The EADGENE Microarray Data Analysis Workshop (open access publication).

Microarray analyses have become an important tool in animal genomics. While their use is becoming widespread, there is still a lot of ongoing research regarding the analysis of microarray data. In the context of a European Network of Excellence, 31 researchers representing 14 research groups from 10 countries performed and discussed the statistical analyses of real and simulated 2-colour microarray data that were distributed among participants.

Genetic variation for egg production, egg quality and bone strength in selected and traditional breeds of laying fowl.

1. A multi-breed experiment was conducted with 25 commercial and traditional lines of laying fowl to determine the extent of between-breed genetic variation for adult body weight, sexual maturity, rate of lay, egg weight and egg composition to 55 weeks of age. The genetic variability for bone strength and eggshell strength was determined at 55 weeks of age and a comparison of commercially selected and traditional breeds was performed.

Age-related changes in fear, sociality and pecking behaviours in two strains of laying hen.

1. Two lines of commercial hybrid layers (Tetra and ISA Brown) were reared from hatch to 30 weeks of age in groups of 8. The objectives of the experiment were to evaluate the significance of the different selection practices involved in the development of the lines and to assess the potential association between selected behavioural states and the potential for feather damage and cannibalism.

Spatial distribution and behaviour of laying hens housed in an alternative system.

The spatial distribution and behaviour of perchery housed laying hens were compared at a constant stocking density (18.5 birds/m(2)) in eight pens with colonies of five different sizes (323 birds (N=1), 374 birds (N=2), 431 birds (N=2), 572 birds (N=1) and 912 birds (N=2)). The birds were placed in the perchery when they were 12 weeks old.

Inhibitory and stimulatory action of porcine follicular fluid on FSH-induced 125I-hCG specific binding in rat granulosa cells.

In previous studies, follicular fluid (FF) of large antral follicles (LFF) manifested a stimulatory effect on granulosa cell (GC) luteinization in domestic livestock, but was found to have an inhibitory effect on rodent GC. In the present study, the type of LFF effect on the luteinizing of rat GC was reevaluated in two different models. GC obtained from immature hypophysectomized rats treated with diethylstilbestrol were cultured with increasing concentrations of FSH alone or FSH + estradiol-17 beta (E2), either for 3 or 4 successive days of culture (model 1) or for 4 days of culture with medium change after 2 days of culture (model 2).

Increasing 125I-human chorionic gonadotrophin specific binding in human granulosa cells by follicle-stimulating hormone and follicular fluid.

It has been reported that success at in-vitro fertilization/embryo transfer (IVF/ET) treatment is increased by follicular fluid (FF) re-injected into the abdomen. In the present study a possible direct effect of FF on human granulosa cell (GC) progesterone (P4) secretion and LH/human chorionic gonadotrophin (HCG) receptor content was studied in the presence and absence of FSH. Human GC cultured for 8 days in medium alone showed a 40-fold decrease in P4 secretion.

Characterization of follicular fluid stimulatory factor upon FSH-induced granulosa cell differentiation.

The modulatory role of follicular fluid (FF) upon gonadotropin-induced granulosa cell (GC) differentiation has been previously demonstrated. In the present study, the stimulatory factor of large-antral FF (LFF) was partially characterized. Addition of ovine FSH to porcine GC culture increased 125I-hCG specific binding about 6-fold and progesterone (P4) secretion 15-fold.

Effect of calcium ion on the maturation of cumulus-enclosed pig follicular oocytes isolated from medium-sized graafian follicles.

Porcine follicular oocytes from medium-sized follicles (3-5 mm in diameter) were cultured in modified Hank's balanced salts solution (MHBS) to which pyruvate, lactate, and glucose were added as an energy source. Bovine serum albumin (0.4%) was added as a protein source and the oocytes were cultured for 42 h at 37 degrees C in 5% CO2 in air.

Follicular fluid modulation of functional LH receptor induction in pig granulosa cells.

Follicular fluid was collected from small (1-2 mm), medium (3-5 mm) and large (6-12 mm) follicles of pigs, treated with charcoal to remove steroids, and tested for effects on the induction of functional LH/hCG receptors in cultures of granulosa cells from small antral pig follicles. Granulosa cells were cultured for 2, 4 or 6 days in Medium 199 + 10% pig serum. Granulosa cells cultured in the presence of purified human FSH (0.

Direct evidence for de-novo synthesis of LH receptors in cultured pig granulosa cells in response to FSH.

FSH plus insulin, cortisol, and thyroxine (IFT) stimulated incorporation of dense isotope-containing (2H, 13C, 15N) amino acids into soluble 125I-labelled hCG binding sites. Evidence of new synthesis of binding sites appeared as early as 3 h after the beginning of the pulse-labelling period. By 48 h the majority of detectable soluble 125I-labelled hCG binding sites appeared to be newly synthesized.

Demonstration of a gradient in inhibin activity, estrogen, progesterone, and delta 4-androstenedione in follicular fluid, ovarian vein blood, and peripheral blood of normal women.

Ovarian vein serum from 3 subjects during the late follicular phase of the menstrual cycle had detectable inhibin activity, whereas ovarian vein serum of 12 other subjects during the early follicular phase and luteal phase had no detectable inhibin activity in a rat anterior pituitary cell culture assay. Subjects having detectable inhibin activity (102 +/- 47 U/100 microliters) had 1257 +/- 582 U/100 microliters inhibin activity in FF, whereas subjects having no detectable inhibin activity had FF levels of 711 +/- 203 U/100 microliters of inhibin activity. Estrogen levels of FF and ovarian vein serum of the group having detectable inhibin activity in ovarian vein serum were 282 +/- 239 ng/ml and 4.

Stimulatory action of follicular fluid components on maturation of granulosa cells from small porcine follicles.

Follicular fluid obtained from large (6-12 mm) porcine follicles (LFF) was investigated to determine its stimulatory activity on progesterone secretion and on follicle stimulating hormone (FSH) induction of 125I-human chorionic gonadotropin (hCG)-luteinizing hormone (LH) binding sites in porcine granulosa cells in a 4-day culture. Incubation of granulosa cells harvested from small porcine follicles (1-2 mm) with 50% LFF led to stimulation of LH/hCG binding sites and progesterone secretion. After partial purification of pooled LFF or proteins precipitated with 90% ethanol on Sephadex G-100 the greatest stimulatory activity was found in the second protein peak eluted from the column.

Follicular fluid inhibin activity and steroid levels in ovarian tissue obtained at autopsy from human infants from 18 to 200 days of age.

The ovaries of 25 human infants from 18 to 200 days of age were obtained at autopsy, and their follicular fluid was subjected to measurement of inhibin activity, estrogen, progesterone, androstenedione, and testosterone. Significant inhibin activity was present in all samples of follicular fluid (charcoal-treated) (138 +/- 19 U/10 microliter follicular fluid; 10,545 +/- 2758 U/ovary). There was a tendency for greater inhibin activity, follicular volume, and estrogen in infants from 18 to 59 days than in older infants.

Stimulatory effects of follicle-stimulating hormone and luteinizing hormone upon secretion of progesterone and inhibin activity by cultured infant human ovarian granulosa cells.

Ovarian tissue obtained from three human infants (60, 120, and 210 days of age) was separated into cell types and cultured. Granulosa cells from two of three subjects were viable and grew in culture. The cells had the potential to secrete low levels of progesterone and responded to luteinizing hormone and follicle-stimulating hormone added in culture with greatly enhanced ability to secrete progesterone.

Inhibin activity and steroid hormone levels in ovarian extracts and ovarian vein plasma of female monkeys during postnatal development.

Inhibin activity, follicle-stimulating hormone (FSH)-suppressing substance, estrogen, progesterone, and androstenedione were measured in charcoal-treated ovarian tissue and ovarian venous and peripheral blood of eight rhesus monkeys ranging from 12 to 48 months of age. All of the monkeys demonstrated inhibin activity in ovarian tissue, which, if expressed per milligram protein, was relatively constant throughout development. However, if the activity was expressed per ovary, the amount of ovarian FSH-suppressing substance increased between 26 and 48 months; it was present in detectable amounts in ovarian venous blood only in one 26-month-old monkey.

Effect of follicle-stimulating hormone upon estrogen and progesterone secretion by cultures of monkey granulosa cells recovered during the periovulatory period.

For determination of how exposure of the monkey follicle to the preovulatory luteinizing hormone (LH)/follicle-stimulating hormone (FSH) surge alters its responsiveness to FSH in terms of estrogen and progesterone secretory ability, monkey thecal tissue and granulosa cells were harvested prior to and during the midcycle LH/FSH surge and cultured for 8 days with testosterone and with and without 100 ng human FSH. The addition of FSH enhanced estrogen secretion in culture (6.8 and 7 times on the average after 6 and 8 days, respectively; P less than 0.

Inhibin activity of preovulatory follicles of gonadotropin-treated and untreated women.

In order to determine whether treatment of endocrinologically normal women with human menopausal gonadotropin (hMG) and human chorionic gonadotropin (hCG) led to alternations in follicular fluid inhibin activity and steroids, both inhibin activity and steroid levels were measured in follicular fluid of 10 follicles of 9 women treated with hMG/hCG and in follicular fluid of 15 preovulatory follicles of 15 untreated women. The women were given 150 IU hMG daily from day 3 until the serum estrogen levels were greater than 300 pg/ml, at which time hMG was discontinued and 10,000 IU hCG was given 50 hours later. Follicular fluid was aspirated from all visible follicles 36 hours after hCG.

Porcine inhibin: initial fractionation as a high molecular weight complex.

A high molecular weight complex or aggregate of inhibition was obtained by chromatography of porcine follicular fluid in Fractogel TSK65F. Recovery of activity was good (usually 80-100%), but only 30-60% was recovered as a high molecular weight complex (greater than 160,000) free of albumin and gamma globulin (the two major proteins in follicular fluid). The balance of the activity was distributed in the gamma globulin-albumin region of the chromatogram (i.

Changes in inhibin activity, and progesterone, oestrogen and androstenedione concentrations, in rat follicular fluid throughout the oestrous cycle.

Follicular fluid was aspirated from all visible surface follicles of rats at selected times of the oestrous cycle. Fluids from a pair of rat ovaries were pooled and assayed for inhibin activity by the rat anterior pituitary cell culture assay. Serum LH, FSH and progesterone as well as follicular fluid progesterone, total oestrogens and androstenedione were also measured.

Purification of porcine granulosa cells by continuous Percoll gradient.

Granulosa cells purified on a continuous Percoll gradient have considerably increased responsiveness to FSH in formation of cAMP, secretion of progesterone, and 125I[hCG] binding to cells incubated for 4 days in culture.

Decline of follicular oocyte maturation inhibitor coincident with maturation and achievement of fertilizability of oocytes recovered at midcycle of gonadotropin-treated women.

To examine whether a decline in follicular oocyte maturation inhibitor (OMI) is associated with attainment of oocyte maturation and fertilizability, OMI was measured in follicular fluid (FF) of 39 follicles of 20 normal women given human menopausal gonadotrophin and human chorionic gonadotrophin to induce follicular growth and maturation. Oocytes were aspirated per laparoscope, the fluid was saved, and the egg was observed, incubated, and inseminated with the husband's sperm. Concepti that developed to the 4- to 8-cell stage were transferred to the uterus and the women were followed for pregnancy.

Levels of inhibin-F activity and steroids in human follicular fluid from normal women and women with polycystic ovarian disease.

To examine inhibin-F activity (FSH-suppressing activity) in human follicular fluid of polycystic ovary (PCO) patients, 13 follicles from 5 documented PCO patients and an additional 31 follicles from normal women in the follicular phase of the menstrual cycle were sampled, and inhibin-F activity was measured in rat anterior pituitary cell cultures. Inhibin-F activity was measured in follicular fluid after stripping steroids from the fluids using treatment with dextran and activated charcoal. Estrogen, progesterone, and delta 4-androstenedione in the follicular fluid were also determined by RIA.