93 research outputs found
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Comparison of bioactivities, binding properties and intra-follicular levels of bovine follistatins
Five isoforms of follistatin (FST) (Mr 31, 33, 35, 37, 41kDa) were purified from bovine follicular fluid (bFF). Comparison of their activin- and heparan sulphate proteoglycan (HSP)-binding properties and bio-potencies in neutralization of activin-A action in vitro revealed that all five isoforms bound activin-A, but with different affinities. Only the 31kDa isoform (FST-288) bound to HSP. FST-288 also showed the greatest biopotency with 35 and 41kDa isoforms being least potent. To determine whether bovine follicle development is associated with changing intrafollicular FST and activin profiles, we analyzed bFF from dominant (DF) and subordinate (SF) follicles collected at strategic times during a synchronized estrous cycle. Total FST, activin-A and activin-AB were measured by immunoassay while individual FST isoforms were quantified by immunoblotting. Follicle diameter was positively correlated with estrogen:progesterone ratio (r=0.56) in bFF but negatively correlated with activin-A (r=-0.34), activin-AB (r=-0.80) and ‘total’ FST (r=-0.70) levels. Follicle diameter was positively correlated with abundance of the 41 kDa isoform (r=0.59) but negatively correlated with abundance of 33 and 31 kDa isoforms (r=-0.56, -0.41). Both follicle status (DF vs SF) and cycle stage affected total FST, activin-A, activin-B levels while follicle status, but not cycle stage, affected abundance of 41, 37, 33 and 31kDa FST isoforms. Collectively, these findings indicate that intrafollicular FST isoforms that differ in their ability to bind and neutralise activins and associate with cell-surface proteoglycans, show divergent changes during follicle development. Enhanced FST production may have an important negative role, either directly or via inhibition of the positive effects of activins, on follicle growth and function during follicular waves
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The global effect of follicle-stimulating hormone and tumour necrosis factor α on gene expression in cultured bovine ovarian granulosa cells
Background
Oocytes mature in ovarian follicles surrounded by granulosa cells. During follicle growth, granulosa cells replicate and secrete hormones, particularly steroids close to ovulation. However, most follicles cease growing and undergo atresia or regression instead of ovulating. To investigate the effects of stimulatory (follicle-stimulating hormone; FSH) and inhibitory (tumour necrosis factor alpha; TNFα) factors on the granulosa cell transcriptome, bovine ovaries were obtained from a local abattoir and pools of granulosa cells were cultured in vitro for six days under defined serum-free conditions with treatments present on days 3–6. Initially dose–response experiments (n = 4) were performed to determine the optimal concentrations of FSH (0.33 ng/ml) and TNFα (10 ng/ml) to be used for the microarray experiments. For array experiments cells were cultured under control conditions, with FSH, with TNFα, or with FSH plus TNFα (n = 4 per group) and RNA was harvested for microarray analyses.
Results
Statistical analysis showed primary clustering of the arrays into two groups, control/FSH and TNFα/TNFα plus FSH. The effect of TNFα on gene expression dominated that of FSH, with substantially more genes differentially regulated, and the pathways and genes regulated by TNFα being similar to those of FSH plus TNFα treatment. TNFα treatment reduced the endocrine activity of granulosa cells with reductions in expression of FST, INHA, INBA and AMH. The top-ranked canonical pathways and GO biological terms for the TNFα treatments included antigen presentation, inflammatory response and other pathways indicative of innate immune function and fibrosis. The two most significant networks also reflect this, containing molecules which are present in the canonical pathways of hepatic fibrosis/hepatic stellate cell activation and transforming growth factor β signalling, and these were up regulated. Upstream regulator analyses also predicted TNF, interferons γ and β1 and interleukin 1β.
Conclusions
In vitro, the transcriptome of granulosa cells responded minimally to FSH compared with the response to TNFα. The response to TNFα indicated an active process akin to tissue remodelling as would occur upon atresia. Additionally there was reduction in endocrine function and induction of an inflammatory response to TNFα that displays features similar to immune cells
Polarization observables in deuteron photodisintegration below 360 MeV
High precision measurements of induced and transferred recoil proton polarization in d((gamma) over right arrow, (p) over right arrow )n have been performed for photon energies of 277-357 MeV and theta(cm) = 20 degrees-120 degrees. The measurements were motivated by a longstanding discrepancy between meson-baryon model calculations and data at higher energies. At the low energies of this experiment, theory continues to fail to reproduce the data, indicating that either something is missing in the calculations and/or there is a problem with the accuracy of the nucleon-nucleon potential being used. (C) 2011 Elsevier B.V. All rights reserved
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The anti-epileptic drug Valproic Acid (VPA) inhibits steroidogenesis in bovine theca and granulosa cells in vitro
Valproic acid (VPA) is used widely to treat epilepsy and bipolar disorder. Women undergoing VPA treatment reportedly have an increased incidence of polycystic ovarian syndrome (PCOS)-like symptoms including hyperandrogenism and oligo- or amenorrhoea. To investigate potential direct effects of VPA on ovarian steroidogenesis we used primary bovine theca (TC) and granulosa (GC) cells maintained under conditions that preserve their 'follicular' phenotype. Effects of VPA (7.8-500 µg/ml) on TC were tested with/without LH. Effects of VPA on GC were tested with/without FSH or IGF analogue. VPA reduced (P99% decrease; P<0.0001) with lesser effects on LHR, STAR, CYP11A1 and HSD3B1 mRNA (<90% decrease; P<0.05). VPA only reduced TC progesterone secretion induced by the highest (luteinizing) LH dose tested; TC number was unaffected by VPA. At higher concentrations (125-500 µg/ml) VPA inhibited basal, FSH- and IGF-stimulated estradiol secretion (P<0.0001) by GC without affecting progesterone secretion or cell number. VPA reversed FSH-induced upregulation of CYP19A1 and HSD17B1 mRNA abundance (P<0.001). The potent histone deacetylase (HDAC) inhibitors trichostatin A and scriptaid also suppressed TC androstenedione secretion and granulosal cell oestrogen secretion suggesting that the action of VPA reflects its HDAC inhibitory properties. In conclusion, these findings refute the hypothesis that VPA has a direct stimulatory action on TC androgen output. On the contrary, VPA inhibits both LH-dependent androgen production and FSH/IGF-dependent estradiol production in this in vitro bovine model, likely by inhibition of HDAC
A precise extraction of the induced polarization in the 4He(e,e'p)3H reaction
We measured with unprecedented precision the induced polarization Py in
4He(e,e'p)3H at Q^2 = 0.8 (GeV/c)^2 and 1.3 (GeV/c)^2. The induced polarization
is indicative of reaction-mechanism effects beyond the impulse approximation.
Our results are in agreement with a relativistic distorted-wave impulse
approximation calculation but are over-estimated by a calculation with strong
charge-exchange effects. Our data are used to constrain the strength of the
spin independent charge-exchange term in the latter calculation.Comment: submitted to Physical Review Letter
Polarization Transfer in the 4He(e,e'p)3H Reaction at Q^2 = 0.8 and 1.3 (GeV/c)^2
Proton recoil polarization was measured in the quasielastic 4He(e,e'p)3H
reaction at Q^2 = 0.8 (GeV/c)^2 and 1.3 (GeV/c)^2 with unprecedented precision.
The polarization-transfer coefficients are found to differ from those of the
1H(e,e' p) reaction, contradicting a relativistic distorted-wave approximation,
and favoring either the inclusion of medium-modified proton form factors
predicted by the quark-meson coupling model or a spin-dependent charge-exchange
final-state interaction. For the first time, the polarization-transfer ratio is
studied as a function of the virtuality of the proton
Bone Morphogenetic Protein-6 (BMP-6) induces atresia in goat primordial follicles cultured in vitro
Low Q^2 measurements of the proton form factor ratio
We present an updated extraction of the proton electromagnetic form factor
ratio, mu_p G_E/G_M, at low Q^2. The form factors are sensitive to the spatial
distribution of the proton, and precise measurements can be used to constrain
models of the proton. An improved selection of the elastic events and reduced
background contributions yielded a small systematic reduction in the ratio mu_p
G_E/G_M compared to the original analysis.Comment: 12 pages, 5 figures, archival paper for proton form factor extraction
from Jefferson Lab "LEDEX" experimen
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