Changes in the peripheral concentrations of inhibin, follicle-stimulating hormone, luteinizing hormone, progesterone and estradiol-17beta during turnover of cystic follicles in dairy cows with spontaneous follicular cysts

Several studies have clarified that the follicular cysts degenerate and are replaced by newly growing follicles that develop into new follicular cysts without ovulation, i.e., turnover of ovarian follicular cysts in cows. However, the relativity of endocrinological changes, including the inhibin profile during turnover of spontaneous follicular cysts in dairy cows, is still unclear. In the present study, the relationship between turnover of follicular cysts and changes in the peripheral blood concentrations of progesterone (P), estradiol-17beta (E(2)), luteinizing hormone (LH), follicle stimulating hormone (FSH) and inhibin were examined in lactating dairy cows. Five cows diagnosed with follicular cysts (follicles of more than 25 mm in diameter in the absence of a corpus luteum) were investigated. Their ovarian dynamics were monitored using ultrasonography, and blood samples were collected at 2- or 3- day intervals throughout the experiment. The day when a follicle fated to become a follicular cyst reached more than 8 mm in diameter was defined as the start of a cystic follicular wave. Four of the 5 cows exhibited a similar patterns of cystic follicular changes and hormone profiles. The data from the 4 cows was used for analysis of the relationships between turnover of cystic follicles and the hormone profiles. Two or three new cystic follicular waves occurred in each cow during the experimental period. The mean diameter of the cystic follicles was more than 25 mm 13 to 15 days after the start of the cystic follicular wave, and it began to decrease 1 to 6 days before the start of the subsequent cystic follicular wave. The levels of E(2) and inhibin tended to decrease for 7 to 9 days before the start of a new cystic follicular wave and to increase concomitantly with new follicular cyst growth. The levels of FSH rose for 1 to 3 days before the start of a new cystic follicular wave. The present study clarified the relationship between FSH and inhibin during turnover of spontaneous follicular cysts in dairy cows and found that it was very similar to previous results for cows. The present results suggest that an increase in FSH secretion following a reduction in inhibin secretion triggers turnover of cystic follicles in cows with spontaneous follicular cysts.     

Ovarian follicular cysts in dairy cows: an abnormality in folliculogenesis

Ovarian follicular cysts are a major reproductive problem in lactating dairy cows. The primary physiological defect leading to the formation of ovarian follicular cysts is a failure of the hypothalamus to trigger the preovulatory surge of luteinizing hormone (LH) in response to estradiol. The factor responsible for this hypothalamic defect may be progesterone. Intermediate levels of progesterone have been shown to prevent ovulation and promote persistence of dominant follicles in normal cycling cows. Recently, we found that 66% of cows with ovarian follicular cysts had progesterone concentrations in an unusual, intermediate range (0.1-1.0 ng/mL) at the time of their detection. A majority of new follicles (76%) that develop in the presence of these intermediate progesterone concentrations became cysts. Only 10% ovulated. Based on these observations, a novel model for the formation and turnover of ovarian follicular cysts is proposed.     

Effect of a single injection of progesterone on ovarian follicular cysts in lactating dairy cows

The objective of this experiment was to evaluate the effect of a single injection of progesterone on the lifespan of ovarian follicular cysts and to examine the fate of follicles that mature following treatment. Lactating Holstein and Jersey cows with ovarian follicular cysts were identified by rectal palpation. The ovaries of cystic cows were then examined by transrectal ultrasonography three times weekly to monitor formation of new follicular cysts. Cows with newly formed follicular cysts were treated either with a single injection of progesterone (200 mg, IM, n = 11) or corn oil vehicle (n = 7). Venous blood samples were collected daily for quantification of progesterone. Blood sampling and ultrasonography continued until ovulation or a new follicular cyst formed. Treatment reduced the lifespan of the cyst by 12 days, from 29.8 +/- 2.3 days in control cows to 17.2 +/- 1.8 days in progesterone-treated cows (P = 0.01). Progesterone treatment also tended to alter the frequency of subsequent follicular events. Ovulation occurred in 4/11 cows that were treated with progesterone whereas none of the vehicle treated cows ovulated (P = 0.07). In conclusion, a single injection of 200mg of progesterone, administered early in the life of an ovarian follicular cyst, shortened its lifespan and in some cases was followed by ovulation of a new follicle.     

Estrogen receptors alpha and beta and progesterone receptors in normal bovine ovarian follicles and cystic ovarian disease

The purpose of this study was to determine by immunohistochemistry the expression of estrogen and progesterone receptors in ovarian follicular structures from cows with cystic ovarian disease (COD) and to compare these with normal ovarian structures. Secondary, tertiary, atretic, and cystic follicles were evaluated. The follicular cysts of animals with COD presented a significantly higher expression of estrogen receptor alpha in all follicular layers than secondary, tertiary, and atretic follicles in both groups (P < .05). The intensity of estrogen receptor beta in the granulosa cell layer was stronger in tertiary than in secondary and atretic follicles in normal animals (P < .05) and in growing and cystic follicles in animals with COD (P < .05). Theca cells were scarcely stained in the 2 groups. Growing follicles and cysts from COD animals were less stained than tertiary follicles from normal animals (P < .05). Differences did not exist between the 2 groups with regard to the progesterone receptor. Ovaries of animals with COD exhibited altered estrogen receptors expression compared with that in normal animals.     

Pregnancy-associated plasma protein-A and insulin-like growth factor binding protein mRNAs in granulosa cells of dominant and subordinate follicles of preovulatory cattle

To determine if (1) levels of pregnancy-associated plasma protein-A (PAPP-A) mRNA and insulin-like growth factor binding protein (IGFBP) (-2, -3, -4 and -5) mRNAs differ between the dominant and subordinate follicles during the follicular phase of an estrous cycle, and (2) these differences are associated with differences in follicular fluid (FFL) concentrations of steroids (estradiol, androstenedione, and progesterone), total and free IGF-I, or IGFBPs, estrous cycles of non-lactating Holstein dairy cows (n = 16) were synchronized with two injections of prostaglandin (PGF2 alpha) 11 days apart. Granulosa cells and FFL were collected either 24 h or 48 h after the second injection of PGF2 alpha. FFL from dominant follicles had lower concentrations of progesterone (P < 0.08) and higher concentrations of estradiol (P < 0.05), androstenedione (P < 0.0001), estradiol:progesterone ratio (P < 0.0001), free IGF-I (P < 0.0001), and calculated percentage free IGF-I (P < 0.01) than large subordinate follicles. Levels of IGFBP-2, -4, and -5 in FFL were 3.0- (P < 0.05), 2.4- (P < 0.06), and 3.4-fold (P < 0.05) greater, respectively, in subordinate than in dominant follicles. IGFBP-3, IGFBP-4 and PAPP-A mRNA expression and IGF-II concentration did not differ (P > 0.10) between dominant or subordinate follicles. Levels of IGFBP-2 and -5 mRNA were severalfold greater (P < 0.05) in subordinate than dominant follicles. IGFBP-5 mRNA in granulosa cells decreased (P < 0.05) 62% to 92%, between 24h and 48 h post-PGF2 alpha. We conclude that decreased levels of IGFBP-2 and -5 mRNA in granulosa cells may contribute to the decrease in FFL IGFBP-2 and -5 protein levels of preovulatory dominant follicles, and that changes in granulosa cell IGFBP-3 and -4 mRNA and PAPP-A mRNA levels do not occur during final preovulatory follicular development in cattle.     

Effects of intermittent injections of LHRH on specific binding of 125I-labeled gonadotropins to granulosa and theca, and concentrations of steroids in serum and ovarian follicles during postpartum anovulation in suckled beef cows

To examine ovarian follicular response to low-dose injections of luteinizing hormone-releasing hormone (LHRH), 32 anovulatory, suckled beef cows were allotted to one of four treatment groups and injected with either saline or 500 ng LHRH every 2 h for 48 or 96 h, starting 21.4 +/- .4 d after parturition. Two hours after the last injection of LHRH, cows were ovariectomized and 10 to 15 ovarian follicles per pair of ovaries were removed and categorized by diameter as small (1.0 to 3.9 mm), medium (4.0 to 7.9 mm) or large (greater than or equal to 8.0 mm). Injections of LHRH did not affect (P greater than .10) steroid levels in small follicles or numbers of gonadotropin receptors in small and medium follicles. Concentrations of progesterone in fluid of medium follicles increased 1.5-fold (P less than .05) after 96 h of LHRH, whereas concentrations of estradiol and androstenedione were unchanged. In fluid of large follicles, concentrations of progesterone were fourfold greater (P less than .05) in LHRH-treated than in control cows at 48 h, but by 96 h progesterone was twofold greater (P less than .05) in control than LHRH-treated cows. In large follicles, concentrations of estradiol were unchanged (P greater than .10) after 48 h of LHRH injections but after 96 h estradiol was twofold greater (P less than .05) in LHRH-treated than control cows. Increased concentrations of estradiol in large follicles coincided with increased numbers of binding sites for human chorionic gonadotropin (hCG) but not follicle stimulating hormone (FSH) in granulosa and theca.(ABSTRACT TRUNCATED AT 250 WORDS)     

Insulin-like growth factor I in sera, ovarian follicles and follicular fluid of cows with spontaneous or induced cystic ovarian disease

The objective of this research was to determine changes in IGF-I levels in serum and follicular fluid, and immunoreactivity of the follicle wall of cows with spontaneous (slaughter specimens) or ACTH-induced follicular cysts, and to compare results to normal cycling (control) cows after selection of the ovulatory follicle. Concentrations of IGF-I in serum did not differ between control and cystic animals (p=0.76). Fluid from the ovulatory follicle in control cows had 41% higher concentrations of IGF-I than that from cystic follicles collected at slaughter (spontaneous cysts; p<0.05) and 70% higher than that in induced follicular cysts (p<0.05). An intense positive immunostaining with anti-IGF-I was observed in granulosa cells (p<0.05) and in the theca interna (p<0.05) of secondary and tertiary follicles in all three groups of animals, but staining was less intense in cystic (p<0.05) and atretic follicles (p<0.05). This study provides evidence to suggest that cystic ovarian disease in cattle is associated with decreased concentrations of IGF-I in follicular fluid, but not in serum, and decreased production of IGF-I in the follicular wall. These data support the notion that IGF-I plays a role in the regulation of folliculogenesis, and may participate in the pathogenesis of cystic ovarian disease in cattle.     

Ovarian follicular development in cattle selected for twin ovulations and births

Comparisons of numbers of antral ovarian follicles and corpora lutea (CL), of blood hormone concentrations, and of follicular fluid steroid concentrations and IGFBP activity were conducted between cows selected (twinner) and unselected (control) for twin births to elucidate genetic differences in the regulation of ovarian follicular development. Ovarian follicular development was synchronized among cows by a single i.m. injection of PGF2alpha on d 18 of the estrous cycle; six cows per population were slaughtered at 0, 24, 48, and 72 h after PGF2alpha. Jugular vein blood was collected from each animal at PGF2alpha injection and at 24-h intervals until slaughter. Ovaries of twinner cows contained more small (< or = 5 mm in diameter, P < 0.05), medium (5.1 to 9.9 mm, P < 0.05), and large (> or = 10.0 mm, P < 0.01) follicles and more (P < 0.01) CL than ovaries of controls. Follicular fluid concentrations of estradiol, androstenedione, testosterone, and progesterone reflected the stage of follicular development and were similar for twinner and control follicles at the same stage. Earlier initiation of follicular development and/or selection of twin-dominant follicles in some twinner cows resulted in greater concentrations of estradiol in plasma at 0, 24, and 48 h and of estradiol, androstenedione, and testosterone in follicular fluid of large follicles at 0 h after PGF2alpha for twinner vs. control cows (follicular status x time x population, P < 0.01). Binding activities of IGFBP-5 and -4 were absent or reduced (P < 0.01) in follicular fluid of developing medium and large estro-gen-active (estradiol:progesterone ratio > 1) follicles but increased with atresia. Only preovulatory Graafian follicles lacked IGFBP-2 binding, suggesting a possible role for IGFBP-2 in selection of the dominant follicle. Concentrations of IGF-I were twofold greater (P < 0.01), but GH (P = 0.10) and cholesterol (P < 0.05) were less in blood of twinners. Three generations of selection of cattle for twin ovulations and births enhanced ovarian follicular development as manifested by increased numbers of follicles within a follicular wave and subsequent selection of twin dominant follicles. Because gonadotropin secretion and ovarian steroidogenesis were similar for control and twinner cattle, enhanced follicular development in twinners may result from decreased inhibition by the dominant follicle(s), increased ovarian sensitivity to gonadotropins, and/or increased intragonadal stimulation, possibly by increased IGF-I.     

Use of milk progesterone enzyme immunoassay for differential diagnosis of follicular cyst, luteal cyst, and cystic corpus luteum in cows

In 160 cows with ovarian cysts as determined by rectal palpation, differentiation was made of follicular cyst, luteal cyst, and cystic corpus luteum on the basis of milk progesterone concentrations estimated by an enzyme immunoassay before and at 10 days after cows were treated with gonadotropin-releasing hormone. Cows having a progesterone concentration in skim milk less than 1.0 ng/ml were considered to have follicular cysts and those with concentrations of 1.0 ng/ml or higher were regarded as the cases of luteal cyst or cystic corpus luteum. Luteal cyst was characterized by progesterone values remaining high in the cows for 10 days after treatment, and cystic corpus luteum was characterized by a decrease in progesterone concentration after cows were treated. By the rectal palpation procedure it was impossible to differentiate luteal cyst and cystic corpus luteum from follicular cyst. The frequencies of follicular cyst, luteal cyst, and cystic corpus luteum were 65%, 19%, and 16%, respectively. Of 104 cows with follicular cysts as defined by milk progesterone assay result, 73 (70%) responded to the treatment with gonadotropin-releasing hormone, the milk progesterone concentration increasing from 0.7 +/- 0.2 ng/ml (mean +/- SD) to 1.8 +/- 1.1 ng/ml. The accuracy of rectal palpation 10 days after treatment for judgment of luteinization of follicular cyst confirmed by milk progesterone analysis was only 30% (48 cows of 160).     

Effects of intraovarian infusion of insulin-like growth factor-I on ovarian follicular function in cattle

The objective of this study was to determine if increased secretion of intraovarian insulin-like growth factor-I (IGF-I), experimentally induced via minipumps, affects follicular function in cattle. Fourteen cycling Holstein cows were divided equally into two groups: Control, osmotic minipumps (containing vehicle) surgically inserted into each ovary, or IGF-I treated, osmotic minipumps as in Controls but pumping 2.0 microg of recombinant human IGF-I per hr for 7 days. All cows were synchronized with prostaglandin F(2alpha) 0.10) between Control and IGF-I-treated cows during Days 2 to 6 of treatment. IGF-I treatment increased (P<0.05) estradiol concentrations in follicular fluid of small follicles, but had no effect (P<0.10) on estradiol concentrations in follicular fluid of large follicles, or on progesterone, androstenedione, or IGF binding protein concentrations in small or large follicles. We conclude that a 7-day infusion of IGF-I directly into the stroma of the ovary altered follicular growth and follicular fluid estradiol concentrations.     

Steroids and cAMP in follicles of postpartum beef cows treated with norgestomet.

To determine time of occurrence of follicular changes that may be associated with the length of the subsequent luteal phase, follicles were collected at different times before ovulation from cows expected to have corpora lutea of short (control) or normal (norgestomet-treated) life span. Beginning on d 20 to 23 postpartum (d 0 of study), 34 crossbred beef cows received either a 6-mg implant of norgestomet for 9 d or served as untreated controls. Ovaries were removed from norgestomet-treated cows on d 6 (N6; n = 9), d 8 (N8; n = 8), or the day after implant removal (N10; n = 8). Control cows were ovariectomized on d 6 (C6; n = 4) or d 10 (C10; n = 5). The largest and second largest follicles greater than 8 mm (F1 and F2, respectively) were dissected from the ovaries. Granulosal and thecal layers and follicular fluid were separated and assayed for estradiol-17 beta, progesterone, androstenedione, and testosterone. Cyclic 3'5'adenosine monophosphate (cAMP) was determined in thecal and granulosal tissue. Diameters of the F1 (14.6 +/- .4 mm) and F2 (10.6 +/- .4 mm) did not differ due to treatment. A greater proportion (P less than .05) of the F1 (20/33) than of the F2 (4/27) had estradiol:progesterone ratios of greater than 1 in follicular fluid. Contents of estradiol, androstenedione, and testosterone in theca and granulosa and follicular fluid, androstenedione in theca, and testosterone in theca and follicular fluid (all P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Plasma progesterone concentrations in dairy cows with cystic ovaries and clinical responses following treatment with fenprostalene

Sixty-two dairy cows diagnosed as having cystic ovarian degeneration were used to study the correlation between rectal palpation findings and plasma progesterone concentrations and the response of cysts to treatment using fenprostalene, a luteolytic agent. Rectal palpation accurately determined the presence of luteal cysts as confirmed by plasma progesterone concentrations of 3 ng/mL or more. Treatment with fenprostalene was very effective for luteal cysts: a high percentage of treated cows exhibited estrus within seven days after treatment. The conception rate following artifical insemination during the induced estrus was 87.5% (21/24). Rectal palpation was much less accurate for the diagnosis of follicular cysts. Cows diagnosed as having follicular cysts had wide variations in plasma progesterone concentrations. Response to fenprostalene treatment was poor in cows with nonluteinized cystic follicles associated with low progesterone concentrations. However, cows diagnosed as having follicular cysts, but with progesterone concentrations of 1 ng/mL or more, responded better to fenprostalene treatment than cows with low progesterone concentrations.

It was concluded that, if correctly diagnosed, luteal cysts can be successfully treated with fenprostalene, and conception rates following treatment can be expected to be normal.

     

Elevated temperature (heat stress) in vitro reduces androstenedione and estradiol and increases progesterone secretion by follicular cells from bovine dominant follicles

Dairy cattle are susceptible to heat stress-induced reductions in fertility; however, direct effects of hyperthermia on specific reproductive functions are difficult to determine in vivo. The objective of this experiment was to examine the effect of elevated temperature in vitro on follicular steroidogenesis, to gain insight into specific follicular responses associated with heat stress. Dominant follicles were obtained from Holstein heifers on day 6 post-estrus (luteal phase; n = 4) or day 8, 36 h after an injection with 25 mg PGF(2alpha) to induce regression of the corpus luteum (follicular phase; n = 4). Pieces of follicle wall were isolated from dominant follicles and cultured for 96 h with 0, 2 or 100 ng/ml LH or FSH at 37, 39 or 41 degrees C. Concentrations of androstenedione, estradiol and progesterone were determined in culture media collected every 24h. During the last 48 h of culture, basal secretion of androstenedione and estradiol by pieces of follicle wall was lower at 41 degrees C than at 37 or 39 degrees C (P < 0.05). In contrast, cumulative secretion of progesterone by pieces of follicle wall in medium alone was higher at 41 degrees C than at 37 or 39 degrees C (P < 0.05). Pieces of follicle wall responded to treatment with both low (2 ng/ml) and high (100 ng/ml) doses of gonadotropins at all temperatures. However, gonadotropin-induced secretion of androstenedione and estradiol was generally lower, whereas gonadotropin-induced secretion of progesterone was higher at 41 degrees C and sometimes at 39 than at 37 degrees C. The changes in basal steroidogenesis and in responses to gonadotropins suggest that follicular cells begin to luteinize at elevated temperatures in vitro. Premature luteinization of follicular cells in vivo has been associated with reduced fertility in cattle with persistent follicles, suggesting that the premature differentiation of follicular cells observed in the current study may be responsible, in part, for the reduced fertility of dairy cattle under heat-stressed conditions.     

Fate of cystic ovarian follicles and the subsequent fertility of early postpartum dairy cows

The fate of cystic ovarian follicles that developed spontaneously during the early postpartum period of 50 lactating dairy cows was traced by ultrasonography to characterise the follicular dynamics in relation to the fertility of the cows. The absence of postpartum ovulations caused by repeated waves of anovulatory large follicles was also characterised and evaluated. Fifteen of the 50 cows developed cystic follicles, and these follicles became follicular cysts in five of the 15 cows. Most of the cystic follicles emerged before the first postpartum ovulation of the cows. The transition from cystic follicles to follicular cysts delayed the cows' first ovulation, oestrus and insemination, but had less influence on their fertility after they had recovered spontaneously. In addition to the 15 cows that developed cystic follicles or follicular cysts, six of the cows had five to 13 waves of follicles before their first ovulation. These repeated waves of follicles caused a more severe delay in the early postpartum reproductive events but did not affect the cows' fertility.     

Steroid concentrations in cows with corticotropin-induced cystic ovarian follicles and the effect of prostaglandin F2alpha and indomethacin given by intrauterine injection.

In 7 instances, cystic ovarian follicles resulted when adrenocorticotropin (ACTH) was administered daily during the follicular phase of the estrous cycle in cows. Two cows given daily injections of hydrocortisone (cortisol) during the follicular phase of the estrous cycle did not develop cystic ovaries. Plasma concentrations of estradiol in cows with induced cystic ovarian follicles were similar to the peak values observed at estrus and were between 6 and 12 pg/ml. Progesterone concentrations in plasma of cows with cystic ovaries were low, between 1 and 2 ng/ml. Ovulation occurred when 2 cows were given human chorionic gonadotropin (HCG) during the period of ovarian cyst development with ACTH administration. Several days of administration of ACTH was required to cause cyst development. Ovulation occurred at the expected time in 1 cow when injections began on day 19, that is, late in the follicular period. In another cow, when treatment was stopped on day 3, after the expected time of estrus a delayed ovulation occurred. In 2 cows with induced cystic ovarian follicles, cyst atresia occurred spontaneously about day 13 to 17 of the cycle. In these cows, new follicular growth and ovulation followed (although delayed in 1 cow). The time of atresia of cystic follicles was not influenced by the intrauterine injection of 10 ml of sterile saline solution on days 8, 9, and 10 in 1 cow. When 5 mg of prostaglandin F2alpha in 10 ml of sterile saline solution was given (uterine injection) in 2 cows on days 8, 9, and 10, cyst atresia occurred earlier than the time of spontaneous atresia. Intrauterine administration of 100 mg of indomethacin in 10 ml of sterile saline solution daily for 13 or 14 days to 2 cows, starting on day 12 or 13 of the cycle, resulted in persistence of the induced cystic ovarian follicles. After cessation of indomethacin treatment, atresia of cysts followed and new follicular growth and ovulation occurred.     

Involvement of PAPP‐A and IGFR1 in Cystic Ovarian Disease in Cattle

Cystic ovarian disease (COD) is one of the main causes of infertility in dairy cattle. It has been shown that intra‐ovarian factors, such as members of the insulin‐like growth factor (IGF) system, may contribute to follicular persistence. The bioavailability of IGF to initiate its response by binding to specific receptors (IGFRs) depends on interactions with related compounds, such as pregnancy‐associated plasma protein A (PAPP‐A). The aim of this study was to determine IGFR1 and PAPP‐A expression both in follicles at different stages of development and in cysts, to evaluate the roles in the etiopathogenesis of COD in cattle. The mRNA expression of PAPP‐A was higher in granulosa cells of large tertiary follicles than in cysts, whereas the protein PAPP‐A present in the follicular fluid from these follicles showed no differences. Although no PAPP‐A mRNA expression was detected in smaller tertiary follicles, in their follicular fluid, this protease was detected in lesser concentration than in cysts. The mRNA expression of IGFR1 was lower in granulosa cells from cystic follicles than in those from tertiary ones. However, the protein expression of this receptor presented the highest levels in cystic structures, probably to increase the possibility of IGF response. The data obtained would indicate that animals with COD have an altered regulation of the IGF system in the ovary, which could be involved in the pathogenesis of this disease in cattle.     

Bovine cystic ovarian disease: plasma hormone concentrations and treatment.

Ovarian function in 91 dairy cows with cystic ovarian disease was assessed by rectal palpation and by plasma hormone analysis before and after treatment. Plasma analysis showed that 84% of the cysts were correctly classified clinically and only these cows are considered further. Luteinised cysts occurred in 59 cows whereas only 18 had non-luteinised cysts. The mean plasma concentrations of luteinising hormone (LH), follicular stimulating hormone (FSH), progesterone, oestradiol and testosterone were not significantly different when compared with values at relevant stages of the oestrous cycle in normal cows. Success of treatment with progesterone, a synthetic prostaglandin, human gonadotrophin (HCG), or gonadotrophin releasing hormone (GnRH) was not dependent upon prior hormone concentrations, except for the prostaglandin which required active luteal tissue. LH and FSH concentrations in cows with luteinised cysts were not significantly different before and after successful treatment with GnRH or progesterone. Normal luteal function was not always established after treatment of non-luteinised cysts with GnRH.     

Characteristics of developing prolonged dominant follicles in cattle

In cattle, sub-luteal circulating progesterone induces an increase in the frequency of LH pulses, prolonged growth of the dominant follicle, increased peripheral estradiol and reduced fertility. The objective of this study was to examine the earliest stages of development of prolonged dominant follicles, to gain insight into the etiology of this aberrant condition. Heifers were treated with an intravaginal progesterone-releasing device (CIDR) from Day 4-8 post-estrus and PGF2alpha was injected on Day 6 and again 12h later (early prolonged dominant group). Follicular phase (CIDR: Day 4-6, with PGF2alpha) and luteal phase (CIDR: Day 4-8, without PGF2alpha) groups served as controls. As expected, peripheral progesterone in heifers of the early prolonged dominant group was intermediate between luteal and follicular phase groups after luteal regression (P<0.05). On Day 7, the frequency of LH pulses was higher in heifers of the follicular phase and early prolonged dominant groups than the luteal phase group (P<0.05). Dominant follicles (n = 4 per group) were collected by ovariectomy on Day 8 and were similar in size among groups (P>0.05). Estradiol and androstenedione concentrations in the follicular fluid at ovariectomy were higher in the follicular phase and early prolonged dominant groups versus the luteal phase group (P<0.01), whereas progesterone did not differ among groups (P>0.05). Granulosa cells and theca interna isolated from dominant follicles were incubated for 3h with or without gonadotropins or frozen for later analysis of mRNA for steroidogenic enzymes. Luteinizing doses (128 ng/ml) of LH and FSH increased secretion of progesterone (P<0.05) but did not affect secretion of estradiol by granulosa cells in all groups. Low (2 or 4 ng/ml) and luteinizing doses of LH increased secretion of androstenedione by theca interna to a similar extent among groups. Expression of mRNA for P450 side chain cleavage (P450scc), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), P450 aromatase (aromatase) and Steroidogenic Acute Regulatory (StAR) protein by granulosa cells did not differ among groups (P>0.05). Levels of mRNA for P450scc, 3beta-HSD, 17alpha-hydroxylase (17alpha-OH) and StAR protein in theca interna were similar in the follicular phase and early prolonged dominant groups (P>0.05), but lower in the luteal phase group (P<0.05-0.1). In summary, the premature follicular luteinization observed in previous studies after prolonged periods of sub-luteal progesterone was absent in early prolonged dominant follicles, exposed to sub-luteal progesterone for 36 h, and their characteristics resembled those of control follicles during the follicular phase.     

Supra-basal progesterone concentrations during the follicular phase are associated with development of cystic follicles in dairy cows

The objective of this study was to test the hypothesis that supra-basal concentrations of progesterone during the follicular phase are associated with the development of follicular cysts. Twenty-five non-lactating dairy cows were used in the study, which was performed over five identical replicate trials. Luteolysis was induced during the mid-luteal phase. Transrectal ultrasonography was performed daily to determine the occurrence/timing of ovulation. Plasma samples were collected for progesterone, oestradiol and luteinizing hormone (LH) analysis. Three cows failed to ovulate (cystic anovulatory) but did ovulate in a subsequent replicate (cystic ovulatory). Eight cows from the appropriate replicates were used as control cows (normal group). Follicular growth patterns and plasma oestradiol concentrations were similar between the three groups. However, the plasma progesterone concentrations during the follicular phase were twofold higher in the cystic anovulatory group (P < 0.01). Furthermore, no LH surge was detected in these animals. While LH pulse amplitude was similar between groups, LH pulse frequency in the cystic anovulatory group was attenuated (P < 0.05). In conclusion, the formation of follicular cysts were preceded by elevated plasma progesterone concentrations and the suppression of the LH surge.