Nitric oxide induces apoptosis in bovine luteal cells

We previously showed in in vivo and in vitro studies that nitric oxide (NO) is engaged in luteolysis in cattle. Nitric oxide produced locally in the bovine corpus luteum (CL) inhibits progesterone (P4) synthesis and is suggested to be a component of the luteolytic cascade induced by uterine prostaglandin (PG) F2alpha. In the present study, the molecular mechanisms of NO action during structural luteolysis were studied in cultured bovine luteal cells (Days 15-17 of the estrous cycle). The effects of the NO donor (NONOate; 10(-4)M) on DNA fragmentation, cell viability, P4 production and caspase-3 activity were compared with those of PGF2alpha (10(-6)M). Moreover, mobilization of intracellular calcium [Ca2+]i and gene expressions of Fas-L, Fas, bcl-2, bax, and caspase-3 in the cells were determined by semi-quantitative RT-PCR after NONOate treatment. Caspase-3 activity was examined calorimetrically. Contrary to PGF2alpha NONOate decreased cell viability. DNA fragmentation after NONOate treatment increased by more than with PGF22alpha. NONOate increased mobilization of [Ca2+]i in the cells. Although the NO donor did not affect Fas-L and bcl-2 gene expression, it stimulated Fas and bax mRNA and caspase-3 expression. The ratio of bcl-2 to bax mRNA level decreased in the cells treated with NONOate. Moreover, NONOate stimulated caspase-3 activity more effectively than PGF2alpha. The overall results suggest that NO is a luteolytic factor that plays a crucial role in regulation of the estrous cycle in structural luteolysis by inducing apoptosis of luteal cells in cattle.     

Leukotrienes are Auto‐/Paracrine Factors in the Bovine Corpus Luteum: An In Vitro Study

The aim of this study was to determine leukotrienes (LTs) functions in the bovine corpus luteum (BCL) during the oestrous cycle. In steroidogenic CL cells we examined the effect of luteotropic [LH, prostaglandin E₂ (PGE₂)] and luteolytic (PGF_2α, cytokines) factors on: the levels of LTB₄ and C₄, the expression of 5‐lipoxygenase (LO), LT receptors type I (LTR‐I) and LTR‐II, and the effects of LTB₄ and C₄ stimulations on the levels of progesterone (P4), PGE₂, F_2α and nitric oxide (NO) metabolites. Both luteolytic and luteotropic factors stimulated 5‐LO expression on days 2–4 and 17–19 of the cycle. Leukotriene receptors type I expression increased after PGE₂ and tumour necrosis factor α with interferon γ (TNF/IFN) stimulation on days 2–4 of the cycle. Leukotriene receptor type II expression increased after PGE_2α and TNF/IFN stimulation on days 2–4 and 17–19 of the cycle, and LTR‐II expression on days 8–10 of the cycle was unchanged after cell stimulation with any factor. Leukotriene B₄ level increased after BSC incubation with luteotropic factors during all examined days of the cycle and after cytokine stimulation at early‐ and mid‐luteal stages, whereas luteolytic factors stimulated LTC₄ secretion over the entire cycle. Leukotriene B₄ stimulated P4 secretion at the mid‐luteal stage and stimulated NO secretion during all examined phases. Leukotriene B₄ stimulated PGE₂ secretion at the early‐ and mid‐luteal stage. Leukotriene C₄ inhibited P4 secretion at the mid‐ and regressing‐luteal stage, stimulated NO (entire cycle) and PGF_2α at mid‐ and regressing‐luteal phases. Leukotrienes modulate steroidogenic cells functions, depending on the stage of the cycle. Leukotriene B₄ plays a luteotropic role stimulating P4 and PGE₂ secretions; LTC₄ stimulates the secretion of luteolytic factors and enhances the luteolytic cascade within BCL.     

Do the impacts of alien invasive plants differ from expansive native ones? An experimental study on arbuscular mycorrhizal fungi communities

No studies have compared so far the effects of alien invasive and expansive native (widespread, mono-dominant) plants on arbuscular mycorrhizal fungi (AMF). Four global or European most successful invaders (Impatiens glandulifera, Reynoutria japonica, Rudbeckia laciniata, Solidago gigantea) and two expansive plants native to Europe (Artemisia vulgaris, Phalaris arundinacea) were grown in pots to elucidate the magnitude and direction of changes in AMF abundance, species richness, and species composition in soils from under multispecies native vegetation. In a second stage, the effects of these changes on a native plant, Plantago lanceolata, were assessed. Plant species identity had larger impact on AMF abundance, species richness, and species composition as well as on P. lanceolata than origin of the species (alien vs. native). This could be due to the character of AMF relationships with the plants, i.e., their mycorrhizal status and dependency on AMF. However, the alterations induced by the plant species in soil chemical properties rather than in AMF community were the major drivers of differences in shoot mass and photosynthetic performance of P. lanceolata. We determined that the plants produced species-specific effects on soil properties that, in turn, resulted in species-specific soil feedbacks on the native plant. These effects were not consistent within groups of invaders or natives.     

Effects of Exogenous Tumour Necrosis Factor-α on the Secretory Function of the Bovine Reproductive Tract Depend on Tumour Necrosis Factor-α Concentrations

The aim of study was to correlate tumour necrosis factor-α (TNF) infused doses used with the TNF concentrations achieved and with the secretory function of both the ovary and the uterus in cows. We evaluated the concentrations of progesterone (P4), prostaglandin (PG)F_2α, PGE₂ nitric oxide (NO) and TNF in the jugular vein and vena cava caudalis as parameters of exogenous TNF action on the female reproductive tract. Aortae abdominalis of cows (n = 18) were infused with saline or two doses of TNF (luteolytic – 1 μg or luteotrophic – 10 μg). In the peripheral blood, 1 μg TNF concentrations achieved within the range of 30–45 pg/ml, and 10 μg TNF provoked a sharp increase in achieved concentrations at a range of 250–450 pg/mL). The TNF concentrations achieved in vena cava caudalis were five to six times higher than that in peripheral blood (p < 0.001). One microgram TNF increased PGF_2α and NO (p < 0.001) and decreased P4 (p < 0.05). The higher TNF dose stimulated P4 and PGE₂ (p < 0.01). TNF infusion at luteolytic dose achieved its concentrations at the physiological range previously observed in cows. Luteotrophic TNF dose achieved the concentrations in vena cava caudalis that are much higher than physiological level and were previously noted in pathological circumstances (i.e. mastitis , metritis ).     

Luteolytic Effect of Prostaglandin F2α on Bovine Corpus Luteum Depends on Cell Composition and Contact

Prostaglandin F_2α (PGF_2α) is a main luteolytic factor in vivo; however, its direct luteolytic influence on steroidogenic cells of bovine corpus luteum (CL) is controversial and not fully understood. The aim of the study was to clarify PGF_2α action on bovine CL in different in vivo and in vitro conditions and to examine whether the contact among all main types of CL cells is necessary for luteolytic PGF_2α action. In experiment 1, the bovine CL (day 15 of the oestrous cycle) was perfused using in vivo microdialysis system with dinoprost (an analogue of PGF_2α) for 0.5 h. Dinoprost caused a short‐time increase in progesterone (P4), whose concentration decreased thereafter (at 6‐, 10‐, 12‐ and 24‐h after treatment). In experiment 2, the direct effect of PGF_2α on P4 accumulation in CL steroidogenic cells cultured in monolayer (day 15 of the cycle) was determined. PGF_2α after 24 h of incubation increased P4 accumulation in steroidogenic CL cells. In experiment 3 steroidogenic, endothelial CL and immune cells (day 15 of the cycle) were incubated with PGF_2α in cocultures for 24 h in glass tubes and the levels of P4, stable metabolites of nitric oxide (NO) and leukotriene (LT) C₄ were determined. Although PGF_2α treatment increased P4 secretion in homogeneous steroidogenic CL cell culture, the decrease in P4 secretion in cocultures of all types of CL cells was observed. The secretion of NO and LTC₄ increased after the treatment of PGF_2α both in pure cultures of CL cells and in cocultures. The interactions between endothelial and immune cells with steroidogenic CL cells are needed for luteolytic PGF_2α action within the bovine CL. Our results indicate that the cell coculture model, including the main types of CL cells, is the most approximate to study PGF_2α role in vitro.     

Differential influence of four invasive plant species on soil physicochemical properties in a pot experiment

Purpose

This study compared the effects of four invasive plants, namely Impatiens glandulifera, Reynoutria japonica, Rudbeckia laciniata, and Solidago gigantea, as well as two native species—Artemisia vulgaris, Phalaris arundinacea, and their mixture on soil physicochemical properties in a pot experiment.

Materials and methods

Plants were planted in pots in two loamy sand soils. The soils were collected from fallows located outside (fallow soil) and within river valley (valley soil) under native plant communities. Aboveground plant biomass, cover, and soil physicochemical properties such as nutrient concentrations, pH, and water holding capacity (WHC) were measured after two growing seasons. Discriminant analysis (DA) was used to identify soil variables responsible for the discrimination between plant treatments. Identified variables were further compared between treatments using one-way ANOVA followed by Tukey’s HSD test.

Results and discussion

Plant biomass, cover, and soil parameters depended on species and soil type. DA effectively separated soils under different plant species. DA on fallow soil data separated R. laciniata from all other treatments, especially I. glandulifera, native species and bare soil, along axis 1 (related mainly to exchangeable K, N-NH₄, total P, N-NO_3, and WHC). Large differences were found between R. laciniata and S. gigantea as indicated by axis 2 (S-SO₄, exchangeable Mg, total P, exchangeable Ca, and total Mg). DA on valley soil data separated R. japonica from all other treatments, particularly S. gigantea, R. laciniata, and native mixture, along axis 1 (N-NO₃, total N, S-SO₄, total P, pH). Along axis 2 (N-NO₃, N-NH₄, Olsen P, exchangeable K, WHC), large differences were observed between I. glandulifera and all other invaders.

Conclusions

Plant influence on soil differed both among invasive species and between invasive and native species. Impatiens glandulifera had a relatively weak effect and its soil was similar to both native and bare soils. Multidirectional effects of different invaders resulted in a considerable divergence in soil characteristics. Invasion-driven changes in the soil environment may trigger feedbacks that stabilize or accelerate invasion and hinder re-colonization by native vegetation, which has implications for the restoration of invaded habitats.

     

Nitric oxide stimulates progesterone and prostaglandin E2 secretion as well as angiogenic activity in the equine corpus luteum

Cytokines and nitric oxide (NO) are potential mediators of luteal development and maintenance, angiogenesis, and blood flow. The aim of this study was to evaluate (i) the localization and protein expression of endothelial and inducible nitric oxide synthases (eNOS and iNOS) in equine corpora lutea (CL) throughout the luteal phase and (ii) the effect of a nitric oxide donor (spermine NONOate, NONOate) on the production of progesterone (P4) and prostaglandin (PG) E(2) and factor(s) that stimulate endothelial cell proliferation using equine luteal explants. Luteal tissue was classified as corpora hemorrhagica (CH; n = 5), midluteal phase CL (mid-CL; n = 5) or late luteal phase CL (late CL; n = 5). Both eNOS and iNOS were localized in large luteal cells and endothelial cells throughout the luteal phase. The expression of eNOS was the lowest in mid-CL (P < 0.05) and the highest in late CL (P < 0.05). However, no change was found for iNOS expression. Luteal explants were cultured with no hormone added or with NONOate (10(-5) M), tumor necrosis factor-α (TNFα; 10 ng/mL; positive control), or equine LH (100 ng/mL; positive control). Conditioned media by luteal tissues were assayed for P4 and PGE(2) and for their ability to stimulate proliferation of bovine aortic endothelial cells (BAEC). All treatments stimulated release of P4 in CH, but not in mid-CL. TNFα and NONOate treatments also increased PGE(2) levels and BAEC proliferation in CH (P < 0.05). However, in mid-CL, no changes were observed, regardless of the treatments used. These data suggest that NO and TNFα stimulate equine CH secretory functions and the production of angiogenic factor(s). Furthermore, in mares, NO may play a role in CL growth during early luteal development, when vascular development is more intense.     

Predicting the effects of whale population recovery on Northeast Pacific food webs and fisheries: an ecosystem modelling approach

The recovery of whale populations from historical depletion may have the potential to noticeably affect Northeast Pacific ecosystems and fisheries. Surplus production models based on whaling catch records were used to reconstruct the historical abundances of five large whale species in the waters surrounding Haida Gwaii, British Columbia. The results suggest that the local abundances of all five species were vastly higher before the onset of modern whaling. A comparison of ecosystem models representing the states of the local marine food web before and after full whale recovery indicates that abundant whales could consume large proportions of the annual production of their principal prey, ranging up to 87% for Pacific herring (Clupea pallasii) and 72% for piscivorous rockfish (Sebastes spp.). Dynamic modelling of the food web effects of whale recovery, including simulations of simultaneous top‐down and bottom‐up forcing and a Monte Carlo sensitivity analysis, revealed noticeable (～6–12%) top‐down effects on Pacific herring biomass owing to increased predation by humpback and fin whales. However, these effects cannot explain the magnitude of recent declines in local herring biomass. The dynamic modelling results also suggest that top‐down effects of whale recovery could result in reduced biomasses of large rockfish as a result of predation by sperm whales, as well as potential cascading effects on many demersal fish groups. These findings have numerous practical implications for ecosystem‐based fisheries management and whale conservation strategies in Northeast Pacific waters.