Limits...
Identification and Characterization of a Novel Microvitellogenin from the Chinese Oak Silkworm Antheraea pernyi.

Liu Y, Chen M, Su J, Ma H, Zheng X, Li Q, Shi S, Qin L - PLoS ONE (2015)

Bottom Line: This gene does not contain introns.ApmVg decreased dramatically during embryonic development.These results represent the first study of mVg outside M. sexta and B. mori and provide insight into the physiological role and evolution of mVgs.

View Article: PubMed Central - PubMed

Affiliation: Department of Sericulture, Shenyang Agricultural University, Shenyang, Liaoning, China; Sericultural Institute of Liaoning Province, Fengcheng, Liaoning, China.

ABSTRACT
Microvitellogenin (mVg) is a relatively small vitellogenic protein only characterized in the eggs of the lepidopteran insects Manduca sexta and Bombyx mori. In the present study, we report a novel mVg (ApmVg) isolated from the Chinese oak silkworm Antheraea pernyi. The obtained ApmVg cDNA sequence contains an open reading frame of 783 bp encoding a protein of 260 amino acids with a predicted molecular weight of 29.96 kDa. This gene does not contain introns. Structural analysis revealed that this protein shares putative conserved domains with the lepidopteran low-molecular weight lipoprotein, which belongs to the lipoprotein_11 superfamily. The protein sequence of ApmVg exhibits 48% sequence identity with mVg from M. sexta and 40-47% sequence identity with the 30K lipoproteins from B. mori. Phylogenetic analysis suggests that ApmVg is a novel member of the lepidopteran low-molecular weight lipoproteins. Transcriptional analysis indicated that ApmVg mRNA is mainly expressed in the fat body (both female and male) during post-diapause development of the pupal stage, and it was also detected in ovaries and spermaries in smaller amounts. RT-PCR and Western blot analyses revealed that ApmVg is synthesized by the fat body and secreted into hemolymph and ultimately accumulates in eggs. The ApmVg transcript can be detected in the fat bodies of female pupae four days after treatment with 20-hydroxyecdysone and shows an expression pattern distinct from that of vitellogenin (Vg), which is detectable throughout diapausing and in post-diapause development. ApmVg decreased dramatically during embryonic development. These results represent the first study of mVg outside M. sexta and B. mori and provide insight into the physiological role and evolution of mVgs.

No MeSH data available.


Expression of the mVg gene.(A) The developmental stages 1–4 represent eggs, larvae, pupae, and adults, respectively. (B) Tissue distributions in the pupal stage. The labels 5–10 indicate the brain, hemolymph (♀), fat body (♀), ovaries (♀), spermaries (♂), and fat body (♂), respectively. (C) The diapause stages 11–14 represent the diapausing stage, diapause-activated stage, post-diapause (1) stage, and post-diapause (2) stage, respectively. Post-diapause pupae (1) and (2) represent diapause-activated pupae without, and with, respectively, developing ovarian follicles undergoing vitellogenesis. The expression patterns were analyzed by RT-PCR using a gene-specific primer pair. The eIF-4A gene was used as control.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4488348&req=5

pone.0131751.g003: Expression of the mVg gene.(A) The developmental stages 1–4 represent eggs, larvae, pupae, and adults, respectively. (B) Tissue distributions in the pupal stage. The labels 5–10 indicate the brain, hemolymph (♀), fat body (♀), ovaries (♀), spermaries (♂), and fat body (♂), respectively. (C) The diapause stages 11–14 represent the diapausing stage, diapause-activated stage, post-diapause (1) stage, and post-diapause (2) stage, respectively. Post-diapause pupae (1) and (2) represent diapause-activated pupae without, and with, respectively, developing ovarian follicles undergoing vitellogenesis. The expression patterns were analyzed by RT-PCR using a gene-specific primer pair. The eIF-4A gene was used as control.

Mentions: We first used RT-PCR to examine the mRNA expression profile. Sequencing the positive RT-PCR product confirmed that it was derived from the targeted gene. Investigation of the expression patterns during the four developmental stages (egg, larva, pupa, and adult) showed that the A. pernyi mVg mRNA was only expressed in the pupal stage (Fig 3A). The tissue distribution indicated that the A. pernyi mVg mRNA was specifically expressed in the fat body (female and male), spermary, or ovary (Fig 3B).


Identification and Characterization of a Novel Microvitellogenin from the Chinese Oak Silkworm Antheraea pernyi.

Liu Y, Chen M, Su J, Ma H, Zheng X, Li Q, Shi S, Qin L - PLoS ONE (2015)

Expression of the mVg gene.(A) The developmental stages 1–4 represent eggs, larvae, pupae, and adults, respectively. (B) Tissue distributions in the pupal stage. The labels 5–10 indicate the brain, hemolymph (♀), fat body (♀), ovaries (♀), spermaries (♂), and fat body (♂), respectively. (C) The diapause stages 11–14 represent the diapausing stage, diapause-activated stage, post-diapause (1) stage, and post-diapause (2) stage, respectively. Post-diapause pupae (1) and (2) represent diapause-activated pupae without, and with, respectively, developing ovarian follicles undergoing vitellogenesis. The expression patterns were analyzed by RT-PCR using a gene-specific primer pair. The eIF-4A gene was used as control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488348&req=5

pone.0131751.g003: Expression of the mVg gene.(A) The developmental stages 1–4 represent eggs, larvae, pupae, and adults, respectively. (B) Tissue distributions in the pupal stage. The labels 5–10 indicate the brain, hemolymph (♀), fat body (♀), ovaries (♀), spermaries (♂), and fat body (♂), respectively. (C) The diapause stages 11–14 represent the diapausing stage, diapause-activated stage, post-diapause (1) stage, and post-diapause (2) stage, respectively. Post-diapause pupae (1) and (2) represent diapause-activated pupae without, and with, respectively, developing ovarian follicles undergoing vitellogenesis. The expression patterns were analyzed by RT-PCR using a gene-specific primer pair. The eIF-4A gene was used as control.
Mentions: We first used RT-PCR to examine the mRNA expression profile. Sequencing the positive RT-PCR product confirmed that it was derived from the targeted gene. Investigation of the expression patterns during the four developmental stages (egg, larva, pupa, and adult) showed that the A. pernyi mVg mRNA was only expressed in the pupal stage (Fig 3A). The tissue distribution indicated that the A. pernyi mVg mRNA was specifically expressed in the fat body (female and male), spermary, or ovary (Fig 3B).

Bottom Line: This gene does not contain introns.ApmVg decreased dramatically during embryonic development.These results represent the first study of mVg outside M. sexta and B. mori and provide insight into the physiological role and evolution of mVgs.

View Article: PubMed Central - PubMed

Affiliation: Department of Sericulture, Shenyang Agricultural University, Shenyang, Liaoning, China; Sericultural Institute of Liaoning Province, Fengcheng, Liaoning, China.

ABSTRACT
Microvitellogenin (mVg) is a relatively small vitellogenic protein only characterized in the eggs of the lepidopteran insects Manduca sexta and Bombyx mori. In the present study, we report a novel mVg (ApmVg) isolated from the Chinese oak silkworm Antheraea pernyi. The obtained ApmVg cDNA sequence contains an open reading frame of 783 bp encoding a protein of 260 amino acids with a predicted molecular weight of 29.96 kDa. This gene does not contain introns. Structural analysis revealed that this protein shares putative conserved domains with the lepidopteran low-molecular weight lipoprotein, which belongs to the lipoprotein_11 superfamily. The protein sequence of ApmVg exhibits 48% sequence identity with mVg from M. sexta and 40-47% sequence identity with the 30K lipoproteins from B. mori. Phylogenetic analysis suggests that ApmVg is a novel member of the lepidopteran low-molecular weight lipoproteins. Transcriptional analysis indicated that ApmVg mRNA is mainly expressed in the fat body (both female and male) during post-diapause development of the pupal stage, and it was also detected in ovaries and spermaries in smaller amounts. RT-PCR and Western blot analyses revealed that ApmVg is synthesized by the fat body and secreted into hemolymph and ultimately accumulates in eggs. The ApmVg transcript can be detected in the fat bodies of female pupae four days after treatment with 20-hydroxyecdysone and shows an expression pattern distinct from that of vitellogenin (Vg), which is detectable throughout diapausing and in post-diapause development. ApmVg decreased dramatically during embryonic development. These results represent the first study of mVg outside M. sexta and B. mori and provide insight into the physiological role and evolution of mVgs.

No MeSH data available.