Indomethacin protects rats from neuronal damage induced by traumatic brain injury and suppresses hippocampal IL-1β release through the inhibition of Nogo-A expression

BACKGROUND: Nogo-A is a member of the reticulon family of membrane-associated proteins and plays an important role in axonal remodeling. The present study aimed to investigate alterations in Nogo-A expression following traumatic brain injury (TBI)-induced inflammation and neuronal damage. METHODS: A weight-drop device was used to deliver a standard traumatic impact to rats. Western blot, RT-PCR and ELISA were used to analyze the expression of Nogo-A and IL-1β. Nogo-A antisense, and an irrelevant control oligonucleotide was intracerebroventricularly infused. We also performed H & E staining and luxol fast blue staining to evaluate the neuronal damage and demyelination resulting from TBI and various treatments. RESULTS: Based on RT-PCR and western blot analyses, the expression of Nogo-A was found to be significantly upregulated in the hippocampus beginning eight hours after TBI. In addition, TBI caused an apparent elevation in IL-1β levels and severe neuronal damage and demyelination in the tested animals. All of the TBI-associated molecular and cellular consequences could be effectively reversed by treating the animals with the anti-inflammatory drug indomethacin. More importantly, the TBI-associated stimulation in the levels of both Nogo-A and IL-1β could be effectively inhibited by a specific Nogo-A antisense oligonucleotide. CONCLUSIONS: Our findings suggest that the suppression of Nogo-A expression appears to be an early response conferred by indomethacin, which then leads to decreases in the levels of IL-1β and TBI-induced neuron damage

The genomes of two key bumblebee species with primitive eusocial organization

Background: The shift from solitary to social behavior is one of the major evolutionary transitions. Primitively eusocial bumblebees are uniquely placed to illuminate the evolution of highly eusocial insect societies. Bumblebees are also invaluable natural and agricultural pollinators, and there is widespread concern over recent population declines in some species. High-quality genomic data will inform key aspects of bumblebee biology, including susceptibility to implicated population viability threats. Results: We report the high quality draft genome sequences of Bombus terrestris and Bombus impatiens, two ecologically dominant bumblebees and widely utilized study species. Comparing these new genomes to those of the highly eusocial honeybee Apis mellifera and other Hymenoptera, we identify deeply conserved similarities, as well as novelties key to the biology of these organisms. Some honeybee genome features thought to underpin advanced eusociality are also present in bumblebees, indicating an earlier evolution in the bee lineage. Xenobiotic detoxification and immune genes are similarly depauperate in bumblebees and honeybees, and multiple categories of genes linked to social organization, including development and behavior, show high conservation. Key differences identified include a bias in bumblebee chemoreception towards gustation from olfaction, and striking differences in microRNAs, potentially responsible for gene regulation underlying social and other traits. Conclusions: These two bumblebee genomes provide a foundation for post-genomic research on these key pollinators and insect societies. Overall, gene repertoires suggest that the route to advanced eusociality in bees was mediated by many small changes in many genes and processes, and not by notable expansion or depauperation

Cloning, expression and characterization of low-molecular-weight acid phosphatases from bovine heart and human placenta

Acid phosphatases (orthophosphoric-monoester phosphohydrolyases; EC 3.1.3.2) catalyze the hydrolysis of orthophosphate monoesters and the reaction has a pH optimum in the dilute acid range. Based upon the published amino acid sequence of a low-molecular-weight (18 kDa) bovine liver acid phosphatase, a full length cDNA clone coding for bovine heart low-molecular-weight acid phosphatase (BHPTP) was isolated and sequenced. By using a T7 expression system, the native bovine heart enzyme was overproduced in E. coli. The expressed BHPTP was nearly identical to the authentic enzyme in molecular weight, immunological reactivity and kinetic properties. A DNA fragment corresponding to the coding region of the isolated bovine heart cDNA clone was subsequently used as a probe to screen a human placental $\lambda$gt11 cDNA library. Several overlapping cDNA clones coding for two distinct human cytoplasmic, low molecular weight, phosphotyrosyl protein phosphatases (HCPTPs) were thus obtained. The two longest clones, HCPTP1-1 and HCPTP2-1, were found to have identical nucleotide sequences, with the exception of 108 base pair segment in the middle of the open reading frame. The gene coding for these HCPTPs was assigned to chromosome 2, which is also known to be the location of the human red cell acid phosphatase locus ACP1. Studies using genomic DNA indicate that the variable region of HCPTP1-1 and HCPTP2-1 does not result from the alternative RNA splicing. Based on RNA blotting experiments, the gene coding for both types of HCPTPs is expressed in all human tissues studied. The isoenzymes encoded by HCPTP1-1 and HCPTP2-1 were overexpressed in E. coli and the resulting recombinant enzymes (i.e. HCPTP-A and HCPTP-B) were characterized by SDS-PAGE, immunoblotting and phosphatase activity assay. The expressed proteins were strongly active towards the phosphatase substrates, p-nitrophenyl phosphate, $\beta$-naphthyl phosphate, O-phospho-L-tyrosine, phosphotyrosyl peptides and phosphotyrosyl myelin basic protein, but not $\alpha$-naphthyl phosphate, threonine phosphate, O-phospho-L-serine, or phosphoseryl and phosphothreonyl casein. HCPTP-A and HCPTP-B exhibited different substrate specificities and sensitivities towards Zn\sp{2+}. It is concluded that HCPTPs are in fact two members of a phosphotyrosyl protein phosphatase family, and that they may have different physiological substrates in the cell

Effects of resveratrol or L-NAME on escape latency in the training trials of the water maze task.

<p>The different treated rats were subjected to the analysis: the rats were infused icv with vehicle (sham), the rats with Aβ_25–35 administration for seven days (Aβ), the rats with the injection of Aβ_25–35 and resveratrol (Aβ+Res), the rats administered with resveratrol only (Res), and the rats with Aβ_25–35 and L-NAME administration (Aβ+L-NAME). Data were represented as mean ± SEM values (n = 5). **<i>p</i><0.01 was considered significantly different from sham values by Mann-Whiney U test.</p

Drug effects on the hippocampal iNOS expression.

<p>(A) Western blot analysis on sham rats (sham), the rats with Aβ_25–35 administration for seven days (Aβ), the rats with injection of Aβ_25–35 and resveratrol (Aβ+Res), the rats administered with resveratrol only (Res), and the rats injected with Aβ_25–35 and L-NAME (Aβ+L-NAME); (B) Relative iNOS level quantified as compared with sham group (normalized to 100%). Data were represented as mean ± SEM values (n = 5). *<i>p</i><0.05 was considered significantly different from sham values by Mann-Whiney U test. #<i>p</i><0.05 was considered significantly different from Aβ values by Mann-Whiney U test.</p

Effects of Aβ, resveratrol and L-NAME administration on the retention time of rota-rod.

<p>The training procedure is as described in Materials and Method. Bars represent mean ± SEM values (n = 5).</p

Drug effects on the hippocampal HO-1 expression.

<p>(A) Western blot analysis on sham rats (sham), the rats with Aβ_25–35 administration for seven days (Aβ), the rats with injection of Aβ_25–35 and resveratrol (Aβ+Res), the rats administered with resveratrol only (Res), and the rats injected with Aβ_25–35 and L-NAME (Aβ+L-NAME); (B) Relative iNOS level quantified as compared with sham group (normalized to 100%). Data were represented as mean ± SEM values (n = 5). *<i>p</i><0.05 was considered significantly different from sham values by Mann-Whiney U test. #<i>p</i><0.05 was considered significantly different from Aβ values by Mann-Whiney U test.</p