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WSSV particles were detected in separated granular cells (GCs) and semigranular cells (SGCs) by in situ hybridisation from WSSV-infected crayfish and the prevalence of WSSV-infected GCs was 5%, whereas it was 22% in SGCs. This indicates that SGCs are more susceptible to WSSV and that this virus replicated more rapidly in SGCs than in GCs and as a result the number of SGCs gradually decreased from the blood circulation. The effect of haemocyte lysate supernatant (HLS), containing the degranulation factor (peroxinectin), phorbol 12-myristate 13-acetate (PMA), the Ca(2+) ionophore A23187 on GCs from WSSV-infected and sham-injected crayfish was studied. The results showed that the percentage of degranulated GCs of WSSV-infected crayfish treated with HLS or PMA was significantly lower than that in the control, whereas no significant difference was observed when treated with the Ca(2+) ionophore. It was previously shown that peroxinectin and PMA have a degranulation effect via intracellular signalling involving protein kinase C (PKC), whereas the Ca(2+) ionophore uses an alternative pathway. HLS treatment of GCs and SGCs from WSSV-infected crayfish results in three different morphological types: non-spread, spread and degranulated cells. The non-spread cell group from both GCs and SGCs after treatment with HLS had more WSSV positive cells than degranulated cells, when detected by in situ hybridisation. Taken together, it is reasonable to speculate that the PKC pathway might be affected during WSSV infection. Another interesting phenomenon was that GCs from non-infected crayfish exhibited melanisation, when incubated in L-15 medium, while no melanisation was found in GCs of WSSV-infected crayfish. However, the phenoloxidase activities of both sham- and WSSV-injected crayfish in HLS were the same as well as proPO expression as detected by RT-PCR. This suggests that the WSSV inhibits the proPO system upstream of phenoloxidase or simply consumes the native substrate for the enzyme so that no activity is shown. The percentage of apoptotic haemocytes in WSSV-infected crayfish was very low, but it was significantly higher than that in the sham-injected crayfish on day 3 or 5 post-infection. The TEM observation in haematopoietic cells (hpt cells) suggests that WSSV infect specific cell types in haematopoietic tissue and non-granular hpt cells seem more favourable to WSSV infection.  相似文献   
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Background

Mitochondrial biogenesis is crucial for myogenic differentiation and regeneration of skeletal muscle tissue and is tightly controlled by the peroxisome proliferator-activated receptor-γ co-activator 1 (PGC-1) signaling network. In the present study, we hypothesized that inactivation of glycogen synthase kinase (GSK)-3β, previously suggested to interfere with PGC-1 in non-muscle cells, potentiates PGC-1 signaling and the development of mitochondrial biogenesis during myogenesis, ultimately resulting in an enhanced myotube oxidative capacity.

Methods

GSK-3β was inactivated genetically or pharmacologically during myogenic differentiation of C2C12 muscle cells. In addition, m. gastrocnemius tissue was collected from wild-type and muscle-specific GSK-3β knock-out (KO) mice at different time-points during the reloading/regeneration phase following a 14-day hind-limb suspension period. Subsequently, expression levels of constituents of the PGC-1 signaling network as well as key parameters of mitochondrial oxidative metabolism were investigated.

Results

In vitro, both knock-down as well as pharmacological inhibition of GSK-3β not only increased expression levels of important constituents of the PGC-1 signaling network, but also potentiated myogenic differentiation-associated increases in mitochondrial respiration, mitochondrial DNA copy number, oxidative phosphorylation (OXPHOS) protein abundance and the activity of key enzymes involved in the Krebs cycle and fatty acid β-oxidation. In addition, GSK-3β KO animals showed augmented reloading-induced increases in skeletal muscle gene expression of constituents of the PGC-1 signaling network as well as sub-units of OXPHOS complexes compared to wild-type animals.

Conclusion

Inactivation of GSK-3β stimulates activation of PGC-1 signaling and mitochondrial biogenesis during myogenic differentiation and reloading of the skeletal musculature.  相似文献   
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Agri-environment schemes (AES) are increasingly being employed to mitigate biodiversity loss in agricultural environments. The European Turtle Dove Streptopelia turtur is an obligate granivorous bird in rapid decline within both the UK (−96% since 1970) and across continental Europe (−77% since 1980), despite widespread uptake of AES. Here, we assess the efficacy of a potentially new, sown agri-environment option designed to provide abundant, accessible seed for S. turtur during the breeding season. During summer 2011 we compared vegetation structure and seed provision on trial plots to control habitat types (existing agri-environment options thought to potentially provide S. turtur foraging habitat) to assess whether trial plots performed better for foraging S. turtur than control habitats. In September 2011 all trial plots were topped (cut) and half of a subset of trial plots were then scarified (60% of soil surface disturbed). Vegetation structure on topped, and topped and scarified trial plots was measured during summer 2012 to determine which management regime was most effective in maintaining suitable sward structure and seed provision into the second year. No control habitat type produced as much seed important in S. turtur diet as trial plots at any point during year one. Trial plots provided accessible vegetation structure early in the season with no difference in vegetation metrics between trial plots and previously published data on S. turtur foraging locations. However, to allow later access, management is required during mid-June to open up the sward through localized topping or scarification. Vegetation structure during year two was generally too dense to attract foraging S. turtur. However, scarifying trial plots during the September following sowing encouraged self-seeding of Fumaria. officinalis (a plant species historically forming a significant proportion of S. turtur diet during the breeding season) into the second year, with this species present in 16% of scarified trial plots compared to only 4% of topped trial plots during year two. Thus, autumn scarification, possibly followed by topping or scarification of part of the trial plots in June, is necessary for trial plots to provide more seed and access for S. turtur than existing agri-environment options during year two. We recommend modifications to our original seed mix in order to reduce vegetation density and improve vegetation structure. The study provides an example of the need to strike the right balance between food abundance and accessibility, through vegetation structure, when designing agri-environment scheme management options that provide food for birds.  相似文献   
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