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PubMed | Czech University of Life Sciences, Research Institute of Animal Production, French National Center for Scientific Research, Charles University and Academy of Sciences of the Czech Republic
Type: | Journal: Nitric oxide : biology and chemistry | Year: 2015

Hydrogen sulfide, one of three known gasotransmitters, is involved in physiological processes, including reproductive functions. Oocyte maturation and surrounding cumulus cell expansion play an essential role in female reproduction and subsequent embryonic development. Although the positive effects of exogenous hydrogen sulfide on maturing oocytes are well known, the role of endogenous hydrogen sulfide, which is physiologically released by enzymes, has not yet been described in oocytes. In this study, we observed the presence of Cystathionine -Synthase (CBS), Cystathionine -Lyase (CTH) and 3-Mercaptopyruvate Sulfurtransferase (3-MPST), hydrogen sulfide-releasing enzymes, in porcine oocytes. Endogenous hydrogen sulfide production was detected in immature and matured oocytes as well as its requirement for meiotic maturation. Individual hydrogen sulfide-releasing enzymes seem to be capable of substituting for each other in hydrogen sulfide production. However, meiosis suppression by inhibition of all hydrogen sulfide-releasing enzymes is not irreversible and this effect is a result of M-Phase/Maturation Promoting Factor (MPF) and Mitogen-Activated Protein Kinase (MAPK) activity inhibition. Futhermore, cumulus expansion expressed by hyaluronic acid (HA) production is affected by the inhibition of hydrogen sulfide production. Moreover, quality changes of the expanded cumuli are indicated. These results demonstrate hydrogen sulfide involvement in oocyte maturation as well as cumulus expansion. As such, hydrogen sulfide appears to be an important cell messenger during mammalian oocyte meiosis and adequate cumulus expansion.


PubMed | Constantine the Philosopher University, Research Institute of Animal Production, King Saud University, Copenhagen University and University of Trnava
Type: | Journal: Reproductive biology | Year: 2016

Leptin is a hormone that mediates the effect of the metabolic state on several biological functions, including reproduction. Leptin affects reproductive functions via alterations in the release of hormonal regulators. However, the extent to which caloric restriction (CR) can affect the complex processes of reproduction by other mechanisms, such as altering ovarian functions via direct binding/response to leptin, is unknown. Therefore, the aim of the present study was to show basic ovarian cell functions and CR on the response of ovarian cells to leptin. Female rabbits were subjected to 50% CR restriction for 10days before ovulation. On the day of ovulation, both control and CR animals were sacrificed. Isolated granulosa cells were cultured for 2days with and without leptin (100ng/ml), and the accumulation of various markers was evaluated using immunocytochemistry; i.e., cell proliferation (PCNA and cyclin B1), apoptosis (bax), MAP/ERK1,2 kinase (MAPK), protein kinase A (PKA), and IGF-I. In addition, the release of IGF-I and estradiol (E


Sirotkin A.V.,Constantine the Philosopher University | Sirotkin A.V.,Research Institute of Animal Production | Mertin D.,Research Institute of Animal Production | Vegova K.S.,Research Institute of Animal Production | And 2 more authors.
Biology Open | Year: 2016

The aim of our study was to understand whether ovarian steroid hormones, and their response to the metabolic hormones leptin and IGF-I leptin, could be involved in the control of mink reproductive aging via changes in basal release of ovarian progesterone and estradiol. For this purpose, we compared the release of progesterone and estradiol by ovarian fragments isolated from young (yearlings) and old (3-5 years of age) minks cultured with and without leptin and IGF-I (0, 1, 10 or 100 ng/ml). We observed that isolated ovaries of older animals produced less progesterone but not less estradiol than the ovaries of young animals. Leptin addition stimulated estradiol release by the ovarian tissue of young animals but inhibited it in older females. Leptin did not influence progesterone output by the ovaries of either young or older animals. IGF-I inhibited estradiol output in young but not old animals, whereas progesterone release was inhibited by IGF-I irrespective of the animal age. Our observations demonstrate the involvement of both leptin and IGF-I in the control of mink ovarian steroid hormones release. Furthermore, our findings suggest that reproductive aging in minks can be due to (a) reduction in basal progesterone release and (b) alterations in the response of estradiol but not of progesterone to leptin and IGF-I. © 2016. Published by The Company of Biologists Ltd.


Petr J.,Research Institute of Animal Production | Krejcova M.,Czech University of Life Sciences | Rajmon R.,Czech University of Life Sciences | Jilek F.,Czech University of Life Sciences
Animal | Year: 2011

When cultured for an extended time, pig oocytes that matured in vitro to the stage of metaphase II undergo the complex process designated as ageing. Under our conditions, some pig oocytes aged 3 days remained at the stage of metaphase II (22%), but others underwent spontaneous parthenogenetic activation (45%), and still others perished through fragmentation (28%) or lysis (5%). Activation of protein kinases C (PKCs) using phorbol-12-myristate-13-acetate (PMA) protects oocytes from fragmentation. None of the oocytes were fragmented after 3 days of aging in 50 nM of PMA. A similar effect (8% of fragmented oocytes) was observed after a 3-day treatment of aging oocytes with 100 M of 1-stearoyl-2arachidonoyl-sn-glycerol (STEAR). PMA and STEAR activate both calcium-dependent and calcium-independent PKCs. This combined effect on PKCs seems to be essential for the protection of oocytes from fragmentation. Neither the specific activator of calcium-dependent PKCs 1-oleoyl-2-acetyl-sn-glycerol (OLE) nor the specific activator of calcium-independent PKCs dipalmitoyl-l - phosphatidylinositol-3,4,5-triphosphate heptaammonium salt (DIPALM) suppressed the fragmentation of aging pig oocytes. Twenty-one percentage of oocytes fragmented when aged for 3 days in 10 M OLE and 26% of aged oocytes fragmented in 100 nM of DIPALM. However, fragmentation was significantly suppressed to 7% when the oocytes were exposed to the combination of both 10 M OLE and 100 nM DIPALM. Aging pig oocytes cultured for 1 day with PMA maintained a high capability of being parthenogenetically activated (86% of activated oocytes), using calcium ionophore with 6-dimethylaminopurine. Ageing oocytes treated with PMA also had high capability of cleavage (82%) after their artificial parthenogenetic activation. However, their ability to develop to the stage of blastocyst (12%) was suppressed when compared with oocytes activated immediately after their maturation (29%). © The Animal Consortium 2010.


Sirotkin A.V.,Constantine the Philosopher University | Sirotkin A.V.,Research Institute of Animal Production | Harrath A.H.,King Saud University | Grossmann R.,Friedrich Loeffler Institute
Poultry Science | Year: 2016

The aim of the present study was to examine the role and interrelationships between calorie restriction and obestatin in the control of hormone release by chicken ovarian tissue. For this purpose, we compared the release of progesterone (P), testosteron (T), estradiol (E), and arginine-vasotocin (AVT) by ovarian fragments isolated from chicken subjected and not subjested to food restriction, as well as the response of these ovarian fragments to obestatin additions. It was observed that food restriction promoted release of P, reduced output of T, but did not affect basal E and AVT release. Obestatin addition reduced E, promoted AVT, and did not alter P and T release by ovarian tissue isolated from ad libitum fed chicken. In ovarian fragments of fasted hens it reduced E, promoted T, and did not influence P and AVT release. The present observations demonstrate (1) that obestatin can directly control the release of avian ovarian hormones - regulators of reproduction, (2) that metabolic state can control the release of these hormones, and (3) metabolic state can alter the response of ovarian hormones to obestatin. © 2016 Poultry Science Association Inc.


Varadyova Z.,Slovak Academy of Sciences | Jalc D.,Slovak Academy of Sciences | Laukova A.,Slovak Academy of Sciences | Mihalikova K.,Slovak Academy of Sciences | Homolka P.,Research Institute of Animal Production
Turkish Journal of Agriculture and Forestry | Year: 2013

Survival of inoculants in grass silages may enable them to improve the quality of silages through enrichment with polyunsaturated fatty acids (PUFAs). The effects of 2 inoculants, Enterococcus faecium 2/3s (EF2/3s) and E. faecium 26/42 (EF26/42), on nutrient composition, fermentation parameters, and fatty acid content in grass silages during ensiling (111 days) of fresh grass (G) were examined under laboratory conditions. The G [285 g of dry matter (DM) kg-1] was ensiled in 36 polyethylene jars (1 L) divided into 3 × 12 sets per treatment and ensiled at 21 °C for 111 days. The 3 silage treatments used were: 1) grass without inoculant (GS, control), 2) grass inoculated by the strain EF2/3s (GS+EF2/3s), and 3) grass inoculated by the strain EF26/42 (GS+EF26/42). The inoculant strains were sufficiently established during ensiling and reached 4.62 log10 cfu g-1 for EF2/3s and 3.76 log10 cfu g-1 for EF26/42 on day 111 of ensiling. Crude protein contents were G > GS+EF2/3s > GS+EF26/42 > GS (129, 110, 109, and 100 g kg-1 of DM, respectively). The lactate-to-acetate ratios were GS < GS+EF26/42 < GS+EF2/3s (2.82, 4.20, and 4.70, respectively). Concentrations of α-linolenic acid and γ-linolenic acid were highest in grass before ensiling (P < 0.001). Higher isomer C18:2 (9,11) content (P < 0.01) was detected in GS+EF2/3s and GS+EF26/42 than in GS. Nutritional manipulation associated with Enterococcus faecium EF2/3s and EF26/42 inoculation of GS resulted in better quality of silages based on lower lactate (GS+EF26/42) and a greater lactic-to-acetic acid ratio (GS+EF2/3s and GS+EF26/42). This might positively affect PUFAs and their isomers. © TÜBİTAK.


In total, 360 one-day-old, Ross 308 broiler chickens were allocated to 9 groups in 5 replicates of 8 birds (4♂ and 4♀). From the age of 1 to 42 days, the chickens were kept in wire-floored cages in order to facilitate excreta collection, and were provided with water and feed ad libitum. The basal starter (1-14 d) and grower (15-42 d) diets contained 10.7 g and 10.8 g of potassium, 1.7 g and 1.6 g of sodium and 1.7 g and 1.9 g · kg-1 of chloride, respectively. The experimental diets were supplemented with potassium hydrogen carbonate (KHCO3), sodium hydrogen carbonate (NaHCO3), and ammonium chloride (NH4Cl) to contain as analyzed: 11.8/12.2 g or 12.5/12.7 g · kg-1 potassium, 2.0/2.0 g · kg-1 sodium, and 2.7/2.6 g or 3.1/3.0 g · kg-1 chloride for starter/grower feeding periods, respectively. The dietary electrolyte balance (DEB) values in the diets ranged from 271 to 349 and 272 to 336, respectively in the two feeding periods. Chicken performance, carcass analysis, dry matter content in excreta and nitrogen and sodium balance were investigated. Used electrolytes levels had a significant effect on performance only in first feeding period (1-14 days). Increased body weight gain was found in the chickens fed diets with higher (12.2 g and 12.7 g) potassium levels (DEB 309-336) or with a lower potassium level (10.7 g) but higher levels of sodium (2.0 g) and chloride (2.7 g and 3.1 g · kg-1) at 274-325 DEB. In starter period, the excreta of the chickens fed the diet with the highest levels of potassium (12.7 g) and adequate levels of sodium (1.7/1.6 g) and chloride (1.7 g) · kg-1 was characterised by a lower dry matter content than in the remaining groups. An increase in dietary potassium (12.5 g) or chloride (2.6 g · kg-1) significantly lowered nitrogen excretion and elevated nitrogen retention and nitrogen utilization. The lowest sodium intake and Na excretion was found in chickens fed a diet containing a lower level of electrolytes (10.8 g K; 1.6 g Na and 1.9 g Cl · kg-1). Sodium retention and sodium utilization were generally lower for diets supplemented with higher level of chloride (2.6 g and 3.0 g · kg-1). © Verlag Eugen Ulmer, Stuttgart.


PubMed | King Saud University, Research Institute of Animal Production, Constantine the Philosopher University and Polish Academy of Sciences
Type: Journal Article | Journal: Biology open | Year: 2016

The aim of our study was to understand whether ovarian steroid hormones, and their response to the metabolic hormones leptin and IGF-I leptin, could be involved in the control of mink reproductive aging via changes in basal release of ovarian progesterone and estradiol. For this purpose, we compared the release of progesterone and estradiol by ovarian fragments isolated from young (yearlings) and old (3-5years of age) minks cultured with and without leptin and IGF-I (0, 1, 10 or 100ng/ml). We observed that isolated ovaries of older animals produced less progesterone but not less estradiol than the ovaries of young animals. Leptin addition stimulated estradiol release by the ovarian tissue of young animals but inhibited it in older females. Leptin did not influence progesterone output by the ovaries of either young or older animals. IGF-I inhibited estradiol output in young but not old animals, whereas progesterone release was inhibited by IGF-I irrespective of the animal age. Our observations demonstrate the involvement of both leptin and IGF-I in the control of mink ovarian steroid hormones release. Furthermore, our findings suggest that reproductive aging in minks can be due to (a) reduction in basal progesterone release and (b) alterations in the response of estradiol but not of progesterone to leptin and IGF-I.


PubMed | King Saud University, Research Institute of Animal Production, Constantine the Philosopher University and Polish Academy of Sciences
Type: Journal Article | Journal: Theriogenology | Year: 2016

The endocrine mechanisms of mink ovarian hormones release and reproductive aging are poorly investigated. The aims of our study were to: (1) identify hormones produced by mink ovaries (the steroids progesterone [P] and estradiol [E], the peptide hormone oxytocin [OT], and the prostaglandin F [PGF] and prostaglandin E [PGE]); (2) examine the effect of FSH and ghrelin on the release of the hormones listed previously; and (3) understand whether these hormones can be involved in the control of mink reproductive aging, i.e., whether aging can be associated with changes (a) in the basal release of P, E, OT, PGF, or PGE and (b) their response to FSH and ghrelin. Fragments of ovaries of young (yearlings) and old (3-5years of age) minks were cultured with and without FSH and ghrelin (0, 1, 10, or 100ng/mL), and the release of hormones was analyzed by EIA/RIA. We found that isolated ovaries were able to release P, E, OT, PGF, and PGE, and the levels of P produced in the ovaries of old animals were lower than those produced in the ovaries of young animals, whereas the levels of other hormones did not differ. FSH was able to stimulate P and E and suppress OT and PGF and did not affect PGE release. Aging was associated with the inhibition of the effect of FSH on ovarian P and E, the appearance of the inhibitory action of FSH on OT, and the disappearance of this action on ovarian PGF. PGE was not affected by FSH, irrespective of animal age. Ghrelin was able to promote E (but not P) and suppress OT, PGF, and PGE output. Aging was associated with the appearance of an inhibitory influence of ghrelin on ovarian OT and PGE and with the disappearance of this influence on PGF output. Aging did not affect the action of ghrelin on ovarian P and E. Our observations (1) confirm the production of P and E and show that OT, PGF, and PGE are released from mink ovaries, (2) confirm the involvement of FSH and demonstrate the involvement of ghrelin in the control of mink ovarian hormone release, and (3) suggest that reproductive aging in minks is due to a reduction in basal P release and alterations in the response of E, OT, PGF (but not of PGE) to FSH and ghrelin.


PubMed | Research Institute of Animal Production
Type: Journal Article | Journal: Functional & integrative genomics | Year: 2015

MicroRNAs (miRNAs) are known to influence ovarian cell proliferation, apoptosis and hormone release, but it remains unknown whether miRNAs affect ovarian functions via transcription factors. We examined the effect of miRNAs on nuclear factor-appaB (NF-kB) (p65) expression in human ovarian luteinized granulosa cells. We transfected cultured primary human ovarian luteinized granulosa cells with 80 different constructs encoding human pre-miRNAs and then evaluated NF-kB (p65) expression (percentage of cells containing p65) by immunocytochemistry. We found that 21 of the constructs stimulated NF-kB (p65) expression and 18 of the constructs inhibited NF-kB (p65) expression. This is the first direct demonstration that miRNAs affect NF-kB (p65) expression and the first genome-scale miRNA screen to identify upregulation and downregulation of NF-kB accumulation by miRNAs in the ovary. Novel miRNAs that affect the NF-kB signalling pathway could be useful for the control of NF-kB-dependent reproductive processes and the treatment of NF-kB-dependent reproductive disorders.

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