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1.
Theriogenology ; 147: 192-196, 2020 Apr 15.
Article in English | MEDLINE | ID: mdl-31767185

ABSTRACT

Apelin is a potent inotropic agent causing endothelium-mediated vasodilation and is involved in vessel formation by interacting with a specific receptor. Its cardiovascular profile suggests a role in the regulation of gestational hemodynamic changes. The expression of apelin and its receptor has been reported in some portions of the reproductive tract of different mammalian species. As far as we know, there are no reports describing the expression of apelin and apelin receptor in bitch's placenta. Therefore, the aim of this study was to investigate, for the first time, the presence and distribution of apelin and apelin receptor in bitch placenta by molecular biology and immunohistochemical techniques. Sixteen adult female half-breed bitches were used. The animals were divided into two groups based on the stage of pregnancy: group 1 (mid-gestation n = 8) and group 2 (end gestation n = 8). These bitches were subjected to ovariohysterectomy (group1) or non-conservative caesarean section (group 2). The immunohistochemical technique revealed the presence of positive immune reaction for apelin and apelin receptor in all the samples examined at 30 days and at the end of pregnancy. In particular, apelin and apelin receptor staining was evident in the cytoplasms of cytotrophoblasts and in epithelial cells of the maternal portion. Even if not included into the structure of the placenta, the uterine glands also exhibited a positive immune reaction for apelin and apelin receptor. The RT-PCR analysis showed the presence of transcripts for apelin and apelin receptor in all the placenta samples examined. On the basis of our results it was also possible to hypothesize a potential role of apelin in the control of local placenta blood flow during pregnancy development in bitches.


Subject(s)
Apelin Receptors/metabolism , Apelin/metabolism , Dogs/metabolism , Placenta/metabolism , Animals , Apelin/genetics , Apelin Receptors/genetics , Female , Gene Expression Regulation/physiology , Placenta/blood supply , Pregnancy
3.
Theriogenology ; 115: 1-8, 2018 Jul 15.
Article in English | MEDLINE | ID: mdl-29698886

ABSTRACT

Kisspeptin and its receptor KISS1R are involved in the neuroendocrine regulation of mammalian reproduction and their role on follicular development and function can be hypothesized. The present work was designed to confirm the immunopresence of kisspeptin and its receptor in the ovary of swine and to study the effects of kisspeptin 10 and its antagonist, kisspeptin 234, on main functional parameters of granulosa cells (i.e. cell proliferation, steroid production, and redox status) as well as their modulatory action on angiogenesis. The immunopresence of kisspeptin and KISS1R were detected in granulosa cells. Kisspeptin 10 stimulated progesterone in vitro production, thus indirectly suggesting that it can have a role in the luteinization process of granulosa cells. Kisspeptin 10 displayed potentiating effects on non-enzymatic scavenging activity, thus supporting its involvement in the control of the antioxidant defense system of ovarian follicles. In addition, results from the angiogenesis bioassay suggest that kisspeptin may have a role in the physiological development of new ovarian vessels. Additional studies are needed to confirm the functional significance of the kisspeptin/KISS1R system within the swine ovary.


Subject(s)
Kisspeptins/physiology , Ovarian Follicle/chemistry , Ovarian Follicle/physiology , Receptors, Kisspeptin-1/physiology , Swine , Animals , Cell Proliferation/drug effects , Female , Granulosa Cells/chemistry , Granulosa Cells/drug effects , Granulosa Cells/physiology , Kisspeptins/analysis , Kisspeptins/antagonists & inhibitors , Kisspeptins/pharmacology , Neovascularization, Physiologic/drug effects , Ovary/blood supply , Ovary/physiology , Oxidation-Reduction , Progesterone/biosynthesis , Receptors, Kisspeptin-1/analysis
4.
Folia Morphol (Warsz) ; 76(4): 603-607, 2017.
Article in English | MEDLINE | ID: mdl-28553853

ABSTRACT

The present research used immunohistochemistry to analyse the detection and localisation of nitric oxide synthase (NOS) isoforms in the ductuli efferentes and epididymis of prepubertal and adult alpaca. In the ductuli efferentes and epididymis of prepubertal and adult animals, nNOS and eNOS were similarly expressed in epithelial lining cells, conversely differences were observed in the immunopresence of iNOS. Our data provide evidence that NOS isoforms may have roles in reproductive functions and in the developmental processes of the excurrent duct system in the alpaca.

5.
Domest Anim Endocrinol ; 56: 20-8, 2016 07.
Article in English | MEDLINE | ID: mdl-26986844

ABSTRACT

The aim of the present study was to evaluate: (1) the presence of nerve growth factor (NGF), neurotrophic tyrosine kinase receptor 1 (NTRK1), and nerve growth factor receptor (NGFR) in the rabbit uterus; and (2) the in vitro effects of NGF on PGF2α and PGE2 synthesis and on the PGE2-9-ketoreductase (PGE2-9-K) activity by the rabbit uterus. Nerve growth factor, NTRK1, and NGFR were immunolocalized in the luminal and glandular epithelium and stroma cells of the endometrium. reverse transcriptase polymerase chain reaction indicated the presence of messenger RNA for NGF, NTRK1, and NGFR in the uterus. Nerve growth factor increased (P < 0.01) in vitro secretions of PGF2α and PGE2 but coincubation with either NTRK1 or oxide nitric synthase (NOS) inhibitors reduced (P < 0.01) PGF2α production and blocked (P < 0.01) PGE2 secretion. Prostaglandins releases were lower (P < 0.01) than control when uterine samples were treated with NGF plus cyclooxygenase inhibitor. However, addition of NGFR inhibitor reduced (P < 0.01) PGF2α secretion less efficiently than NTRK1 or NOS inhibitors but had no effect on PGE2 yield. Nerve growth factor increased (P < 0.01) the activity of PGE2-9-K, whereas coincubation with NTRK1 or NOS inhibitors abolished (P < 0.01) this increase in PGE2-9-K activity. However, cotreatment with either cyclooxygenase or NGFR inhibitors had no effect on PGE2-9-K activity. This is the first study to document the distribution of NGF/NTRK1 and NGFR systems and their effects on prostaglandin synthesis in the rabbit uterus. NGF/NTRK1 increases PGF2α and PGE2 productions by upregulating NOS and PGE2-9-K activities, whereas NGF/NGFR augments only PGF2α secretion, through an intracellular mechanism that is still unknown.


Subject(s)
Gene Expression , Nerve Growth Factor/genetics , Prostaglandins/biosynthesis , Rabbits/metabolism , Receptors, Nerve Growth Factor/genetics , Uterus/metabolism , Animals , Dinoprost/biosynthesis , Female , Hydroxyprostaglandin Dehydrogenases/metabolism , Immunohistochemistry , Nerve Growth Factor/analysis , Nerve Growth Factor/pharmacology , RNA, Messenger/analysis , Receptor, trkA/analysis , Receptor, trkA/genetics , Receptor, trkA/physiology , Receptors, Nerve Growth Factor/analysis , Receptors, Nerve Growth Factor/physiology , Uterus/chemistry
6.
Reprod Domest Anim ; 50(6): 918-25, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26392300

ABSTRACT

Experiments were devised to characterize the expression of nerve growth factor, beta polypeptide (NGF), and its cognate receptors neurotrophic tyrosine kinase receptor type 1 (NTRK1) and nerve growth factor receptor (NGFR) in rabbit male sex organs, as well as the concentrations of NGF in both seminal and blood plasma of sexually mature male rabbits. Immunoreactivity and gene expression for NGF and cognate receptors were detected in testis, prostate gland and seminal vesicle. The highest levels of NGF and NTRK1 transcripts were found in the prostate, while intermediate expressions were found in the testis. NGFR transcripts were expressed at the same levels in both testis and prostate and were more abundant than in seminal vesicles. The widespread distribution of NGF in all prostate glandular cells, together with its relative high mRNA abundance, confirms that the prostate of rabbits is the main source of this neurotrophin. In conclusion, the present data suggest that the NGF system is involved in the testicular development and spermatogenesis of rabbits and that NGF may act as a potential ovulation-inducing factor being abundantly present in the seminal plasma.


Subject(s)
Nerve Growth Factor/genetics , RNA, Messenger/genetics , Rabbits/genetics , Receptor, trkA/genetics , Receptors, Nerve Growth Factor/genetics , Animals , Epithelial Cells/metabolism , Gene Expression , Male , Prostate/metabolism , Testis/metabolism
7.
Res Vet Sci ; 98: 7-12, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25499747

ABSTRACT

This study examined the effects of food restriction during rabbit pregnancy on hormones and metabolites involved in energy homeostasis and metabolic programming. Pregnant does were assigned to four groups: the control group was fed a standard ration while the others received a restricted amount of food (30% restriction) during early (0-9 days), mid (9-18 days), and late (19-28 days) pregnancy. The pregnancy induced a coordinated range of adaptations to fulfil energy requirements of both mother and foetus, such as hyperleptinaemia and hyperinsulinaemia, reduced insulin sensitivity, increased cortisol and non-esterified fatty acid. Food restriction altered leptin, insulin, T3, non-esterified fatty acids and glucose concentrations depending on the gestational phase in which it was applied. Collectively, present data confirm that the endocrinology of pregnancy and the adaptive responses to energy deficit make the rabbit an ideal model for studying nutritional-related disorders and foetal programming of metabolic disease.


Subject(s)
Diet/veterinary , Energy Metabolism/physiology , Homeostasis/physiology , Hormones/metabolism , Malnutrition/veterinary , Pregnancy, Animal/metabolism , Prenatal Exposure Delayed Effects/veterinary , Rabbits/metabolism , Animal Nutritional Physiological Phenomena/physiology , Animals , Fatty Acids, Nonesterified/metabolism , Female , Fetus/physiology , Glucose/metabolism , Insulin/metabolism , Insulin Resistance/physiology , Leptin/metabolism , Models, Animal , Pregnancy , Prenatal Exposure Delayed Effects/metabolism
8.
Anim Reprod Sci ; 147(3-4): 128-34, 2014 Jun 30.
Article in English | MEDLINE | ID: mdl-24838082

ABSTRACT

Systemic and local infections and inflammations are known to cause infertility in humans and animals. However, the mechanisms by which infection/inflammation induces infertility are only partially known. The objectives of this study were: (i) to provide models of systemic (acute) and local (sub-acute) inflammation by intra-peritoneal injection or intra-cervical deposition of lipopolysaccharide (LPS) in the rabbit and (ii) to assess their effects on uterine tissues and sperm transport in the genital tract of rabbit does. Intra-peritoneal administration of different doses of LPS induced systemic effects such as fever, anorexia and changes in white blood cells (WBC) count. In our study, LPS inoculation (100µg/kg) produced an inflammation-like status that lasted for about 3 days, with minimal distress for the animals. Intra-peritoneal administration of LPS 60h before artificial insemination induced a rapid increase of IL-1ß concentrations. The intra-cervical inoculation of LPS did not show any systemic effects, as confirmed by the lack of changes in body temperature, feed intake and WBC count. Histological examination of uterine tissues showed an endometritis-like inflammation status in LPS-treated does, more severe in those inoculated intra-cervically. The number of spermatozoa recovered from uterine horns and oviducts of intra-cervically treated does was less than that retrieved from intra-peritoneally treated animals and controls. These results suggest (i) that sub-acute or acute inflammation may cause infertility by compromising the uterine environment and/or impairing sperm transport and (ii) that the LPS-induced -infection/inflammation experimental model is useful for studying the mechanisms involved in reproductive dysfunctions in the rabbit.


Subject(s)
Lipopolysaccharides/pharmacology , Rabbits/physiology , Reproduction/drug effects , Acute Disease , Animals , Biological Transport/drug effects , Female , Infertility, Female/etiology , Infertility, Female/veterinary , Inflammation/complications , Inflammation/immunology , Inflammation/veterinary , Interleukin-1beta/blood , Male , Reproduction/physiology , Semen/drug effects , Semen/physiology , Sperm Motility/drug effects , Uterine Diseases/complications , Uterine Diseases/immunology , Uterine Diseases/veterinary
9.
Microsc Res Tech ; 77(3): 201-10, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24375763

ABSTRACT

To investigate the mechanisms by which caloric restriction affects reproductive function in female rabbits, we measured, in animals intact or ovariectomized (OVX) estrogen-primed and fed ad libitum or fasted for 48 h, the adenohypophysial expression of estrogen receptor-alpha (ESR1) and gonadotropin releasing hormone receptor (GnRHR) and the dynamic secretion of LH following GnRH stimulation. Fasting increased the number of GnRHR-immunoreactive (-IR) cells in intact animals, whereas reduced the density of ESR1-IR cells in OVX rabbits. Estrogen priming decreased the number of ESR1-IR cells in fasted and OVX animals. Ovariectomy increased the number of ESR1-IR cells in fed rabbits, but caused an opposite effect in both fed and fasted animals treated with estrogen. Fasting down regulated the mRNA levels for ESR1 and GnRHR. Estrogen-priming reduced the abundance for ESR1 mRNA in both fed and fasted rabbits, and that for GnRHR in fasted rabbits. Ovariectomy halved ESR1 mRNA levels independently of treatment and feeding condition, whereas increased (P < 001) that for GnRHR in estrogen-primed rabbits. In all rabbits, an LH surge occurred 30 min after GnRH injection but the lowest levels were found in intact fasted rabbits and the highest in fasted, estrogen-primed animals. The LH profile was similar in intact and OVX rabbits and neither fasting nor estrogen priming modified it. In conclusion, fasting differentially modifies the ESR1 and GnRHR expression in the pituitary, depending on the presence of gonadal hormones, indicating complex interactions between metabolic signals and ovarian steroids.


Subject(s)
Estrogen Receptor alpha/metabolism , Fasting , Gonadotropin-Releasing Hormone/metabolism , Pituitary Gland/metabolism , Receptors, LHRH/metabolism , Animals , Estrogen Receptor alpha/genetics , Estrogens/physiology , Female , Gene Expression , Ovariectomy , Pituitary Gland, Anterior/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rabbits , Receptors, LHRH/genetics
10.
Domest Anim Endocrinol ; 45(4): 171-9, 2013 Nov.
Article in English | MEDLINE | ID: mdl-24099736

ABSTRACT

The aim of this study was to elucidate the possible direct regulatory role of the endocannabinoids in the modulation of LH secretion in rabbits, a reflex ovulator species. The cannabinoid receptor type 1 (CB1) was characterized by RT-PCR techniques in the anterior pituitary of intact and ovariectomized does treated with GnRH and primed with estrogen and CB1 antagonist, rimonabant. Cannabinoid receptor type 1 immune reaction was evidenced by immunohistochemistry in the cytoplasm of approximately 10% of the pituitary cells with a density of 8.5 ± 1.9 (per 0.01 mm(2)), both periodic acid-Schiff positive (30%) and negative (70%). All CB1-immunoreactive cells were also immune reactive for estrogen receptor type 1. Ovariectomy, either alone or combined with estrogen priming, did not modify the relative abundances of pituitary CB1 mRNA, but decreased (P < 0.01) the expression of estrogen receptor type 1 mRNA. Treatment with CB1 antagonist (rimonabant) inhibited (P < 0.01) LH secretory capacity by the pituitary after GnRH injection, and estrogen priming had no effect. The present findings indicate that the endocannabinoid system is a potential candidate for the regulation of the hypothalamic-pituitary-ovarian axis in reflex ovulatory species.


Subject(s)
Luteinizing Hormone/metabolism , Pituitary Gland/physiology , Rabbits/physiology , Receptor, Cannabinoid, CB1/genetics , Receptor, Cannabinoid, CB1/physiology , Animals , Cannabinoid Receptor Antagonists/pharmacology , Estrogen Receptor alpha/analysis , Estrogen Receptor alpha/genetics , Estrogens/pharmacology , Gene Expression , Gonadotropin-Releasing Hormone/pharmacology , Male , Ovariectomy , Piperidines/pharmacology , Pituitary Gland/chemistry , Pituitary Gland/drug effects , Pyrazoles/pharmacology , RNA, Messenger/analysis , Receptor, Cannabinoid, CB1/analysis , Rimonabant
11.
Reprod Domest Anim ; 48(4): 673-80, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23320475

ABSTRACT

In six German Shepherds dogs, GnRH agonist implants (Deslorelin) were inserted subcutaneously one month after histological confirmation of benign prostatic hyperplasia (BPH). Prostatic volume (PV), characteristics of ejaculate, serum testosterone concentrations and Doppler parameters of prostatic and subcapsular arteries were detected at different time intervals, for 6 month. The prostatic volume showed a significantly reduction starting at day 37. The decrease in sperm concentration, motility and increase in morphological abnormal sperm were observed from day 22 to day 37, when it was no longer possible to obtain the ejaculate. The values of peak systolic velocity and end-diastolic velocity in prostatic and subcapsular arteries showed from day 11 a gradual decrease, significant at day 22 until day 37 and reaching the lowest values at day 52 until the end of observation. The power Doppler pixel intensity of both arteries showed a gradual decrease from day 5 until day 52. In particular, a significant decrease was observed for both arteries from day 11. Testosterone serum concentration decreased to undetectable levels by day 11 until the end of the observations. All these Doppler parameters and testosterone values were positively correlated with the prostatic volume. Furthermore, testosterone values were positively correlated with peak systolic velocity, end diastolic velocity and pixel numbers. The use of implants containing GnRH analogues, even in asymptomatic subjects, is effective for the control of BPH and the application of Doppler exam of prostatic blood flow represent an non-invasive tool for monitoring the response of medical treatment.


Subject(s)
Dog Diseases/drug therapy , Gonadotropin-Releasing Hormone/agonists , Prostate/blood supply , Prostatic Hyperplasia/veterinary , Triptorelin Pamoate/analogs & derivatives , Animals , Arteries/physiopathology , Blood Flow Velocity , Dogs , Drug Implants , Male , Prostate/pathology , Prostatic Hyperplasia/drug therapy , Prostatic Hyperplasia/physiopathology , Semen Analysis/veterinary , Testosterone/blood , Triptorelin Pamoate/administration & dosage , Triptorelin Pamoate/therapeutic use , Ultrasonography, Doppler/veterinary
12.
Microsc Res Tech ; 75(12): 1682-90, 2012 Dec.
Article in English | MEDLINE | ID: mdl-22865504

ABSTRACT

Immunopresence, gene expression, and enzymatic activity of cyclooxygenase 1 (COX1), COX2, PGE2-9-ketoreductase (PGE2-9-K), endothelial (eNOS), and inducible nitric oxide synthases (iNOS), and hormone in vitro production were examined in early, mid, late, and regressive buffalo corpora lutea (CL). COX1 immunosignals were detected in the cytoplasm of small luteal cells, COX2 in large luteal cells, and PGE2-9-K in all luteal cells. COX2 and PGE2-9-K immunosignals were greater in late CL. Immunopresence of both NOS types were evidenced in the nuclei and cytoplasm of all luteal cells, as well as in the nuclei of endothelial cells, during all stages studied. The eNOS and iNOS immunosignals increased during the early stage. COX1 transcripts were lower in late and regressive CL, COX2 in late, PGE2-9-K higher in regressive, and iNOS higher in early and lower in regressive CL. COX1 enzymatic activity was lower in regressive CL, COX2 increased in mid and late stages, and PGE2-9-K was higher in late CL. Endothelial NOS activity was higher during mid and late stages and lower in regressive, whereas iNOS was greater in late and lower in early. Progesterone in vitro release was higher in mid and lower in late phase, while PGF2α synthesis was higher in late CL and lower in regressive, and PGE2 was higher during regressive stage. These results support the idea that COX, NOS, and PGE2-9-K regulate buffalo CL life span. In particular, regressive CL seems involved in the development of the contralateral early CL, through the production of the luteotrophic PGE2.


Subject(s)
Buffaloes/physiology , Corpus Luteum/enzymology , Diestrus , Gene Expression , Hydroxyprostaglandin Dehydrogenases/analysis , Nitric Oxide Synthase/analysis , Prostaglandin-Endoperoxide Synthases/analysis , Animals , Cell Nucleus/chemistry , Cytoplasm/chemistry , Female , Gene Expression Profiling , Hydroxyprostaglandin Dehydrogenases/genetics , Immunohistochemistry , Microscopy , Nitric Oxide Synthase/genetics , Prostaglandin-Endoperoxide Synthases/genetics
13.
Anim Reprod Sci ; 129(3-4): 106-17, 2011 Dec.
Article in English | MEDLINE | ID: mdl-22169022

ABSTRACT

The profitability of rabbit farms has increased in recent years due primarily to improvements in the management of reproduction and genetic selection. This review summarizes the most important scientific papers relating to ovulation in rabbit does dealing in particular with: (a) studies from 1905 to the present day relating to ovulatory mechanisms in rabbit does; (b) research on the primary gonadotrophin-releasing hormone (GnRH), its analogues and their functions; and (c) descriptions of parenteral and intravaginal (iv.) treatments for induction of ovulation in does and their reported efficacies. The addition of GnRH analogues via the seminal dose (iv.) fulfils the need for a welfare-orientated method of inducing ovulation in rabbits. The structure, tissues, secretions, contractions, and innervations of the vagina in rabbits that can affect absorption profiles are reviewed in the context of recent reports of the achievement of high ovulation rates obtained by adding GnRH analogues directly to the seminal dose. This review demonstrates the possibility of ovulation induction in rabbits by the addition of GnRH synthetic analogues to the seminal doses and provides new perspectives for simplifying the AI technique.


Subject(s)
Ovulation Induction/methods , Ovulation Induction/veterinary , Rabbits/physiology , Administration, Intravaginal , Animals , Female , Gonadotropin-Releasing Hormone/administration & dosage , Gonadotropin-Releasing Hormone/analogs & derivatives , Ovulation/drug effects , Vagina/blood supply , Vagina/drug effects , Vagina/microbiology
14.
Reprod Fertil Dev ; 23(6): 759-68, 2011.
Article in English | MEDLINE | ID: mdl-21791177

ABSTRACT

Food deprivation affects female reproduction. The goal of the present study was to elucidate in the rabbit model the effects of acute energy restriction on ovarian function (follicle development, atresia rate and in vitro oocyte maturation) and embryonic development and gene expression of some candidate genes. Serum metabolic parameters (non-esterified fatty acids (NEFA), triglycerides, glucose, insulin and leptin concentrations) and endocrine markers (oestradiol-17ß and progesterone concentrations) were also studied. A control group of nulliparous does fed ad libitum and a 72-h fasted group were used. At the end of the nutritional treatment, the ovaries of half of the animals were retrieved while the other animals were re-fed and artificially inseminated to recover embryos at 84 h after insemination, during the luteal phase. At the end of fasting, increased serum NEFA and decreased leptin concentrations were observed in the fasted group, but no differences appeared in serum steroid concentrations, follicle population and atresia rate or nuclear and cytoplasmic oocyte maturation. In the luteal phase, insulin concentrations increased notably in the fasted group. The number of recovered embryos per female and the speed of embryo development were reduced in the food-deprived group. Acute fasting altered both metabolic and endocrine markers and embryo development, but follicle and oocyte development and embryo gene expression were not affected.


Subject(s)
Endocrine System/physiology , Energy Metabolism/physiology , Fasting/physiology , Gene Expression Regulation, Developmental/physiology , Maternal-Fetal Exchange/physiology , Oocytes/growth & development , Ovarian Follicle/physiology , Analysis of Variance , Animals , Blood Glucose/analysis , Body Composition , DNA Primers/genetics , Embryo, Mammalian/physiology , Estradiol/blood , Fatty Acids/blood , Female , Gene Expression Profiling , In Situ Nick-End Labeling , Insulin/blood , Leptin/blood , Microscopy, Confocal , Pregnancy , Progesterone/blood , Rabbits , Reverse Transcriptase Polymerase Chain Reaction , Triglycerides/blood
15.
Domest Anim Endocrinol ; 40(1): 51-9, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20961724

ABSTRACT

The main objective of this study was to examine the modulatory in vitro effects of gonadotropin-releasing hormone (GnRH) on isolated Leydig cells of adult alpaca (Lama pacos) testis. We first evaluated the presence of GnRH receptor (GnRHR) and cyclooxygenase (COX) 1 and COX2 in alpaca testis. We then studied the in vitro effects of buserelin (GnRH analogue), antide (GnRH antagonist), and buserelin plus antide or inhibitor of phospholipase C (compound 48/80) and COXs (acetylsalicylic acid) on the production of testosterone, PGE(2), and PGF(2α) and on the enzymatic activities of COX1 and COX2. Immunoreactivity for GnRHR was detected in the cytoplasm of Leydig cells and in the acrosomal region of spermatids. COX1 and COX2 immunosignals were noted in the cytoplasm of spermatogonia, spermatocytes, spermatids, Leydig cells, and Sertoli cells. Western blot analysis confirmed the GnRHR and COX1 presence in alpaca testis. The in vitro experiments showed that buserelin alone increased (P < 0.01) and antide and buserelin plus acetylsalicylic acid decreased (P < 0.01) testosterone and PGF(2α) production and COX1 activity, whereas antide and compound 48/80 counteracted buserelin effects. Prostaglandin E(2) production and COX2 activity were not affected by buserelin or antide. These data suggest that GnRH directly up-regulates testosterone production in Leydig cells of adult alpaca testis with a postreceptorial mechanism that involves PLC, COX1, and PGF(2α).


Subject(s)
Camelids, New World/metabolism , Gonadotropin-Releasing Hormone/pharmacology , Leydig Cells/drug effects , Prostaglandin-Endoperoxide Synthases/metabolism , Prostaglandins/biosynthesis , Testosterone/biosynthesis , Animals , Buserelin/pharmacology , Cyclooxygenase 1/analysis , Cyclooxygenase 1/metabolism , Cyclooxygenase 2/analysis , Cyclooxygenase 2/metabolism , Dinoprost/biosynthesis , Dinoprostone/biosynthesis , Gonadotropin-Releasing Hormone/analogs & derivatives , Leydig Cells/chemistry , Leydig Cells/metabolism , Male , Oligopeptides/pharmacology , Prostaglandin-Endoperoxide Synthases/analysis , Receptors, LHRH/analysis , Type C Phospholipases/antagonists & inhibitors , Type C Phospholipases/metabolism
16.
Theriogenology ; 74(4): 672-81, 2010 Sep 01.
Article in English | MEDLINE | ID: mdl-20452002

ABSTRACT

The effect of the antiprogestagen aglepristone (10 mg/kg bw), administered at days 29 and 30 following the estimated day of LH surge (day 0), on corpora lutea (CL) function was examined during the diestrus phase of non-pregnant bitches. Aglepristone shortened (P < 0.01) the luteal phase and complete luteolysis (progesterone <2 ng/mL) was observed at days 40.8 +/- 3.5 and 71.5 +/- 4.6 (means +/- SD; n = 9/group) in treated and control bitches, respectively. Peripheral estradiol-17beta concentrations declined from 91.5 +/- 14.3 pg/mL at day 9 to 50 pg/mL at day 18, remaining at approximately the same levels thereafter in both treated and control bitches. Intraluteal in vitro synthesis of progesterone and estradiol-17beta released by CL explanted at day 38 from control bitches (511.9 +/- 285.6 and 40.7 +/- 17.2 pg/mg protein, respectively) did not differ from that of treated. From day 38, intraovarian hemodynamic variables (arterial blood flow, systolic peak, and end-diastolic velocities), monitored by color-coded and pulsed Doppler, decreased more steeply (P < 0.01) in aglepristone-treated (n = 4) than in control (n = 4) bitches, whereas the resistance index increased (P < 0.01) in treated animals. All the blood flow parameters were undetectable at 60 +/- 3.6 and 68 +/- 2.0 days (medians +/- SD) after LH peak in treated and control bitches, respectively. In conclusion, aglepristone administration to dogs during the mid-luteal phase markedly accelerates the luteolytic process which is accompanied by a parallel decline in ovarian blood flow supply with a shift from approximately 8 to 10 days.


Subject(s)
Dogs/physiology , Estrenes/pharmacology , Hormone Antagonists/pharmacology , Luteolysis/drug effects , Animals , Corpus Luteum/drug effects , Corpus Luteum/metabolism , Estradiol/blood , Estrenes/adverse effects , Female , Hormone Antagonists/adverse effects , Ovary/blood supply , Ovary/diagnostic imaging , Progesterone/antagonists & inhibitors , Progesterone/metabolism , Regional Blood Flow , Ultrasonography
17.
Domest Anim Endocrinol ; 39(2): 116-30, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20427144

ABSTRACT

The dynamic expression for estrogen receptor subtype-1 (ESR1), interleukin-1beta (IL1B), and apoptosis-associated genes, as well as nitric oxide synthase activity, were examined in corpora lutea (CL) of rabbits after prostaglandin F(2alpha) (PGF(2alpha)) administration on either day 4 or day 9 of pseudopregnancy. By reverse transcriptase polymerase chain reaction, the steady-state level of ESR1 transcript was lower (P < 0.01) and that of anti-apoptotic B-cell CLL/lymphoma 2 (BCL2) -like 1 (BCL2L1) was greater in day 4 (P < 0.01) than in day 9 CL. Western blot analysis revealed that BCL2-associated X protein (BAX) abundance was greater in day 4 (P < 0.01) than in day 9 CL, whereas BCL2L1 protein was undetectable at both luteal stages. After PGF(2alpha), ESR1 transcript decreased (P < 0.01) in day 9 CL, whereas IL1B mRNA showed a transitory increase (P < 0.01) at both stages. The pro-apoptotic tumor protein p53 (TP53) gene had diminished (P < 0.01) on day 4 and on day 9 after a transitory increase (P < 0.01), whereas the BAX/BCL2L1 expression ratio increased (P < 0.01) in day 9 CL 24 h after treatment. Following PGF(2alpha), TP53 protein increased (P < 0.01) at both luteal stages, and BAX decreased (P < 0.01) in day 4 CL but increased (P < 0.01) 24 h later in day 9 CL; BCL2L1 became detectable 6 h later in day 4 CL. Nitric oxide synthase activity temporarily increased (P < 0.01) following PGF(2alpha). These findings suggest that PGF(2alpha) regulates luteolysis by ESR1 mRNA down-regulation and modulation of pro- and anti-apoptotic pathways in CL that have acquired a luteolytic capacity.


Subject(s)
Apoptosis Regulatory Proteins/metabolism , Dinoprost/physiology , Estrogen Receptor alpha/metabolism , Interleukin-1/metabolism , Luteolysis/metabolism , Pseudopregnancy/metabolism , Animals , Apoptosis Regulatory Proteins/genetics , Corpus Luteum/metabolism , Estrogen Receptor alpha/genetics , Female , Gene Expression Regulation/physiology , Interleukin-1/genetics , Nitric Oxide Synthase/genetics , Nitric Oxide Synthase/metabolism , RNA, Messenger/analysis , Rabbits , Signal Transduction/physiology , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Protein p53/metabolism , bcl-2-Associated X Protein/genetics , bcl-2-Associated X Protein/metabolism , bcl-X Protein/genetics , bcl-X Protein/metabolism
18.
Reproduction ; 133(5): 1005-16, 2007 May.
Article in English | MEDLINE | ID: mdl-17616729

ABSTRACT

The objective of the present study was to investigate in rabbit corpora lutea (CL), at both the cellular and molecular level, intraluteal cyclooxygenase (COX)-1, COX-2 and prostaglandin (PG) E2-9-ketoreductase (PGE2-9-K) enzymatic activities as well as in vitro PGE2 and PGF2alpha synthesis following PGF2alpha treatment at either early- (day-4) or mid-luteal (day-9) stage of pseudopregnancy. By immunohistochemistry, positive staining for COX-2 was localized in luteal and endothelial cells of stromal arteries at both the stages. In CL of both stages, basal COX-2 mRNA levels were poorly expressed, but rose (P < 0.01) 4- to 10-fold 1.5-6 h after treatment and then gradually decreased within 24 h. Compared to mid-stage, day-4 CL had lower (P < 0.01) COX-2 and PGE2-9-K basal activities, and PGF2alpha synthesis rate, but higher (P < 0.01) PGE2 production. Independent of luteal stage, PGF2alpha treatment did not affect COX-1 activity. In day-4 CL, PGF2alpha induced an increase (P < 0.01) in both COX-2 activity and PGF2alpha synthesis, whereas that of PGE2 remained unchanged. In day-9 CL, PGF2alpha up-regulated (P < 0.01) both COX-2 and PGE-9-K activities, and PGF2alpha production, but decreased (P < 0.01) PGE2 synthesis. All changes in gene expression and enzymatic activities occurred within 1.5 h after PGF2alpha challenge and were more marked in day-9 CL. Our data suggest that PGF2alpha directs intraluteal PG biosynthesis in mature CL, by affecting the CL biosynthetic machinery to increase the PGF2alpha synthesis in an auto-amplifying manner, with the activation of COX-2 and PGE-9-K; this may partly explain their differentially, age-dependent, luteolytic capacity to exogenous PGF2alpha in rabbits.


Subject(s)
Corpus Luteum/metabolism , Dinoprost/pharmacology , Dinoprostone/biosynthesis , Luteolysis , Pseudopregnancy/metabolism , Up-Regulation , Animals , Corpus Luteum/drug effects , Cyclooxygenase 1/biosynthesis , Cyclooxygenase 2/biosynthesis , Dinoprost/biosynthesis , Dinoprostone/analysis , Female , Gene Expression/drug effects , Hydroxyprostaglandin Dehydrogenases/analysis , Hydroxyprostaglandin Dehydrogenases/metabolism , Progesterone/analysis , RNA, Messenger/analysis , Rabbits , Reverse Transcriptase Polymerase Chain Reaction , Time Factors
19.
J Endocrinol ; 185(2): 319-25, 2005 May.
Article in English | MEDLINE | ID: mdl-15845924

ABSTRACT

In this study, we have examined the presence and the distribution of receptors for leptin (Ob-R) in the oviduct of rabbits, and the effects of leptin on the release of prostaglandin (PG) F2alpha and PGE2 and on the activity of nitric oxide (NO) synthase (NOS) by oviducts cultured in vitro. Rabbits were killed during the follicular phase and the oviducts were incubated in vitro with leptin, PGF2alpha, PGE2, NO donor and inhibitors of NOS and cyclo-oxigenase (COX). Using immunohistochemistry, Ob-R-like positive reaction was observed only in the cytoplasm of secretory cells, having stronger intensity in the infundibulum and ampulla tracts than in the isthmus. Both leptin and NO donor inhibited PGE2 release, whereas they enhanced PGF2alpha release; NOS inhibitor alone or with leptin increased PGE2 and decreased PGF2alpha production; NOS activity was enhanced by leptin, while PGs did not affect this enzyme. This study suggests that the oviduct could be a potential target for endocrine regulation by leptin, whose circulating levels may act as a metabolic signal modulating oviductal PG release through mediation of the NOS/NO system.


Subject(s)
Leptin/pharmacology , Nitric Oxide Synthase/metabolism , Oviducts/metabolism , Prostaglandins/metabolism , Receptors, Cell Surface/metabolism , Animals , Aspirin/pharmacology , Cyclooxygenase Inhibitors/pharmacology , Dinoprost/metabolism , Dinoprost/pharmacology , Dinoprostone/metabolism , Dinoprostone/pharmacology , Female , Immunohistochemistry/methods , NG-Nitroarginine Methyl Ester/pharmacology , Nitric Oxide Donors/pharmacology , Nitric Oxide Synthase/antagonists & inhibitors , Nitroprusside/pharmacology , Organ Culture Techniques , Oviducts/chemistry , Rabbits , Radioimmunoassay , Receptors, Cell Surface/analysis , Receptors, Leptin
20.
Reproduction ; 125(6): 807-16, 2003 Jun.
Article in English | MEDLINE | ID: mdl-12773103

ABSTRACT

Total activity of nitric oxide synthase (NOS) and the gene expression of both endothelial NOS (eNOS) and inducible NOS (iNOS) isoforms in corpora lutea of pseudopregnant rabbits were examined during prostaglandin F(2alpha) (PGF(2alpha))-induced luteolysis. Corpora lutea were collected at 0, 6, 12, 24 and 48 h after an injection of PGF(2alpha) at day 9 of pseudopregnancy. At 12 h after PGF(2alpha) administration, luteal mRNA encoding eNOS decreased (P

Subject(s)
Corpus Luteum/metabolism , Dinoprost/pharmacology , Luteolysis/drug effects , Nitric Oxide Synthase/metabolism , Nitric Oxide/physiology , Animals , Blotting, Western/methods , Corpus Luteum/chemistry , Enzyme Inhibitors/pharmacology , Female , NG-Nitroarginine Methyl Ester/pharmacology , Nitric Oxide Synthase/antagonists & inhibitors , Nitric Oxide Synthase/genetics , Nitric Oxide Synthase Type II , Nitric Oxide Synthase Type III , Progesterone/analysis , Progesterone/blood , Pseudopregnancy/metabolism , RNA, Messenger/analysis , Rabbits , Reverse Transcriptase Polymerase Chain Reaction
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