Fluctuations in arylsulphatase activity in the rabbit endometrium during the sexual cycle

Summary Histochemical and biochemical studies were performed to verify the presence of arylsulphatase A (ASA) and B (ASB) in the rabbit uterus. Fluctuations in the activity of these sulphatases during the sexual cycle were also studied. Some structural and functional properties of purified ASA were determined. The results indicate that arylsulphatases are active in the endometrium during both the estrogenic and progesteronic phases. The activity of ASA was much more intense than that of ASB; it increased during estrus and decreased during the post-ovulatory phase. ASB activity, however, decreased during estrus and increased during the post-ovulatory phase. The significance of these fluctuations is discussed in relation to the action of sexual hormones and physiological substrates of arylsulphatases.     

The effect of administering a dopamine agonist (Cabergoline) on follicular and luteal development during pro-estrus and estrus in the female greyhound

To study the effect of the dopamine agonist Cabergoline, on ovarian activity in the female dog during pro-estrus and estrus, 6 greyhounds were treated with 5 microg/kg per os of Cabergoline for 20 days beginning on the first day of pro-estrus; 6 animals were left untreated (controls). Ovarian morphology was determined by ultrasound examination once a day during pro-estrus and twice a day during estrus. Follicles were divided into three classes on the basis of their diameter: F1 (<3mm), F2 (3-6mm) and F3 (>6mm). The presence and diameters of post-ovulatory follicles (F-POST) and corpora lutea (CL) were also recorded. Blood samples were taken once a day during pro-estrus and twice a day during estrus. The plasma was assayed for LH, prolactin and progesterone by radioimmunoassay. There were no differences between Cabergoline-treated and control dogs in the duration of pro-estrus or estrus. There was a progressive increase in follicle diameter from the start of pro-estrus when follicles were mainly class F1 to the day of estrus when follicles were mainly class F3. Three days after the start of estrus, the first F-POST follicles were detected. This pattern of development continued and on day 5 the first CLs were detected. By day 9, only CLs were detected. The peak of pre-ovulatory LH was within 3 days of estrus and ovulation was detected in all animals within 3 days of the LH peak. There were no differences in LH concentrations between groups. Plasma prolactin levels varied between animals and were reduced in treated dogs, however, this was not statistically significant (P=0.12). Plasma progesterone levels were below 1.0 ng/ml before the LH surge and thereafter gradually increased. There were no differences in plasma progesterone concentrations between treated and control dogs. In conclusion these results show that the administration of the dopamine agonist Cabergoline during pro-estrus and estrus did not affect the duration of pro-estrus or estrus or the pattern of follicular and luteal development in female dogs.     

Sexual behavior in ovariectomized and seasonally anovulatory pony mares (Equus caballus)

Ten ovariectomized (OVEX) and ten intact, but seasonally anovulatory (ANOV), pony mares were observed for sexual activity with five stallions, using a “harem group” social testing paradigm (two OVEX and two ANOV mares plus one stallion per group) for 15 consecutive daily tests lasting 20 min each. All mares in both conditions showed proceptive behavior in at least one test, all mares but one were mounted, and 14 of 20 mares received ejaculations. No statistical differences were found between the two conditions for any measure of proceptivity, copulatory activity, or days in estrus. The quality of estrus was judged to be equivalent to that displayed by periovulatory mares during their initial and terminal days of estrus, but less intense than that seen near ovulation. Mares in both groups were in estrus during approximately 60–70% of the tests and only 3 of the 20 mares were sexually refractory for more than five consecutive tests. Thus, the typical 2-week phase of sexual refractoriness seen in intact diestrous mares was absent in OVEX and ANOV mares, suggesting that the ovary plays a major role in actively suppressing estrous responses during the luteal phase of the cycle.     

Steroid sulfatase, arylsulfatases A and B, galactose-6-sulfatase, and iduronate sulfatase in mammary cells and effects of sulfated and non-sulfated estrogens on sulfatase activity

Sulfatase enzymes have important roles in metabolism of steroid hormones and of glycosaminoglycans (GAGs). The activity of five sulfatase enzymes, including steroid sulfatase (STS; arylsulfatase C), arylsulfatase A (ASA; cerebroside sulfatase), arylsulfatase B (ASB; N-acetylgalactosamine-4-sulfatase), galactose-6-sulfatase (GALNS), and iduronate-2-sulfatase (IDS), was compared in six different mammary cell lines, including the malignant mammary cell lines MCF7, T47D, and HCC1937, the MCF10A cell line which is associated with fibrocystic disease, and in primary epithelial and myoepithelial cell lines established from reduction mammoplasty. The effects of estrogen hormones, including estrone, estradiol, estrone 3-sulfate, and estradiol sulfate on activity of these sulfatases were determined. The malignant cell lines MCF7 and T47D had markedly less activity of STS, ASB, ASA, and GAL6S, but not IDS. The primary myoepithelial cells had highest activity of STS and ASB, and the normal epithelial cells had highest activity of GALNS and ASA. Greater declines in sulfatase activity occurred in response to estrone and estradiol than sulfated estrogens. The study findings demonstrated marked variation in sulfatase activity and in effects of exogenous estrogens on sulfatase activity among the different mammary cell types.     

ENZYMIC STUDIES ON SULPHATIDE METABOLISM IN DIFFERENT STAGES OF EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS

Abstract— The activities of three enzymes–cerebroside sulphotransferase, 3'-phospho-adenosine 5'-phosphosulphate synthesizing enzyme and arylsulphatases A and B have been studied in various developmental and recovery stages of experimental allergic encephalomyelitis. The concentrations of cerebroside and sulphatide were also analysed during these stages. It was observed that the sulphatide concentration decreased during the development of the disease, with a concurrent increase in the activity of arylsulphatase and vice versa during the recovery stages. 3'-Phosphoadenosine 5'-phosphosulphate synthesis as well as sulphotransferase activity increased during the pre-acute stage of the disease, reached a maximum at the acute stage and decreased during recovery stages.     

Hormonal control of endometrial glycogen metabolism in the Macaca arctoides

Endometrial biopsies obtained throughout the menstrual cycle of the Macaca arctoides show the glycogen content paralleling the serum progesterone fluctuations which occur during the menstrual cycle. Secretory phase samples contained a three-fold higher concentration of glycogen when compared to follicular phase tissue. Changes in the activity levels of the glycogen metabolizing enzymes, glycogen phosphorylase and glycogen synthetase, during various stages of the menstrual cycle are in accord with the concept that the post-ovulatory increase in endometrial metabolism is a function of progesterone influence on this tissue. Endometrial glycogen synthetase activity remains low during the early proliferative phase of the cycle and becomes significantly elevated (two-to three-fold) during the early secretory phase of the cycle. Glycogen phosphorylase shows a similar cyclicity later in the luteal phase, reaching maximal activity between the seventeenth to nineteenth day of the cycle and remaining elevated through the twenty-sixth day of the cycle. The coincident nature of the rise in peripheral progesterone to increases in uterine glycogen metabolism suggest that progesterone may be the prime modulator of uterine endometrial metabolism during the post-ovulatory phase.     

Increased expression of kisspeptin and GnRH forms in the brain of scombroid fish during final ovarian maturation and ovulation

ABSTRACT: BACKGROUND: Kisspeptins (Kiss) are prime players in the control of reproductive function through their regulation of gonadotropin-releasing hormone (GnRH) expression in the brain. The experimental scombroid fish, chub mackerel (Scomber japonicus) expresses two kiss (kiss1 and kiss2) and three gnrh (gnrh1, gnrh2, and gnrh3) forms in the brain. In the present study, we analyzed expression changes of kiss and gnrh mRNAs in the brain and corresponding GnRH peptides in the brain and pituitary during final ovarian maturation (FOM) and ovulation. METHODS: Female fish possessing late vitellogenic oocytes were injected with GnRH analogue to induce FOM and ovulation. Fish were observed for daily spawning activities and sampled one week post-injection at germinal vesicle migration (GVM), oocyte hydration, ovulation, and post-ovulatory time periods. Changes in relative mRNA levels of kiss and gnrh forms in the brain were determined using quantitative real-time PCR. Changes in GnRH peptides in the brain and pituitary were analyzed using time-resolved fluoroimmunoassay. RESULTS: Both kiss1 and kiss2 mRNA levels in the brain were low at late vitellogenic stage and increased significantly during the GVM period. However, kiss1 mRNA levels decreased during oocyte hydration before increasing again at ovulatory and post-ovulatory periods. In contrast, kiss2 mRNA levels decreased at ovulatory and post-ovulatory periods. Levels of gnrh1 mRNA in the brain increased only during post-ovulatory period. However, levels of gnrh2 and gnrh3 mRNAs were elevated during GVM and then, decreased during oocyte hydration before increasing again at ovulatory period. During post-ovulatory period, both gnrh2 and gnrh3 mRNA levels declined. Peptide levels of all three GnRH forms in the brain were elevated during GVM and oocyte hydration; their levels were significantly lower during late vitellogenic, ovulatory, and post-ovulatory periods. In contrast, pituitary GnRH peptide levels did not show any significant fluctuations, with the GnRH1 peptide levels being many-fold higher than the GnRH2 and GnRH3 forms. CONCLUSION: The results indicate increased expression of multiple Kiss and GnRH forms in the brain and suggest their possible involvement in the regulation of FOM and ovulation in captive female chub mackerel.     

Autophagic and proteolytic processes in the Harderian gland are modulated during the estrous cycle

The Syrian hamster Harderian gland (HG) is an organ that undergoes physiological autophagy in response to oxidative stress induced by porphyrin production. Porphyrin production in the HG has marked sex differences and is closely linked to reproductive function. In the present study, we observed that the estrous cycle and associated estrogen variations may affect oxidative-stress-induced proteolytic processes. In particular, significant changes in autophagic activity were detected during the estrous cycle. Notably, increased activation of macroautophagy as well as chaperone-mediated autophagy in the estrus phase coincided with a minimal antioxidant capability and the highest protein damage levels. By contrast, autophagic machinery was found to be blocked in the diestrus phase, likely due to mammalian target of rapamycin activation, which could be corroborated by the subsequent pS6K activation. Analogous results were observed regarding proteasome activity, which also showed maximal activity in the estrus phase. Interestingly, all these mechanisms were associated with important morphological changes in the HG during the estrous cycle. We observed statistically significant increases in Type II cells, which may be related to extensive autophagy in the estrus phase. Physiologically, this would result in a significant release of porphyrins specifically when females are more receptive. These data support the role of porphyrins as pheromones, as other authors have previously suggested, thus making the HG a scent organ. In addition, these results suggest a porphyrin-based approach to the treatment of porphyria during pregnancy, a condition for which no treatment is currently known.     

Water temperature and pH influence olfactory sensitivity to pre-ovulatory and post-ovulatory ovarian pheromones in male Barilius bendelisis

The attractive response and sexual activity elicited by pre-ovulatory steroid sulphate and post-ovulatory 15K-PGF pheromones are greater in wild caught tubercular males and immature males which express breeding tubercles on the snout (at 12–13 days post androgen implant) than in non-tubercular and non-androgen implanted males of freshwater fishBarilius bendelisis. This shows that circulatory androgens exert an activational effect on olfactory receptors of male fish. Wild caught tubercular males and androgen implanted juvenile males exhibit a high responsiveness to steroid sulphate at the water temperature and pH which fish experience during the pre-spawning phase. The male’s sensitivity to 15K-PGF is almost equally high at the water temperature and pH which they experience in wild during the both pre-spawning and spawning periods. This suggests that the differential olfactory sensitivity to the two classes of pheromones in androgen implanted males is due to the varied temperature and pH of water, and that during the breeding season the male’s olfactory sensitivity to PGF pheromone is more widespread than to the steroidal pheromone. An increased and decreased olfactory sensitivity in mature males to sex pheromones and L-alanine respectively during the breeding phase is in agreement with the hypothesis that pheromonal stimuli dominate over feeding stimuli to promote spawning success.     

抗坏血酸对花生原生质体分离过程中膜损伤的保护作用

酶解处理使花生叶肉细胞原生质体膜脂质过氧化产物丙二醛(MDA)积累,添加抗坏血酸(ASA)能降低MDA的积累。未添加ASA时,酶解初期,过氧化物酶(POD)、过氧化氢酶(CAT)活性上升,说明原生质体存在各种抵御不良变化的机制,但酶解时间加长,POD活性下降。添加ASA能使超氧化物歧化酶(SOD)活性明显上升。试验结果说明:在原生质体分离过程中会导致膜损伤,细胞自身存在一个响应的防御机制,添加ASA能降低酶解过程对原生质膜的损伤。     

Dynamics of Male–Female Multimodal Signaling Behavior across the Estrous Cycle in Giant Pandas (Ailuropoda melanoleuca)

Giant panda courtship behavior includes multimodal signaling assemblages consisting of olfactory, vocal, and postural elements. While signaling is generally conspicuous, successful copulation is inconsistently achieved in captivity, even when female behavioral and physiological data indicate that ovulation is imminent. We set out to characterize these complex patterns of social behavior by observing interactions between 26 unique pairs of giant pandas housed in adjoining pens throughout the females' reproductive cycle. We categorized social behaviors from a transactional perspective and examined social exchanges via analyses of the relative frequency of social behaviors, and via the sequential relationship between male and female social behavior. From non‐estrus to peak‐estrus, we found that the relative frequency of female affiliative and sexual behavior increased and that the relative frequency of ambivalent and aggressive behavior decreased. Male behavior was fairly constant, except for sexual behavior, which increased during peak‐estrus, when it was facilitated by female sexual behavior. Sequential analysis of social interactions showed that preceding behavior had a significant influence over the other panda's response behavior primarily during peak‐estrus, suggesting that pandas are most responsive to conspecific signaling during the peri‐ovulatory period. However, behavioral momentum was a dominant feature of the intra‐individual transitions. Females maintained sexual, ambivalent, and neutral behavior during interactions significantly more than would be expected by chance, with male behavior bearing little influence once the behavior was initiated. A similar pattern was also observed in males, who maintained affiliative, interested, and neutral behaviors. Overall, our data suggest that the multimodal signals used by giant pandas during courtship do not consistently evoke a discrete, immediate response from receivers. Instead, signals appear to advertise reproductive condition and influence potential mates over longer timeframes, suggesting the potential tonic role of communication.     

Progesterone profiles around the time of insemination do not show clear differences between of pregnant and not pregnant dairy cows

In this study, features of progesterone profiles were examined in relation to the outcome of insemination. Three groups of estrous cycles were analyzed: resulting in pregnancy, not resulting in pregnancy and resulting in lost pregnancy. The aim of the study was to identify a complex of progesterone profile features associated with successful insemination. The features used were (1) from the estrous cycle preceding the artificial insemination: estrus progesterone concentration, post-estrus maximum rate of increase in progesterone, luteal phase peak, pre-estrus maximum rate of decline in progesterone and the length of follicular and luteal phase and (2) from the estrous cycle following insemination: estrus progesterone concentration, post-estrus maximum rate of increase in progesterone and days from estrus to post-estrus maximum rate of increase in progesterone. A discriminant analysis did not reveal clear differences between the groups. However, the analysis correctly classified 75% of true pregnant cows. Conversely, only 60% of not pregnant animals were classified as such by the discriminate analysis. Individual analysis of progesterone profile features in pregnant and not pregnant groups of estrous cycles showed that a shorter follicular phase preceding insemination is associated with proper timing of post-ovulatory luteinisation and therefore is more likely to result in pregnancy.     

Investigations on spermicidal activity in the sheep vagina.

This study was conducted to elucidate some of the effects of a synthetic progestagen and natural ovarian hormones on spermicidal activity in the sheep vagina. In the first experiment, parous ewes were treated for 17 days either intravaginally with medroxyprogesterone acetate (MAP) or subcutaneously with progesterone. They were inseminated artificially either on the last day of progestagen treatment or during estrus after progestagen withdrawal. Their vulvovaginal junctions were ligated to prevent the loss of sperm cells by drainage to the exterior. Untreated control ewes were inseminated during either estrus or the luteal phase of the estrous cycle. The ewes were killed 22 hr. after insemination, their vaginas flushed, and intact sperm cells and tailless sperm heads counted. In the second and third experiments, some of the ewes were bilaterally ovariectomized and inseminated several weeks later. Other ewes were ovariectomized and given subcutaneous injections of estradiol, progesterone, or both hormones.In the first experiment, most sperm cells were recovered intact from estrous or luteal phase control ewes. The intravaginal administration of MAP increased both the breakage of sperm cells into heads and tails and the disappearance of sperm cells. The spermicidal effects of MAP were just as great in ewes inseminated on the last day of treatment. as in those inseminated during the ensuing estrus; these results indicated that the peak estrogen secretion that occurs near the beginning of estrus was not necessary for the intensification of spermicidal activity.In the second experiment, ovariectomized ewes were compared to estrous and luteal phase ewes in regard to vaginal spermicidal activity. Sperm breakage and disappearance occurred least in estrous ewes, to a somewhat greater degree in luteal phase ewes, and to the greatest extent in ovariectomized ewes. The results suggested that endogenous ovarian hormones, particularly those in estrous ewes, suppress spermicidal mechanisms in the vagina.In the third experiment, the administration of estradiol and progesterone to ovariectomized ewes prevented the increase in sperm cell disappearance. Neither hormone alone prevented the increase.     

Prolonged effect of stress at weaning on the brain serotonin metabolism and sexuality of female rats.

Weanling female rats were stressed (by water and food deprivation for two days) and three months later the following indexes were studied: 5-HT and 5-HIAA levels in five brain regions, blood plasma and cerebrospinal fluid (CSF), sexual activity and nocistatin level of the plasma and CSF. The 5-HIAA content of hypothalamus and brainstem was significantly decreased (in the brainstem with one third) and in the striatum significantly increased. Plasma nocistatin level was significantly increased. Meyerson index and lordosis quotient were similar to control, but the estrus frequency almost doubled in the stressed animals. Much more defense reactions were observed in the stressed females during trials of mating. The results demonstrate that, 1) the perinatal period is not only sensitive to the remote-effects of stress but later could also be stress-sensitive critical periods, and 2) the continuously differentiating (e.g. bone marrow) cells are sensitive to late imprinting by stress, as well as to the brain and the sexual system.     

Uterine motility in the cow during the estrous cycle. I. Spontaneous activity

This study describes a method for measuring intrauterine pressure (IUP) changes and uterine motility in cows. Spontaneous uterine motility was recorded during the estrous cycle in stanchioned, nonlactating dairy cows using a pair of miniature pressure transducers mounted 15 cm apart at the distal end of a dacron catheter placed in one uterine horn via the cervix. Clinical examination of ovarian status and determination of the peripheral plasma levels of estradiol-17beta and progesterone were used to determine the stages of the cycle. The pressure sensors recorded variations in muscular resting tension (tone) and the occurrence, spatial distribution, and force of the uterine contractions. Both tone and uterine activity varied significantly during the cycle. They were minimal during diestrus, increased during proestrus, reached maximal values at estrus, and then decreased. The highest synchronized motor activity with presence of peristaltic-antiperistaltic movements occurred during estrus. The prevailing direction of the uterine contractions during late estrus (immediate preovulatory period) was cervico-tubal.     

Activity of some hepatic enzymes in schistosomiasis and concomitant alteration of arylsulfatase B

The levels of arylsulfatases A and B, alpha-amylase, aspartate transcarbamylase, and gamma-glutamyl transpeptidase were investigated during the infection of mice with schistosoma mansoni. This infection caused a significant (p < 0.001) increase in the activity of hepatic arylsulfatase B (ASB), aspartate transcarbamylases and gamma-glutamyl transpeptidase. A non-significant difference occurred for alpha-amylase (p < 0.3) and arylsulfatase A (p > 0.5) when compared to the control. The specific activity of hepatic ASB was progressively increased with the progression of the Schistosoma-infection. Moreover, the kinetic studies of hepatic ASB in Schistosoma-infection showed that a slight decrease in the value of K(m) and about a 40% increase in V(max) when compared to the control. In addition, the pH optimum of hepatic ASB was altered from 6 to 7 as a result of schistosomiasis. These observations suggest that there are schistosomiasis-associated changes of the catalytic and kinetic properties of hepatic ASB.     

A comparison of the estrous behavior of Holstein-Friesian cows when cubicle-housed and at pasture

This study compared estrous behavior of dairy cows kept in cubicle housing and fed a total mixed ration diet (HOUSED treatment) with that of cows kept at pasture with concentrate supplementation (PASTURE treatment). Behavior was compared both in the 48 h around standing estrus and during the standing estrus period. The 23 spring-calving Holstein-Friesians in each treatment were observed directly three times per day for nine weeks. The occurrence of nine selected behaviors associated with estrus was recorded during 20 min observation sessions. Twelve standing estrus events from each treatment were selected for analysis of the frequency of these nine behaviours over the 48 h around standing estrus. Milk progesterone profiles were used to confirm the dates of standing estrus events. Attempting to mount other cows, sniffing the anogenital region of other cows, resting the chin on other cows, receiving chin rests and head-to-head butts all showed significant changes in frequency in the 48 h around standing estrus in both treatments, reaching a peak during standing estrus (P ≤ 0.05). Mounting other cows increased significantly in the PASTURE treatment around standing estrus (P < 0.001), but not in the HOUSED treatment. The frequency of ano-genital sniffs received by the animals in the PASTURE treatment also increased significantly around standing estrus (P < 0.01) but not in the HOUSED treatment. When the animals were in standing estrus there was a significantly higher frequency of standing to be mounted in PASTURE than in HOUSED cows (median (q1, q3) PASTURE = 2.5 (1.0, 3.0), HOUSED = 0.0 (0.0, 1.0)) (P < 0.01), but no difference in the frequency of the other eight sexual behaviors recorded. HOUSED cows did not exhibit the same increase in mounting during the standing estrus period as PASTURE cows and received fewer mounts in observation sessions during standing estrus. These results have implications for the use of estrus detection systems that rely solely on mounting behavior in cubicle-housed dairy cows.