Effects of pedunculopontine nucleus cholinergic lesion on gait and dyskinesia in hemiparkinsonian rats.

Pedunculopontine nucleus (PPN) cholinergic neurons are implicated in freezing of gait in Parkinson's disease (PD) and motor stereotypy in normal animals, but the causal role of these neurons on specific gait parameters and treatment-induced dyskinesia remains speculative. Therefore, we examined whether selective cholinergic lesion of the rostral PPN affects PD motor and gait deficits, L-DOPA-induced dyskinesia and motor improvement, and DA-agonist-induced dyskinesia. Sprague-Dawley rats were assigned to one unilaterally lesioned group: Sham lesion, PPN cholinergic lesion with diphtheria urotensin II fusion toxin, medial forebrain bundle dopamine lesion with 6-hydroxydopamine, or dual acetylcholine and dopamine lesion. We used gait analysis and forepaw adjusting steps to examine PD gait and motor deficits. Forepaw adjusting steps were also used to assess motor improvement with L-DOPA treatment. The abnormal involuntary movements scale measured L-DOPA and dopamine D1- and D2-receptor agonist-induced dyskinesia. Lesions, verified via tyrosine hydroxylase and choline acetyltransferase immunohistochemistry reduced an average of 95% of nigral dopamine neurons and 80% of PPN cholinergic neurons, respectively. Rats receiving acetylcholine and dual lesion demonstrated enhanced freezing, and acetylcholine lesioned rats exhibited increased print area and stand index. Dopamine and dual lesion produced similar forepaw adjusting steps task on and off L-DOPA. Relative to DA lesioned rats, dual lesioned rats displayed reduced L-DOPA and DA agonist-induced dyskinesia at specific time points. Our results indicate that PPN cholinergic neurons affect gait parameters related to postural stability. Therefore, therapeutically targeting PPN cholinergic neurons could reduce intractable postural instability in PD without affecting motor benefits or side effects of L-DOPA treatment.

Sex-dependent effects of preconception exposure to arsenite on gene transcription in parental germ cells and on transcriptomic profiles and diabetic phenotype of offspring.

Chronic exposure to inorganic arsenic (iAs) has been linked to diabetic phenotypes in both humans and mice. However, diabetogenic effects of iAs exposure during specific developmental windows have never been systematically studied. We have previously shown that in mice, combined preconception and in utero exposures to iAs resulted in impaired glucose homeostasis in male offspring. The goal of the present study was to determine if preconception exposure alone can contribute to this outcome. We have examined metabolic phenotypes in male and female offspring from dams and sires that were exposed to iAs in drinking water (0 or 200 µg As/L) for 10 weeks prior to mating. The effects of iAs exposure on gene expression profiles in parental germ cells, and pancreatic islets and livers from offspring were assessed using RNA sequencing. We found that iAs exposure significantly altered transcript levels of genes, including diabetes-related genes, in the sperm of sires. Notably, some of the same gene transcripts and the associated pathways were also altered in the liver of the offspring. The exposure had a more subtle effect on gene expression in maternal oocytes and in pancreatic islets of the offspring. In female offspring, the preconception exposure was associated with increased adiposity, but lower blood glucose after fasting and after glucose challenge. HOMA-IR, the indicator of insulin resistance, was also lower. In contrast, the preconception exposure had no effects on blood glucose measures in male offspring. However, males from parents exposed to iAs had higher plasma insulin after glucose challenge and higher insulinogenic index than control offspring, indicating a greater requirement for insulin to maintain glucose homeostasis. Our results suggest that preconception exposure may contribute to the development of diabetic phenotype in male offspring, possibly mediated through germ cell-associated inheritance. Future research can investigate role of epigenetics in this phenomenon. The paradoxical outcomes in female offspring, suggesting a protective effect of the preconception exposure, warrant further investigation.

Evaluating the dynamics of groundwater, lakebed transport, nutrient inflow and algal blooms in Upper Klamath Lake, Oregon, USA.

Transport of nutrients to lakes can occur via surface-water inflow, atmospheric deposition, groundwater (GW) inflow and benthic processes. Identifying and quantifying within-lake nutrient sources and recycling processes is challenging. Prior studies in hypereutrophic Upper Klamath Lake, Oregon, USA, indicated that ~60% of the early summer phosphorus (P) load to the lake was internal and hypothesized to be lakebed sediment release. Dynamic nutrient transport processes were examined to better characterize the nutrient sources. One-dimensional heat transport models calibrated to observed lakebed temperatures and a cross-sectional GW flow model provided estimates of GW-inflow rates that were greatest in spring and decreased through summer. One-dimensional solute transport models calibrated to observed lakebed pore-water dissolved silica (Si) and dissolved phosphate-phosphorus (DP) concentrations indicated that nutrients were transported from the lakebed by advection, diffusion, and enhanced mixing by benthic organisms and waves, and that DP removal occurred near the lakebed interface. Estimated water, Si, DP and total-phosphorus (TP) budgets indicated that GW contributed 21% of lake water inflow and at least 26, 20 and 16% of total Si, DP and TP inflow, respectively, when conservatively assuming background GW nutrient concentrations. However, lakebed GW (LGW) is enriched in nutrients during flow through lakebed sediment and the estimated GW contribution increased to 29 (33), 49 (67) and 43% (61%) of total Si, DP and TP inflow, respectively, if 20% (50%) of GW inflow to the lake was assumed to have LGW concentrations. Net nutrient inflow to the lake was greatest in spring and coincident with the annual diatom bloom. Inflowing dissolved nutrients appear to be assimilated by diatoms during the spring and become available for the summer Aphanizomenon flos-aquae bloom when the diatoms senesce. Thus, nutrient-enriched GW inflow and nutrient recycling by successive algal blooms must be considered when evaluating internal nutrient loading to lakes.

Unintended consequences of management actions in salt pond restoration: cascading effects in trophic interactions.

Salt evaporation ponds have played an important role as habitat for migratory waterbirds across the world, however, efforts to restore and manage these habitats to maximize their conservation value has proven to be challenging. For example, salinity reduction has been a goal for restoring and managing former salt evaporation ponds to support waterbirds in the South Bay Salt Pond Restoration Project in San Francisco Bay, California, USA. Here, we describe a case study of unexpected consequences of a low-dissolved oxygen (DO) event on trophic interactions in a salt pond system following management actions to reduce salinity concentrations. We document the ramifications of an anoxic event in water quality including salinity, DO, and temperature, and in the response of the biota including prey fish biomass, numerical response by California Gulls (Larus californicus), and chick survival of Forster's Tern (Sterna forsteri). Management actions intended to protect receiving waters resulted in decreased DO concentrations that collapsed to zero for ≥ 4 consecutive days, resulting in an extensive fish kill. DO depletion likely resulted from an algal bloom that arose following transition of the pond system from high to low salinity as respiration and decomposition outpaced photosynthetic production. We measured a ≥ 6-fold increase in biomass of fish dropped on the levee by foraging avian predators compared with weeks prior to and following the low-DO event. California Gulls rapidly responded to the availability of aerobically-stressed and vulnerable fish and increased in abundance by two orders of magnitude. Mark-recapture analysis of 254 Forster's Tern chicks indicated that their survival declined substantially following the increase in gull abundance. Thus, management actions to reduce salinity concentrations resulted in cascading effects in trophic interactions that serves as a cautionary tale illustrating the importance of understanding the interaction of water quality and trophic structure when managing restoration of salt ponds.

Changes in benthic nutrient sources within a wetland after hydrologic reconnection.

Removing dams and levees to restore hydrologic connectivity and enhance ecosystem services such as nutrient removal has been an increasingly common management practice. In the present study, the authors assessed geochemical and biological changes following engineered levee breaches that reconnected eutrophic Upper Klamath Lake and Agency Lake, Oregon, USA, to an adjacent, historic wetland that had been under agricultural use for the last seven decades. Over the three-year study, the reconnected wetland served as a benthic source for both macronutrients (dissolved organic carbon [DOC], soluble reactive phosphorus [SRP], and ammonia) and micronutrients (dissolved iron and manganese). The magnitude of those benthic sources was similar to or greater than that of allochthonous sources. The highest DOC benthic flux to the water column occurred immediately after rewetting occurred. It then decreased during the present study to levels more similar to the adjacent lake. Dissolved ammonia fluxes, initially negative after the levee breaches, became consistently positive through the remainder of the study. Nitrate fluxes, also initially negative, became negligible two years after the levee breaches. In contrast to previous laboratory studies, SRP fluxes remained positive, as did fluxes of dissolved iron and manganese. Our results indicate that the timescales of chemical changes following hydrologic reconnection of wetlands are solute-specific and in some cases extend for multiple years beyond the reconnection event. During the present study, colonization of the reconnected wetlands by aquatic benthic invertebrates gradually generated assemblages similar to those in a nearby wetland refuge and provided further evidence of the multiyear transition of this area to permanent aquatic habitat. Such timescales should be considered when developing water-quality management strategies to achieve wetland-restoration goals.

Benthic nutrient sources to hypereutrophic upper Klamath Lake, Oregon, USA.

Three collecting trips were coordinated in April, May, and August 2006 to sample the water column and benthos of hypereutrophic Upper Klamath Lake (OR, USA) through the annual cyanophyte bloom of Aphanizomenon flos-aquae. A pore-water profiler was designed and fabricated to obtain the first high-resolution (centimeter-scale) estimates of the vertical concentration gradients of macro- and micronutrients for diffusive-flux determinations. A consistently positive benthic flux for soluble reactive phosphorus (SRP) was observed with solute release from the sediment, ranging between 0.4 and 6.1 mg/m(2)/d. The mass flux over an approximate 200-km(2) lake area was comparable in magnitude to riverine inputs. An additional concern related to fish toxicity was identified when dissolved ammonium also displayed consistently positive benthic fluxes of 4 to 134 mg/m(2)/d, again comparable to riverine inputs. Although phosphorus was a logical initial choice by water quality managers for the limiting nutrient when nitrogen-fixing cyanophytes dominate, initial trace-element results from the lake and major inflowing tributaries suggested that the role of iron limitation on primary productivity should be investigated. Dissolved iron became depleted in the lake water column during the course of the algal bloom, while dissolved ammonium and SRP increased. Elevated macroinvertebrate densities, at least of the order of 10(4) individuals/m(2), suggested that the diffusive-flux estimates may be significantly enhanced by bioturbation. In addition, heat-flux modeling indicated that groundwater advection of nutrients could also significantly contribute to internal nutrient loading. Accurate environmental assessments of lentic systems and reasonable expectations for point-source management require quantitative consideration of internal solute sources.

Mercury speciation in piscivorous fish from mining-impacted reservoirs.

Guadalupe Reservoir (GUA), California, and Lahontan Reservoir (LAH), Nevada, U.S. are both affected either directly or indirectly by the legacy of gold and silver mining in the Sierra Nevada during the nineteenth century. Analysis of total mercury in fish from these lentic systems consistently indicate elevated concentrations (>1 microg x g(-1) wet weight; hereinafter, all concentrations are reported as wet weight unless indicated otherwise) well above the U.S. Environmenal Protection Agency's human consumption advisory level for fish (<0.3 microg x g(-1)). Replicate X-ray absorption near edge structure (XANES) analyses on largemouth bass and hybrid striped bass from GUA and LAH were performed to determine predominant chemical species of mercury accumulated by these high-trophic-level piscivores that are exposed to elevated mercury through trophic transfer in mining-impacted lentic systems. Despite distinct differences in mercury source, the proximity of the source, and concentrations of complexing ligands, results of XANES analysis clearly indicated that mercury accumulated in these individual fish from the two reservoirs were dominated by methylmercury cysteine complexes. These findings are consistent with results from commercial fish species inhabiting marine environments which are presumed to include differing mercury sources (e.g., atmospheric, hydrothermal, or benthic). The dominance of methylmercury cysteine complexes in muscle tissues of fish obtained from such contrasting environments and exposure conditions suggests that a generic toxicological model for the consumption of fish could be applicable over a wide range of ecologic settings.

Free zinc ion and dissolved orthophosphate effects on phytoplankton from Coeur d'Alene Lake, Idaho.

Coeur d'Alene Lake in northern Idaho is fed by two major rivers: the Coeur d'Alene River from the east and the St. Joe River from the south, with the Spokane River as its outlet to the north. This phosphorus-limited lake has been subjected to decades of mining (primarily for zinc and silver) and other anthropogenic inputs. A 32 full-factorial experimental design was used to examine the interactive effects of free (uncomplexed) zinc ion and dissolved-orthophosphate concentrations on phytoplankton that were isolated from two sites along a longitudinal zinc-concentration gradient in Coeur d'Alene Lake. The two sites displayed different dominanttaxa. Chlorella minutissima, a dominant species near the southern St. Joe River inlet, exhibited greater sensitivity to free Zn ions than Asterionella formosa, collected nearer the Coeur d'Alene River mouth with elevated dissolved-zinc concentrations. Empirical phytoplankton-response models were generated to describe phytoplankton growth in response to remediation strategies in the surrounding watershed. If dissolved Zn can be reduced in the water column from >500 nM (i.e., current concentrations near and down stream of the Coeur d'Alene River plume) to <3 nM (i.e., concentrations near the southern St. Joe River inlet) such that the lake is truly phosphorus limited, management of phosphorus inputs by surrounding communities will ultimately determine the limnologic state of the lake.

Stable metal isotopes reveal copper accumulation and loss dynamics in the freshwater bivalve Corbicula.

Characterization of uptake and loss dynamics is critical to understanding risks associated with contaminant exposure in aquatic animals. Dynamics are especially important in addressing questions such as why coexisting species in nature accumulate different levels of a contaminant. Here we manipulated copper (Cu) stable isotopic ratios (as an alternative to radioisotopes) to describe for the first time Cu dynamics in a freshwater invertebrate, the bivalve Corbicula fluminea. In the laboratory, Corbicula uptake and loss rate constants were determined from an environmentally realistic waterborne exposure to 65Cu (5.7 microg L(-1)). That is, we spiked deionized water with Cu that was 99.4% 65Cu. Net tracer uptake was detectable after 1 day and strongly evident after 4 days. Thus, short-term exposures necessary to determine uptake dynamics are feasible with stable isotopes of Cu. In Corbicula, 65Cu depuration was biphasic. An unusually low rate constant of loss (0.0038 d(-1)) characterized the slow component of efflux, explaining why Corbicula strongly accumulates copper in nature. We incorporated our estimates of rate constants for dissolved 65Cu uptake and physiological efflux into a bioaccumulation model and showed that dietary exposure to Cu is likely an important bioaccumulation pathway for Corbicula.

Importance of sediment-water interactions in Coeur d'Alene Lake, Idaho, USA: management implications.

A field study at Coeur d'Alene Lake, Idaho, USA, was conducted between October 1998 and August 2001 to examine the potential importance of sediment-water interactions on contaminant transport and to provide the first direct measurements of the benthic flux of dissolved solutes of environmental concern in this lake. Because of potential ecological effects, dissolved zinc and orthophosphate were the solutes of primary interest. Results from deployments of an in situ flux chamber indicated that benthic fluxes of dissolved Zn and orthophosphate were comparable in magnitude to riverine inputs. Tracer analyses and benthic-community metrics provided evidence that solute benthic flux were diffusion-controlled at the flux-chamber deployment sites. That is, effects of biomixing (or bioturbation) and ground-water interactions did not strongly influence benthic flux. Remediation efforts in the river might not produce desired water-quality effects in the lake because imposed shifts in concentration gradients near the sediment-water interface would generate a benthic feedback response. Therefore, development of water-quality models to justify remediation strategies requires consideration of contaminant flux between the water column and underlying sediment in basins that have been affected by long-term (decadal) anthropogenic activities.