In the present study, zooblooting, ELISA, and whole-mount immunocytochemistry methods were used to identify the FXPRLamide family neuropeptides from the Japanese oak silkworm, Antheraea yamamai. The results showed tha...In the present study, zooblooting, ELISA, and whole-mount immunocytochemistry methods were used to identify the FXPRLamide family neuropeptides from the Japanese oak silkworm, Antheraea yamamai. The results showed that the genomic DNA from A. yamamai showed positive bands after being hybridized with the fragment of DH-PBAN cDNA from Samia cynthia ricini, which was labeled with [α-32p]-dCTP. The SG showed highest FXPRLamide peptides titer in neural organs. Using an antiserum against Helicoverpa armigera PBAN, PBAN-like immunoreactivity was detected in the SG and TG of A. yamamai by whole-mount immunocytochemistry, and there were three cluster cells in the SG which shows positive PBAN-like immunoreactivity. The titers of FXPRLamide peptides immunoreactivity in the hemolymph were kept at a steady level. During pupation, the titer was increased promptly, but then decreased to a low level after the early pupal stage. The above-mentioned results demonstrate the existence of FXPRLamide family peptides in A. yamamai, but its function needs to be further investigated in the future.展开更多
Effects of ecdysteroid and juvenile hormone (JH) on vitellogenesis of the Japanese oak silkworm, Antheraea yamamai are reported in this article. After topical treatment with 20 hydroxyecdysone alone or JH analog ...Effects of ecdysteroid and juvenile hormone (JH) on vitellogenesis of the Japanese oak silkworm, Antheraea yamamai are reported in this article. After topical treatment with 20 hydroxyecdysone alone or JH analog (i.e. methoprene) alone and combined treatment with these two chemicals, vitellogenin (Vg) titers in the fat body and haemolymph at the pupal stage were mostly higher than those of the control, indicating that both ecdysteroid and JH exerted a promoting effect on the synthesis of Vg. In contrast, the Vg uptake was markedly inhibited by JH while stimulating effect of the ecdysteroid could be shown that vitellin (Vt) titer in the ovary was lower after methoprene treatments, but higher after 20 hydroxyecdyson treatments. Meanwhile, effects of these two hormones on Vg synthesis in the fat body were also tested with the incubation in vitro with Grace medium containing 3H leucine and the hormones. The results demonstrated that Vg synthesis was stimulated after treating with methoprene alone or 20 hydroxyecdysone alone and combined treating with these two chemicals, and particularly ecdysteroid had more marked positive effect. To comprehensively concluded our results, it could be regarded that ecdysteroid play the main role in the regulation of vitellogenesis for the Japanese oak silkworm.展开更多
To understand the evolutionary conservation ofthe gene expression mechanism and secretion machinery between Antheraea and Bombyx fibroins, we introduced the genomic A. yamamai fibroin gene into the domesticated silkwo...To understand the evolutionary conservation ofthe gene expression mechanism and secretion machinery between Antheraea and Bombyx fibroins, we introduced the genomic A. yamamai fibroin gene into the domesticated silkworm, B. mori. The spliced A. yamamai fibroin mRNA appeared only in the posterior region of the silk gland of the transgenic silkworm, suggesting that the functions of the fibroin promoter region and the splicing machinery are conserved between these two species. The A. yamarnai fibroin protein was detected in the lumen of the silk gland of the transgenic silkworm, albeit at lower levels compared with the B. mori-type fibroin. We found a strong degeneration of the posterior region of the silk gland of the transgenic silkworm. As a result, the cocoon shell weight was much lower in the transgenic silkworm than in the non-transgenic line. These results indicate that the promoter function and splicing machinery are well conserved between A. yamamai and B. mori but that the secretion mechanism of fibroin is diversified between the two.展开更多
基金Acknowledgements This study was supported by a Grant-in-Aid for the Natural Scientific Foundation (30500374) from the National Natural Science Foundation of China, the Key Research Program of Anhui Province, China (05021003).
文摘In the present study, zooblooting, ELISA, and whole-mount immunocytochemistry methods were used to identify the FXPRLamide family neuropeptides from the Japanese oak silkworm, Antheraea yamamai. The results showed that the genomic DNA from A. yamamai showed positive bands after being hybridized with the fragment of DH-PBAN cDNA from Samia cynthia ricini, which was labeled with [α-32p]-dCTP. The SG showed highest FXPRLamide peptides titer in neural organs. Using an antiserum against Helicoverpa armigera PBAN, PBAN-like immunoreactivity was detected in the SG and TG of A. yamamai by whole-mount immunocytochemistry, and there were three cluster cells in the SG which shows positive PBAN-like immunoreactivity. The titers of FXPRLamide peptides immunoreactivity in the hemolymph were kept at a steady level. During pupation, the titer was increased promptly, but then decreased to a low level after the early pupal stage. The above-mentioned results demonstrate the existence of FXPRLamide family peptides in A. yamamai, but its function needs to be further investigated in the future.
文摘Effects of ecdysteroid and juvenile hormone (JH) on vitellogenesis of the Japanese oak silkworm, Antheraea yamamai are reported in this article. After topical treatment with 20 hydroxyecdysone alone or JH analog (i.e. methoprene) alone and combined treatment with these two chemicals, vitellogenin (Vg) titers in the fat body and haemolymph at the pupal stage were mostly higher than those of the control, indicating that both ecdysteroid and JH exerted a promoting effect on the synthesis of Vg. In contrast, the Vg uptake was markedly inhibited by JH while stimulating effect of the ecdysteroid could be shown that vitellin (Vt) titer in the ovary was lower after methoprene treatments, but higher after 20 hydroxyecdyson treatments. Meanwhile, effects of these two hormones on Vg synthesis in the fat body were also tested with the incubation in vitro with Grace medium containing 3H leucine and the hormones. The results demonstrated that Vg synthesis was stimulated after treating with methoprene alone or 20 hydroxyecdysone alone and combined treating with these two chemicals, and particularly ecdysteroid had more marked positive effect. To comprehensively concluded our results, it could be regarded that ecdysteroid play the main role in the regulation of vitellogenesis for the Japanese oak silkworm.
文摘To understand the evolutionary conservation ofthe gene expression mechanism and secretion machinery between Antheraea and Bombyx fibroins, we introduced the genomic A. yamamai fibroin gene into the domesticated silkworm, B. mori. The spliced A. yamamai fibroin mRNA appeared only in the posterior region of the silk gland of the transgenic silkworm, suggesting that the functions of the fibroin promoter region and the splicing machinery are conserved between these two species. The A. yamarnai fibroin protein was detected in the lumen of the silk gland of the transgenic silkworm, albeit at lower levels compared with the B. mori-type fibroin. We found a strong degeneration of the posterior region of the silk gland of the transgenic silkworm. As a result, the cocoon shell weight was much lower in the transgenic silkworm than in the non-transgenic line. These results indicate that the promoter function and splicing machinery are well conserved between A. yamamai and B. mori but that the secretion mechanism of fibroin is diversified between the two.