Authors: Accialini, Paula Lucia; Hernandez, Silvia Fátima; Bas, Diana Ester; Pazos Maidana, María Camila; Irusta, Griselda; Abramovich, Dalhia Nurit; Tesone, Marta
Publication Date: 2015.
Language: English.
Abstract:
In this study, we investigated the interaction between the Notch pathway and progesterone to maintain the functionality of the corpus luteum(CL).WhenNotch signaling is activated, the g-secretase complex releases the active intracellular domains (NICD) of their receptors, which exert survival effects. We designed studies to analyze whether the in vitro inhibition of Notch affects progesterone production, steroidogenic regulators, apoptotic parameters, and signaling transduction pathways in the cultures of CL isolated from pregnant and superovulated rats. We detected a decrease in progesterone production when corpora lutea (CL) were incubated with N-(N-(3,5- difluorophenacetyl-L-alanyl))-S-phenylglycine t-butyl ester (DAPT), a g-secretase inhibitor. This effect could be in part due to the decrease detected in the CL protein levels of P450scc because STAR and 3b-hydroxysteroid dehydrogenase were not affected by Notch inhibition. Besides, the addition of aminoglutethimide to the CL culture medium decreased NICD of NOTCH1. We observed an increase in the expression of active CASPASE3 (CASP3) after inhibition by Notch,whichwas reversed by the presence of progesterone. The BAX:BCLXL ratio was increased in CL treated with DAPT and the presence of progesterone reversed this effect. In addition, phosphorylation of AKTwas inhibited inCL treated withDAPT, but had no effect on ERK activation. To demonstrate that the action ofDAPTis specifically related with the inhibition of Notch, CLswere incubated with DLL4 antibody and a decrease in progesterone production was detected. These results suggest the existence of a novel link between progesterone and the Notch signaling pathway to maintain the functionality of the CL.
Author affiliation: Accialini, Paula Lucia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Author affiliation: Hernandez, Silvia Fátima. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Author affiliation: Bas, Diana Ester. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Author affiliation: Pazos Maidana, María Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Author affiliation: Irusta, Griselda. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Author affiliation: Abramovich, Dalhia Nurit. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Author affiliation: Tesone, Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Publication Date: 2007.
Language: English.
Abstract:
Studies were designed to examine the expression and activity of four caspases that contribute to the initial (caspases-2, -8, and -9) and final (caspase-3) events in apoptosis in the rat corpus luteum (CL) during pregnancy (days 7, 17, 19, and 21 of gestation), postpartum (days 1 and 4), and after injection (0, 8, 16, 24, and 36 h) of the physiological luteolysin PGF2α. In addition, the temporal relationship of caspase expression/activity relative to steroid production and luteal regression was evaluated. During pregnancy, the activity of all four caspases was significantly greater on day 19, before a decline in CL progesterone (P) and CYP11A1 levels at day 21 of gestation. The levels of the caspase-3 active fragment (p17, measured by Western blot) also increased at days 19 and 21 of pregnancy. Immunohistochemical analyses detected specific staining for the caspases in luteal cells (large and small) as well as in endothelial cells. However, the percentage of apoptotic cells did not increase in the CL until postpartum. Following PGF2α injection, there was a significant decrease in CL P by 24 h, although the activity of all four caspases did not increase until 36 h posttreatment. The active p17 fragment of caspase-3 also significantly increased at 36 h post-PGF2α. These results suggest that an increase in the activity of caspases-2, -8, -9, and -3 is associated with the early events of natural luteolysis at the end of pregnancy. Also, the exogenous administration of the luteolysin PGF2α may regulate members of the caspase family.
Author affiliation: Peluffo, Marina Cinthia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Author affiliation: Stouffer, Richard L.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Author affiliation: Tesone, Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Authors: Vallcaneras, Sandra; Casais, Marilina; Anzulovich Miranda, Ana Cecilia; Delgado, Silvia M.; Sosa, Zulema; Telleria, Carlos Marcelo; Rastrilla, Ana M.
Publication Date: 2011.
Language: English.
Abstract:
Androstenedione can affect luteal function via a neural pathway in the late pregnant rat. Here, we investigate whether androstenedione is capable of opposing to regression of pregnancy corpus luteum that occurs after parturition, indirectly, from the coeliac ganglion. Thus, androstenedione was added into the ganglionar compartment of an ex vivo coeliac ganglion–superior ovarian nerve–ovary system isolated from non-lactating rats on day 4 postpartum. At the end of incubation, we measured the abundance of progesterone, androstenedione and oestradiol released into the ovarian compartment. Luteal mRNA expression and activity of progesterone synthesis and degradation enzymes, 3β-hydroxysteroid-dehydrogenase (3β-HSD) and 20α-hydroxysteroid-dehydrogenase (20α-HSD), respectively, as well as the aromatase, Bcl-2, Bax, Fas and FasL transcript levels, were also determined. Additionally, we measured the ovarian release of norepinephrine, nitric oxide and luteal inducible nitric oxide synthase (iNOS) mRNA expression. The presence of androstenedione in the ganglion compartment significantly increased the release of ovarian progesterone, androstenedione and oestradiol without modifying 3β-HSD and 20α-HSD activities or mRNA expression. The ovarian release of oestradiol in response to the presence of androstenedione in the ganglion compartment declined with time of incubation in accord with a reduction in the aromatase mRNA expression. Androstenedione added to the ganglion compartment decreased FasL mRNA expression, without affecting luteal Bcl-2, Bax and Fas transcript levels; also increased the release of norepinephrine, decreased the release of nitric oxide and increased iNOS mRNA. In summary, on day 4 after parturition, androstenedione can mediate a luteotropic effect acting at the coeliac ganglion and transmitting to the ovary a signaling via a neural pathway in association with increased release of norepinephrine, decreased nitric oxide release, and decreased expression of FasL.
Author affiliation: Vallcaneras, Sandra. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Bioquímica y Ciencias Biológicas. Laboratorio de Biología de la Reproducción; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Casais, Marilina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Bioquímica y Ciencias Biológicas. Laboratorio de Biología de la Reproducción; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Anzulovich Miranda, Ana Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis; Argentina
Author affiliation: Delgado, Silvia M.. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Bioquímica y Ciencias Biológicas. Laboratorio de Biología de la Reproducción; Argentina
Author affiliation: Sosa, Zulema. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Bioquímica y Ciencias Biológicas. Laboratorio de Biología de la Reproducción; Argentina
Author affiliation: Telleria, Carlos Marcelo. University of South Dakota; Estados Unidos
Author affiliation: Rastrilla, Ana M.. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Bioquímica y Ciencias Biológicas. Laboratorio de Biología de la Reproducción; Argentina
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Authors: de la Vega, Magali del Carmen; Delsouc, María Belén; Ponce, Ivana Tamara; Ragusa, Juan Antonio Vicente; Vallcaneras, Sandra; Anzulovich Miranda, Ana Cecilia; Casais, Marilina
Publication Date: 2018.
Language: English.
Abstract:
The rhythm of factors involved in luteal regression is crucial in determining the physiological duration of the oestrous cycle. Given the role of tumour necrosis factor (TNF)-α in luteal function and circadian regulation and that most of the effects of TNF-α are mediated by p55 TNF receptor (TNFRp55), the aims of the present study were to analyse the following during the luteal regression phase in the ovary of mice: (1) whether the pattern of expression of progesterone (P4) and the enzymes involved in the synthesis and degradation of P4 is circadian and endogenous (the rhythm persists in constant conditions, (i.e., constant darkness) with a period of about 24 hours); (2) circadian oscillations in clock gene expression; (3) whether there are daily variations in the expression of key genes involved in apoptosis and antioxidant mechanisms; and (4) the consequences of TNFRp55 deficiency. P4 was found to oscillate circadianally following endogenous rhythms of clock factors. Of note, TNFRp55 deficiency modified the circadian oscillation in P4 concentrations and its enzymes involved in the synthesis and degradation of P4, probably as a consequence of changes in the circadian oscillations of brain and muscle ARNT-Like protein 1 (Bmal1) and Cryptochrome 1 (Cry1). Furthermore, TNFRp55 deficiency modified the circadian rhythms of apoptosis genes, as well as antioxidant enzymes and peroxidation levels in the ovary in dioestrus. The findings of the present study strengthen the hypothesis that dysregulation of TNF-α signalling may be a potential cause for altered circadian and menstrual cycling in some gynaecological diseases.
Author affiliation: de la Vega, Magali del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Bioquímica y Ciencias Biológicas. Laboratorio de Biología de la Reproducción; Argentina
Author affiliation: Delsouc, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Bioquímica y Ciencias Biológicas. Laboratorio de Biología de la Reproducción; Argentina
Author affiliation: Ponce, Ivana Tamara. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
Author affiliation: Ragusa, Juan Antonio Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
Author affiliation: Vallcaneras, Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
Author affiliation: Anzulovich Miranda, Ana Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
Author affiliation: Casais, Marilina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Authors: Irusta, Griselda; Murphy, M. J.; Perez, W. D.; Hennebold J.D.
Publication Date: 2007.
Language: English.
Abstract:
Epoxyeicosatrienoic acids (EpETrEs), produced from arachidonic acid via cytochrome P450 (CYP) epoxygenases, regulate inflammation, angiogenesis, cellular proliferation, ion transport and steroidogenesis. EpETrE actions are regulated through their metabolism to diols (dihydroxyeicosatrienoic acids; DiHETrE) via the enzyme soluble epoxide hydrolase (EPHX2). We set out to determine, therefore, whether EpETrE generating (epoxygenases CYP2C8, 2C9, 2C19, 2J2, 1A2 and 3A4) and metabolizing (EPHX2) enzymes are expressed in the primate corpus luteum (CL). CL were isolated from rhesus macaques during the early (day 3-5 post-LH surge), mid (day 6-8), mid-late (day 10-12), late (day 14-16) and very-late (day 17-19: menses) luteal phase of natural menstrual cycles. EPHX2 mRNA levels peaked in mid-late CL (5-fold when compared with early CL, P<0.05) and remained elevated in the late CL. Ablation of pituitary LH secretion and luteal steroid synthesis significantly reduced (P<0.05) EPHX2 mRNA levels in the mid-late CL, with progestin replacement being insufficient to restore its level of expression to control values. EPHX2 protein was localized to large and small luteal cells, as well as vascular endothelial cells. The EpETrE-generating CYP epoxygenase 2J2, 2C9 and 3A4 genes were also expressed in the macaque CL. While CYP2J2 mRNA levels did not significantly change through the luteal phase, CYP2C9 and CYP3A4 levels were significantly (P<0.05) higher in the mid-late phase when compared with the early phase. CYP2C9, 2J2 and 3A4 proteins were each localized to the large luteal cells, with 2C9 and 2J2 also being present in the small luteal, stromal and endothelial cells. These studies demonstrate for the first time that an EpETrE generating and metabolizing system exists in the primate CL, with the latter being regulated by LH and steroid hormone(s).
Author affiliation: Irusta, Griselda. State University of Oregon; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Author affiliation: Murphy, M. J.. Oregon Health and Science University; Estados Unidos
Author affiliation: Perez, W. D.. Oregon Health and Science University; Estados Unidos
Author affiliation: Hennebold J.D.. Oregon Health and Science University; Estados Unidos
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Authors: Abalos, Marcos; Acuña, Francisco Antonio; Cancino, Andrea Karina; Aller Atucha, Juan Florencio
Publication Date: 2018.
Language: English.
Abstract:
The objectives of the present study were to determine the effects of exogenous GnRH administered7 days after breeding on the formation of an accessory corpus luteum (ACL), plasma progesterone(P4) concentrations and pregnancy rates. Adult females (n=71) having a follicle≥ 7mm in diameter in the ovary were naturally mated (Day 0). On Day 7, ultrasonicexamination was performed to confirm the occurrence of ovulation as evidenced by presence o an induced corpus luteum (ICL). Females with an ICL plus a dominant follicle≥7mm (n=56) were treated with saline solution (SS, n=29) or GnRH analogue (n=27). On Day 14, the formation of an ACL was observed by ultrasonography. Blood samples were collected on Days 7 and 14 to quantify plasma P4 concentrations. On Day 14, 21 of 27 (77.8%) females in the GnRH group developed an ACL, whereas females in the SS group did not. Progesterone concentrations on Day 7 and 14 in those llamas diagnosed as pregnant on Day 30 were not different (P > 0.05) between groups. In addition, P4 concentration was similar for GnRH-treated females having two CL to those with a single CL. Pregnancy rates were similar (P > 0.05) between SS and GnRH groups (55.2% compared with 74.1% respectively) and the pregnancy rate for the GnRH group was notaffected (P > 0.05) by the number of CL observed at Day 14 (66.6% and 75.6% for females with one and two CL respectively). In conclusion, GnRH administration on Day 7 after breeding leads to ACL formation; however, neither the plasma P4 concentration nor pregnancy rate was affected by having an ACL.
EEA Balcarce
Author affiliation: Abalos, Marcos. INTA, Estación Experimental Agropecuaria Abra Pampa; Argentina
Author affiliation: Acuña, Francisco Antonio. INTA, Estación Experimental Agropecuaria Abra Pampa; Argentina
Author affiliation: Cancino, Andrea Karina. INTA. Estación Experimental Agropecuaria Bariloche; Argentina
Author affiliation: Aller Atucha, Juan Florencio. INTA. Estación Experimental Agropecuaria Balcarce; Argentina
Keywords: Llama; Cuerpo Lúteo; Progesterona; Gestación; Hormonas; Llamas; Corpus Luteum; Progesterone; Pregnancy; Hormones.
Repository: INTA Digital (INTA). Instituto Nacional de Tecnología Agropecuaria
Publication Date: 2006.
Language: English.
Abstract:
Apoptosis is associated with the regression of the corpus luteum (CL) in many species. Since caspases play a central role in apoptosis, we studied several initiators (-2, -8, and -9) and the main effector (-3) caspase in the CL during the estrous cycle of the rat. Two different populations of CL (old and new) were identified on ovaries at estrus and diestrus II (DII). Diminished (P < 0.05) luteal progesterone content and P450scc levels suggested that functional luteolysis occurred between the new CL at DII and old CL at estrus, whereas the decline (P < 0.05) in luteal weight indicated that structural regression was occurring between old CL at estrus to DII. Immunostaining for caspase-2 in luteal and endothelial cells appeared to increase as the luteal phase progressed, peaking at DII in the old CL. However, caspase-8 and -9 immunostaining showed little change with a slight increase at estrus in the old population. Notably, caspase-3 staining appeared to peak at DII in the new CL. Enzyme activity of caspase-9 increased (P < 0.05) in the new CL at DII, followed by that of caspase-2 and -3 in old CL at estrus. Caspase-8 activity did not change at any stage. The number of apoptotic cells increased at DII in the old CL. These results suggest an important role for this protease family during early events of luteolysis in the rat estrous cycle
Author affiliation: Peluffo, Marina Cinthia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Author affiliation: Bussmann, L. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Author affiliation: Stouffer, Richard L.. Oregon Health & Science University; Estados Unidos
Author affiliation: Tesone, Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Authors: Fernandez, Jimena Beatriz; Bruno Galarraga, María Macarena; Soto, A. T.; de la Sota, Rodolfo Luzbel; Cueto, M. I.; Lacau, Isabel María; Gibbons, A. E.
Publication Date: 2017.
Language: English.
Abstract:
We determined the effect of GnRH or hCG treatment on day 4 post-time artificial insemination (FTAI) on the formation of accessory corpora lutea (acc-CL) and on the concentration of serum progesterone (P4) in sheep. Multiparous adult Merino ewes (n = 36) were synchronized for estrus using double injection of PGF2α agonist (125 μg Cloprostenol) with an interval of 14 days. At 53–56 h after the second PG application, FTAI was performed. On day 4 post FTAI, ewes were either treated with analogue of GnRH (4 μg buserelin; n = 12) or hCG (300 IU, hCG; n = 12) or saline solution (1 ml; Control; n = 12). Two laparoscopic ovarian examinations were performed on days 4 and 10 post FTAI. In the first observation, we determined the number of post ovulation corpora lutea (po-CL) and the site, number and diameter of follicles present in both ovaries. In the second laparoscopy, we observed the number of po-CL and acc-CL. The sizes of the follicles that generated the acc-CL were determined according to the position of the follicles observed in the first laparoscopy. Serum P4 concentration was determined on days 4, 7, 10, 13, 17 and 21 post FTAI by chemiluminescence. A similar follicular population in size and number was observed in the three experimental groups prior to the beginning of treatments (Follicles 2 mm: 6.4 ± 3.7, 3 mm: 3.0 ± 2.3, 4 mm: 1.1 ± 0.5, 5 mm: 1.4 ± 0.8; P ˃ 0.05). The formation of 1.0 ± 0.4 and 1.1 ± 0.3 acc-CL was observed in the GnRH and hCG groups, respectively (P ˃ 0.05), but was not observed in the Control group (P < 0.05). Follicle sizes from which acc-CL generated were 3, 4 and 5 mm and did not differ between hormonal treatments (P ˃ 0.05). The hCG group had higher mean concentrations of P4 on days 7, 10, 13 and 17 post FTAI compared with the GnRH group and the Control group (P < 0.05), while no differences were observed between these two latter groups (P > 0.05). Mean P4 concentrations in ewes treated with hCG showed no differences according to the size of the follicle from which acc-CL were generated (P ˃ 0.05). In conclusion, administration of hCG or GnRH on day 4 post FTAI induced the formation of one acc-CL from follicles of 3, 4 or 5 mm, indistinctly. However, serum P4 concentration increased significantly only in the hCG group. The serum P4 concentrations of acc-CL that originated from different follicle sizes did not differ.
Author affiliation: Fernandez, Jimena Beatriz. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Bruno Galarraga, María Macarena. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Soto, A. T.. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Author affiliation: de la Sota, Rodolfo Luzbel. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Cueto, M. I.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche; Argentina
Author affiliation: Lacau, Isabel María. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Author affiliation: Gibbons, A. E.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche; Argentina
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Authors: Fernandez, Jimena; Bruno Galarraga, Maria Macarena; Soto, Andrés; de la Sota, Rodolfo Luzbel; Cueto, Marcela Isabel; Lacau, Isabel Maria; Gibbons, Alejandro Eduardo
Publication Date: 2018.
Language: English.
Abstract:
We determined the effect of GnRH or hCG treatment on day 4 post-time artificial insemination (FTAI) onthe formation of accessory corpora lutea (acc-CL) and on the concentration of serum progesterone (P4)insheep. Multiparous adult Merino ewes (n¼36) were synchronized for estrus using double injection ofPGF2aagonist (125mg Cloprostenol) with an interval of 14 days. At 53e56 h after the second PGapplication, FTAI was performed. On day 4 post FTAI, ewes were either treated with analogue of GnRH(4mg buserelin; n¼12) or hCG (300 IU, hCG; n¼12) or saline solution (1 ml; Control; n¼12). Twolaparoscopic ovarian examinations were performed on days 4 and 10 post FTAI. In thefirst observation,we determined the number of post ovulation corpora lutea (po-CL) and the site, number and diameter offollicles present in both ovaries. In the second laparoscopy, we observed the number of po-CL and acc-CL.The sizes of the follicles that generated the acc-CL were determined according to the position of thefollicles observed in thefirst laparoscopy. Serum P4concentration was determined on days 4, 7, 10, 13, 17and 21 post FTAI by chemiluminescence. A similar follicular population in size and number was observedin the three experimental groups prior to the beginning of treatments (Follicles 2 mm: 6.4±3.7, 3 mm:3.0±2.3, 4 mm: 1.1±0.5, 5 mm: 1.4±0.8; P˃0.05). The formation of 1.0±0.4 and 1.1±0.3 acc-CL wasobserved in the GnRH and hCG groups, respectively (P˃0.05), but was not observed in the Control group(P<0.05). Follicle sizes from which acc-CL generated were 3, 4 and 5 mm and did not differ betweenhormonal treatments (P˃0.05). The hCG group had higher mean concentrations of P4on days 7, 10, 13and 17 post FTAI compared with the GnRH group and the Control group (P<0.05), while no differenceswere observed between these two latter groups (P>0.05). Mean P4concentrations in ewes treated withhCG showed no differences according to the size of the follicle from which acc-CL were generated(P˃0.05). In conclusion, administration of hCG or GnRH on day 4 post FTAI induced the formation of oneacc-CL from follicles of 3, 4 or 5 mm, indistinctly. However, serum P4concentration increased signifi-cantly only in the hCG group. The serum P4concentrations of acc-CL that originated from different folliclesizes did not differ.
Author affiliation: Fernández, Jimena. INTA. Estación Experimental Agropecuaria Bariloche. Laboratorio de Reproduccion de Rumiantes Menores; Argentina
Author affiliation: Bruno Galarraga, Maria Macarena. INTA. Estación Experimental Agropecuaria Bariloche. Laboratorio de Reproduccion de Rumiantes Menores; Argentina
Author affiliation: Soto, Andrés. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Author affiliation: de la Sota, Rodolfo Luzbel. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Author affiliation: Cueto, Marcela Isabel. INTA. Estación Experimental Agropecuaria Bariloche. Laboratorio de Reproduccion de Rumiantes Menores; Argentina
Author affiliation: Lacau, Isabel Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental; Argentina
Author affiliation: Gibbons, Alejandro Eduardo. INTA. Estación Experimental Agropecuaria Bariloche. Laboratorio de Reproduccion de Rumiantes Menores; Argentina
Repository: INTA Digital (INTA). Instituto Nacional de Tecnología Agropecuaria
Authors: Hernandez, Silvia Fátima; Peluffo, Marina Cinthia; Bas, Diana Ester; Stouffer, Richard L.; Tesone, Marta
Publication Date: 2009.
Language: English.
Abstract:
Since the regression of the corpus luteum (CL) occurs via a tightly controlled apoptotic process, studies were designed to determine if local administration of the antiapoptotic agent sphingosine 1-phosphate (S1P) effectively blocks the luteolytic action of prostaglandin F-2alpha (PGF-2alpha). On day 19 of pregnancy, 2 hr before systemic PGF-2alpha administration, rats were injected intrabursa with either S1P or vehicle (control). The activity of four caspases, which contribute to the initial (caspase-2, -8, and -9) and final (caspase-3) events in apoptosis was measured in pooled CL from four individual ovaries at 0 and 4 hr after PGF-2alpha injection. The expression of the phosphorylated form of AKT (pAKT) and tumor necrosis factor-alpha (TNF-alpha) was analyzed by ELISA. In addition, cell death was evaluated by electronic microscopy (EM) in CL 4 and 36 hr after PGF-2alpha injection. The activity of caspase-2, -3, and -8 was significantly greater by 4 hr after PGF-2alpha, but not caspase-9 activity. In contrast, expression of pAKT and TNF-alpha decreased significantly. Administration of S1P suppressed (P < 0.05) these effects, decreasing caspase activities and increasing pAKT and TNF-alpha expression. The administration of S1P also significantly decreased the percentage of luteal apoptotic cells induced by PGF-2alpha. PGF-2alpha treatment increased the prevalence of luteal cells with advanced signs of apoptosis (i.e., multiple nuclear fragments, chromatin condensation, or apoptotic bodies). S1P treatment suppressed these changes and increased the blood vessel density. These results suggest that S1P blocks the luteolytic effect of the PGF-2alpha by decreasing caspase-2, -3, and -8 activities and increasing AKT phosphorylation and TNF-alpha expression.
Author affiliation: Hernandez, Silvia Fátima. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Author affiliation: Peluffo, Marina Cinthia. Oregon Health and Science University; Canadá
Author affiliation: Bas, Diana Ester. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Author affiliation: Stouffer, Richard L.. Oregon Health and Science University; Canadá
Author affiliation: Tesone, Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas