Radioimmunoassay of Ovine and Bovine Serum Progesterone Without Extraction and Chromatography

Schanbacher, B. D.; Hruska, Roman L.

doi:10.1080/07435807909061106

Cited by 17 publications

(5 citation statements)

References 18 publications

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“…After centrifugation (3000 g, 4 8C) for 30 min, plasma was stored at 2 20 8C until assayed. The concentrations of progesterone were determined by radioimmunoassay in unextracted plasma as described (Schanbacher 1979), except that charcoal-dextran solution was used instead of polyethylene glycol for the separation of bound and free radioactivity. Tritiated progesterone ([1,2,6,7-3 H]progesterone, specific activity.…”

Section: Blood Sampling and Progesterone Assaymentioning

confidence: 99%

Antioxidant enzymatic defence systems in sheep corpus luteum throughout pregnancy

Al-Gubory¹,

Bolifraud²,

Germain³

et al. 2004

Reproduction

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The activities of copper, zinc-superoxide dismutase (SOD1), manganese SOD (SOD2), glutathione peroxidase (GPX), glutathione reductase (GSSG-R) and glutathione S-transferase (GST) were studied in sheep corpora lutea (CL) obtained on days 15, 40, 60, 80 and 128 of pregnancy. Maintained enzymatic activity of SOD1, SOD2, GPX, GSSG-R and GST were found in the sheep CL throughout pregnancy. Enzymatic activity of SOD1, GPX and GST increased significantly from day 15 to day 40 of pregnancy, and thereafter remained constant until day 128. SOD2 and GSSG-R activities were not different between any days of pregnancy examined. Apoptotic luteal cells identified by the terminal deoxynucleotidyl transferase-mediated fluoresceindUTP nick-end labelling were very rarely observed, and their incidence (less than 0.5%) was not different between days of pregnancy. These results showed that the activities of antioxidant enzymes in the sheep CL are subject to major changes during early pregnancy, suggesting that the CL of early pregnancy may be rescued from luteolysis through increasing activities of key antioxidant enzymes and inhibition of apoptosis. Maintained levels of antioxidant enzymes in the CL throughout pregnancy may be linked to reactive oxygen species continuously generated in the steroidogenically active luteal cells, and may be involved in the maintenance of luteal steroidogenic activity and cellular integrity.

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Section: Blood Sampling and Progesterone Assaymentioning

confidence: 99%

Antioxidant enzymatic defence systems in sheep corpus luteum throughout pregnancy

Al-Gubory¹,

Bolifraud²,

Germain³

et al. 2004

Reproduction

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“…After centrifugation (3000 g, 4 8C) for 30 min, plasma was stored at K20 8C until assayed. The concentrations of P 4 were determined by RIA in unextracted plasma as described (Schanbacher 1979) and validated for sheep jugular venous plasma (Al-Gubory et al 2005) with slight modifications (Al-Gubory et al 2006). Tritiated P 4 (1,2,6, 7-3 H-P 4 , sp act 88 Ci/mmol) was obtained from Amersham, and a specific anti-P 4 antibody was obtained from the Institut Pasteur (Paris, France).…”

Section: P 4 Assaymentioning

confidence: 99%

Ovine corpus luteum proteins, with functions including oxidative stress and lipid metabolism, show complex alterations during implantation

Arianmanesh¹,

McIntosh²,

Lea³

et al. 2011

Journal of Endocrinology

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Progesterone (P 4 ) secreted by the corpus luteum (CL) is critical for in utero embryo survival and development, although CL proteins are key regulatory factors during the luteal phase. We, therefore, characterised protein expression patterns in ovine CL of pregnancy (days 12, 16 and 20) compared with those of controls, CL of oestrous cycle (days 12 and 16), using two-dimensional gel electrophoresis (2DE) gel-based proteomics. Proteins in 24 significantly altered spots were identified by tandem mass spectroscopy. At the time of embryo implantation (day 16), 77 spots were up-regulated and 101 spots were down-regulated in CL of pregnancy compared with regressed CL. Vimentin, lamin A/C (LMNA), [Mn] superoxide dismutase (SOD2), isocitrate dehydrogenase 1, annexin A1 and elongation factor Tu, mitochondrial (TUFM) altered during CL regression, whereas glutathione S-transferase A1, apolipoprotein A-1, myxovirus resistance protein 1, ornithine aminotransferase and enoyl-CoA hydratase, mitochondrial (ECHS1) tended to be altered during CL maintenance. biliverdin reductase B (BLVRB), FDXR, guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-2 (GNB2) and cytochrome bc1 complex subunit 1, mitochondrial (UQCRC1) showed divergent expression during CL regression and maintenance. The expression of two representative proteins, SOD2 and BLVRB, by western blot increased in CL of non-pregnant ewes on day 16 compared with that on day 12. SOD2 and BLVRB were localised in the large and small luteal cells and endothelial cells of CL over peri-implantation periods. 2DE gel and mass spectrometry have been used, for the first time, to study ovine CL function. We have identified proteins involved in key pathways, including oxidative stress, steroidogenesis, signal transduction and apoptosis, which have not previously been associated with changes occurring in the CL during the peri-implantation period. These proteins are most likely involved with mechanisms allowing the CL to produce P 4 during early pregnancy.

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“…Placentome ethanol extracts were analyzed for progesterone content in duplicate by a RIA as previously described (Schanbacher 1979) and validated for sheep jugular venous plasma with slight modifications (Al-Gubory et al 2006). Briefly, dextran-coated charcoal solution was used instead of polyethylene glycol for the separation of bound and free radioactivity.…”

Section: Progesterone Riamentioning

confidence: 99%

Developmental changes in antioxidant enzymatic defences against oxidative stress in sheep placentomes

Garrel¹,

Fowler²,

Al-Gubory³

2010

Journal of Endocrinology

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Early pregnancy is susceptible to oxidative stress, and thus characterisation of antioxidant systems and pro-and antiapoptotic pathways would improve understanding of placental development and function. We aimed, therefore, to determine the activities of the antioxidant enzymes, copper/zinc-superoxide dismutase (SOD1), manganese-SOD (SOD2), catalase (CAT), glutathione (GSH) peroxidase (GPX) and GSH reductase (GSR); and to quantify the expression of BAX and MCL1 proteins in relation to the developmental changes in antioxidant defences in sheep placentomes sampled on days 35, 55 and 80 of pregnancy. Placentome progesterone content was analyzed to determine steroidogenic capacity. Malondialdehyde (MDA) and protein carbonyl were quantified in placentomes as biomarkers of lipid peroxidation and protein damage respectively. Placentome tissues demonstrated significantly increased content of progesterone and MDA at day 80 of pregnancy and protein carbonyl as early as day 50 of pregnancy. Progesterone and MDA contents were not different between days 35 and 55 of pregnancy. While SOD1 and CAT activities did not alter significantly, SOD2 activity decreased from days 35 to 55. GPX activity increased from days 35 to 55 and increased further to day 80 of pregnancy. GSR activity increased from days 35 to 55 of pregnancy. BAX protein expression decreased, while MCL1 increased from days 35 to 55 and 80 of pregnancy. The increased GPX activity was associated with a decrease in the BAX/MCL1 protein expression ratio. Changes in the antioxidant enzymatic defences could be a part of placentome adaptation to reactive oxygen speciesinduced oxidative stress at specific early developmental stages of pregnancy.

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Radioimmunoassay of Ovine and Bovine Serum Progesterone Without Extraction and Chromatography

Cited by 17 publications

References 18 publications

Antioxidant enzymatic defence systems in sheep corpus luteum throughout pregnancy

Antioxidant enzymatic defence systems in sheep corpus luteum throughout pregnancy

Ovine corpus luteum proteins, with functions including oxidative stress and lipid metabolism, show complex alterations during implantation

Developmental changes in antioxidant enzymatic defences against oxidative stress in sheep placentomes

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