FSH ratio, Ttotal, An. Conclusion. Complex pathogenetic therapy of OHA syndrome in girls in puberty leads to significant improvement of hormonal status in this group of patients.


1975 ◽  
Vol 79 (4) ◽  
pp. 625-634 ◽  
Author(s):  
Elwyn M. Grimes ◽  
Irwin E. Thompson ◽  
Melvin L. Taymor

ABSTRACT Thirty-one ovulatory women between 20 and 33 years of age were given 150 μg of synthetic LH-RH during different phases of the menstrual cycle. Five patients were studied during the early follicular phase (days 4–7); 10 patients during the late follicular phase (days 9–12); 6 patients during the "LH Surge"; 5 patients during the early luteal phase (days 14–16); 3 patients during mid-luteal phase (days 17–21); and 2 patients during late luteal phase (days 22–27). Oestrogen, progesterone, FSH and LH levels were determined from 30 min prior to LH-RH administration to 90 min thereafter in all cases. LH response to LH-RH increased progressively during the follicular phase. Enhanced pituitary responsiveness to LH-RH occurred at mid-cycle for both LH and FSH and maximum LH responses occurred during the "LH Surge" and early luteal phase. LH responses during the mid and late luteal phases were similar to late follicular phase responses. There were no significant differences between FSH responses during the early follicular, late follicular, mid-luteal and late luteal phases. Maximum pituitary responsiveness appears to occur in a gonadal steroid milieu of high oestrogen levels in association with rising but low progesterone levels. Progesterone or a crucial oestrogen: progesterone ratio may in fact potentiate pituitary release of LH during the early stages of corpus luteum formation. Pituitary responsiveness to LH-RH correlates positively with basal LH and oestrogen levels during the menstrual cycle and with the oestrogen:progesterone ratio during the luteal phase.


2017 ◽  
Vol 58 (2) ◽  
pp. R87-R106 ◽  
Author(s):  
Emily Tubbs ◽  
Jennifer Rieusset

Beyond the maintenance of cellular homeostasis and the determination of cell fate, ER–mitochondria contact sites, defined as mitochondria-associated membranes (MAM), start to emerge as an important signaling hub that integrates nutrient and hormonal stimuli and adapts cellular metabolism. Here, we summarize the established structural and functional features of MAM and mainly focus on the latest breakthroughs highlighting a crucial role of organelle crosstalk in the control of metabolic homeostasis. Lastly, we discuss recent studies that have revealed the importance of MAM in not only metabolic diseases but also in other pathologies with disrupted metabolism, shedding light on potential common molecular mechanisms and leading hopefully to novel treatment strategies.


2002 ◽  
Vol 1 (2) ◽  
pp. 69-74
Author(s):  
Shuji Takiguchi ◽  
Yasuhiko Nakamura ◽  
Yoshiaki Yamagata ◽  
Hisako Takayama ◽  
Ayako Harada ◽  
...  

1978 ◽  
Vol 89 (1) ◽  
pp. 48-59 ◽  
Author(s):  
M. Ferin ◽  
J. Bogumil ◽  
J. Drewes ◽  
I. Dyrenfurth ◽  
R. Jewelewicz ◽  
...  

ABSTRACT The effects of prolonged gonadotrophin-releasing hormone (GnRH) infusions on LH, FSH, oestrogens and progesterone secretion were studied in female rhesus monkeys at various times of the menstrual cycle and after castration. GnRH was infused at the rate of 15 μg/h for 48 h. This resulted in mean peripheral GnRH levels of 398 ± 31.5 pg/ml (± se) as measured by radioimmunoassay. As expected the pattern of gonadotrophin responses to GnRH varied considerably with the phase of the menstrual cycle. The largest LH increase was seen during the late follicular phase (6-fold over baseline), with a 3-fold increase during the luteal phase and a 2-fold one during the early follicular phase and in the period following the LH surge. Significant FSH increases (4-fold) were seen only during the follicular phase. Oestrogens increased about 2-fold within 4 h of the start of the infusion during the early follicular phase. In the late follicular phase and during the LH surge, they declined within 24 and 1 h, respectively. Large progesterone increases were seen only during the luteal phase. Of special interest is the fact that the increase in gonadotrophin secretion could not be maintained for the entire duration of the experiment even though GnRH continued to be infused at rates sufficient to elicit initial increases of several fold over baseline. Gonadotrophin release declined 4–28 h after the initial stimulation. A further decrease below pre-infusion control levels was particularly evident during the midcycle surge and, for FSH, after ovariectomy. These results indicate that a continuous mode of administration may rapidly induce a desensitization phenomenon at the level of the gonadotroph.


Reproduction ◽  
2001 ◽  
pp. 707-718 ◽  
Author(s):  
A Dhar ◽  
BW Doughton ◽  
E Pruysers ◽  
RW Brown ◽  
JK Findlay

The aims of this study were to investigate the role of inhibin in the distribution of healthy and atretic antral follicles and the secretion patterns of gonadotrophins. Ewes were actively immunized against either alphaN or alphaC of the inhibin alpha subunit with a primary injection and three booster injections. The control ewes received adjuvant only. The ovaries were removed either before or at 24 h after hCG administration in a synchronized follicular phase 48 h after removal of intravaginal progesterone pessaries. Morphological observations were made on every fifth section of the complete ovary (one per ewe) stained with haematoxylin and eosin. The mean number of corpora lutea observed per ewe with corpora lutea was not significantly different in ewes immunized against alphaN (2.4; alphaN-immunized ewes) or alphaC (2.6; alphaC-immunized ewes), and control (2.4) ewes, although some corpora lutea appeared cystic in the immunized ovaries. Compared with luteal phase concentrations, mean basal FSH concentrations in the early follicular phase were significantly increased in the alphaC-immunized ewes, similar in alphaN-immunized ewes and reduced in control ewes. No differences were observed in any of the LH parameters. Before hCG treatment, healthy antral follicles > 1 mm in diameter were not observed in any of the 52 follicles in the aC-immunized ewes and were observed in one of 37 follicles from alphaN-immunized ewes compared with 19 of 28 follicles in control ewes (P < 0.0001). For healthy antral follicles < 1 mm in diameter, there were 72 of 85 follicles in the alphaC-immunized ewes, 79 of 81 follicles in the alphaN-immunized ewes and 81 of 82 follicles in the control ewes. Similar results were obtained in healthy antral follicles < 1 mm in diameter at 24 h after hCG administration. In contrast to the control ewes, no healthy preovulatory follicles (> 6 mm in diameter) were observed in alphaN- and alphaC-immunized ewes either before or 24 h after hCG administration. Two newly formed corpora lutea from alphaC-immunized ovaries contained retained oocytes compared with none in control and alphaN-immunized ovaries. In conclusion, immunization against alphaN and alphaC may result in disruption of the normal processes of antral follicular growth and maturation independent of the concentrations of FSH and LH.


1988 ◽  
Vol 119 (1) ◽  
pp. R5-R8 ◽  
Author(s):  
C. G. Tsonis ◽  
D. T. Baird ◽  
B. K. Campbell ◽  
J. A. Downing ◽  
R. J. Scaramuzzi

ABSTRACT The secretion rates of bioactive inhibin, oestradiol and progesterone were measured during the mid-luteal phase and at various times during the follicular phase of the cycle by a sensitive bioassay using sheep pituitary cells in culture in 12 Booroola ewes with and without copies of the Fecundity (F) gene in which the left ovary had been auto-transplanted to the neck. Inhibin secretion was high during the luteal phase and fell in the early follicular phase in all genotypes (P < 0.01). In Booroola ewes with a F/- genotype, inhibin secretion then increased again, towards luteal rates, in the mid and late follicular phases. In Booroola ewes without a copy of the F gene (+/+) inhibin secretion remained low at all three sampling times in the follicular phase. The secretion rate of inhibin at 36 h (P < 0.1) and 48 h (P < 0.01) were significantly lower in ewes from the +/+ (no copy of the gene) ewes than in F/(one copy of the gene) ewes. Oestradiol secretion was low during the luteal phase and increased steadily during the early (24 h) to a plateau in the mid (36 h; P < 0.01) and late (48 h; P < 0.05) follicular phase. Progesterone secretion was high during the luteal phase, and decreased to a very low rate by 24 h after prostaglandin (PG) treatment (P < 0.001) and remained low. At 24 h after PG the concentration of FSH was significantly lower (P < 0.01) than that during the luteal phase and remained suppressed until the onset of the LH surge. There were no significant differences in LH concentrations. We conclude that (1) the secretion of inhibin by the ovary is highest in the luteal phase and (2) inhibin secretion is significantly raised during the mid to late follicular phase in Booroola ewes with a copy of the Fecundity gene compared with those without.


1990 ◽  
Vol 126 (3) ◽  
pp. 385-393 ◽  
Author(s):  
B. K. Campbell ◽  
G. E. Mann ◽  
A. S. McNeilly ◽  
D. T. Baird

ABSTRACT The pattern of pulsatile secretion of inhibin, oestradiol and androstenedione by the ovary at different stages of the oestrous cycle in sheep was studied in five Finn–Merino ewes in which the left ovary had been autotransplanted to the neck. The ewes had jugular venous blood samples collected at 4-hourly intervals from 42 h before the induction of luteolysis by i.m. injection of cloprostenol (100 μg) on day 10 of the oestrous cycle, until day 3 of the following cycle. There were five periods of intensive blood sampling, when both ovarian and jugular venous blood samples were collected, as follows: (a) mid-luteal phase, before the second injection of cloprostenol on day 10 (15-min intervals for 6 h); (b) early follicular phase, 24 h after the second injection of cloprostenol (10-min intervals for 4 h); (c) late follicular phase, 48 h after the second injection of cloprostenol (10-min intervals for 4 h); (d) after the LH surge on day 1 of the cycle, 76 h after the second injection of cloprostenol (10-min intervals for 4 h); (e) early luteal phase on day 3 of the cycle, 120 h after the second injection of cloprostenol (10-min intervals for 3 h). Plasma was collected and the samples assayed for LH, FSH, progesterone, oestradiol, androstenedione and inhibin. The ovarian secretion rates for oestradiol, androstenedione and inhibin were calculated. All ewes responded normally to the luteolytic dose of cloprostenol with the preovulatory surge of LH occurring within 56·4±1·6 h (mean ± s.e.m.) followed by the establishment of a normal luteal phase. The pulse frequency of LH, oestradiol and androstenedione increased in the transition from the luteal to the follicular phase (P<0·01). On day 1 of the cycle LH secretion consisted of low-amplitude high-frequency pulses (1·0±0·1 pulse/h) to which androstenedione, but not oestradiol, responded. On day 3 of the cycle LH secretion was similar to that on day 1 but both androstenedione and oestradiol secretion were pulsatile in response to LH, indicating the presence of oestrogenic follicles. The stage of the cycle had no significant effects on LH pulse amplitude and nadir but the ovarian secretory response to LH stimulation did vary with the stage of the cycle. Prolactin pulse frequency, amplitude and nadir were higher (P<0·05) during the follicular phase than the luteal phase. Prolactin pulse frequency was depressed (P<0·05) on day 1 of the cycle but increased to follicular phase levels on day 3. Prolactin pulse frequency was significantly correlated to oestradiol pulse frequency (r = 0·54; P<0·01). During the luteal phase there were insufficient oestradiol pulses to obtain an estimate of pulse amplitude and nadir but both these parameters reached their highest level during the late follicular phase, fell to negligible levels on day 1 and increased to early follicular phase levels on day 3. Androstenedione pulse amplitude and nadir exhibited similar but less marked variation. Inhibin secretion was episodic at all stages of the cycle examined but did not exhibit significant variation with stage of cycle in any of the parameters of episodic secretion measured. Inhibin pulses were not related to either LH or prolactin at any stage of the cycle. FSH secretion was not detectably pulsatile but jugular venous concentrations of FSH at each stage of the oestrous cycle were negatively correlated with mean oestradiol (r= −0·52; P<0·01 but not inhibin secretion (r = 0·19). We conclude that (i) LH secretion is pulsatile at all stages of the oestrous cycle but the steroidogenic responses of the ovary varies with the stage of the cycle, reflecting changes in characteristics of the follicle population, (ii) ovarian inhibin secretion is episodic and displays little change with the stage of the oestrous cycle and (iii) episodic inhibin secretion is not related to either pulses of LH or prolactin. The aetiology of these inhibin pulses therefore remains unknown. Journal of Endocrinology (1990) 126, 385–393


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