Correlation of Blood Flow of the Preovulatory Follicle to its Diameter and Endocrine Profile in Dairy Buffalo

Blood flow of the preovulatory follicle (POF) wall can be used as a predictor of the quality of POF. Our aim was to determine the correlation of blood flow of POF with the POF diameter, and intra‐follicular and plasma concentrations of Insulin‐like Growth Factor‐I (IGF‐1) and oestradiol in dairy buffalo. Nine Murrah buffalo subjected to an ovulation synchronization protocol (Ovsynch) were assessed on day 10 of the protocol for diameter and blood flow of POF, followed by the aspiration of follicle fluid. Prior to follicular aspiration, blood samples were obtained from jugular vein for estimation of IGF‐1 and oestradiol. The vascularity of POF was determined (Range: 250–967 pixel²) along with intra‐follicular and plasma concentration of IGF‐1 (Range: 9.3–31.8 ng/ml and 14.7–29.7 ng/ml respectively) and oestradiol (Range: 124.2–447.9 ng/ml and 0.25–1.05 ng/ml respectively). Diameter of the POF was weakly correlated (r = 0.21, p < 0.01) with blood flow to it. As compared to POF diameter, the blood flow of POF had greater positive correlation with intra‐follicular and plasma concentrations of hormones (IGF‐1 and oestradiol). A strong positive correlation was recorded between intra‐follicular IGF‐1 and oestradiol. Also, plasma concentrations of oestradiol and progesterone were negatively correlated In brief, assessment of the blood flow of the POF is a non‐invasive and reliable indicator of its functional competence as compared to the POF diameter.     

Real-time RT-PCR quantification of pregnancy-associated plasma protein-A mRNA abundance in bovine granulosa and theca cells: effects of hormones in vitro

Ovarian follicular growth and dominance are controlled by a series of hormonal and intraovarian events including a decrease in intrafollicular IGF-binding proteins −2, −4 and −5 levels. Proteolytic enzymes such as pregnancy-associated plasma protein-A (PAPP-A) degrade IGFBPs and increase bioavailability of IGF-I and -II during follicular development. The objective of this study was to determine the effect of IGF-I, IGF-II, insulin (INS), LH, FSH, estradiol (E2), leptin or cortisol on ovarian PAPP-A mRNA levels. Granulosa (GC) from small (SM) (1–5 mm) and large (LG) (8–22 mm) follicles as well as theca cells (TC) from LG follicles were collected from bovine ovaries and cultured for 48 h in medium containing 10% FCS and then treated with various hormones in serum-free medium for an additional 24 h. Cells were treated with various concentrations (3–500 ng/ml) and combinations of IGF-I, IGF-II, FSH, LH, E2, INS, leptin and (or) cortisol for 24 h (Experiments 1–10). PAPP-A mRNA levels were measured using quantitative real-time RT-PCR. In SM-GC and LG-GC, none of the treatments significantly affected (P > 0.10) PAPP-A mRNA abundance. In LG-TC, IGF-I, LH or cortisol did not affect (P > 0.10) PAPP-A mRNA levels, whereas INS with or without LH decreased (P < 0.05) PAPP-A mRNA. E2 alone decreased PAPP-A mRNA levels in LG-TC, and E2 amplified the insulin-induced inhibition of PAPP-A mRNA abundance in LG-TC. We conclude that control of PAPP-A mRNA abundance in granulosa and theca cells differs, and that E2 may be part of an intraovarian negative feedback system which may reduce the bioavailable IGFs in the theca layer during growth and selection of follicles.     

Oxytocin mediates some effects of insulin-like growth factor-I on porcine ovarian follicles

The aims of the present study were (1) to investigate the influence of insulin-like growth factor-I (IGF-I) on follicular size, on the secretion of oxytocin (OT), progesterone (P), estradiol (E), IGF binding protein-3 (IGFBP-3), inhibin A, inhibin B and cAMP and on the expression of proliferation-associated peptide PCNA, ERK-related mitogen activated protein kinase (MAPK/ERK1, 2) and protein kinase A (PKA) in cultured porcine ovarian follicles; (2) to examine the effects of OT on IGF-I and on these functions; and (3) to determine whether the effects of IGF-I can be mediated by OT. To define the involvement of OT in mediating IGF-I action, we compared responses of porcine ovarian follicles to IGF-I and OT and examined whether blockade of endogenous OT by specific antiserum can affect IGF-I action. It was observed that IGF-I (1, 10 or 100 ng/ml) was able to prevent a decrease in the size of ovarian follicles during culture and caused an increase in the diameter of some follicles. It also stimulated the secretion of OT, P, IGFBP-3, inhibin A and cAMP, decreased the secretion of E and inhibin B (RIA/EIA/ELISA), and induced the expression of PCNA, PKA, MAPK/ERK1, but not MAPK/ERK2 (Western blotting). Like IGF-1, OT (100 ng/ml) prevented decrease in follicular size and increased the diameter of some follicles. It also stimulated the secretion of P and IGF-I, but not E. Antiserum against OT (1%), when given alone, did not affect the reduction of follicular size but slightly increased the percentage of follicles increasing their diameter during culture. The antiserum also inhibited secretion of OT and cAMP but not the secretion of P, E, IGFBP-3 or the expression of PKA, MAPK/ERK1 or 2. When given together with IGF-I, the antiserum prevented the stimulatory action of IGF-I on the proportion of enlarged follicles and on OT, IGFBP-3 and MAPK/ERK1. It augmented the effect of IGF-I on P, but not the effect on E, cAMP, PKA or MAPK/ERK2. These observations demonstrate the involvement of IGF-I and OT in the control of ovarian follicular size and follicular cell proliferation, progestagen, estrogen, IGFBP-3, inhibin A and B secretion and in cAMP/PKA- and MAPK/ERK1-dependent intracellular mechanisms. Furthermore, the reciprocal stimulation of IGF-I and OT and the similarity of some their effects, together with the prevention or augmentation of some IGF-I effects after OT blockade, suggest that IGF-I action can be mediated by OT.     

Regulation of in vitro growth of preantral follicles by growth factors in goats

Goat preantral follicles were cultured to investigate the effects of insulin-like growth factor-I (IGF-I), epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on the in vitro growth and viability of oocytes. Preantral follicles were isolated mechanically and enzymatically (using collagenase and DNase) from prepuberal goat ovaries. The working medium was composed of Defined Eagle's Minimum Essential Medium (DMEM) supplemented with HEPES (20 mM), 10% fetal calf serum (FCS), hypoxanthine (2 mM), dibutyryl cyclic adenosine 3',5'-monophosphate (dbcAMP) (2 mM), penicillin (75 ng/ml) and streptomycin (50 ng/ml). The culture medium consisted of the working medium with follicle stimulating hormone (FSH) (100 ng/ml) and hydrocortisone (40 ng/ml) added. In the experiment, goat preantral follicles were cultured for 9 days in the culture medium and in the culture medium supplemented with either IGF-I (100 ng/ml), EGF (50 ng/ml), bFGF (50 ng/ml) or IGF-I (100 ng/ml)+EGF (50 ng/ml). The results indicated that IGF-I (100 ng/ml) effectively maintained the survival of oocytes and promoted their growth; EGF (50 ng/ml) enhanced the survival rate of oocytes but had a negative effect on oocyte growth; bFGF (50 ng/ml) stimulated oocyte survival but had no obvious effect on their growth while IGF-I (100 ng/ml) and EGF (50 ng/ml) in combination had a greater effect on both survival and growth rate of oocytes than IGF-I or EGF alone. The supplementation of IGF-1 and EGF to the culture medium is recommended in the culture of goat preantral follicles.     

Anti‐Muellerian hormone concentration in bitches with histopathologically diagnosed ovarian tumours and cysts

Increased concentrations of Anti‐Muellerian hormone (AMH) can indicate a granulosa cell tumour as shown in women, mares and cows. To investigate AMH to differentiate canine granulosa cell tumour from other ovarian pathologies, we evaluated the ovaries of 63 bitches. Blood serum samples were collected before surgery for AMH analysis. Ovaries were submitted for histopathological examination. Fourteen bitches showed normal ovaries. These bitches had AMH values between 0.12 and 0.99 ng/ml. In 20 bitches ovarian cysts i.e., follicular cysts (n = 8), corpora lutea cysts (n = 7), subsurface cysts (n = 5) were diagnosed. These dogs had AMH values of 0.11–2.09 ng/ml. Bitches with small luteinized follicular cysts had slightly higher AMH values than those without ovarian alteration. In 29 cases ovarian neoplasms i.e., granulosa cell tumour (n = 9), epithelial tumours (n = 16), dysgerminomas (n = 3) and one sarcoma were identified. Anti‐Muellerian hormone values of bitches with an ovarian neoplasm except granulosa cell tumour ranged from 0.18 to 1.18 ng/ml. The AMH values of bitches with granulosa cell tumour ranged from 1.12 to ≤23 ng/ml and were significantly higher (p < .05) than in all of the other bitches. The cut‐off of 0.99 ng/ml gave a sensitivity of 100% and a specificity of 94.44% to diagnose granulosa cell tumour. In conclusion, markedly elevated AMH concentrations in bitches are indicative for a granulosa cell tumour. However, negative testing does not rule out the existence of small one. Differentiation of GCT from luteinized follicular cysts may especially be difficult.     

Normal calves produced after transfer of embryos cultured in a chemically defined medium supplemented with epidermal growth factor and insulin-like growth factor I following ovum pick up and in vitro fertilization in Japanese black cows

The objective of this study was to examine whether high concentrations of epidermal growth factor (EGF) and/or insulin-like growth factor I (IGF-I) would have a beneficial effect on bovine embryo development in vitro and to obtain normal calves by using an ovum pick up method and embryo culture in a chemically defined medium. When compared with controls, EGF (100 or 200 ng/ml) or IGF-I (50 or 100 ng/ml) significantly increased the rate of embryos that developed into blastocysts during an 8-day culture after the in vitro fertilization of oocytes obtained from ovaries from a slaughterhouse. IGF-I induced a dose-dependent increase in cell number in both the inner cell mass and the trophectoderm, whereas EGF stimulated proliferation only in the inner cell mass. A combination of EGF (100 ng/ml) and IGF-I (50 ng/ml) produced an additive effect, and embryos developed into blastocysts at a comparatively high rate (27.9%) compared with controls (12.0%). A similar rate of development was achieved using a combination of EGF and IGF-I in the culture of embryos following ovum pick up by ultrasound-guided transvaginal follicular aspiration and in vitro fertilization, and 5 blastocysts that developed after the culture were transferred into uteri; two embryos implanted, and normal calves were born. These results suggest that the combined use of EGF and IGF-I makes bovine embryo culture in a chemically defined medium a practical and useful procedure for producing blastocysts, and its application to embryo culture following ovum pick up and in vitro fertilization could be useful for producing normal calves.     

Follicle-stimulating hormone (FSH) stimulates the expression of Pin1, a peptidyl-prolyl isomerase, in the bovine granulosa cells

A peptidyl-prolyl isomerase, Pin 1, has been shown to play a role in the regulation of cell cycle progression, both in vitro and in vivo. However, the involvement of Pin 1 during follicular development is not well understood. The aim of this study was first to investigate the expression of Pin 1 mRNA in the granulosa and theca cells of the follicle at different developmental stages of follicles in the bovine ovary, and second, to examine the effects of follicle-stimulating hormone (FSH) and estradiol (E2) on the expression of Pin 1 in the cultured bovine granulosa cells. Follicles were classified into four groups based on the diameter (dominant follicles >8.5mm in diameter, subordinate follicles <8.5mm in diameter) and the relative levels of E2 and progesterone (P4) (E2:P4>1, estrogen active; E2:P4<1, estrogen inactive): i.e. preovulatory dominant follicles (POFs); E2 active dominant follicles (EADs); E2 inactive dominant follicles (EIDs); small follicles (SFs). The expression of the Pin 1 gene was significantly increased in the granulosa cells of EADs as compared with those of other follicles, whereas its expression in theca cells did not differ among follicles at different developmental stages. The concentration of 5 ng/ml FSH alone and the combination of 1 ng/ml E2 and 5 ng/ml FSH stimulated the expression of the Pin 1 gene in bovine granulosa cells. Our data provide the first evidence that Pin 1 expression in the granulosa cells but not the theca cells changes during follicular development, and that FSH stimulate the expression of the Pin 1 gene. These results suggest that Pin 1 regulates the timing of cell proliferation and may act as an intracellular signal responder in the granulosa cells during bovine follicle development.     

Effect of epidermal growth factor and insulin-like growth factor I on porcine preantral follicular growth, antrum formation, and stimulation of granulosal cell proliferation and suppression of apoptosis in vitro

The object of this study was to investigate the role of epidermal growth factor (EGF) and IGF-I in the regulation of preantral follicular growth, antrum formation, and granulosal cell proliferation/ apoptosis. Porcine preantral follicles were manually dissected and cultured for up to 8 d in Waymouth's (Exp. 1) or alpha-minimum Eagle's essential medium (Exp. 2 and 3) supplemented with 10 microg/mL of transferrin, 100 microg/mL of L-ascorbic acid, and 2 mU/mL of ovine FSH, in the presence (Exp. 1 and 3) or absence (Exp. 2) of 7.5% fetal calf serum. According to the experimental protocol, IGF-I (0, 1, 10, or 100 ng/mL; Exp. 1), or IGF-I (50 ng/mL), EGF (10 ng/mL) and EGF+IGF-I (Exp. 2 and 3) were added to the culture media. In Exp. 1, follicles exhibited a concentration-dependent response (P < 0.05) to IGF-I, with the highest rates of granulosal cell proliferation, follicular integrity, and recovery rate of cumulus cell-oocyte complexes and lowest incidence of apoptosis occurring at the highest IGF-I dose. In Exp. 2 serum-free medium, granulosal cell proliferation was low (1 to 5%), irrespective of whether EGF and/or IGF-I were present and cellular apoptosis was increased (P < 0.05) on d 4 and 8 in the EGF+IGF-I group compared with the addition of either factor alone. In Exp. 3, granulosal cell proliferation was high in all follicles cultured in serum-containing medium for the first 3 d, but fell sharply (P < 0.05) on d 4, except in media containing IGF-I. Collectively, EGF and IGF-I increased granulosal cell proliferation, decreased apoptosis, and promoted follicular antrum formation. These results may provide useful information for developing a preantral follicular culture system in which the oocytes are capable of fertilization and embryonic development.     

Serum concentrations of IGF-I in postpartum beef cows

Four experiments assessed changes in serum IGF-I under various physiologic conditions in postpartum cows. In Exp. 1, anestrous suckled cows (n = 25) were infused for 6 d with either saline or glucose at two different infusion rates. In Exp. 2, anestrous cows (n = 29) received either a saline (weaned and suckled controls) or 3 g/d phlorizin (weaned phlorizin) infusion for 3 d. Calves from the weaned groups were removed from 15 h before and throughout infusions. In Exp. 3, cycling suckled cows (n = 20) received prostaglandin F2 alpha (PGF2 alpha) when the 5-d saline or phlorizin infusion began. In Exp. 4, suckled cows (n = 20) had ad libitum access to feed or received 50% of control feed consumption from 30 to 40 d postpartum. Increasing glucose availability (Exp. 1) increased (P less than .05) serum IGF-I by 30 to 35%. IGF-I remained stable after weaning (Exp. 2) in phlorizin-infused cows (128.8 +/- 12.7 ng/ml), but increased (P less than .05) by 3 d after calf removal in weaned control cows (152.2 +/- 7.5 ng/ml). IGF-I also remained stable in phlorizin-infused cows following PGF2 alpha injection (Exp. 3), but increased in control cows by 2 d after PGF2 alpha (156.8 +/- 18.3 on d 2 vs. 133.7 +/- 9.8 ng/ml pre-injection; P less than .05) and remained elevated (P less than .05) during the periovulatory period. In cows receiving restricted feed intake (Exp. 4), IGF-I decreased by approximately 50% within 4 d of feed restriction (71.3 +/- 9.4 vs 137.4 +/- 16.6 ng/ml; P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Plasma progesterone levels in sows with induced cystic ovarian follicles.

Large multiple cystic ovarian follicles were induced in three sows and small multiple cystic ovarian follicles were induced in three other sows by injections of adrenocorticotrophic hormone (ACTH) during the follicular phase of the oestrous cycle. Plasma progesterone levels in sows with large cysts were relatively high, while those of sows with small cystic follicles were low. Removal of the ovaries with large cysts from one sow resulted in a precipitous drop in progesterone levels indicating that the large cysts were primarily the source of progesterone. The method by which ACTH may induced cystic ovaries was also investigated. Plasma progesterone levels in two ovariectomised sows during ACTH treatment fluctuated markedly during a 24 h period achieving peak values of 4 or 5 ng/ml. The results suggest that progesterone of adrenal cortical origin may be a factor in the development of the cystic ovarian condition.     

Insulin-like growth factor I in follicular development and function in postpartum beef cows

The objective of this study was to determine whether nutrition affects follicular growth and(or) steroid and insulin-like growth factor I (IGF-I) concentrations in follicular fluid. Beginning 6 d after calving, Hereford-cross cows (n = 28) were fed either 14 (ad libitum) or 7 (restricted) kg.animal1.d-1 of chopped alfalfa-brome hay. Half the cows in each treatment were ovariectomized on d 20 (OVX-20) and the remaining half on d 35 (OVX-35) postpartum. Cow weight and condition score were recorded weekly, and blood was collected thrice weekly for determination of insulin, IGF-I, glucose, and free fatty acid (FFA) concentrations. At ovariectomy, follicular fluid from each follicle greater than or equal to 4 mm in diameter was aspirated for determination of IGF-I, progesterone (P4), and estradiol-17 beta (E2) concentrations. Restricted cows lost more weight after calving than did ad libitum cows (P less than .0001), although all cows lost similar amounts of body condition (time postpartum, P = .008). Concentrations of FFA were elevated (P less than .0001) in restricted cows from wk 2 through 5 after calving but did not change with time in ad libitum cows. Plasma concentrations of glucose were lower in restricted than in ad libitum cows (59.6 +/- .4 vs 61.8 +/- .4 mg/dl; P = .05), but insulin and IGF-I were similar (P greater than .10) between dietary treatments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Expression of mRNA for the angiopoietin-tie system in granulosa cells during follicular development in cows

Recent findings indicate that the changing profile of angiopoietins (ANPT) and their receptor Tie2 are closely associated with development and regression of the vascular network in the cyclic ovary. We previously reported that mRNA expression for the ANPT-Tie system in theca interna changes during bovine follicular development and atresia, and both ANPTs affect steroidogenesis in the preovulatory follicle. The aim of this study was to investigate mRNA expression for ANPT1, ANPT-2 and Tie2 in granulosa cells (GC) during follicular development in the cow. Bovine follicles were classified according to the estradiol-17beta (E(2)) concentration in follicular fluid (FF) as follows: (1) E(2)<0.5, (2) 0.5180 ng/ml FF. Semi-quantitative RT-PCR analysis revealed that the expression of ANPT-1 mRNA was not detected in most of the follicle with E(2)<5 ng/ml (diameter of 5-10 mm), but clearly detected in all follicles with E(2)>5 ng/ml (diameter of >10 mm). The mRNA expression for ANPT-2 was drastically decreased in the follicles with E(2)>5 ng/ml. Tie2 mRNA expression remained unchanged at the different stages of follicular development. The present data show that ANPT-1 becomes predominant in the follicle producing high levels of E(2), indicating the possible switch-over from ANPT-2 (antagonist) to ANPT-1 (agonist). Thus, the result suggests that the ANPT-Tie system in bovine GC may stimulate E(2) secretion rather than angiogenesis in the late stages of follicular development.     

Endocrine and metabolic changes during altered growth rates in beef cattle

Eight steers from a group of 14 were fed ad libitum from 240 to 510 kg live weight, gaining at 1.4 +/- .2 kg/d. The six other steers were diet-restricted and grew at .37 +/- .09 kg/d from 240 to 307 kg, prior to ad libitum realimentation on the same diet to a final weight of 510 kg. Blood samples taken during the growth phases from both treatments were analyzed for insulin-like growth factor-I (IGF-I), triiodothyronine (T3), thyroxine (T4), glucose (GLU), nonesterified fatty acids (NEFA), and blood urea nitrogen (BUN) and (or) growth hormone (GH). During restricted growth, mean serum concentrations of GH were elevated (45.6 vs 23.4 ng/ml; P less than .05), serum concentrations of IGF-I decreased (108 vs 167 ng/ml; P less than .05) compared with control steers with ad libitum access to feed. Levels of T4 and GLU also were lower (P less than .05) during restricted than during normal growth. During early realimentation, levels of GLU (P less than .05), IGF-I (P less than .01), T4 and BUN (P less than .01) increased. Levels of T3 remained unchanged, whereas concentration of NEFA declined (P less than .001). Blood urea nitrogen decreased during early realimentation despite a large increase in diet protein intake and in protein storage, suggesting an increased efficiency of nitrogen use for protein synthesis. During realimentation, IGF-I levels rose above those of control steers and remained higher at the final weight of 510 kg (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of different methods of diagnosis of cystic ovarian disease in cattle and an assessment of its treatment with a progesterone-releasing intravaginal device

The aims of this study were to assess the accuracy of different common methods of differentiating between follicular and luteal ovarian cysts, and to monitor the response of the cysts to 12 days treatment with a progesterone-releasing intravaginal device (PRID). On the basis of agreement between the different methods, 25 of the 46 cases examined were diagnosed as follicular and 14 as luteal cysts; for the other seven cases the methods disagreed. The use of ultrasound was more accurate in diagnosing follicular cysts than luteal cysts, and combined with plasma progesterone concentrations gave the most accurate assessment of cyst type (92 per cent for follicular cysts and 82 per cent for luteal cysts). The mean (se) plasma progesterone concentration was lower in the cows with follicular cysts than in those with luteal cysts (0.29 [0.05] v 3.90 [0.63] ng/ml; P<0.05). Luteal cysts had thicker walls (5.3 [0.04] v 2.5 [0.2] mm; P<0.0001), and the wall thickness of all the cysts was positively correlated with plasma progesterone concentration (r=0.52, P<0.0004). Cows with luteal cysts had more additional follicles greater than 5 mm in diameter (P<0.01). In cows with follicular cysts and other follicles greater than 5 mm in diameter, the mean oestradiol concentration was 7.9 (1.8) pg/ml compared with 24.2 (3.1) pg/ml (P=0.002) in cows without other follicles greater than 5 mm in diameter on either ovary. At the time of PRID removal, plasma progesterone concentration had increased in the cows with follicular cysts to 1.59 (0.06) ng/ml (P<0.05) and decreased in the cows with luteal cysts to 0.87 (0.01) ng/ml (P<0.05), although there was no change in original cyst structure in 45 per cent of the cases. However, new ovarian structures were frequently observed during the treatment. The overall pregnancy rate for cows with both types of cyst after treatment was 50 per cent after three inseminations, but the first service pregnancy rate was only 18 per cent for cows with follicular cysts and 28 per cent for cows with luteal cysts. After treatment, the fertility of cows with follicular cysts was similar to that of paired herdmates, whereas cows with luteal cysts took 40 days longer to calve again than healthy herdmates. However, the culling rate was higher for cows with follicular cysts (41 v 11 per cent).     

Diagnosis of luteal and follicular ovarian cysts by palpation per rectum and linear-array ultrasonography in dairy cows.

The purpose of this study was to determine and compare the accuracy of palpation per rectum and linear-array ultrasonography for diagnosing follicular vs luteal ovarian cysts in cows. Forty-seven examinations of ovarian cysts from 28 cows were diagnosed by palpation per rectum as either a firm, thick-walled structure (luteal cyst) or a soft, thin-walled structure (follicular cyst) during weekly herd examinations. The ovaries of each cow were then examined by ultrasonography. Ultrasonograms of cysts greater than 25 mm in diameter were diagnosed as luteal or follicular cysts and were recorded on videotape for evaluation by a second clinician. Serum progesterone concentrations at the time of examination were determined by radioimmunoassay and used to classify luteal (greater than 0.5 ng/ml) or follicular (less than or equal to 0.5 ng/ml) cysts. Selection of this discriminatory level was based on response of a proportion of cows with luteal cysts that were given 25 mg of prostaglandin F2 alpha at the time of diagnosis by ultrasonography. Sensitivity and specificity of palpation per rectum for diagnosis of type of ovarian cyst were low (43.3 and 64.7%, respectively). In contrast, sensitivity and specificity of ultrasonography were considerably higher (86.7 and 82.3%, respectively). Agreement between the 2 methods of diagnosis was 57.4%. Overall agreement between the 2 clinicians' diagnoses by ultrasonography was 85.1%. On the basis of our findings, we confirm that luteal and follicular cysts cannot be accurately differentiated by palpation per rectum alone. These data suggest that linear-array ultrasonography is more effective than palpation per rectum for diagnosing type of ovarian cyst in cows.     

Effect of gonadotropin-releasing hormone on clinical response and fertility in cows with cystic ovaries, as related to milk progesterone concentration and days after parturition

We gave gonadotropin-releasing hormone (GnRH) or an analog of GnRH to 264 lactating cows with cystic ovaries. The effects of milk progesterone concentration (MPC) and days after parturition (DAP) at diagnosis on clinical response 30 days after treatment and on subsequent fertility were examined. Palpation per rectum revealed ovarian cysts in 264 cows; 118 had true follicular cysts (MPC less than 1 ng/ml). Clinicians with more than 3 years of experience reported significantly more true cysts than did less experienced clinicians. Clinical response 30 days after treatment did not differ in 3 groups of cows (divided on the basis of MPC at treatment), but fewer cows with MPC less than 1 ng/ml were bred or conceived than were cows with MPC greater than or equal to 33 ng/ml. Days after parturition at diagnosis did not affect 30-day clinical response rate, but cows treated less than 35 DAP had significantly more days to first estrus and to conception than did cows treated greater than 90 DAP. Significantly more cows treated less than 35 DAP had MPC less than 1 ng/ml at treatment.     

Insulin-like growth factor (IGF)-I and IGF binding proteins-2, -3, -4, -5 in porcine corpora lutea during the estrous cycle; evidence for inhibitory actions of IGFBP-3

In this study we measured protein concentrations of insulin-like growth factor (IGF)-I and IGF binding proteins (IGFBPs) 2-5 in porcine corpora lutea (CLs) throughout the estrous cycle (Experiment 1), and examined the effects of IGFBP-3 and IGFBP-3 antibody (AB) on luteal progesterone (P4) secretion in vitro (Experiment 2). For Experiment 1, (CLs) and serum were collected on days (D) 4, 7, 10, 13, 15 and 16 of the estrous cycle (n = 5 animals per day). IGF-I was extracted from CLs and sera, and measured by radioimmunoassay (RIA). IGFBPs were measured in CLs by ligand blots. For Experiment 2, CLs (from Experiment 1) were enzyme dissociated and luteal cells cultured (24 h) in Medium 199 (M199) containing (0-500 ng/ml) IGFBP-3 (+/-IGF-I; 100 ng/ml), or (0-10 microg/ml) IGFBP-3 AB. P4 in media was measured by RIA. In Experiment 1, luteal IGF-I concentrations (ng/g tissue) were maximal on day 4 and gradually decreased thereafter. Serum IGF-I concentrations (ng/ml) were highest on days 4 and 7, compared with days 10-15. Peak levels of luteal IGFBP-3 were also seen on days 4 and 7 of the cycle. Luteal IGFBP-2 concentrations showed a tendency to increase on day 16 (P < 0.05 versus day 10), but no significant changes in IGFBP-4 or -5 were seen. In Experiment 2, IGFBP-3 (w IGF) inhibited the steroidogenic actions of IGF-I, but had no significant actions alone (IGFBP-3 w/o IGF). Finally, IGFBP-3 AB stimulated P4 secretion on days 4 and 7, but not on days 10-16. We conclude that IGFBP-3 inhibits IGF-I actions in the porcine CL.     

Cystic Degeneration in Porcine Ovaries - Second Communication: Concentrations of Progesterone, Estradiol-17β, and Testosterone in Cystic Fluid and Plasma; Interpretation of the Results

Contents: The content of progesterone, estradiol-17β, and testosterone of plasma and cystic fluid was determined in 79 sows with ovarian cysts. The average progesterone concentration of sows with dark corpora lutea (CL) was higher than of sows with pale or absent CL (39.4 vs. 8.7 vs. 8.0 ng/ml plasma; p < 0.001; and 7512 vs. 3644 vs. 2723 ng/ml cystic fluid, respectively; p < 0.001). The cystic fluid of animals with oligocystic ovaries (10 cystslanimal) had a significant higher progesterone concentration in comparison to potycystic animals (7200 vs. 3682 ng/ml; p < 0.001). Testosterone and estradwl-17β levels in plasma and in cystic fluid of polycystic animals were significantly higher in comparison to oligocystic animals (Plasma-Testosterone: p < 0.01; Plasma-Estradwl: p < 0.05; Cyst-Testosterone: p < 0.01; Cyst-Estradiol: p < 0.001). In oligocystic ovaries testosterone in cysts exceeded the estradiol-17β levels, whereas in polycystic ovaries the situation was vice versa (p < 0.001).
It is suggested that cystic ovarian degeneration in the sow is not exclusively a gradually progressing process, rather then a complex syndrome with three components, were characterized by a separate course of development (oligocystic, polycystic. oligo-polycystic syndrome).