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Transcriptomic Analysis of Musca domestica to Reveal Key Genes of the Prophenoloxidase-Activating System.

Li D, Liang Y, Wang X, Wang L, Qi M, Yu Y, Luan Y - G3 (Bethesda) (2015)

Bottom Line: Of the 89,842 unigenes, based on a similarity search with known genes in other insects, 24 putative genes related to the proPO system were identified.Eight of the identified genes encoded for peptidoglycan recognition receptors, two encoded for prophenoloxidases, three encoded for prophenoloxidase-activating enzymes, and 11 encoded for serine proteinase inhibitors.Collectively, this study has provided the comprehensive transcriptomic data of an insect and some fundamental basis toward achieving understanding of the activation mechanisms and immune functions of the proPO system in M. domestica.

View Article: PubMed Central - PubMed

Affiliation: Biotechnology Department, The School of Biological Sciences and Biotechnology, University of Jinan, Jinan 250022, People's Republic of China chm_lidx@ujn.edu.cn.

No MeSH data available.


Related in: MedlinePlus

Measurement of PO activity. Phenoloxidase (PO) activity of hemolymph samples were assayed with 96-well microplates method using L-DOPA as a substrate. The PO level in hemolymph from normal larvae remained constant, as marked in the control (blank column). Compared with controls, four sample groups marked 1, 2, 3, and 4 (decorative columns) showed the variable increases in PO activity in the hemolymph. The most significant increase in PO activity appeared in sample 3 from larvae challenged by S. aureus and E. coli for 30 min. The lowest increase in PO activity existed in sample 2 from larvae injected with mdproPO 1 antibodies. Importantly, the marked increase in PO activity in bacteria-infected larvae was reverse of the quietly low level after blocking with mdproPO 1 antibodies in sample 4, which was close to the PO level in sample 1 from PBS challenged larvae. The significant variation between control and tested samples was calculated by t-test (***P < 0.001, **P < 0.01, *P < 0.05).
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fig11: Measurement of PO activity. Phenoloxidase (PO) activity of hemolymph samples were assayed with 96-well microplates method using L-DOPA as a substrate. The PO level in hemolymph from normal larvae remained constant, as marked in the control (blank column). Compared with controls, four sample groups marked 1, 2, 3, and 4 (decorative columns) showed the variable increases in PO activity in the hemolymph. The most significant increase in PO activity appeared in sample 3 from larvae challenged by S. aureus and E. coli for 30 min. The lowest increase in PO activity existed in sample 2 from larvae injected with mdproPO 1 antibodies. Importantly, the marked increase in PO activity in bacteria-infected larvae was reverse of the quietly low level after blocking with mdproPO 1 antibodies in sample 4, which was close to the PO level in sample 1 from PBS challenged larvae. The significant variation between control and tested samples was calculated by t-test (***P < 0.001, **P < 0.01, *P < 0.05).

Mentions: We had previously measured PO activity that could increase rapidly in housefly larvae after challenge by S. aureus and E. coli with the 96-well microplates method (Guo et al. 2014) but did not know whether it was related to mdproPO1 cleavage into mdPO1. Here, prior to immune challenge by the mixture of S. aureus and E. coli, housefly larvae were injected with mdproPO1 antibodies to block the interior mdproPO1 proteins. As an important result, the PO activity increase was reversed to the very low level in infected larvae after blocking with mdproPO1 antibodies (Figure 11). It was confirmed that mdproPO1 can play an indispensable role in the activation of proPO system in M. domestica.


Transcriptomic Analysis of Musca domestica to Reveal Key Genes of the Prophenoloxidase-Activating System.

Li D, Liang Y, Wang X, Wang L, Qi M, Yu Y, Luan Y - G3 (Bethesda) (2015)

Measurement of PO activity. Phenoloxidase (PO) activity of hemolymph samples were assayed with 96-well microplates method using L-DOPA as a substrate. The PO level in hemolymph from normal larvae remained constant, as marked in the control (blank column). Compared with controls, four sample groups marked 1, 2, 3, and 4 (decorative columns) showed the variable increases in PO activity in the hemolymph. The most significant increase in PO activity appeared in sample 3 from larvae challenged by S. aureus and E. coli for 30 min. The lowest increase in PO activity existed in sample 2 from larvae injected with mdproPO 1 antibodies. Importantly, the marked increase in PO activity in bacteria-infected larvae was reverse of the quietly low level after blocking with mdproPO 1 antibodies in sample 4, which was close to the PO level in sample 1 from PBS challenged larvae. The significant variation between control and tested samples was calculated by t-test (***P < 0.001, **P < 0.01, *P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
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fig11: Measurement of PO activity. Phenoloxidase (PO) activity of hemolymph samples were assayed with 96-well microplates method using L-DOPA as a substrate. The PO level in hemolymph from normal larvae remained constant, as marked in the control (blank column). Compared with controls, four sample groups marked 1, 2, 3, and 4 (decorative columns) showed the variable increases in PO activity in the hemolymph. The most significant increase in PO activity appeared in sample 3 from larvae challenged by S. aureus and E. coli for 30 min. The lowest increase in PO activity existed in sample 2 from larvae injected with mdproPO 1 antibodies. Importantly, the marked increase in PO activity in bacteria-infected larvae was reverse of the quietly low level after blocking with mdproPO 1 antibodies in sample 4, which was close to the PO level in sample 1 from PBS challenged larvae. The significant variation between control and tested samples was calculated by t-test (***P < 0.001, **P < 0.01, *P < 0.05).
Mentions: We had previously measured PO activity that could increase rapidly in housefly larvae after challenge by S. aureus and E. coli with the 96-well microplates method (Guo et al. 2014) but did not know whether it was related to mdproPO1 cleavage into mdPO1. Here, prior to immune challenge by the mixture of S. aureus and E. coli, housefly larvae were injected with mdproPO1 antibodies to block the interior mdproPO1 proteins. As an important result, the PO activity increase was reversed to the very low level in infected larvae after blocking with mdproPO1 antibodies (Figure 11). It was confirmed that mdproPO1 can play an indispensable role in the activation of proPO system in M. domestica.

Bottom Line: Of the 89,842 unigenes, based on a similarity search with known genes in other insects, 24 putative genes related to the proPO system were identified.Eight of the identified genes encoded for peptidoglycan recognition receptors, two encoded for prophenoloxidases, three encoded for prophenoloxidase-activating enzymes, and 11 encoded for serine proteinase inhibitors.Collectively, this study has provided the comprehensive transcriptomic data of an insect and some fundamental basis toward achieving understanding of the activation mechanisms and immune functions of the proPO system in M. domestica.

View Article: PubMed Central - PubMed

Affiliation: Biotechnology Department, The School of Biological Sciences and Biotechnology, University of Jinan, Jinan 250022, People's Republic of China chm_lidx@ujn.edu.cn.

No MeSH data available.


Related in: MedlinePlus