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Anopheles gambiae miRNAs as actors of defence reaction against Plasmodium invasion.

Winter F, Edaye S, Hüttenhofer A, Brunel C - Nucleic Acids Res. (2007)

Bottom Line: Twelve of them are expressed ubiquitously across the body, independently of gender, while the other six exhibited an expression pattern restricted to the digestive system.Strikingly, the expression patterns of four miRNAs, including the three unique to mosquito, are affected by the presence of Plasmodium.Altogether, these data support an involvement of miRNAs as new layers in the regulation of Anopheles defence reaction.

View Article: PubMed Central - PubMed

Affiliation: Architecture et Réactivité de l'ARN, Université Louis Pasteur, CNRS, Institut de Biologie Moléculaire et Cellulaire, 15 rue Descarte, 67084 Strasbourg, France.

ABSTRACT
The path Plasmodium takes across the Anopheles midgut constitutes the major bottleneck during the malaria transmission cycle. In the present study, using a combination of shot-gun cloning and bioinformatic analysis, we have identified 18 miRNAs from Anopheles gambiae including three miRNAs unique to mosquito. Twelve of them are expressed ubiquitously across the body, independently of gender, while the other six exhibited an expression pattern restricted to the digestive system. Strikingly, the expression patterns of four miRNAs, including the three unique to mosquito, are affected by the presence of Plasmodium. We also show that knocking down Dicer1 and Ago1 mRNAs led to an increased sensitivity to Plasmodium infection. Altogether, these data support an involvement of miRNAs as new layers in the regulation of Anopheles defence reaction.

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Incidence of regular and infectious blood feeding on miRNA expression profiles. The intensity of the reverse transcriptase stops was quantified (see Materials and Methods section). (A) Effect of blood feeding on miRNA expression in leftovers samples. The female leftovers signal was set as the internal reference. (B) Effect of P. berghei presence on miRNA expression in leftovers samples. The blood-fed female leftovers signal was set as the internal reference. (C) Effect of blood feeding on miRNA expression in midguts samples. The female midguts signal was set as the internal reference. (D) Effect of P. berghei presence on miRNA expression in midguts samples. The blood-fed female midguts signal was set as the internal reference. (E) Representative examples of autoradiographies of the primer-extension assays. Total RNAs were extracted from leftovers or from midguts, 24–48 h after blood feeding on a regular mouse or after blood feeding (BF) on P. berghei-infected mouse.
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Figure 2: Incidence of regular and infectious blood feeding on miRNA expression profiles. The intensity of the reverse transcriptase stops was quantified (see Materials and Methods section). (A) Effect of blood feeding on miRNA expression in leftovers samples. The female leftovers signal was set as the internal reference. (B) Effect of P. berghei presence on miRNA expression in leftovers samples. The blood-fed female leftovers signal was set as the internal reference. (C) Effect of blood feeding on miRNA expression in midguts samples. The female midguts signal was set as the internal reference. (D) Effect of P. berghei presence on miRNA expression in midguts samples. The blood-fed female midguts signal was set as the internal reference. (E) Representative examples of autoradiographies of the primer-extension assays. Total RNAs were extracted from leftovers or from midguts, 24–48 h after blood feeding on a regular mouse or after blood feeding (BF) on P. berghei-infected mouse.

Mentions: The miRNA expression profiles were determined during the course of Plasmodium invasion of the midgut, using reverse transcription assays (Figure 2). We have chosen to consider Plasmodium invasion to have an effect if changes in the level of the reverse transcription stops between two samples differed by more than 33% (see Materials and Methods section). Three types of expression profiles were encountered.Figure 2.


Anopheles gambiae miRNAs as actors of defence reaction against Plasmodium invasion.

Winter F, Edaye S, Hüttenhofer A, Brunel C - Nucleic Acids Res. (2007)

Incidence of regular and infectious blood feeding on miRNA expression profiles. The intensity of the reverse transcriptase stops was quantified (see Materials and Methods section). (A) Effect of blood feeding on miRNA expression in leftovers samples. The female leftovers signal was set as the internal reference. (B) Effect of P. berghei presence on miRNA expression in leftovers samples. The blood-fed female leftovers signal was set as the internal reference. (C) Effect of blood feeding on miRNA expression in midguts samples. The female midguts signal was set as the internal reference. (D) Effect of P. berghei presence on miRNA expression in midguts samples. The blood-fed female midguts signal was set as the internal reference. (E) Representative examples of autoradiographies of the primer-extension assays. Total RNAs were extracted from leftovers or from midguts, 24–48 h after blood feeding on a regular mouse or after blood feeding (BF) on P. berghei-infected mouse.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2175301&req=5

Figure 2: Incidence of regular and infectious blood feeding on miRNA expression profiles. The intensity of the reverse transcriptase stops was quantified (see Materials and Methods section). (A) Effect of blood feeding on miRNA expression in leftovers samples. The female leftovers signal was set as the internal reference. (B) Effect of P. berghei presence on miRNA expression in leftovers samples. The blood-fed female leftovers signal was set as the internal reference. (C) Effect of blood feeding on miRNA expression in midguts samples. The female midguts signal was set as the internal reference. (D) Effect of P. berghei presence on miRNA expression in midguts samples. The blood-fed female midguts signal was set as the internal reference. (E) Representative examples of autoradiographies of the primer-extension assays. Total RNAs were extracted from leftovers or from midguts, 24–48 h after blood feeding on a regular mouse or after blood feeding (BF) on P. berghei-infected mouse.
Mentions: The miRNA expression profiles were determined during the course of Plasmodium invasion of the midgut, using reverse transcription assays (Figure 2). We have chosen to consider Plasmodium invasion to have an effect if changes in the level of the reverse transcription stops between two samples differed by more than 33% (see Materials and Methods section). Three types of expression profiles were encountered.Figure 2.

Bottom Line: Twelve of them are expressed ubiquitously across the body, independently of gender, while the other six exhibited an expression pattern restricted to the digestive system.Strikingly, the expression patterns of four miRNAs, including the three unique to mosquito, are affected by the presence of Plasmodium.Altogether, these data support an involvement of miRNAs as new layers in the regulation of Anopheles defence reaction.

View Article: PubMed Central - PubMed

Affiliation: Architecture et Réactivité de l'ARN, Université Louis Pasteur, CNRS, Institut de Biologie Moléculaire et Cellulaire, 15 rue Descarte, 67084 Strasbourg, France.

ABSTRACT
The path Plasmodium takes across the Anopheles midgut constitutes the major bottleneck during the malaria transmission cycle. In the present study, using a combination of shot-gun cloning and bioinformatic analysis, we have identified 18 miRNAs from Anopheles gambiae including three miRNAs unique to mosquito. Twelve of them are expressed ubiquitously across the body, independently of gender, while the other six exhibited an expression pattern restricted to the digestive system. Strikingly, the expression patterns of four miRNAs, including the three unique to mosquito, are affected by the presence of Plasmodium. We also show that knocking down Dicer1 and Ago1 mRNAs led to an increased sensitivity to Plasmodium infection. Altogether, these data support an involvement of miRNAs as new layers in the regulation of Anopheles defence reaction.

Show MeSH
Related in: MedlinePlus