Can environmental factors increase oxidative responses in fish exposed to polycyclic aromatic hydrocarbons (PAHs)?

Relations among polycyclic aromatic hydrocarbons (PAHs), biomarkers of oxidative stress (lipid peroxidation, glutathione, and glutathione S-transferase activity), and the possible influence of environmental factors (temperature, pH, and salinity) were assessed in situ for specimens of Ramnogaster arcuata, a native estuarine fish. PAH levels found in the muscular tissue of R. arcuata ranged from 0.7 to 293.4 ng g-1 wet weight with petrogenic and pyrolytic inputs. Lipid peroxidation in the liver showed positive correlations (P < 0.05) with total PAHs (r = 0.66), 3-ring (r = 0.66) and 4-ring PAHs (r = 0.52) and glutathione in muscle (r = 0.58). Significant positive correlations (P < 0.05) were also evidenced between muscular glutathione with total (r = 0.62) and 3-ring PAHs (r = 0.75). Hepatic glutathione S-transferase negatively correlated with 4-ring PAHs (r = -0.58). These correlations suggest that lipid peroxidation and muscular glutathione could be good biomarkers for complex mixtures of PAHs, and hepatic glutathione S-transferase could be a suitable biomarker for 4-ring PAHs. Furthermore, significant correlations (P < 0.05) of environmental factors with PAH levels and biomarkers were observed, especially pH with 3-ring PAHs (r = -0.65), lipid peroxidation (r = -0.6), glutathione in the liver (r = -0.73) and muscle (r = -0.75); and temperature with 2-ring PAHs (r = -0.75) and glutathione in muscle (r = 0.51). The data suggest an influence of physicochemical parameters which could be driving a shift in PAH toxicity in R. arcuata. These results are essential for an integrated understanding of ecotoxicology and could help to predict environmental effects in present and future scenarios of ocean warming and acidification.

Microplastics and anthropogenic debris in rainwater from Bahia Blanca, Argentina.

Concern about atmospheric microplastic (MP) contamination has increased in recent years. This study assessed the abundance of airborne anthropogenic particles, including MPs, deposited in rainfall in Bahia Blanca, southwest Buenos Aires, Argentina. Rainwater samples were collected monthly from March to December 2021 using an active wet-only collector consisting of a glass funnel and a PVC pipe that is only open during rain events. Results obtained show that all rain samples contained anthropogenic debris. The term "anthropogenic debris" is used to refer to the total number of particles as not all the particles found could be determined as plastic. Among all the samples, an average deposition of 77 ± 29 items (anthropogenic debris) m-2d-1 was found. The highest deposition was observed in November (148 items m-2d-1) while the lowest was found in March (46 items m-2d-1). Anthropogenic debris ranged in size from 0.1 mm to 3.87 mm with the most abundant particles being smaller than 1 mm (77.8%). The dominant form of particles found were fibers (95%), followed by fragments (3.1%). Blue color predominated (37.2%) in the total number of samples, followed by light blue (23.3%) and black (21.7%). Further, small particles (<2 mm), apparently composed of mineral material and plastic fibers, were recognized. The chemical composition of suspected MPs was examined by Raman microscopy. The analysis of µ-Raman spectra confirmed the presence of polystyrene, polyethylene terephthalate, and polyethylene vinyl acetate fibers and provided evidence of fibers containing industrial additives such as indigo dye. This is the first assessment of MP pollution in rain in Argentina.

Effects of microplastic ingestion on feeding activity in a widespread fish on the southwestern Atlantic coast: Ramnogaster arcuata (Clupeidae).

The presence of microplastics (MPs) in the fish gastrointestinal tract has been documented in several studies. However, it is unclear whether this ingestion is active or passive and whether it affects feeding activity in natural environments. In this study, three sites with different anthropogenic pressure were selected from the Bahia Blanca estuary, Argentina, and the small zooplanktivorous pelagic fish Ramnogaster arcuata was used to evaluate MP ingestion and its effect on specie's trophic activity. We analyzed the zooplanktonic composition, levels, and types of MPs in both the environment and the stomach contents of R. arcuata. Moreover, we assessed the trophic behavior of R. arcuata to determine electivity, stomach fullness, and vacuity indexes. The results showed that despite the availability of prey in the environment, 100 % of the specimens ingested MPs, and their levels and characteristics were different according to the site. Stomach contents at sites related to harbor activities presented the lowest MPs concentrations, most of them were paint fragments, with the smallest sizes and a low diversity of colors. The highest MP ingestions were found near the principal sewage discharge, being mostly microfibers, followed by microbeads, and with a greater variety of colors. Electivity indices showed that R. arcuata has a passive or active ingestion according to the size and shape of MPs. In addition, the lowest stomach fullness index and the highest vacuity index values were associated with the highest level of MP ingestion near the sewage discharge. Altogether, these results demonstrate a negative effect of MPs in the feeding activity of R. arcuata and help to elucidate some mechanisms by which these particles are ingested by a bioindicator fish used in South America.

Insights on metal pollution of a Patagonia watershed: A case study in the lower course of the Negro river, Argentina.

This study evaluated the occurrence and distribution of largely known pollutants (Ag, Cd, Cu, Cr, Hg, Ni, Pb, Pd, and Zn), as well as emerging ones (Li, and V) in the water dissolved fraction, suspended particulate matter, and surface sediments from the lower course of the Negro River, Argentina. There are scarce preceding data on inorganic pollution in the entire watershed and, in the case of the emerging pollutants, there are almost no studies performed worldwide. Sampling was conducted in 2019 at six sampling sites, three of them mostly river dominated and the rest under marine domain. The samples were subjected to an acid digestion in a microwave digester, and analyzed using an Inductively Coupled Plasma Atomic Emission Spectrometer. Results: revealed that Cu, Li, V, and Zn were always on the top four of the highest average metal concentrations in water and sediment fractions. The pollution assessment indicated that the watershed might be exposed to anthropogenic pollution, as over 60% of Cu and Zn, and over 85% of Hg in water dissolved samples from the marine dominated sites were above the maximum recommended values from guidelines. The multivariate analyses characterized the watershed into two clusters, with metals in the sediment fraction mainly contributing to the uppermost sites. Indeed, sedimentary Cu and Zn background enrichment indices pointed out a moderate pollution of the river dominated sites. This study highlights the relevance of an integrative approach in metal pollution evaluation, as the results denoted a progressive deterioration of the watershed, affecting the water quality of the lower course of the Negro River and its adjacent coastal zone. Overall, these results contribute to a more complete evaluation of the potential to fulfill the Sustainable Development Goals, with implications for future treatment strategies to enhance the environmental quality of the area.

Large-scale spatiotemporal variations, sources, and risk assessment of banned OCPs and PAHs in suspended particulate matter from the Negro river, Argentina.

Organochlorine pesticides (OCPs) and polycyclic aromatic hydrocarbons (PAHs) threaten the environment due to their wide environmental resistance. Environmental paradigms coexist along the Negro River (NR) in Argentina, South America, which flows to the sea below the latitude of 40o S; however, this is the first environmental assessment of OCPs and PAHs in water of the NR for more than 15 years. With 21 sampling sites covering a range of 600 km of river extension, we assessed 16 OCPs and 16 PAHs in suspended particulate material (SPM) with regard to their levels, seasonality, sources, and potential biological risk assessment. Using gas chromatography-mass spectrometry and gas chromatography coupled with electron capture detection, we found an overall mean value for Σ16 OCPs of 648.56 ng. g-1, d.w. Despite a ban spanning 25 years, an increasing trend of accumulation of hexachlorocyclohexanes (HCHs) and endosulfan was shown in the lower valley. The ɑ-HCH/ɤ-HCH and ß-HCH/(ɑ + É¤)-HCH ratios indicated a prevalent usage of technical HCH over lindane and recent HCH inputs. The most abundant compound, α-endosulfan, averaged 141.64 ng. g-1, d.w. and DDX (Σ 4,4'-DDE, 4,4'-DDD, and 4,4'-DDT) averaged 99.98 ng. g-1, d.w. Winter OCP loads in the NR reflected the runoff of the heaviest pesticide application period. We estimated the total discharge of DDT into the Atlantic ocean was 96 g.day-1, added to 458 g of HCHs and 257 g of endosulfans (ɑ + ß + epoxide) adsorbed by the SPM. PAHs occurred widely along the river (38.83 ± 43.52 µg. g-1) and the highest levels coincided with locations with marked anthropogenic-related activity, such as petroleum/gas exploitation facilities. Risk quotient analysis showed a low risk posed by OCPs, but a high risk of potential effects on biota posed by the PAHs, highlighting the need for mitigation measures.

First assessment of debris pollution in the gastrointestinal content of juvenile Magellanic penguins (Spheniscus magellanicus) stranded on the west south Atlantic coasts.

This paper provides the first evidence of debris pollution, including plastic, in juvenile Magellanic penguins (Spheniscus magellanicus) found stranded on the Atlantic coast of southern Buenos Aires Province, Argentina. Macro-, meso- and microparticles of anthropogenic origin were observed in 100 % of the studied birds, with debris abundance ranging between 33 and 200 items/bird. Microparticles represented 91 % of the total debris and 97 % of them were fibers. Black particles were the most abundant (30 %), followed by transparent (26 %), blue (14 %), yellow (10.3 %), and red (10 %). Infrared and Raman spectroscopy identified 62.7 % of the total particles as plastics, with polypropylene (27.8 %) and polyester (21.6 %) being the most abundant polymers. Semi-synthetic cellulosic fibers, metallic particles, and pigments were also found. The presence of metallic microparticles was suggested for the first time in penguins. Stranded juvenile Magellanic penguins are proposed as promising bioindicators of plastic pollution in the South Atlantic.

Occurrence, behavior and the associated health risk of organochlorine pesticides in sediments and fish from Bahía Blanca Estuary, Argentina.

Organochlorine pesticides (OCPs) were assessed for their occurrence, behavior and the associated human health and ecological risks in four fish species (Micropogonias furnieri, Cynoscion guatucupa, Mustelus schmitti, and Ramnogaster arcuata) and sediments from the Bahía Blanca estuary, Argentina, an important coastal environment of South America. Total OCPs values ranged from 0.86 to 6.23 ng/g dry weight in sediments and from

Spatial and seasonal dynamics of phosphorous and physicochemical variables in the Negro River Estuary (Argentina): a preliminary approach.

Nutrient discharge into rivers and estuaries and the factors that control it need to be further understood to decrease the risk of harmful algae blooms on these ecosystems. Preliminary seasonal physicochemical parameters at six stations along the Negro River Estuary (Argentina) were studied during 2019 (Austral summer, winter, and spring) with high- and low-frequency data. Three of the stations were mainly estuarine-influenced and three were marine-influenced ones. The concentration of phosphate (P), river discharge, meteorological conditions, seasonality, and physicochemical variables were analyzed. Total phosphorus (TP) showed seasonal variations and was higher than previously reported for the upper watershed in all stations in the warmer months, except for the marine control one. Orthophosphate values were also high compared to previous watershed data and changed independently of TP fluctuations. Changing turbidity, water temperature, pH, and conductivity did not appear to have an essential role in phosphorus variations. An unexplained high TP spike in the late spring sample shows the need for further research in the area, while the seawater mixing with P-rich river water could be acting as a dilution agent at the mouth of the river.

Seasonal distribution pattern and bioaccumulation of Polycyclic aromatic hydrocarbons (PAHs) in four bioindicator coastal fishes of Argentina.

Polycyclic aromatic hydrocarbons (PAHs) are pollutants of global concern in coastal environments. They have a wide range of biological toxicity and due to their inherent properties, can easily bioaccumulate in organisms and concentrate in the environment. This work evaluated, in an integrated way, the seasonal PAH distribution patterns in sediments and four bioindicators fish species in a highly impacted estuary of Argentina; besides, their bioaccumulation patterns were assessed for the first time as indicator of ecological risk. The highest PAH levels in fish were found for Ramnogaster arcuata with an average of 64 ng g-1 w.w., followed by Micropogonias furnieri (45 ng g-1 w.w.), Cynoscion guatucupa (28 ng g-1 w.w.), and Mustelus schmitti (16 ng g-1 w.w.). Fish presented the highest PAH levels in fall with a predominance of petrogenic PAHs in colder seasons and pyrolytic PAHs in warmer seasons. Sediments presented an average of 233 ng g-1 d.w. with the same seasonal composition pattern of the fish tissues. Additionally, the data suggested that the main source of PAHs are wastewater discharges. The bioaccumulation factor (BAF) of PAHs in the tested fishes were found to range from 0.3 to 8. The highest values were observed during fall and winter, while bioaccumulation did not occur in moist spring and summer samples, which would suggest a high biotransformation process during these seasons. Results suggested that class III of juvenile C. guatucupa and M. furnieri, and adults R. arcuata are more sensitive bioindicators of chronic PAH contamination and that their bioaccumulation is independent of the compound hydrophobicity; this could have a positively influence on the criteria used for biological monitoring programs along the Atlantic coast. In addition, the presented BAF data on the target species will serve as a useful pollution indicator for South Atlantic coastal fish.

Plastic Impacts in Argentina: a Critical Research Review Contributing to the Global Knowledge.

PURPOSE OF REVIEW: Plastic pollution research has experienced exponential growth in the last decade; however, Argentina concentrates more than 70% of their research in the last 4 years. This review compiles regional research on plastic pollution in water, soil, sediment, air and organisms in Argentina. It discusses current sampling, quantification, and plastics identification methodologies while analyzing levels, gaps, and opportunities. RECENT FINDINGS: Research in plastic pollution was mainly focused on the biosphere component (52.9%), followed by the hydrosphere component (29.4%), and finally the lithosphere component (17.7%), with less than 20% addressing multiple components simultaneously. Sixty percent of this research was focused around microplastics, and less than 20% have considered multiple plastic debris sizes. Marine coastal species from Argentina had higher levels of microplastics than organisms from other South American studies, while microfibers were identified in 100% of the freshwater organisms studied. The lowest microplastic concentrations were found in lakes and in the Paraná and La Plata rivers, while the maximum concentrations were found in Pampa´s streams. There was a lack of standardization in methodology and unit expression in studies of sediment microplastics, which hinders comparison between reports. Argentine scientists have created the national alliance called SciEnce for Plastic Impacts Argentina (SEPIA). SEPIA is a network which aims to systemize plastic pollution research, coordinate methodologies, and enhance relationships with decision-makers, NGOs, and the general public. A time gap was found between the designation of principal international multilateral agreements and the implementation of national regulations for plastic waste treatment, with a tendency to include advanced concepts as Extended Producer Responsibility and Circular Economy.

Continental microplastics: Presence, features, and environmental transport pathways.

Microplastics (MPs) are ubiquitous contaminants of great concern for the environment. MPs' presence and concentration in the air, soil, marine, and freshwater environments have been reported as a matter of priority in recent years. This review addresses the current knowledge of the main pathways of MPs in air, soil, and freshwater reservoirs in order to provide an integrated understanding of their behaviors in the continental environment. Therefore, MPs' occurrence (as particle counts), sources, and how their features as shape, size, polymer composition, and density could influence their transport and final sink were discussed. Wind resuspension and atmospheric fallout, groundwater migration, runoff from catchments, and water flow from rivers and effluents were pointed as the principal pathways. MPs' size, shape, polymer composition, and density interact with environmental variables as soil structure and composition, precipitation, wind, relative humidity, water temperature, and salinity. Sampling designs for MPs research should further consider soil characteristics, climate variability and extreme events, time lag and grasshopper effects, morphological and hydrological features of aquatic systems, and water currents, among others. Furthermore, long-term monitoring and lab experiments are still needed to understand MPs' behavior in the environment. This information will provide a unified understanding of the continental MPs pathways, including the key main findings, knowledge gaps, and future challenges to understand this emerging contaminant.

Assessing threats, regulations, and strategies to abate plastic pollution in LAC beaches during COVID-19 pandemic.

Plastic use and production have dramatically increased globally over the past 65 years with the improvement of life quality by the daily use of plastic products. Still, around 50% of the plastic produced is disposable products that generate substantial waste. Several reports pointed out the adverse effects of plastic litter in coastal environments in recent years, emphasizing single-use plastics (SUP). In this manner, Latin America and the Caribbean (LAC) coastal environments are highly vulnerable due to wastewater treatment facilities scarcity and poor plastic waste management strategies. Since COVID-19 pandemic, the single-use plastic waste/person rate is expected to rise due to the use of personal protective equipment and SUP as health care measures. Based on literature research and the review of plastic waste regulations, this paper will assess the main COVID-19 plastic pollution threats and LAC beaches' regulations to suggest possible measures to abate this problem. The main findings suggest that unifying the ongoing fragmented and overlapped policies is key to abate plastic pollution, including plastic industry regulations and circular economies. In addition, increasing public risk perception about plastic pollution is critical to reducing plastic waste generation. Research advances in the adverse effects of plastic debris could improve the public's perception of plastic pollution risk, pushing forward global marine plastic governance.

First evidence of polycyclic aromatic hydrocarbons in sediments from a marine protected area within Argentinean Continental Shelf.

This study evaluated for the first time, the distribution, sources, and ecological risk assessment of 16 polycyclic aromatic hydrocarbons (PAHs) in sediments collected along the northern Patagonian shelf in the Argentine Sea -included in a Marine Protected Area (MPA). Total concentration of the PAHs varied from 19.47 to 183.17 ng/g dry weight and low molecular weight compounds, such as Anthracene and Naphthalene were the dominants. The selected PAHs ratios suggested a predominance of petrogenic sources and ecotoxicological risk evaluation based on the sediment quality guidelines model, indicated that some PAHs might cause occasional adverse biological effects in the study area.

Characterization of atmospheric and soil polycyclic aromatic hydrocarbons and evaluation of air-soil relationship in the Southwest of Buenos Aires province (Argentina).

Sixteen polycyclic aromatic hydrocarbons (PAHs) proposed by the US EPA as priority were analyzed in air and soil samples in the Southwest of Buenos Aires, Argentina, in order to study the levels, distribution, sources and fugacity ratios of PAHs, evaluating the relationship between them. For this, 10 passive air samplers (XAD-2® resin) were deployed along the area and replaced three-monthly from January to December 2015. PAHs were analyzed through gas chromatography -mass spectrometry (GC-MS). Results obtained showed that total PAHs levels (∑16) ranged from 27.97 to 1052.99 ng m-3 and from 52.40 to 2118.34 ng. g-1 d.w. for air and soil samples, respectively. The highest air- PAHs levels were registered in Bahía Blanca city (1052.99 ng. m-3, d.w.) an urban-industrial site, while the highest soil-PAHs levels were found in La Vitícola (2118.34 ng. g-1, d.w.), a rural location closed to a high traffic national route. For all sites the highest levels were observed during the winter; however, both spatial and temporal variations were only statistically significant for certain specific PAHs. Diagnostic ratios + PCA, determined dominance of pyrolytic sources. Further, data showed that source of PAHs could be attributed to vehicular and industrial emissions (observed in all periods), biomass combustion (linked mainly to warm period) and domestic emissions (linked mainly to cold period). Finally, fugacity ratios resulted <1, indicating that soil and air samples were not in equilibrium for the majority of PAHs determining a net tendency of air PAHs towards deposition while soil acted principally as a sink.

Evidence of Microplastic Ingestion by Fish from the Bahía Blanca Estuary in Argentina, South America.

The aim of this study was to assess - for the first time - the occurrence and distribution of microplastics (MPs) in the gastrointestinal tracts (GITs) of a marine commercial fish species (Micropogonias furnieri) from the Bahía Blanca Estuary (BBE) in Argentina, and to evaluate fish potential associated stress. In order to do this, juveniles were sampled using artisanal fishing arts at two sampling locations. Basic measurements of individual fish were taken (total length, total weight, weight of the liver) and GITs were subsequently removed, digested with 30% H2O2 for 5 days at 60°C, filtered on Whatman paper and then dried. Samples were observed with a stereomicroscope, and it was demonstrated that 100% of the individuals contained microplastic particles in their GITs. In total, 241 microplastic particles were removed from the GITs of all fish. They were categorized as fibers (60.8%), pellets (28.9%), fragments (8.6%) and laminas (1.4%), and they ranged in size from 0.98 to > 5 mm. The average number of particles per fish was higher than that reported in previous global marine studies. Moreover, a positive correlation between the number of MPs per fish and hepatosomatic index was found, suggesting a probable stress in their health condition. These findings provide the first and southernmost evidence of microplastic contamination in biota from the Argentinean sea, which is found in the South Atlantic sea.

In vitro effects of 1α,25(OH)2D3-glycosides from Solbone A (Solanum glaucophyllum leaves extract; Herbonis AG) compared to synthetic 1α,25(OH)2D3 on myogenesis.

The presence of glycoside derivatives of 1α,25(OH)2D3 endows plants to gradual release of the free bioactive form of 1α,25(OH)2D3 from its glycoconjugates by endogenous animal tissue glycosidases. This results in increased half-life of the hormone in blood when purified plant fractions are administered for therapeutic purposes. In this work, we evaluated the role 1α,25(OH)2D3-glycosides enriched natural product (Solbone A) from Solanum glaucophyllum leaf extract compared with synthetic 1α,25(OH)2D3 on myogenic differentiation in C2C12 myoblasts. For these, differentiation markers and myogenic parameters were studied in C2C12 myoblasts. Results showed that Solbone A, likewise the synthetic hormone, increased creatine kinase activity at day 2 after differentiation induction (60%, p<0.05). Solbone A and synthetic 1α,25(OH)2D3 increased vitamin D3 receptor protein expression at 10nM (50% and 30%, respectively) and the transcription factor myogenin (80%, p<0.05). However, tropomyosin expression was not affected by both compounds. In addition, myosin heavy chain (MHC) protein expression was increased 30% at day 2 of differentiation. Solbone A or synthetic 1α,25(OH)2D3 had no effects on myogenin nor MHC cell localization. Cellular mass increased with myogenesis progression, being Solbone A more effective than synthetic 1α,25(OH)2D3. Finally, Solbone A, as well as synthetic 1α,25(OH)2D3, augmented the index fusion of cultured muscle fibers. In conclusion, these results demonstrated that Solbone A exhibit at least equal or greater effects on early myoblast differentiation as synthetic hormone, suggesting that plant glycosides could be an effective, accessible and cheaper substitute for synthetic 1α,25(OH)2D3 to promote muscle growth.

Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells.

G-protein-coupled receptor 30 (GPR30) is an estrogen receptor that initiates several rapid, non-genomic signaling events triggered by E2. GPR30 has recently been identified in C2C12 cells; however, little is known about the intracelular distribution and its role in C2C12 myoblasts and myotubes. By western blotting and immunohistochemistry, we evidenced expression of GPR30. While in C2C12 myoblasts, the receptor was present in nucleus, mitochondria, and endoplasmic reticulum, in C2C12 myotubes, it was additionally found in cytoplasm. Using trypan blue uptake assay to determine cellular death and fluorescent microscopy to evaluate picnotic nuclei and mitochondrial distribution, we demonstated that treatment of C2C12 myoblasts with G1 (GPR30 agonist) did not protect the cells against apoptosis induced by H2O2 as E2. However, when G15 (GPR30 antagonist) was used, E2 could not prevent the damage caused by the oxidative stress. Further, some of the molecular mechanisms involved were investigated by wertern blot assays. Thus, E2 was able to induce AKT phosphorylation in apoptotic conditions and ERK phosphorylation in proliferating C2C12 cells but not when the cultures were incubated with G15. Additionally, using G15 antagonist we have found that GPR30 participates in the myogenin expression and creatine kinase activity stimulated by E2 in the first steps of C2C12 differentiation. Althogether these findings provide evidences showing that GPR30 is expressed in diverse intracellular compartments in undifferentiated and differentiated C2C12 cells and mediates E2 actions.

17ß-estradiol protects mitochondrial functions through extracellular-signal-regulated kinase in C2C12 muscle cells.

BACKGROUND/AIMS: We have previously shown that exposure to 17ß-estradiol (E2) prior to induction of apoptosis with H2O2 protects skeletal muscle cells against oxidative damage. However, the mechanism involved in the protective action of the hormone is poorly understood. In the present study, we focused on the mechanism by which ERK mediates this survival effect in connection with COXIV activity and mitochondrial membrane potential. METHODS: Immunocytochemistry, Western blot, cytochrome c oxidase complex IV (COXIV) activity, coimmunoprecipitation and JC-1 dye by flow cytometry were carried out using C2C12 myoblasts as experimental model. RESULTS: E2 is able to activate ERK and then induces its translocation to mitochondria. Using the pharmacological inhibitor of ERK activation U0126 we show that E2, through ERK activation, is able to enhance COXIV activity. Moreover, the hormone increases the interaction between COXIV and ERK. Also, we found that hydrogen peroxide decreases COXIV activity and that preincubation of the cells with E2 prior to induction of apoptosis prevents this effect. In addition, we observe that the estrogen inhibits the collapse of mitochondrial membrane potential induced by H2O2, involving ERK and COXIV. CONCLUSION: Our data demonstrate that E2 promotes ERK activation and translocation to mitochondria preventing the decline in COXIV activity and in turn, alteration of mitochondrial membrane potential by oxidative stress, in C2C12 myoblasts.

Resorption controls bone anabolism driven by parathyroid hormone (PTH) receptor signaling in osteocytes.

The contribution of remodeling-based bone formation coupled to osteoclast activity versus modeling-based bone formation that occurs independently of resorption, to the anabolic effect of PTH remains unclear. We addressed this question using transgenic mice with activated PTH receptor signaling in osteocytes that exhibit increased bone mass and remodeling, recognized skeletal effects of PTH elevation. Direct inhibition of bone formation was accomplished genetically by overexpressing the Wnt antagonist Sost/sclerostin; and resorption-dependent bone formation was inhibited pharmacologically with the bisphosphonate alendronate. We found that bone formation induced by osteocytic PTH receptor signaling on the periosteal surface depends on Wnt signaling but not on resorption. In contrast, bone formation on the endocortical surface results from a combination of Wnt-driven increased osteoblast number and resorption-dependent osteoblast activity. Moreover, elevated osteoclasts and intracortical/calvarial porosity is exacerbated by overexpressing Sost and reversed by blocking resorption. Furthermore, increased cancellous bone is abolished by Wnt inhibition but further increased by blocking resorption. Thus, resorption induced by PTH receptor signaling in osteocytes is critical for full anabolism in cortical bone, but tempers bone gain in cancellous bone. Dissecting underlying mechanisms of PTH receptor signaling would allow targeting actions in different bone compartments, enhancing the therapeutic potential of the pathway.

PTH receptor signaling in osteocytes governs periosteal bone formation and intracortical remodeling.

The periosteal and endocortical surfaces of cortical bone dictate the geometry and overall mechanical properties of bone. Yet the cellular and molecular mechanisms that regulate activity on these surfaces are far from being understood. Parathyroid hormone (PTH) has profound effects in cortical bone, stimulating periosteal expansion and at the same time accelerating intracortical bone remodeling. We report herein that transgenic mice expressing a constitutive active PTH receptor in osteocytes (DMP1-caPTHR1 mice) exhibit increased cortical bone area and an elevated rate of periosteal and endocortical bone formation. In addition, DMP1-caPTHR1 mice display a marked increase in intracortical remodeling and cortical porosity. Crossing DMP1-caPTHR1 mice with mice lacking the Wnt coreceptor, LDL-related receptor 5 (LRP5), or with mice overexpressing the Wnt antagonist Sost in osteocytes (DMP1-Sost mice) reduced or abolished, respectively, the increased cortical bone area, periosteal bone formation rate, and expression of osteoblast markers and Wnt target genes exhibited by the DMP1-caPTHR1 mice. In addition, DMP1-caPTHR1 lacking LRP5 or double transgenic DMP1-caPTHR1;DMP1-Sost mice exhibit exacerbated intracortical remodeling and increased osteoclast numbers, and markedly decreased expression of the RANK decoy receptor osteoprotegerin. Thus, whereas Sost downregulation and the consequent Wnt activation is required for the stimulatory effect of PTH receptor signaling on periosteal bone formation, the Wnt-independent increase in osteoclastogenesis induced by PTH receptor activation in osteocytes overrides the effect on Sost. These findings demonstrate that PTH receptor signaling influences cortical bone through actions on osteocytes and defines the role of Wnt signaling in PTH receptor action.