Comprehensive Behavioral Testing in the R6/2 Mouse Model of Huntington's Disease Shows No Benefit from CoQ10 or Minocycline

Previous studies of the effects of coenzyme Q10 and minocycline on mouse models of Huntington''s disease have produced conflicting results regarding their efficacy in behavioral tests. Using our recently published best practices for husbandry and testing for mouse models of Huntington''s disease, we report that neither coenzyme Q10 nor minocycline had significant beneficial effects on measures of motor function, general health (open field, rotarod, grip strength, rearing-climbing, body weight and survival) in the R6/2 mouse model. The higher doses of minocycline, on the contrary, reduced survival. We were thus unable to confirm the previously reported benefits for these two drugs, and we discuss potential reasons for these discrepancies, such as the effects of husbandry and nutrition.     

Evolution of thermotolerance in seasonal environments: the effects of annual temperature variation and life-history timing in Wyeomyia smithii

In organisms with complex life cycles, the adaptive value of thermotolerance depends on life-history timing and seasonal temperature profiles. We illustrate this concept by examining variation in annual thermal environments and thermal acclimation among four geographic populations of the pitcher plant mosquito. Only diapausing larvae experience winter, whereas both postdiapause and nondiapause adults occur only during the growing season. Thus, adults experience transient cold stress primarily during the spring. We show that adult cold tolerance (chill coma recovery) is enhanced in spring-like conditions via thermal acclimation but is unaffected by diapause state. Moreover, adult mosquitoes from northern populations were more cold tolerant than those from southern populations largely because acclimation responses were steeper in the north. In contrast to cold tolerance, there was no significant acclimation of heat tolerance (heat knockdown), and no significant differences in heat tolerance between northern and southern populations. Field temperature data show that because of evolved differences in diapause timing, adult exposure to cold stress is remarkably consistent across geography. This suggests that geographic variation in cold tolerance may not be the result of direct selection on adults. Our results illustrate the importance of the interplay between phenological and thermal adaptation for understanding variation along climatic gradients.     

Reiterated Wnt signaling during zebrafish neural crest development

While Wnt/beta-catenin signaling is known to be involved in the development of neural crest cells in zebrafish, it is unclear which Wnts are involved, and when they are required. To address these issues we employed a zebrafish line that was transgenic for an inducible inhibitor of Wnt/beta-catenin signaling, and inhibited endogenous Wnt/beta-catenin signaling at discrete times in development. Using this approach, we defined a critical period for Wnt signaling in the initial induction of neural crest, which is distinct from the later period of development when pigment cells are specified from neural crest. Blocking Wnt signaling during this early period interfered with neural crest formation without blocking development of dorsal spinal neurons. Transplantation experiments suggest that neural crest precursors must directly transduce a Wnt signal. With regard to identifying which endogenous Wnt is responsible for this initial critical period, we established that wnt8 is expressed in the appropriate time and place to participate in this process. Supporting a role for Wnt8, blocking its function with antisense morpholino oligonucleotides eliminates initial expression of neural crest markers. Taken together, these results demonstrate that Wnt signals are critical for the initial induction of zebrafish neural crest and suggest that this signaling pathway plays reiterated roles in its development.     

Feldspar Tunneling by Fungi along Natural Productivity Gradients

Recently, it was hypothesized that ectomycorrhizas hyphae are involved in mineral tunneling. We evaluated the role of ectomycorrhizas in mineral weathering and the ecosystem influx of basic cations by correlating mineral tunnel density to ectomycorrhizas density in two forest productivity gradients. The gradients, two gentle slopes in northern Sweden, are the result of groundwater movement and are characterized by reduced productivity upslope due to lower nitrogen availability. As expected, ectomycorrhizas density in the O horizon was higher upslope, where nitrogen availability was lower and where the vegetation was dominated by ectomycorrhizas plant species. We consistently found that tunnel formation in mineral grains was more intense in nutrient-poor sites, indicating a higher contribution of fungi to ecosystem influx of potassium and calcium. ectomycorrhizas density was positively correlated with feldspar tunnel density in the upper 2 cm of the E horizon. This suggests that ectomycorrhizas are involved in mineral tunneling. We discuss the possible involvement of ericoid mycorrhizas and saprotrophic fungi in feldspar tunneling and the role of the weathering status of mineral grains as additional factors controlling mineral funneling.     

Avian model for 13-cis-retinoic acid embryopathy: demonstration of neural crest related defects

The effects of 13-cis-retinoic acid on the developing chick embryo were investigated. Fertilized eggs were injected via the yolk sac with single 50 microliters doses of either 1.5 micrograms, 15 micrograms, or 150 micrograms of 13-cis-retinoic acid in dimethyl sulfoxide on varying days of incubation (embryonic days 2, 3, 4, 5, or 6). Control embryos were given solvent alone or a mock injection. The embryos were examined on day 14 of incubation. The effects of retinoic acid on mortality and total malformations were both dose and developmental-stage responsive. The defects caused by 13-cis-retinoic acid occurred in mesenchymal tissues derived in part from the cranial neural crest ectomesenchyme. The craniofacial and cardiovascular malformations produced in the chick are analogous to those seen in animal models of retinoid teratogenesis and in human fetuses exposed to 13-cis-retinoic acid during maternal therapy for cystic acne. Following 13-cis-retinoic acid treatment, craniofacial and specific cardiovascular malformations were increased significantly compared to those in matched solvent and mock treated controls. The greatest number of malformations occurred when 13-cis-retinoic acid was given after cranial neural crest cell migration was complete. We propose that the primary effect of 13-cis-retinoic acid is on region-specific localization and differentiation of the mesenchymal subpopulation of cranial neural crest cells.     

Salivaricin A2 and the novel lantibiotic salivaricin B are encoded at adjacent loci on a 190-kilobase transmissible megaplasmid in the oral probiotic strain Streptococcus salivarius K12

The commercial probiotic Streptococcus salivarius strain K12 is the prototype of those S. salivarius strains that are the most strongly inhibitory in a standardized test of streptococcal bacteriocin production and has been shown to produce the 2,368-Da salivaricin A2 (SalA2) and the 2,740-Da salivaricin B (SboB) lantibiotics. The previously uncharacterized SboB belongs to the type AII class of lantibiotic bacteriocins and is encoded by an eight-gene cluster. The genetic loci encoding SalA2 and SboB in strain K12 have been fully characterized and are localized to nearly adjacent sites on pSsal-K12, a 190-kb megaplasmid. Of 61 strongly inhibitory strains of S. salivarius, 19 (31%) were positive for the sboB structural gene. All but one (strain NR) of these 19 strains were also positive for salA2, and in each of these cases of double positivity, the two loci were separated by fewer than 10 kb. This is the first report of a single streptococcus strain producing two distinct lantibiotics.     

Influence of seasonal timing on thermal ecology and thermal reaction norm evolution in Wyeomyia smithii

Evolutionary changes in the seasonal timing of life-history events can alter a population's exposure to seasonally variable environmental factors. We illustrate this principle in Wyeomyia smithii by showing that: (1) geographic divergence in diapause timing reduces differences among populations in the thermal habitat experienced by nondiapause stages; and (2) the thermal habitat of the growing season is more divergent at high compared with low temperatures with respect to daily mean temperatures. Geographic variation in thermal reaction norms for development time was greater in a warm compared with a cool rearing treatment, mirroring the geographic trend in daily mean temperature. Geographic variation in body size was unrelated to geographic temperature variation, but was also unrelated to development time or fecundity. Our results suggest that proper interpretation of geographic trends may often require detailed knowledge of life-history timing.     

Two lytic transglycosylases in Neisseria gonorrhoeae impart resistance to killing by lysozyme and human neutrophils

Symptomatic infection by Neisseria gonorrhoeae (Gc) produces a potent inflammatory response, resulting in a neutrophil‐rich exudate. A population of Gc can survive the killing activities of neutrophils for reasons not completely understood. Unlike other Gram‐negative bacteria, Gc releases monomeric peptidoglycan (PG) extracellularly, dependent on two nonessential, nonredundant lytic transglycosylases (LTs), LtgA and LtgD. PG released by LtgA and LtgD can stimulate host immune responses. We report that ΔltgAΔltgD Gc were decreased in survival in the presence of primary human neutrophils but otherwise grew equally to wild‐type Gc. Adding PG monomer failed to alter ΔltgAΔltgD Gc survival. Thus, LTs protect Gc from neutrophils independently of monomer release. We found two reasons to explain decreased survival of the double LT mutant. First, ΔltgAΔltgD Gc was more sensitive to the neutrophil antimicrobial proteins lysozyme and neutrophil elastase, but not others. Sensitivity to lysozyme correlated with decreased Gc envelope integrity. Second, exposure of neutrophils to ΔltgAΔltgD Gc increased the release of neutrophil granule contents extracellularly and into Gc phagosomes. We conclude that LtgA and LtgD protect Gc from neutrophils by contributing to envelope integrity and limiting bacterial exposure to select granule‐localized antimicrobial proteins. These observations are the first to link bacterial degradation by lysozyme to increased neutrophil activation.