Additional file 2 of The ontogenic gonadal transcriptomes provide insights into sex change in the ricefield eel Monopterus albus

Additional file 2: Table S1. Quality control parameters of the sequencing data. TableS2. Summary of high-quality reads mapped to the ribosome database. Table S3. Overview of mapping status for gonadal transcriptomes during sexual change of ricefield eel. Table S4. Number of reference genes and new genes in sequencing data. Table S5. Annotation of novel genes.Table S6. GO Enrichment Analysis of DEGs between EI vs F. Table S7. GO Enrichment Analysis of DEGs between MI vs EI. Table S8. GO Enrichment Analysis of DEGs between LI vs MI. Table S9. KEGG enrichment analysis of DEGs between EI vs F. Table S10. KEGG enrichment analysis of DEGs between MI vs EI. Table S11. KEGG enrichment analysis of DEGs between LI vs MI. Table S12. The expression of down-regulated DEGs for EI vs F during sex change. Table S13. The expression of up-regulated for DEGs for EI vs F during sex change

Superconducting magnetic separation of phosphate using freshly formed hydrous ferric oxide sols

<p>Paramagnetic materials, such as ferric hydroxides, which are cost-effective and highly-efficient, have been little studied in relation to the magnetic separation process. In this study, freshly formed hydrous ferric oxide (HFO) sols were used to remove aqueous phosphate, followed by superconducting magnetic separation. The magnetization of HFO was determined to be 5.7 emu/g in 5.0 T. The particle size distributions ranged from 1 to 80 μm. Ferrihydrite was the primary mineral phase according to XRD analysis. Dissolved P (DP) was first adsorbed on HFO, and second, the P-containing HFO were separated by high gradient superconducting magnetic separation (HGSMS) to remove the Total P (TP). To obtain a P concentration of <0.05 mg/l in the effluent, 0.3, 1.0 and 1.3 g/l HFO were added to 2.5, 5 and 10 mg/l P solutions. The capacity of the HGSMS canister for capturing P-adsorbed HFO depends on the magnetic intensity and flow rate. In the 5.0 T HGSMS at a 1.0 cm/s flow rate, there were 75 column volumes in a single HGSMS cycle. The P concentration increased by 37.5 times after regeneration. Approximately 170 mg/l TP was measured in the backwash water.</p

Data_Sheet_5.PDF

<p>The neurohypophyseal hormone oxytocin (Oxt) has been shown to stimulate prolactin (Prl) synthesis and release from the adenohypophysis in rats. However, little is known about the functional roles of Oxt-like neuropeptides in the adenohypophysis of non-mammalian vertebrates. In this study, cDNAs encoding ricefield eel oxytocin-like receptors (Oxtlr), namely isotocin (Ist) receptor 1 (Istr1) and 2 (Istr2), were isolated and specific antisera were generated, respectively. RT-PCR and Western blot analysis detected the presence of both Istr1 and Istr2 in the brain and pituitary, but differential expression in some peripheral tissues, including the liver and kidney, where only Istr1 was detected. In the pituitary, immunoreactive Istr1 and Istr2 were differentially distributed, with the former mainly in adenohypophyseal cell layers adjacent to the neurohypophysis, whereas the latter in peripheral areas of the adenohypophysis. Double immunofluorescent images showed that immunostaining of Istr1, but not Istr2 was localized to growth hormone (Gh) cells, but neither of them was expressed in Prl cells. Ist inhibited Gh release in primary pituitary cells of ricefield eels and increased Gh contents in the pituitary gland of ricefield eels at 6 h after in vivo administration. Ist inhibition of Gh release is probably mediated by cAMP, PKC/DAG, and IP3/Ca²⁺ pathways. In contrast, Ist did not affect either prl gene expression or Prl contents in primary pituitary cells. Results of this study demonstrated that Ist may not be involved in the regulation of Prl, but inhibit Gh release via Istr1 rather than Istr2 in ricefield eels, and provided evidence for the direct regulation of Gh cells by oxytocin-like neuropeptides in the pituitary of non-mammalian vertebrates.</p

Data_Sheet_1.DOCX

<p>The neurohypophyseal hormone oxytocin (Oxt) has been shown to stimulate prolactin (Prl) synthesis and release from the adenohypophysis in rats. However, little is known about the functional roles of Oxt-like neuropeptides in the adenohypophysis of non-mammalian vertebrates. In this study, cDNAs encoding ricefield eel oxytocin-like receptors (Oxtlr), namely isotocin (Ist) receptor 1 (Istr1) and 2 (Istr2), were isolated and specific antisera were generated, respectively. RT-PCR and Western blot analysis detected the presence of both Istr1 and Istr2 in the brain and pituitary, but differential expression in some peripheral tissues, including the liver and kidney, where only Istr1 was detected. In the pituitary, immunoreactive Istr1 and Istr2 were differentially distributed, with the former mainly in adenohypophyseal cell layers adjacent to the neurohypophysis, whereas the latter in peripheral areas of the adenohypophysis. Double immunofluorescent images showed that immunostaining of Istr1, but not Istr2 was localized to growth hormone (Gh) cells, but neither of them was expressed in Prl cells. Ist inhibited Gh release in primary pituitary cells of ricefield eels and increased Gh contents in the pituitary gland of ricefield eels at 6 h after in vivo administration. Ist inhibition of Gh release is probably mediated by cAMP, PKC/DAG, and IP3/Ca²⁺ pathways. In contrast, Ist did not affect either prl gene expression or Prl contents in primary pituitary cells. Results of this study demonstrated that Ist may not be involved in the regulation of Prl, but inhibit Gh release via Istr1 rather than Istr2 in ricefield eels, and provided evidence for the direct regulation of Gh cells by oxytocin-like neuropeptides in the pituitary of non-mammalian vertebrates.</p

Figure 2

<p>A. Cell growth curve measured with MTT assay indicated siRNA mediated knockdown of CHRNA3 increased the growth rate of KYSE510 cells. B. Transwell assay indicated that knockdown of CHRNA3 increased the invasion and migration of KYSE30 cells. C. Western blot analysis indicated 3 Stealth siRNA constructs effectively knocked down CHRNA3. Depletion of CHRNA3 led to a decrease of phosphorylated YAP1 and an increased of dephosphorylated YAP1, particularly a decreased S127 phosphorylation of YAP1. D. Observation of YAP1 subcellular localization with confocal fluorescence microscopy following depletion of CHRNA3. Translocation of YAP1 (green) from the cytoplasm to the nucleus was observed after siRNA mediated knockdown of CHRNA3 in KYSE510 cells for 48 h. E. Real-time PCR test indicated YAP1 targeted genes were induced by CHRNA3 knockdown in KYSE510 cell.</p

Figure 3

<p>A. Endogenous IP test showed positive protein interactions between YAP1 and CHRNA3/CHRNB4/CHRNA5 in KYSE510 cells. B. Positive interaction between tGFP-CHRNA3 and YAP1 was detected in cells transfected with tGFP-CHRNA3. C. GST Pull assay indicated positive interaction of exogenously expressed GST-YAP1 with CHRNA3 in KYSE510 cell. D. Confocal immunofluorescence microscopy observed colocalization of YAP1 and CHRNA3 in KYSE510 cell. The KYSE510 cells were transiently expressed tGFP labeled CHRNA3 of TureClone vector (OriGene). YAP1 was labeled with red-FITC conjugated goat anti rabbit IgG. Cell nuclei were visualized with DAPI. E. endogenous IP detected positive interactions between 14-3-3 and CHRNA3/CHRNB4/CHRNA5.</p

Data_Sheet_4.PDF

<p>The neurohypophyseal hormone oxytocin (Oxt) has been shown to stimulate prolactin (Prl) synthesis and release from the adenohypophysis in rats. However, little is known about the functional roles of Oxt-like neuropeptides in the adenohypophysis of non-mammalian vertebrates. In this study, cDNAs encoding ricefield eel oxytocin-like receptors (Oxtlr), namely isotocin (Ist) receptor 1 (Istr1) and 2 (Istr2), were isolated and specific antisera were generated, respectively. RT-PCR and Western blot analysis detected the presence of both Istr1 and Istr2 in the brain and pituitary, but differential expression in some peripheral tissues, including the liver and kidney, where only Istr1 was detected. In the pituitary, immunoreactive Istr1 and Istr2 were differentially distributed, with the former mainly in adenohypophyseal cell layers adjacent to the neurohypophysis, whereas the latter in peripheral areas of the adenohypophysis. Double immunofluorescent images showed that immunostaining of Istr1, but not Istr2 was localized to growth hormone (Gh) cells, but neither of them was expressed in Prl cells. Ist inhibited Gh release in primary pituitary cells of ricefield eels and increased Gh contents in the pituitary gland of ricefield eels at 6 h after in vivo administration. Ist inhibition of Gh release is probably mediated by cAMP, PKC/DAG, and IP3/Ca²⁺ pathways. In contrast, Ist did not affect either prl gene expression or Prl contents in primary pituitary cells. Results of this study demonstrated that Ist may not be involved in the regulation of Prl, but inhibit Gh release via Istr1 rather than Istr2 in ricefield eels, and provided evidence for the direct regulation of Gh cells by oxytocin-like neuropeptides in the pituitary of non-mammalian vertebrates.</p

Data_Sheet_2.PDF

<p>The neurohypophyseal hormone oxytocin (Oxt) has been shown to stimulate prolactin (Prl) synthesis and release from the adenohypophysis in rats. However, little is known about the functional roles of Oxt-like neuropeptides in the adenohypophysis of non-mammalian vertebrates. In this study, cDNAs encoding ricefield eel oxytocin-like receptors (Oxtlr), namely isotocin (Ist) receptor 1 (Istr1) and 2 (Istr2), were isolated and specific antisera were generated, respectively. RT-PCR and Western blot analysis detected the presence of both Istr1 and Istr2 in the brain and pituitary, but differential expression in some peripheral tissues, including the liver and kidney, where only Istr1 was detected. In the pituitary, immunoreactive Istr1 and Istr2 were differentially distributed, with the former mainly in adenohypophyseal cell layers adjacent to the neurohypophysis, whereas the latter in peripheral areas of the adenohypophysis. Double immunofluorescent images showed that immunostaining of Istr1, but not Istr2 was localized to growth hormone (Gh) cells, but neither of them was expressed in Prl cells. Ist inhibited Gh release in primary pituitary cells of ricefield eels and increased Gh contents in the pituitary gland of ricefield eels at 6 h after in vivo administration. Ist inhibition of Gh release is probably mediated by cAMP, PKC/DAG, and IP3/Ca²⁺ pathways. In contrast, Ist did not affect either prl gene expression or Prl contents in primary pituitary cells. Results of this study demonstrated that Ist may not be involved in the regulation of Prl, but inhibit Gh release via Istr1 rather than Istr2 in ricefield eels, and provided evidence for the direct regulation of Gh cells by oxytocin-like neuropeptides in the pituitary of non-mammalian vertebrates.</p

Table_2.DOCX

<p>The neurohypophyseal hormone oxytocin (Oxt) has been shown to stimulate prolactin (Prl) synthesis and release from the adenohypophysis in rats. However, little is known about the functional roles of Oxt-like neuropeptides in the adenohypophysis of non-mammalian vertebrates. In this study, cDNAs encoding ricefield eel oxytocin-like receptors (Oxtlr), namely isotocin (Ist) receptor 1 (Istr1) and 2 (Istr2), were isolated and specific antisera were generated, respectively. RT-PCR and Western blot analysis detected the presence of both Istr1 and Istr2 in the brain and pituitary, but differential expression in some peripheral tissues, including the liver and kidney, where only Istr1 was detected. In the pituitary, immunoreactive Istr1 and Istr2 were differentially distributed, with the former mainly in adenohypophyseal cell layers adjacent to the neurohypophysis, whereas the latter in peripheral areas of the adenohypophysis. Double immunofluorescent images showed that immunostaining of Istr1, but not Istr2 was localized to growth hormone (Gh) cells, but neither of them was expressed in Prl cells. Ist inhibited Gh release in primary pituitary cells of ricefield eels and increased Gh contents in the pituitary gland of ricefield eels at 6 h after in vivo administration. Ist inhibition of Gh release is probably mediated by cAMP, PKC/DAG, and IP3/Ca²⁺ pathways. In contrast, Ist did not affect either prl gene expression or Prl contents in primary pituitary cells. Results of this study demonstrated that Ist may not be involved in the regulation of Prl, but inhibit Gh release via Istr1 rather than Istr2 in ricefield eels, and provided evidence for the direct regulation of Gh cells by oxytocin-like neuropeptides in the pituitary of non-mammalian vertebrates.</p