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Apolipophorin-III Acts as a Positive Regulator of Plasmodium Development in Anopheles stephensi

View Article: PubMed Central - PubMed

ABSTRACT

Apolipophorin III (ApoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune responses of insects. Here we report the molecular and functional characterization of Anopheles stephensi Apolipophorin-III (AsApoLp-III) gene. This gene consists of 679 nucleotides arranged into two exons of 45 and 540 bp that give an ORF encoding 194 amino acid residues. Excluding a putative signal peptide of the first 19 amino acid residues, the 175-residues in mature AsApoLp-III protein has a calculated molecular mass of 22 kDa. Phylogenetic analysis revealed the divergence of mosquitoes (Order Diptera) ApoLp-III from their counterparts in moths (Order: Lepidoptera). Also, it revealed a close relatedness of AsApoLp-III to ApoLp-III of An. gambiae. AsApoLp-III mRNA expression is strongly induced in Plasmodium berghei infected mosquito midguts suggesting its crucial role in parasite development. AsApoLp-III silencing decreased P. berghei oocysts numbers by 7.7 fold against controls. These effects might be due to the interruption of AsApoLp-III mediated lipid delivery to the developing oocysts. In addition, nitric oxide synthase (NOS), an antiplasmodial gene, is also highly induced in AsApoLp-III silenced midguts suggesting that this gene acts like an agonist and protects Plasmodium against the mosquito immunity.

No MeSH data available.


Time kinetics of AsApoLp-III gene expression in Plasmodium berghei infected females. (A) Relative AsApoLp-III mRNA levels in female midguts fed on an uninfected (control) or P. berghei-infected mouse. Samples were collected at different time points after the blood feeding. (B) Same as (A) but carcass samples were collected from the same mosquitoes. Ribosomal S7 protein mRNA levels were used as internal loading reference. The asterisk denoted statistically significant difference in relative mRNA levels of control and infected samples.
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Figure 5: Time kinetics of AsApoLp-III gene expression in Plasmodium berghei infected females. (A) Relative AsApoLp-III mRNA levels in female midguts fed on an uninfected (control) or P. berghei-infected mouse. Samples were collected at different time points after the blood feeding. (B) Same as (A) but carcass samples were collected from the same mosquitoes. Ribosomal S7 protein mRNA levels were used as internal loading reference. The asterisk denoted statistically significant difference in relative mRNA levels of control and infected samples.

Mentions: To find out the role of AsApoLp-III in immunity, AsApoLp-III mRNA expression in the adult female midgut after P. berghei infected blood feeding at varied time points was investigated by qRT-PCR. As shown in Figure 5A, AsApoLp-III mRNA expression in midguts was significantly upregulated (~4 times) at 3 h after infected blood feeding compared to the controls. This time point represents Plasmodium pre-ookinete stage when fertilization and zygote formation takes place in the gut lumen of infected mosquitoes. Following up-regulation at 3 h, a gradual reduction of AsApoLp-III transcript levels was observed till 12 h of post-infection. Furthermore, the AsApoLp-III transcript levels increased significantly at 18 and 24 h post-infection against controls (Figure 5A). The significance of 24 h time point corresponds to ookinetes invasion of the midgut epithelium (Kumar et al., 2004; Gupta et al., 2010).


Apolipophorin-III Acts as a Positive Regulator of Plasmodium Development in Anopheles stephensi
Time kinetics of AsApoLp-III gene expression in Plasmodium berghei infected females. (A) Relative AsApoLp-III mRNA levels in female midguts fed on an uninfected (control) or P. berghei-infected mouse. Samples were collected at different time points after the blood feeding. (B) Same as (A) but carcass samples were collected from the same mosquitoes. Ribosomal S7 protein mRNA levels were used as internal loading reference. The asterisk denoted statistically significant difference in relative mRNA levels of control and infected samples.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5383653&req=5

Figure 5: Time kinetics of AsApoLp-III gene expression in Plasmodium berghei infected females. (A) Relative AsApoLp-III mRNA levels in female midguts fed on an uninfected (control) or P. berghei-infected mouse. Samples were collected at different time points after the blood feeding. (B) Same as (A) but carcass samples were collected from the same mosquitoes. Ribosomal S7 protein mRNA levels were used as internal loading reference. The asterisk denoted statistically significant difference in relative mRNA levels of control and infected samples.
Mentions: To find out the role of AsApoLp-III in immunity, AsApoLp-III mRNA expression in the adult female midgut after P. berghei infected blood feeding at varied time points was investigated by qRT-PCR. As shown in Figure 5A, AsApoLp-III mRNA expression in midguts was significantly upregulated (~4 times) at 3 h after infected blood feeding compared to the controls. This time point represents Plasmodium pre-ookinete stage when fertilization and zygote formation takes place in the gut lumen of infected mosquitoes. Following up-regulation at 3 h, a gradual reduction of AsApoLp-III transcript levels was observed till 12 h of post-infection. Furthermore, the AsApoLp-III transcript levels increased significantly at 18 and 24 h post-infection against controls (Figure 5A). The significance of 24 h time point corresponds to ookinetes invasion of the midgut epithelium (Kumar et al., 2004; Gupta et al., 2010).

View Article: PubMed Central - PubMed

ABSTRACT

Apolipophorin III (ApoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune responses of insects. Here we report the molecular and functional characterization of Anopheles stephensi Apolipophorin-III (AsApoLp-III) gene. This gene consists of 679 nucleotides arranged into two exons of 45 and 540 bp that give an ORF encoding 194 amino acid residues. Excluding a putative signal peptide of the first 19 amino acid residues, the 175-residues in mature AsApoLp-III protein has a calculated molecular mass of 22 kDa. Phylogenetic analysis revealed the divergence of mosquitoes (Order Diptera) ApoLp-III from their counterparts in moths (Order: Lepidoptera). Also, it revealed a close relatedness of AsApoLp-III to ApoLp-III of An. gambiae. AsApoLp-III mRNA expression is strongly induced in Plasmodium berghei infected mosquito midguts suggesting its crucial role in parasite development. AsApoLp-III silencing decreased P. berghei oocysts numbers by 7.7 fold against controls. These effects might be due to the interruption of AsApoLp-III mediated lipid delivery to the developing oocysts. In addition, nitric oxide synthase (NOS), an antiplasmodial gene, is also highly induced in AsApoLp-III silenced midguts suggesting that this gene acts like an agonist and protects Plasmodium against the mosquito immunity.

No MeSH data available.