Localization of serotonin/tryptophan-hydroxylase-immunoreactive cells in the brain and suboesophageal ganglion of Drosophila melanogaster.

We previously demonstrated that tryptophan hydroxylase (TPH), the rate-limiting enzyme of serotonin (5-HT) synthesis, was commonly present in the brains of some insects. The current study was aimed at determining the number of serotonergic neurons in the brain and suboesophageal ganglion of adult Drosophila melanogaster and to investigate further the differences in immunoreactivity between 5-HT and TPH. Brain sections of Drosophila were immunostaind with sheep anti-TPH polyclonal antibody and rabbit anti-5-HT antiserum. The 5-HT-like immunoreactive neurons were also immunoreactive for TPH and bilaterally symmetrical; 83 neurons were found in each hemisphere of the brain and suboesophageal ganglion of adult Drosophila. This technique of colocalizing 5-HT and TPH revealed a larger number of serotonergic neurons in the brain and suboesophageal ganglion than that previous reported, thus updating our knowledge of the 5-HT neuronal system of Drosophila.

S100A4 contributes to colitis development by increasing the adherence of Citrobacter rodentium in intestinal epithelial cells.

S100A4 has been implicated in cancer and several inflammatory diseases, but its role in inflammatory bowel disease has not been well investigated. Here, upon infection with Citrobacter rodentium, a model for enteropathogenic Escherichia coli infection in humans, induced the infiltration of a large number of S100A4+ cells into the colon in wild type (WT) mice. Deficiency of S100A4 reduced weight loss, bacterial colonization and colonic pathology. Furthermore, the expression of inflammatory cytokines and the recruitment of macrophages and neutrophils also decreased significantly in S100A4 knock out (S100A4 -/-) mice. In vitro, soluble S100A4 directly up-regulated expression of integrin ß-1 in intestinal epithelial cells and significantly increased the adherence of C. rodentium to intestinal epithelial cells. Additionally, the effects of S100A4 on the adherence of C. rodentium to epithelial cells could be abolished by a receptor for advanced glycation end products (RAGE)-specific inhibitor (FPS-ZM1). Therefore, these data indicate a novel mechanism for S100A4 that promotes colitis development by enhancing host adhesion and colonization of Citrobacter rodentium through the S100A4-mediated host inflammatory responses.

Identification and Expression Analysis of Tryptophan Hydroxylase in the Brain and Ventral Nerve Cord of Ragworm Neanthes japonica (Polychaeta, Annelida).

Tryptophan hydroxylase (TPH) was stained in the central nervous system of the Neanthes japonica (Polychaeta, Annelida), using sheep anti-tryptophan hydroxylase antibody by the Streptavidin-Peroxidase immunohistochemical method and Colophony-Paraffin embedded section technique. The immunohistochemistry results revealed that the TPH is distributed in the brain and ventral nerve cord, which is consistent with that of serotonin (5-hydroxytryptamine, 5-HT) that labeled by anti-serotonin antibody. Using the rapid amplification of cDNA ends (RACE) technique, TPH cDNA cloned from Neanthes japonica's central nervous system was 1778bp, which encodes predicted protein of 463 amino acid residues. The co-localization of TPH and 5-HT indicated that the specific TPH was responsible for the central serotonin synthesis in the central nervous system of annelida, TPH and 5-HT not only could be as the novel mutual corroboration marker to detect serotonergic neurons, but also provides the evidences for the evolution of aromatic amino acid hydroxylase genes. Anat Rec, 300:415-424, 2017. © 2016 Wiley Periodicals, Inc.

Discovery of specific tryptophan hydroxylase in the brain of the beetle Harmonia axyridis.

Rabbit anti-serotonin and mouse monoclonal anti-tryptophan hydroxylase antisera were applied on the brain sections of the beetle Harmonia axyridis, butterfly Childrena zenobia, moth Antheraea pernyi and ant Camponotus japonicus, using the Streptavidin-Peroxidase immunohistochemical method and Colophony-Paraffin embedded section technique. Results revealed that all the experimental insects showed notable serotonin-like immunoreactivity in the brain. However, only the brain sections of the beetle H. axyridis were strongly labeled by mouse monoclonal anti-tryptophan hydroxylase antiserum, with the distribution pattern matching that of serotonin. These results demonstrate that specific tryptophan hydroxylase may exist in the brain of the beetle and likely reflect the diversity of serotonin synthetic mechanisms as well as the evolution of aromatic amino acid hydroxylase genes.

[The expression of phenylalanine hydroxylase in the brain of ragworm Neanthes japonica (Polychaeta, Annelida)].

Phenylalanine hydroxylase (PAH) is a member of aromatic amino acid hydroxylase (AAAHs) family, and catalyze phenylalanine (Phe) into tyrosine (Tyr). Using immunological and RT-PCR methods to prove the existence of phenylalanine hydroxylase (PAH) gene in the brain of Neanthes japonica in protein and nucleic acid level. Using Western blotting to detect the pah immunogenicity of Neanthes japonica. Making paraffin sections and using immunohistochemical technique to identify the presence and distribution of the phenylalanine hydroxylase gene in the brain of Neanthes japonica. Clone pah gene from the brain of Neanthes japonica by RT-PCR, constructing plasmid and transferring into E. coli to amplification, picking a single homogeneous colony, double digesting then making sequence and comparing homology. Western blotting results showed that the expression of the protein is present in Neanthes japonica brain, immunohistochemistry technique results showed that phenylalanine hydroxylase mainly expressed in abdominal of forebrain, dorsal and sides of midbrain. RT-PCR technique results showed that the phenylalanine hydroxylase exist in the brain of Neanthes japonica and has a high homology with others animals. PAH is present in the lower organisms Neanthes japonica, in protein and nucleic acid level. Which provide the foundation for further study the evolution of aromatic amino acid hydroxylase genes in invertebrate.

Different roles of the posterior inferior frontal gyrus in Chinese character form judgment differences between literate and illiterate individuals.

In the present study, we used event-related functional magnetic resonance imaging (fMRI) to explore the different roles of the posterior inferior frontal gyrus (pIFG) in Chinese character form judgment between literate and illiterate subjects. Using event-related fMRI, 24 healthy right-handed Chinese subjects (12 literates and 12 illiterates) were asked to perform Chinese character and figure form judgment tasks. The blood oxygen level-dependent (BOLD) differences in pIFG were examined with general linear modeling (GLM). We found differences in reaction times and accuracy between subjects as they performed these tasks. These behavioral differences reflect the different cognitive demands of character form judgment for literate and illiterate individuals. The results showed differences in the BOLD response patterns in the pIFG between the two discrimination tasks and the two subject groups. A comparison of the character and figure tasks showed that literate and illiterate subjects had similar BOLD responses in the inferior frontal gyrus. However, differences in behavioral performance suggest that the pIFG plays a different role in Chinese character form judgment for each subject group. In literate subjects, the left pIFG mediated access to phonology in achieving Chinese character form judgment, whereas the right pIFG participated in the processing of the orthography of Chinese characters. In illiterate subjects, the bilateral frontal gyrus participated in the visual-spatial processing of Chinese characters to achieve form judgment.

Immunohistochemical evidence for the presence of tryptophan hydroxylase in the brains of insects as revealed by sheep anti-tryptophan hydroxylase polyclonal antibody.

Immediately following the discovery of tryptophan hydroxylase in Drosophila, we demonstrated the presence of tryptophan hydroxylase in the brain of the beetle Harmonia axyridis (Coleoptera: Coccinellidae). However, whether tryptophan hydroxylase is present in the brains of other insects is still a matter of discussion. In the current study, sheep anti-tryptophan hydroxylase polyclonal antibody has been applied to test for tryptophan hydroxylase immunoreactivity in a broader taxonomic range of insect brains, including holometabolous and hemimetabolous insects: one species each of Coleoptera, Hymenoptera, Diptera, and Blattaria, and two species of Lepidoptera. All species show consistent tryptophan hydroxylase immunoreactivity with distribution patterns matching that of serotonin. The immuno-positive results of such an antibody in brains from diverse orders of insects suggest that specific tryptophan hydroxylase responsible for central serotonin synthesis is probably present in the brains of all insects.