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Mucosal immunity in the female reproductive tract: correlation of immunoglobulins, cytokines, and reproductive hormones in human cervical mucus around the time of ovulation.

Mucosal surfaces serve as the portal of entry for many viral, bacterial, and parasitic infections. Understanding the immunity at mucosal membranes is essential to enhancing protection and decreasing infections. To evaluate the humoral and cellular immunity in the female reproductive tract, 15 reproductive-age women with a history of regular, cyclic monthly menses were recruited for this study. The presence of immunoglobulins and cytokines in cervical mucus was correlated with the production of reproductive hormones in sera. Cervical mucus specimens were collected at each daily visit beginning on cycle day 8 and continuing for 5 days postovulation. Volunteers were monitored by daily urinary LH testing coupled with transvaginal ultrasonography to ascertain follicular collapse. The cervix was washed in sterile saline before aspirating the cervical mucus from the cervical canal. Collection volumes ranged between 50 and 800 microl and were considered to represent the total mucus produced. Estradiol displayed the characteristic biphasic pattern with a peak before ovulation and in the luteal phase. Both IgG (30 mg/dl) and IgA (15 mg/dl) had a biphasic pattern with peak immunoglobulin levels detected 1 day before the estradiol peak and increasing again just after ovulation. Peak interleukin 10 (40 pg/ml) levels corresponded precisely with estradiol peak levels just before ovulation. Peak interleukin 1beta (1.3 ng/ml) levels occurred approximately 1 day before the estradiol peak. No apparent pattern in interleukin 6 (150 pg/ml) could be ascertained. Our data suggest a correlation between the IgG and IgA immunoglobulin levels, interleukin 1beta and interleukin 10, in the female reproductive tract and estradiol levels in the circulation. The increase in immunoglobulins and cytokines occurs approximately 1 day before the peak estradiol production before ovulation. These data suggest a role for cytokines and hormones in the regulation of reproductive tract immunity.

Adult↗

Distribution of delta-5 -3beta-hydroxysteroid dehydrogenase activity in the Graafian follicle of the sheep.

The localization of delta-5 -3beta-hydroxysteroid dehydrogenase (3 beta-HSD) has been examined in ovarian follicles in vivo and in vitro, and related to oestrogen and progesterone production. In vivo, during the oestrous cycle, enzyme activity was restricted to the theca interna of the one or two most advanced follicles in each animal, but was present only between Day 2 and 5 and between Day 13 and ovulation. High levels of oestrogen were found in the ovarian venous blood only when follicles containing 3 beta-HSD were present. When sheep were injected with PMSG, the theca interna in a number ofsmall follicles acquired 3 beta-HSD activity and began to secrete oestrogen within 12 hr of the injection. The enzyme was not detected in the membrana granulosa of any follicles before ovulation but within a few hours of ovulation, 3 beta-HSD activity was present in the granulosa lutein cells. In vitro, large activated follicles exhibited 3 beta-HSD activity in the theca interna and secreted high levels of oestrogen into the culture medium. When LH was added to the medium oestrogen secretion was inhibited; within 48 hr, the follicles were secreting high levels of progesterone, and 3 beta-HSD activity was present in both the membrana granulosa and the theca interna. Dibutyryl cyclic adenosine monophosphate mimicked the effect of LH in suppressing oestrogen secretiion, but did not induce production of progesterone; the distribution of 3 beta-HSD activity infollicles treated with this nucleotide was the same as in those cultured in control medium.

Animals↗

Gonadotropin regulation of tissue-type and urokinase-type plasminogen activators in rat granulosa and theca-interstitial cells during the periovulatory period.

Plasminogen activators (PAs) are believed to be involved in ovulation. Because both tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA) are secreted by cultured rat granulosa cells, we have examined the activities of these proteins in ovarian homogenates as well as granulosa and theca-interstitial (TI) cells during gonadotropin-induced ovulation. Immature rats were injected with 20 IU pregnant mare serum gonadotropin (PMSG) to initiate follicle development, followed by treatment with 10 IU hCG 48 h later to induce ovulation. Ovarian proteins were separated by SDS-PAGE and PA activity determined by fibrin overlay. The activity of tPA, but not uPA, was stimulated following PMSG treatment in ovarian homogenates. Subsequent hCG injection further increased the tPA activity in a time-dependent manner, reaching a maximum (12 h after hCG treatment) immediately prior to ovulation and declined thereafter. Similar preovulatory increases in tPA activity were detected in isolated granulosa cells. Although both tPA and uPA activities were increased in TI cells after PMSG administration, no further increases were detected after hCG treatment. To estimate enzyme secretion, ovarian cells obtained at various preovulatory periods were incubated for 24 h in vitro. The ability of granulosa cells to secrete tPA, but not uPA, increased following in vivo PMSG and hCG treatment in a time-dependent manner, reaching a maximum immediately prior to ovulation. During the preovulatory period, an abrupt increase in tPA secretion by TI cells was also detected. Using immunohistochemical staining for tPA, it was found that ovarian sections from preovulatory rats at 12 h after hCG injection stained positively in granulosa, theca interna, and interstitial gland cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Membrane-bound carboxypeptidase-M is expressed on human ovarian follicles and corpora lutea of menstrual cycle and early pregnancy.

We have previously reported that membrane-bound aminopeptidases were expressed on human follicles and corpora lutea (CL) and we showed that these aminopeptidases are involved in follicular growth, probably by regulating extracellular peptide concentrations. In this study, the expression of membrane-bound carboxypeptidase-M (CP-M), which cleaves carboxyl-terminal amino acids from peptides extracellularly, on human follicles and CL was examined. In growing and pre-ovulatory follicles, CP-M was immunohistochemically detected with weak or moderate intensity on theca interna cells. Although CP-M was not detected on granulosa cells in growing and pre-ovulatory follicles, it was strongly detected on the cell surface of luteinizing granulosa cells isolated from patients undergoing in-vitro fertilization treatment, indicating that CP-M was rapidly expressed on granulosa cells during ovulation. In menstrual and pregnant CL, CP-M was clearly detected on luteal cells. In menstrual CL, the expression of CP-M mRNA was observed by reverse transcription-polymerase chain reaction (RT-PCR). Western blotting analysis revealed that the molecular mass of the CP-M extracted from mid-luteal CL was 62 kDa. These results indicate that CP-M is a cell surface differentiation-related molecule of human granulosa, theca, and luteal cells. The rapid expression on granulosa cells during ovulation strongly suggests the involvement of CP-M in the ovulation and CL formation processes.

Blotting, Western↗

Corpus luteum function and pregnancy rates with clomiphene citrate therapy: comparison of human chorionic gonadotrophin-induced versus spontaneous ovulation.

A total of 508 clomiphene citrate cycles with intra-uterine insemination (IUI) performed in 233 consecutive patients were studied. In 247 cycles insemination was performed 36-38 h after human chorionic gonadotrophin (HCG)-triggered ovulation; in the remaining 261 cycles IUI was performed 18-20 h after urinary luteinizing hormone (LH) kit detection of a spontaneous LH surge. Corpus luteum function, as determined by luteal phase length and mid-luteal progesterone concentrations, together with pregnancy rates were analysed. There was no difference in luteal phase parameters between spontaneous and HCG-triggered cycles when adjusting for patient age. Furthermore, the pregnancy rates did not differ between the HCG and LH kit groups, even after adjusting for patient age and number of motile spermatozoa inseminated. Additionally, the large numbers of cycles analysed provided sufficient power to detect increases in clinical pregnancy rates in spontaneous ovulatory cycles and HCG-induced ovulation of 10.1 and 2.4% respectively, using the customary significance level (alpha-type error) of 0.05. These findings indicate that pregnancy rates and corpus luteum function in carefully monitored clomiphene citrate/IUI cycles do not differ between HCG-triggered and spontaneous ovulatory cycles.

Adult↗

Expression of prepro-GnRH and GnRH receptor messengers in rainbow trout ovary depends on the stage of ovarian follicular development.

Gonadotropin-Releasing Hormones (GnRHs) are decapeptides well known to regulate the reproductive cycle. They are expressed not only in the brain, but also in other tissues including the gonads. It is believed that they may be involved in the endocrine and paracrine regulation of the reproductive cycle. To date, two forms of GnRH have been identified in salmonids: salmon (sGnRH) and chicken II (cGnRH-II). In the present study, the temporal expression of sGnRH-1, sGnRH-2, cGnRH-II, and rtGnRH receptor genes was studied in rainbow trout ovary during the reproductive cycle according to the stages of follicular development. Using RT-PCR coupled with Southern-blot hybridization, sGnRH-1, sGnRH-2, cGnRH-II, and rtGnRH-R transcripts were detected in morphologically nondifferentiated ovaries as early as 55-65 days post-fertilization and throughout all stages of vitellogenesis. Using Northern blot analysis, cGnRH-II mRNA was detected only in immature previtellogenic ovary, whereas sGnRH mRNA was detected also during early and mid-exogenous vitellogenesis. No sGnRH mRNA was detected at the end of vitellogenesis. In maturing pre-ovulated ovary, sGnRH transiently reappeared before germinal vesicle breakdown (GVBD) and decreased thereafter. A few days after ovulation, a strong sGnRH mRNA expression was found in ovarian tissue as the eggs were kept in the body cavity of females. However, in females stripped just after ovulation, sGnRH mRNA levels remained low in ovary during several weeks. Fully spliced sGnRH-1 and sGnRH-2 messengers were mostly expressed during the reproductive cycle; however different sGnRH-1 and sGnRH-2 splicing variants containing intronic sequences were also detected. Some of these messengers may encode prepro-GnRH precursors with truncated GnRH-associated peptides. The stage-dependent expression and different cell localization of sGnRH, cGnRH-II, and rtGnRH-R transcripts suggest that GnRH-like peptides may have different roles in the paracrine regulation of ovarian follicular development.

Amino Acid Sequence↗

Effect of frequency of boar exposure and adjusted mating times on measures of reproductive performance in weaned sows.

In weaned sows, reduced reproductive performance can result from failure or delayed return to estrus or improper timing of insemination. This experiment evaluated the effect of increased frequency of boar exposure and adjusted mating times on reproductive performance. Sows of mixed parity were weaned approximately 18.7 d after parturition and allotted by genotype, parity, and lactation length to boar exposure frequency of once daily (1X, n = 66), twice daily every 12 h (2X, n = 61), or three times daily exposure at 8-h intervals (3X, n = 60). Sows were weaned into crates and boar exposure was initiated 3 d after weaning. Once estrus was detected, ultrasound was performed every 8 h to determine time of ovulation. All sows were artificially inseminated twice in the 1X group at 0 and 24 h, in the 2X group at 12 and 24 h, and in the 3X group at 16 and 32 h after onset of estrus. The weaning-to-estrus interval was not influenced by treatment and averaged 4.5 d. The percentage of sows expressing estrus in 8 d was higher (P < 0.05) for 1X (97.3%) compared with 2X (92.8%) but not the 3X group (94.0%). The percentage of sows ovulating after estrus was not influenced by treatment (P > 0.10) and averaged 96.5%. Estrus-to-ovulation interval was not affected by treatment (44.7 h) but was influenced by weaning-to-estrus interval (P < 0.0001). Length of estrus was influenced by treatment (P < 0.001), with estrus in the 1X (46.6 h) shorter than in the 2X (60.0 h) and 3X (67.0 h) treatments, and also by weaning-to-estrus interval (P < 0.001). The percentage of first inseminations occurring within 24 h before ovulation was increased (P < 0.001) in the 2X (62%) and 3X (66%) groups compared with the 1X group (28%) and was also influenced by parity (P < 0.001) and weaning-to-estrus interval (P < 0.05). The percentage of second services within 24 h before ovulation was not increased by any factor and averaged 78%. Farrowing rates were not increased (P > 0.10) for 2X (87.2%) and 3X (83.1%) treatments compared with 1X (75.0%). Total pigs born was also not affected by treatment, although 2X (11.2) and 3X (10.7) numbers were greater than 1X (10.0). It appears that once-daily estrus detection combined with delayed mating could achieve optimal reproductive performance.

Animals↗

Protocols for synchronizing estrus and ovulation in buffalo (Bubalus bubalis): a review.

Poor estrus expression and a prolonged intercalving interval compromise the reproductive efficiency of female buffaloes. These limitations are exacerbated during the hot season, when fertility decreases dramatically. Pregnancy rate decrease further because difficulties in detecting estrus. To improve reproductive efficiency, several protocols of estrus and ovulation synchronization have been developed. These procedures are based on manipulating the CL, either to induce premature luteolysis using prostaglandins or to prolong the luteal phase using progestagens. However, it has recently emerged that a more precise manipulation of follicular development may be needed to achieve better synchrony of ovulation and improve fertility. Researchers have therefore turned their attention to evaluating programs in which hormones such as GnRH, FSH, LH, eCG, hCG, prostaglandins, progesterone and estradiol are administered. This review considers the impacts of estrus and ovulation synchronization protocols on fertility in the buffalo. In general, it may be stated that buffaloes respond well to the exogenous administration of hormones, and artificial insemination is possible at a pre-established time after synchronizing ovulation. Most combined hormone protocols give satisfactory pregnancy rates, comparable to those achieved in animals inseminated at natural estrus.

Animals↗

Immunohistochemical study of distribution of estrogen receptors in corpus and cervix uteri.

An immunohistochemical assay based on monoclonal antiestrophilin antibodies has been used to localize estrogen receptor (ER) in frozen sections of normal human endometrial, myometrial and cervical tissues from menstruating, hormonally treated, pregnant and postmenopausal women. Specific staining was confined to the cellular nuclei. In proliferative phase endometrium, postmenopausal emdometrium, and endometrium from patients treated with hormone ERs were easily detected in most glandular and stromal cells. After ovulation and in early pregnancy a quick and distinct decrease of ER expression was noted. This was especially the case with the more superficial layers of endometrium (endometrium functionalis), the majority of whose cells had either weak localization of ER or none at all. In the endometrium basalis, however, the reduction of ER localization turned out to be more moderate. More then half of the epithelial and stromal cells displayed nuclear staining, partly strong. The myometrium of the corpus uteri showed a similar ER localization and dependence on hormonal stage when compared with the endometrium functionalis. The endocervical mucosa displayed a high degree of ER expression in the proliferative phase, in postmenopausal women and in women who had been treated with hormones. Unlike the endometrium and myometrium, the endocervical glands underwent minimal changes in nuclear ER content during the menstrual cycle. Although the endocervical stroma showed cyclic alterations in ER levels, their reduction after ovulation was less marked than in the corresponding endometria. In cervical squamous epithelium ER localization was predominantly confined to the basal layers. In the course of cellular maturation, specific nuclear staining vanished. In the proliferative phase, after the menopause and in early pregnancy, the basal, parabasal and intermediate cells were specifically stained. In the postovulatory phase, However, nuclear staining was confined to the basal and parabasal cells. Hormonally treated squamous epithelia almost completely lacked nuclear ER localization.

Antibodies, Monoclonal↗

Extending the duration of the pill-free interval during combined oral contraception.

The effect of increasing the duration of the pill-free interval from 7 to 9 and 11 days during the first 4 months of combined oral contraceptive therapy was studied by hormonal, ovarian follicle and mucus changes. Increasing the pill-free interval allowed more follicular development and more estradiol production. In some cases, pre-ovulatory values were achieved although ovulation did not occur. No differences could be detected between different types of combined oral contraceptive, including monophasic, triphasic, levonorgestrel-containing and gestodene-containing. The variation between individuals in the group of 28 was far greater than any variation in dose.

Adult↗

Changes in bone turnover during the menstrual cycle in cynomolgus monkeys.

It is well established that estrogen deficiency at menopause results in increased bone turnover, which is reflected in increased concentrations of markers of bone formation and bone resorption in serum and urine. Since serum 17beta-estradiol concentrations vary markedly throughout the menstrual cycle, one would expect to see changes in bone turnover as well. Studies in humans have not yielded consistent results, perhaps because of differences in diet and activity throughout the test period. Therefore, we examined changes in bone biomarkers throughout the menstrual cycle in cynomolgus macaques. Seven intact female cynomolgus macaques (Macaca fascicularis) were evaluated. Vaginal swabs for menstrual blood were performed 3 times/week to determine the stage of the reproductive cycle. Blood and urine were collected at weekly or biweekly intervals for a total of eight samples per monkey for analysis of serum 17beta-estradiol, progesterone, parathyroid hormone (PTH), osteocalcin, bone-specific alkaline phosphatase (BSAP), and urinary CrossLapstrade mark. Cycle lengths were determined, and collection days were adjusted to a standardized length of 28 days for all monkeys. Values for bone biomakers were evaluated as % mean for each monkey cycle. By fitting the data to a sine wave (cosinor analysis), serum osteocalcin, BSAP, and urinary CrossLaps demonstrated significant cycles with peaks at days 2.6, 7.3, and 27.8, respectively. Serum osteocalcin and urinary CrossLaps were inversely correlated to serum 17beta-estradiol. Urinary CrossLaps were significantly lower in the week just prior to and during ovulation when estradiol was elevated. No rhythm was detected in serum PTH. A peak in bone resorption when serum 17beta-estradiol is at its nadir is consistent with the hypothesis that estrogen decreases bone turnover.

Alkaline Phosphatase↗

Qualitative variation in the immune response to ovarian follicular fluid proteins in cattle.

In two experiments 44 heifers were immunised with inhibin preparations of partially purified ovarian follicular fluid (PPFF) of ovine, porcine and equine origin in non-ulcerative Freund's adjuvant. In the first experiment 20 cattle were immunised with ovine PPFF1. In the second, a further 24 were immunised with either a different sheep PPFF (ovine PPFF2) or PPFF from pigs or horses. Large molecules of over 30 kDa initiated a strong immune response in some animals, but none in others, although there was no obvious relationship with the MHC Class I (BoLA) phenotype. Immunisation with ovine partly purified follicular fluid affected ovulation rate in some cattle, and only these cows produced detectable antibodies to an approximately 40 kDa protein. These data show that the immunological response of cattle to a mixture of antigens is highly variable between individuals and may explain the variation in the ovarian response to immunological manipulations of ovulation rate seen in this species.

Animals↗

Ovarian features of the wild collared peccary (Tayassu tajacu) from the northeastern Peruvian Amazon.

In this study, the ovaries of 27 wild collared peccaries (Tayassu tajacu) from the Amazonian region of northeastern Peru were examined macroscopically and microscopically, and expression of major steroidogenic enzymes was detected by immunohistochemistry. Our observations suggest a mean ovulation rate of 2.3 +/- 0.6 follicles and a low rate of reproductive wastage (0.4 +/- 0.6 oocytes or embryos per pregnancy). The collared peccary seems to exhibit follicular waves involving the synchronous growth of a cohort of follicles, several of which seem to attain selection. The presence of antral follicles in pregnant females suggests that follicular turnover continues during pregnancy. In cyclic animals, corpora lutea were characterised by the presence of distinct large and small luteal cell populations. The luteal volume in pregnant females was larger than that recorded for non-pregnant females. Through immunohistochemistry, it was observed that luteal cells from active corpora lutea exhibit intensive 3beta-HSD expression in advanced stages of pregnancy. This suggests that the corpora lutea seems to remain steroidogenically active throughout pregnancy and likely contribute to progesterone production during pregnancy.

3-Hydroxysteroid Dehydrogenases↗

Estrogen and progesterone receptor content in breast epithelial cells from healthy women during the menstrual cycle.

OBJECTIVE: Our objective was to analyze the presence of estrogen and progesterone receptors in breast epithelial cells from healthy women during the follicular and luteal phase of the menstrual cycle. STUDY DESIGN: We analyzed estrogen receptor and progesterone receptor variations in breast epithelial cells procured through fine-needle aspiration biopsy from 42 healthy volunteers during the menstrual cycle using immunocytochemical receptor analysis. Differences were assessed by chi 2 test and the Wilcoxon rank sum tests. RESULTS: Estrogen receptor was detected more often in women aspirated in the follicular (68%) than in the luteal (32%) phase (p < 0.001); progesterone receptor was detected in around 80% in both phases. In ovulating women who were aspirated twice during the same menstrual cycle the proportion of estrogen receptor-positive cells was reduced from 20% to 4% (p < 0.02); the progesterone receptor values were 17% and 24%. CONCLUSION: Our data indicate an important difference in progesterone receptor variation in the breast as compared with the endometrium.

Adult↗

Reproductive performance of dairy cows with ovarian cysts after different GnRH and cloprostenol treatments.

Cystic ovarian disease is an important cause of reproductive failure and economic loss for the dairy industry. This report describes two consecutive studies. The objective of the first was to evaluate the response of cows with ovarian cysts to two therapeutic treatments. In the second study, we compared the effectiveness of the best treatment established in Study 1 with that of the Ovsynch protocol. For Study 1, cows were considered to have an ovarian cyst if it was possible to observe a single follicular structure with a follicular antrum diameter > 25 min in the absence of a corpus luteum in three ultrasonographic examinations performed at 7 days intervals. At diagnosis (Day 0), cows were assigned to one of two treatment groups. Cows in Group GnRH/CLP (n = 31) were treated with 100 microg GnRH i.m. and 500 microg cloprostenol (CLP) i.m. on Day 14. Cows in Group GnRH-CLP/CLP(n = 32) were treated with 100 microg GnRH i.m. plus 500 microg CLP i.m. on Day 0, and 500 microg CLP i.m. on Day 14. The animals were inseminated at observed estrus and monitored weekly by ultrasonography for 4 weeks or until Al. Cows in the GnRH-CLP/CLP group showed a lower cystic persistence rate (15.6% < 45.2%; P = 0.01); a higher estrus detection rate (84.4% > 41.9%; P < 0.0001); a higher ovulation rate (75% versus 32.3%; P < 0.0001) and a higher early response rate (31% > 3%; P = 0.02) than those in the GnRH/CLP group. For the second study, 128 cows with ovarian cysts were randomly assigned to one of two treatment groups: cows in Group Ovsynch (n = 64) were treated with 100 microg GnRH i.m. on Day 0, 500 microg CLP on Day 7, and 100 microm GnRH i.m. 36 h later. Cows in this group were inseminated 24 h after the second GnRH dose (Ovsynch protocol). Cows in Group GnRH-CLP/CLP/GnRH (n = 64)were treated as those in the GnRH-CLP/CLP group of Study 1 but received GnRH 32 h after the second CLP treatment and were inseminated 24 h after this. A further group of cows without ovarian cysts inseminated at natural estrus served as the Group Control (n = 64). Cows in the GnRH-CLP/CLP/ GnRH group showed a lower cystic persistence rate (10.9% < 46.9%; P < 0.0001); higher ovulation rate (79.7% > 17.2%; P < 0.0001); higher return to estrus rate (34.3% > 12.5%; P < 0.01) and higher pregnancy rate (28.1% > 3.1%; P < 0.01) than those in Ovsynch; and a similar pregnancy rate (28.1% versus 35.9%) to Control cows. These findings indicate that lactating cows with ovarian cysts can be successfully synchronized and time inseminated using a protocol that combines GnRH and CLP, starting treatment by simultaneously administering both products. This protocol also allows the insemination of cows showing estrus within the first week of treatment. Ovarian cysts were less responsive when treatment was started with GnRH alone.

Animals↗

Effects of presynchronization during the preservice period on subsequent ovarian activity in lactating dairy cows.

Among the strategies aimed at overcoming difficulties in estrus detection in dairy herds, presynchronization with two PGF2alpha treatments 14 days apart before a timed AI protocol has been related to a significant increase in pregnancy rates. The aim of the present study was to evaluate the effects of presynchronization during the preservice period on subsequent ovarian activity in clinically normal lactating dairy cows. A second objective was to evaluate the incidence of reproductive disorders on Day 50 postpartum. Depending on the chronological order of parturition, cows were alternately assigned to a control (n=102) or treatment (n=101) group. Animals in the treatment group were administered two cloprostenol treatments 14 days apart, beginning on Day 22 postpartum. The reproductive tract of each animal was examined ultrasonographically on Days 43 and 50 postpartum to monitor ovarian structures and uterine contents. Blood samples were collected on Day 50 for progesterone determination. Cows were inspected for signs of estrus between Days 50 and 71 postpartum and were then inseminated. Follicular persistence rates were similar in the presynchronized (14.9%) and control (13.7%) groups. Cows in the presynchronized group showed a lower metritis-pyometra rate (0% < 3.9%; P=0.045); a lower ovarian cyst rate (3% < 10.8%; P=0.03); a higher luteal activity rate (progesterone concentrations>/=1ng/ml) on Day 50 postpartum (76.2% > 52.9%; P=0.0005); a higher estrus detection rate (73.3% > 47.1%; P<0.0001); a higher ovulation rate (72% > 44%; P<0.0001) and a higher pregnancy rate (29.7% > 15.7%; P=0.02) than controls. Our results indicate that presynchronization during the preservice period reduces the incidence of ovarian cysts and metritis-pyometra determined on Day 50, and improves ovarian activity from Days 50 to 71 postpartum along with pregnancy rates in clinically normal lactating dairy cows.

Animals↗

Pattern of lactate production and steroidogenesis during growth and maturation of mouse ovarian follicles in vitro.

A model culture system for the study of follicular metabolism has been developed from existing methods of whole-follicle culture. The modified system mimics the in vivo growth and maturation of mouse ovarian follicles from primary to preovulatory stages and the modulating influences of the LH and FSH. This is the first study to demonstrate ovulation in vitro from individual ovarian follicles. The pattern of follicular lactate production relative to steroidogenesis was studied throughout in vitro follicular development over a period of 6 days. Twenty-four-hour samples of medium from individual follicles were analyzed for lactate and pyruvate by an automated analytical technique, and for estradiol and progesterone by an immunoenzymatic method. Follicles produced remarkably large quantities of lactate and estradiol during FSH-stimulated development in vitro. LH further stimulated lactate production but resulted in a significant decrease in estradiol secretion and an ovulation rate of 30%. Progesterone production was not detectable throughout the culture period, and follicles showed no evidence of pyruvate uptake. These findings demonstrate the validity of using this model culture system for the study of follicular metabolism and provide new information on the pattern of carbohydrate metabolism relative to steroidogenesis during follicular growth and maturation.

Animals↗

Sites of expression of DnaJ homologs and Hsp70 in male and female flowers of the Japanese willow Salix gilgiana Seemen.

We did in situ hybridization to identify the sites at which DnaJ homologs (SGJ1 and SGJ3) and Hsp70 were expressed in male and female flowers of the Japanese willow, Salix gilgiana Seemen. Transcripts of SGJ1, SGJ3, and Hsp70 were detected in microspores and in the tapetal layers of immature anthers, in the inner integument of ovules, and in the funicular epidermal layers of ovaries. Transcripts of SGJ3 and Hsp70 were also detected in the nectaries of male and female flowers, in which transcripts of SGJ1 were undetectable.

HSP40 Heat-Shock Proteins↗