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  • 1.
    A Hulten, Maj
    et al.
    University Warwick, Warwick Med Sch, Coventry CV4 7AL, W Midlands England .
    Patel, Suketu
    University Warwick, Department Biol Science, Coventry CV4 7AL, W Midlands England .
    Jonasson, Jon
    Linköping University, Department of Clinical and Experimental Medicine, Molecular and Immunological Pathology . Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Iwarsson, Erik
    Karolinska University Hospital, Karolinska Institute, Department Mol Med and Surg, Clin Genet Unit, S-17176 Stockholm, Sweden .
    On the origin of the maternal age effect in trisomy 21 Down syndrome: the Oocyte Mosaicism Selection model2010In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 139, no 1, p. 1-9Article, review/survey (Refereed)
    Abstract [en]

    We have recently documented that trisomy 21 mosaicism is common in human foetal ovaries. On the basis of this observation we propose that the maternal age effect in Down syndrome (DS) is caused by the differential behaviour of trisomy 21 in relation to disomy 21 oocytes during development from foetal life until ovulation in adulthood. in particular, we suggest that trisomy 21 oocytes, lagging behind those that are disomic, may escape the timed pruning of the seven million in foetal life to the 300-400 finally selected for ovulation. The net effect of this preferential elimination will be an accumulation of trisomy 21 oocytes in the ovarian reserve of older women. We here highlight the implications of this Oocyte Mosaicism Selection (OMS) model with respect to the prevalent view that the maternal age effect is complex, dependent on many different biological and environmental factors. We examine conclusions drawn from recent large-scale studies in families, tracing DNA markers along the length of chromosome 21q between parents and DS children, in comparison to the OMS model. We conclude that these family linkage data are equally compatible with the maternal age effect originating from the accumulation of trisomy 21 oocytes with advancing maternal age. One relatively straightforward way to get to grips with what is actually going on in this regard would be to compare incidence of trisomy 21 oocytes (and their pairing configurations) in foetal ovaries with that in oocytes at the meiosis I stage from adult women.

  • 2.
    Belen Poretti, Maria
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Functional Pharmacology. Consejo Nacl Invest Cient & Tecn, FCM, Inst Fisiol, INICSA, Cordoba, Argentina.
    Frautschi, Camila
    Univ Nacl Cordoba, Fac Ciencias Med, Catedra Fisiol Humana, Cordoba, Argentina..
    Luque, Eugenia
    Consejo Nacl Invest Cient & Tecn, FCM, Inst Fisiol, INICSA, Cordoba, Argentina..
    Bianconi, Santiago
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Functional Pharmacology. Univ Nacl Cordoba, Fac Ciencias Med, Catedra Fisiol Humana, Cordoba, Argentina..
    Carolina Martini, Ana
    Consejo Nacl Invest Cient & Tecn, FCM, Inst Fisiol, INICSA, Cordoba, Argentina..
    Stutz, Graciela
    Univ Nacl Cordoba, Fac Ciencias Med, Catedra Fisiol Humana, Cordoba, Argentina..
    Vincenti, Laura
    Univ Nacl Cordoba, Fac Ciencias Med, Catedra Fisiol Humana, Cordoba, Argentina..
    Emilia Santillan, Maria
    Univ Nacl Cordoba, Fac Ciencias Med, Catedra Fisiol Humana, Cordoba, Argentina..
    Ponzio, Marina
    Consejo Nacl Invest Cient & Tecn, FCM, Inst Fisiol, INICSA, Cordoba, Argentina..
    Schiöth, Helgi B.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Functional Pharmacology.
    Fiol de Cuneo, Marta
    Univ Nacl Cordoba, Fac Ciencias Med, Catedra Fisiol Humana, Cordoba, Argentina..
    Paola Carlini, Valeria
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Functional Pharmacology. Consejo Nacl Invest Cient & Tecn, FCM, Inst Fisiol, INICSA, Cordoba, Argentina.
    Reproductive performance of male mice after hypothalamic ghrelin administration2018In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 156, no 2, p. 121-132Article in journal (Refereed)
    Abstract [en]

    It has been demonstrated that food intake and reproductive physiology are both simultaneously modulated to optimize reproductive success under fluctuating metabolic conditions. Ghrelin (GHRL) is an orexigenic peptide identified as the endogenous ligand of the growth hormone secretagogue receptor that is being investigated for its potential role on reproduction. Considering that data available so far are still limited and characterization of GHRL action mechanism on the reproductive system has not been fully elucidated, we studied the participation of hypothalamus in GHRL effects on sperm functional activity, plasma levels of gonadotropins and histological morphology in mice testes after hypothalamic infusion of 0.3 or 3.0 nmol/day GHRL or artificial cerebrospinal fluid (ACSF) at different treatment periods. We found that GHRL 3.0 nmol/day administration for 42 days significantly reduced sperm concentration (GHRL 3.0 nmol/day =14.05 +/- 2.44 x 10(6)/mL vs ACSF =20.33 +/- 1.35 x10(6)/mL, P< 0.05) and motility (GHRL 3.0 nmol/day =59.40 +/- 4.20% vs ACSF =75.80 +/- 1.40%, P< 0.05). In addition, histological studies showed a significant decrease percentage of spermatogonia (GHRL 3.0 nmol/day =6.76 +/- 0.68% vs ACSF =9.56 +/- 0.41%, P< 0.05) and sperm (GHRL 3.0 nmol/day =24.24 +/- 1.92% vs ACSF =31.20 +/- 3.06%, P< 0.05). These results were associated with a significant reduction in luteinizing hormone and testosterone plasma levels (P < 0.05). As GHRL is an orexigenic peptide, body weight and food intake were measured. Results showed that GHRL increases both parameters; however, the effect did not last beyond the first week of treatment. Results presented in this work confirm that central GHRL administration impairs spermatogenesis and suggest that this effect is mediated by inhibition of hypothalamic-pituitary-gonadal axis.

  • 3.
    Bourlev, Vladimir
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Volkov, N.
    Pavlovitch, S.
    Lets, N.
    Larsson, Anders
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical Chemistry.
    Olovsson, Matts
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    The relationship between microvessel density, proliferative activity and expression of vascular endothelial growth factor-A and its receptors in eutopic endometrium and endometriotic lesions2006In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 132, no 3, p. 501-509Article in journal (Refereed)
    Abstract [en]

    Studies were performed to elucidate the possible relationship between microvessel density, proliferative activity and angiogenesis in eutopic endometrium from women with and without endometriosis and peritoneal endometriotic lesions. The question whether changes in these parameters in endometriotic lesions were reflected by the level of vascular endothelial growth factor-A (VEGF-A) in serum and peritonea fluid was also studied. Biopsy specimens of both eutopic endometrium and peritoneal endometriotic lesions from women with endometriosis (n=25) as well as eutopic endometrium from women without endometriosis (n=14) were analysed immunohistochemically regarding microvessel density, proliferative activity, and expression of VEGF-A and its receptors vascular endothelial growth factor receptors 1 and 2 (VEGFR-1 and VEGFR-2) in stroma, glands and blood vessels. The VEGF-A concentration was measured in peritoneal fluid and serum. Secretory phase eutopic endometrium from women with endometriosis had significantly higher microvessel density, expression of VEGF-A in glandular epithelium and VEGFR-2 in endometrial blood vessels than those from women without endometriosis. Endometriotic lesions with high proliferative activity had a higher microvessel density and showed higher vascular expression of VEGFR-2 as well as being accompanied by higher levels of VEGF-A in peritoneal fluid and serum, compared with lesions with low proliferative activity. In conclusion, there seems to be a dysregulation of angiogenic activity in the eutopic endometrium of women with endometriosis and endometriotic lesions with high proliferative activity were accompanied by higher local angiogenic activity and higher levels of VEGF in serum and peritoneal fluid.

  • 4. Coward, K.
    et al.
    Ponting, C. P.
    Chang, H. Y.
    Hibbitt, O.
    Savolainen, Peter
    KTH, School of Biotechnology (BIO), Gene Technology.
    Jones, K. T.
    Parrington, J.
    Phospholipase C zeta, the trigger of egg activation in mammals, is present in a non-mammalian species2005In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 130, no 2, p. 157-163Article in journal (Refereed)
    Abstract [en]

    The activation of the egg to begin development into an embryo is triggered by a sperm-induced increase in intracellular egg Ca2+. There has been much controversy about how the sperm induces this fundamental developmental event, but recent studies suggest that, in mammals, egg activation is triggered by a testis-specific phospholipase C: PLC zeta. Since the discovery of PLC zeta, it has been unclear whether its role in triggering egg activation is common to all vertebrates, or is confined to mammals. Here, we demonstrate for the first time that PLC zeta is present in a non-mammalian vertebrate. Using genomic and cDNA databases, we have identified the cDNA encoding a PLC zeta orthologue in the domestic chicken that, like the mammalian isoforms, is a testis-specific gene. The chicken PLC zeta cDNA is 2152 bp in size and encodes an open reading frame of 639 amino acids. When injected into mouse oocytes, chicken PLC zeta cRNA triggers Ca2+ oscillations, indicating that it has functional properties similar to those of mammalian PLC zeta. Our findings suggest that PLC zeta may have a universal role in triggering egg activation in vertebrates.

  • 5.
    Gallardo Bolanos, Juan M.
    et al.
    University of Extremadura, Spain .
    Balao da Silva, Carolina M.
    University of Extremadura, Spain .
    Martin Munoz, Patricia
    University of Extremadura, Spain .
    Morillo Rodriguez, Antolin
    University of Extremadura, Spain .
    Plaza Davila, Maria
    University of Extremadura, Spain .
    Rodriguez-Martinez, Heriberto
    Linköping University, Department of Clinical and Experimental Medicine, Division of Microbiology and Molecular Medicine. Linköping University, Faculty of Health Sciences. University of Extremadura, Spain .
    Aparicio, Ines M.
    University of Extremadura, Spain .
    Tapia, Jose A.
    University of Extremadura, Spain .
    Ortega Ferrusola, Cristina
    University of Extremadura, Spain .
    Pena, Fernando J.
    University of Extremadura, Spain .
    Phosphorylated AKT preserves stallion sperm viability and motility by inhibiting caspases 3 and 72014In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 148, no 2, p. 221-235Article in journal (Refereed)
    Abstract [en]

    AKT, also referred to as protein kinase B (PKB or RAC), plays a critical role in controlling cell survival and apoptosis. To gain insights into the mechanisms regulating sperm survival after ejaculation, the role of AKT was investigated in stallion spermatozoa using a specific inhibitor and a phosphoflow approach. Stallion spermatozoa were washed and incubated in Biggers-Whitten-Whittingham medium, supplemented with 1% polyvinyl alcohol (PVA) in the presence of 0 (vehicle), 10, 20 or 30 mu M SH5, an AKT inhibitor. SH5 treatment reduced the percentage of sperm displaying AKT phosphorylation, with inhibition reaching a maximum after 1 h of incubation. This decrease in phosphorylation was attributable to either dephosphorylation or suppression of the active phosphorylation pathway. Stallion spermatozoa spontaneously dephosphorylated during in vitro incubation, resulting in a lack of a difference in AKT phosphorylation between the SH5-treated sperm and the control after 4 h of incubation. AKT inhibition decreased the proportion of motile spermatozoa (total and progressive) and the sperm velocity. Similarly, AKT inhibition reduced membrane integrity, leading to increased membrane permeability and reduced the mitochondrial membrane potential concomitantly with activation of caspases 3 and 7. However, the percentage of spermatozoa exhibiting oxidative stress, the production of mitochondrial superoxide radicals, DNA oxidation and DNA fragmentation were not affected by AKT inhibition. It is concluded that AKT maintains the membrane integrity of ejaculated stallion spermatozoa, presumably by inhibiting caspases 3 and 7, which prevents the progression of spermatozoa to an incomplete form of apoptosis.

  • 6.
    Gunnarsson, David
    et al.
    Umeå University, Faculty of Science and Technology, Molecular Biology (Faculty of Science and Technology).
    Leffler, Per
    Umeå University, Faculty of Science and Technology, European CBRNE Center.
    Ekwurtzel, Emelie
    Martinsson, Gunilla
    Liu, Kui
    Umeå University, Faculty of Medicine, Medical Biochemistry and Biophsyics.
    Selstam, Gunnar
    Umeå University, Faculty of Medicine, Molecular Biology (Faculty of Medicine).
    Mono-(2-ethylhexyl) phthalate stimulates basal steroidogenesis by a cAMP-independent mechanism in mouse gonadal cells of both sexes2008In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 135, no 5, p. 693-703Article in journal (Refereed)
    Abstract [en]

    Phthalates are widely used as plasticizers in a number of daily-life products. In this study, we investigated the influence of mono-(2-ethylhexyl) phthalate (MEHP), the active metabolite of the frequently used plasticizer di-(2-ethylhexyl) phthalate (DEHP), on gonadal steroidogenesis in vitro. MEHP (25–100 µM) stimulated basal steroid synthesis in a concentration-dependent manner in immortalized mouse Leydig tumor cells (MLTC-1). The stimulatory effect was also detected in KK-1 granulosa tumor cells. MEHP exposure did not influence cAMP or StAR protein levels and induced a gene expression profile of key steroidogenic proteins different from the one induced by human chorionic gonadotropin (hCG). Simultaneous treatment with MEHP and a p450scc inhibitor (aminoglutethimide) indicated that MEHP exerts its main stimulatory effect prior to pregnenolone formation. MEHP (10–100 µM) up-regulated hormone-sensitive lipase and 3-hydroxy-3-methylglutaryl coenzyme A reductase, suggesting that MEHP increases the amount of cholesterol available for steroidogenesis. Our data suggest that MEHP, besides its known inhibitory effect on hCG action, can directly stimulate gonadal steroidogenesis in both sexes through a cAMP- and StAR-independent mechanism. The anti-steroidogenic effect of DEHP has been proposed to cause developmental disorders such as hypospadias and cryptorchidism, whereas a stimulation of steroid synthesis may prematurely initiate the onset of puberty and theoretically affect the hypothalamic–pituitary–gonadal axis.

  • 7.
    Hay-Schmidt, Anders
    et al.
    Univ Copenhagen.
    Finkielman, Olivia T. Ejlstrup
    Univ Copenhagen.
    Jensen, Benjamin A. H.
    Univ Copenhagen.
    Hogsbro, Christine F.
    Univ Copenhagen.
    Holm, Jacob Bak
    Univ Copenhagen.
    Johansen, Kristoffer Haurum
    Univ Copenhagen.
    Jensen, Tina Kold
    Univ Copenhagen.
    Andrade, Anderson Martino
    Univ Fed Parana, Dept Physiol, Curitiba, Parana, Brazil.
    Swan, Shanna H.
    Icahn Sch Med Mt Sinai, Dept Environm Med & Publ Hlth, New York, NY .
    Bornehag, Carl-Gustaf
    Karlstad University, Faculty of Health, Science and Technology (starting 2013), Department of Health Sciences (from 2013).
    Brunak, Soren
    Univ Copenhagen.
    Jegou, Bernard
    INSERM, IRSET, U1085, Rennes, France; EHESP Sch Publ Hlth, Rennes, France.
    Kristiansen, Karsten
    Univ Copenhagen.
    Kristensen, David Mobjerg
    INSERM, IRSET Rennes ;Univ Copenhagen.
    Prenatal exposure to paracetamol/acetaminophen and precursor aniline impairs masculinisation of male brain and behaviour2017In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 154, no 2, p. 145-152Article in journal (Refereed)
    Abstract [en]

    Paracetamol/acetaminophen (N-Acetyl-p-Aminophenol; APAP) is the preferred analgesic for pain relief and fever during pregnancy. It has therefore caused concern that several studies have reported that prenatal exposure to APAP results in developmental alterations in both the reproductive tract and the brain. Genitals and nervous system of male mammals are actively masculinised during foetal development and early postnatal life by the combined actions of prostaglandins and androgens, resulting in the male-typical reproductive behaviour seen in adulthood. Both androgens and prostaglandins are known to be inhibited by APAP. Through intrauterine exposure experiments in C57BL/6 mice, we found that exposure to APAP decreased neuronal number in the sexually dimorphic nucleus (SDN) of the preoptic area (POA) in the anterior hypothalamus of male adult offspring. Likewise, exposure to the environmental pollutant and precursor of APAP, aniline, resulted in a similar reduction. Decrease in neuronal number in the SDNPOA is associated with reductions in male sexual behaviour. Consistent with the changes, male mice exposed in uteri to APAP exhibited changes in urinary marking behaviour as adults and had a less aggressive territorial display towards intruders of the same gender. Additionally, exposed males had reduced intromissions and ejaculations during mating with females in oestrus. Together, these data suggest that prenatal exposure to APAP may impair male sexual behaviour in adulthood by disrupting the sexual neurobehavioral programming. These findings add to the growing body of evidence suggesting the need to limit the widespread exposure and use of APAP by pregnant women.

  • 8.
    Kumaresan, Arumugam
    et al.
    Swedish Univ Agr Sci, Div Reprod, Dept Clin Sci, Uppsala, Sweden.;Natl Dairy Res Inst, Theriogenol Lab, Anim Reprod Gynaecol & Obstet, Karnal, Haryana, India..
    Johannisson, Anders
    Swedish Univ Agr Sci, Div Reprod, Dept Clin Sci, Uppsala, Sweden..
    Nordqvist, Sarah
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health, Obstetrics and Gynaecology. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology.
    Kårehed, Karin
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health, Obstetrics and Gynaecology. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology.
    Åkerud, Helena
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Medicinsk genetik och genomik.
    Lindgren, Karin E.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health, Obstetrics and Gynaecology. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology.
    Morrell, Jane M.
    Swedish Univ Agr Sci, Div Reprod, Dept Clin Sci, Uppsala, Sweden..
    Relationship of DNA integrity to HRG C633T SNP and ART outcome in infertile couples2017In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 153, no 6, p. 865-876Article in journal (Refereed)
    Abstract [en]

    The status of sperm DNA fragmentation, protamine deficiency, free thiols and disulphide bonds in colloid-selected samples and its relationship to ART outcome or HRG C633T SNP is not known. The objective of this study was to determine these relationships in spermatozoa from men with male factor or unknown factor infertility (n=118) undergoing in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI). Sperm DNA integrity was analysed by flow cytometry using three fluorescent probes (acridine orange, monobromobimane and chromomycin A3). Principal component analysis (PCA) was used to identify the parameters that most influenced fertility. The relationships of sperm DNA integrity with seminal parameters, HRG C633T SNP and ART outcome were established using ANOVA and t-test. Sperm concentration and yield after preparation accounted for 27% of the total variance; sperm DNA integrity (% DFI and disulphide bonds) accounted for 16% of the variance in men from infertile couples. Sperm % DFI was significantly higher (P < 0.05) in older men than in younger men. A significant difference (P < 0.01) was observed in % DFI between smokers and non-smokers. Sperm % DFI was significantly higher (P < 0.01) in male factor infertility compared to either female factor or unknown factor infertility while free thiols were significantly higher (P < 0.01) in unknown infertility factor. No significant difference was observed between IVF success/failure in any of the seminal parameters studied. There was a tendency for protamine deficiency to be higher and disulphide concentration to be lower in men with HRG 633T. Such assessments may provide additional useful information about the prognosis for ART outcome, although more research is needed before clinical guidelines can be provided.

  • 9.
    Mattsson, Anna
    et al.
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Environmental Toxicology.
    Olsson, Jan A
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Environmental Toxicology.
    Brunström, Björn
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Environmental Toxicology.
    Selective estrogen receptor alpha activation disrupts sex organ differentiation and induces expression of vitellogenin II and very low-density apolipoprotein II in Japanese quail embryos2008In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 136, no 2, p. 175-186Article in journal (Refereed)
    Abstract [en]

    The Japanese quail (Coturnix japonica) is a widely used model species for studying the roles of steroid hormones in avian sex differentiation. The aim of the present study was to elucidate the significance of estrogen receptors alpha and beta (ER alpha and ER beta) in normal sex differentiation of the reproductive organs in the Japanese quail and in xenoestrogen-induced disruption of reproductive organ differentiation. Real-time PCR indicated that ER alpha (ESR1) mRNA is expressed in both right and left gonads and Mullerian ducts (MDs) in both sexes during early morphological differentiation. ER beta (ESR2) transcripts were also detected in gonads and MDs, but at very low levels. Both receptor subtypes were expressed in the liver and may therefore mediate the expression of estrogen-regulated egg-yolk proteins. Aromatase mRNA was expressed at much higher levels in female than male gonads as early as embryonic day 5, indicating early sex differences in estrogen synthesis. Treatment with the ER alpha-selective agonist propyl pyrazole triol showed that frequently reported xenoestrogen effects, such as ovotestis formation, abnormal MD development, and hepatic expression of egg-yolk proteins, were induced by selective activation of ER alpha. Taken together, our results suggest that activation of ER alpha is crucial for estrogen-dependent sex differentiation of the reproductive organs and that ER alpha mediates xenoestrogen-induced toxicity during reproductive development in birds.

  • 10.
    Nordqvist, Sarah
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health, Obstetrics and Gynaecology.
    Åkerud, Helena
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health, Obstetrics and Gynaecology.
    Kårehed, Karin
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health, Obstetrics and Gynaecology.
    The Effect of SHB Gene Polymorphisms on Ovarian Response and Oocyte Maturation in IVF2014In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990Article in journal (Refereed)
  • 11.
    Ortega Ferrusola, C.
    et al.
    Vet Teaching Hospital, Spain .
    Gonzalez Fernandez, L.
    University of Extremadura, Spain .
    Morrel, J.M.
    Swedish University of Agriculture Science, Sweden .
    Salazar Sandoval, C.
    Vet Teaching Hospital, Spain .
    Macias Garcia, B.
    Vet Teaching Hospital, Spain .
    Rodriguez-Martinez, Heriberto
    Swedish University of Agriculture Science, Sweden .
    Tapia, J.A.
    University of Extremadura, Spain .
    J. Pena, F.
    Vet Teaching Hospital, Spain .
    Lipid peroxidation, assessed with BODIPY-C-11, increases after cryopreservation of stallion spermatozoa, is stallion-dependent and is related to apoptotic-like changes2009In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 138, no 1, p. 55-63Article in journal (Refereed)
    Abstract [en]

    Lipid peroxidation (LPO) of stallion spermatozoa was assessed in fresh semen and in samples of the same ejaculates after freezing and thawing. Particular attention was paid to individual differences in the susceptibility to LPO and its possible relationship with freezability. Innate levels of LPO were very low in fresh spermatozoa but increased after thawing, a change that was largely stallion-dependent. The level of LPO in fresh spermatozoa was not correlated with that of the thawed spermatozoa. Negative correlations existed between LPO and intact membranes post-thaw (r= -0.789, Pless than0.001), and also between LPO and spermatozoa with high mitochondrial membrane potential (Delta psi m) post-thaw (r= -0.689, Pless than0.001). LPO was also highly and significantly correlated with caspase activity. The correlation between caspase activity in ethidium positive cells and LPO was r=0.772, Pless than0.001. This LPO is unlikely to represent, per se, a sign of cryopreservation-induced injury, but it is apparently capable of triggering apoptotic-like changes that could result in the sub-lethal cryodamage often seen among surviving spermatozoa. Reproduction (2009) 138 55-63

  • 12.
    Sancho, S.
    et al.
    University of Girona, Campus de Montilivi, s/n, 17071 Girona, Spain.
    Casas, I.
    University of Girona, Campus de Montilivi, s/n, 17071 Girona, Spain.
    Ekwal, H.
    Swedish University of Agricultural Sciences (SLU), SE-750 07, Uppsala, Sweden .
    Saravia, F.
    Swedish University of Agricultural Sciences (SLU), SE-750 07, Uppsala, Sweden .
    Rodriguez-Martinez, Heriberto
    Swedish University of Agricultural Sciences (SLU), SE-750 07, Uppsala, Sweden .
    E. Rodriguez-Gil, J.
    Autonomous University of Barcelona, Barcelona E-08193, Spain .
    Flores, E.
    Autonomous University of Barcelona, Barcelona E-08193, Spain .
    Pinart, E.
    University of Girona, Campus de Montilivi, s/n, 17071 Girona, Spain.
    Briz, M.
    University of Girona, Campus de Montilivi, s/n, 17071 Girona, Spain.
    Garcia-Gil, N.
    University of Girona, Campus de Montilivi, s/n, 17071 Girona, Spain.
    Bassols, J.
    University of Girona, Campus de Montilivi, s/n, 17071 Girona, Spain.
    Pruneda, A.
    University of Girona, Campus de Montilivi, s/n, 17071 Girona, Spain.
    Bussalleu, E.
    University of Girona, Campus de Montilivi, s/n, 17071 Girona, Spain.
    Yeste, M.
    University of Girona, Campus de Montilivi, s/n, 17071 Girona, Spain.
    Bonet, S.
    University of Girona, Campus de Montilivi, s/n, 17071 Girona, Spain.
    Effects of cryopreservation on semen quality and the expression of sperm membrane hexose transporters in the spermatozoa of Iberian pigs2007In: Reproduction, ISSN 1470-1626, E-ISSN 1476-3990, Vol. 134, no 1, p. 111-121Article in journal (Refereed)
    Abstract [en]

    This study evaluated the effects of cooling, freezing and thawing on the plasma membrane integrity, kinetics and expression of two sugar transporters glucose transporter-3 and -5 (GLUT-3 and GLUT-5) in spermatozoa from Iberian boars. Semen samples were collected twice weekly from eight young, fertile Iberian boars of the Entrepelado and Lampino breeds. The samples were suspended in a commercial extender and refrigerated to 17 degrees C for transport to the laboratory (step A), where they were further extended with a lactose-egg yolk-based extender and chilled to 5 degrees C (step B) prior to freezing in the presence of glycerol (3%). Spermatozoa were assessed for plasma membrane integrity and sperm motility at each of the steps, including post-thaw (step C). Aliquots were also prepared for immunocytochemical localisation of the sugar transporters (fixed and thin smears for transmission and scanning electron microscopy levels respectively) and for SDS-PAGE electrophoresis and subsequent western blotting, using the same antibodies (rabbit anti-GLUT-3 and anti-GLUT-5 polyclonal antibodies). The results showed lower percentages of progressively motile spermatozoa at step C in both breeds, while the percentage of live spermatozoa was significantly lower only in the Entrepelado breed. The results obtained from electron microscopy clearly showed that Iberian boar spermatozoa expressed the hexose transporters, GLUT-3 and GLUT-5. The pattern of expression, in terms of location and concentration, was characteristic in each case but, in the case of isoform GLUT-5, it remained constant during the different steps of freezing-thawing protocol. These results indicate that cryopreservation affects the status of sperm cells of Iberian boars by altering the distribution of some membrane receptors and decreasing the percentage values of parameters linked to sperm quality.

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