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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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Expression profiling of Anopheles miRNAs across gender and body parts. (A) Representative examples of the autoradiographies of primer-extension assays performed on total RNA samples from Anopheles. (B) Profiling of the miRNA expression in Anopheles body parts using primer-extension assays. The intensities of the reverse transcriptase stops were quantified (see Materials and Methods section). The stop intensities obtained using total RNAs extracted from the female gut was set as the internal reference. F is used for female and M for male. As stated in Materials and Methods section, ratios are required to be outside of the estimated confidence interval [0.67; 1.33] to be deemed as significant.
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Figure 3: Expression profiling of Anopheles miRNAs across gender and body parts. (A) Representative examples of the autoradiographies of primer-extension assays performed on total RNA samples from Anopheles. (B) Profiling of the miRNA expression in Anopheles body parts using primer-extension assays. The intensities of the reverse transcriptase stops were quantified (see Materials and Methods section). The stop intensities obtained using total RNAs extracted from the female gut was set as the internal reference. F is used for female and M for male. As stated in Materials and Methods section, ratios are required to be outside of the estimated confidence interval [0.67; 1.33] to be deemed as significant.

Mentions: The expression profiles of the miRNAs were determined using reverse transcription assays (Figure 3). Experiments were run on total RNAs extracted from different body parts of male or female Anopheles: head, thorax, gut and leftover. Based on the observations, we could define three classes: (i) miRNAs or clusters of miRNAs expressed only in one body part; (ii) clusters for which the miRNAs have distinct patterns with preferential expression in one body part and (iii) clusters for which all the miRNAs encoded are expressed evenly and ubiquitously throughout the body.Figure 3.


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

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

Expression profiling of Anopheles miRNAs across gender and body parts. (A) Representative examples of the autoradiographies of primer-extension assays performed on total RNA samples from Anopheles. (B) Profiling of the miRNA expression in Anopheles body parts using primer-extension assays. The intensities of the reverse transcriptase stops were quantified (see Materials and Methods section). The stop intensities obtained using total RNAs extracted from the female gut was set as the internal reference. F is used for female and M for male. As stated in Materials and Methods section, ratios are required to be outside of the estimated confidence interval [0.67; 1.33] to be deemed as significant.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

Figure 3: Expression profiling of Anopheles miRNAs across gender and body parts. (A) Representative examples of the autoradiographies of primer-extension assays performed on total RNA samples from Anopheles. (B) Profiling of the miRNA expression in Anopheles body parts using primer-extension assays. The intensities of the reverse transcriptase stops were quantified (see Materials and Methods section). The stop intensities obtained using total RNAs extracted from the female gut was set as the internal reference. F is used for female and M for male. As stated in Materials and Methods section, ratios are required to be outside of the estimated confidence interval [0.67; 1.33] to be deemed as significant.
Mentions: The expression profiles of the miRNAs were determined using reverse transcription assays (Figure 3). Experiments were run on total RNAs extracted from different body parts of male or female Anopheles: head, thorax, gut and leftover. Based on the observations, we could define three classes: (i) miRNAs or clusters of miRNAs expressed only in one body part; (ii) clusters for which the miRNAs have distinct patterns with preferential expression in one body part and (iii) clusters for which all the miRNAs encoded are expressed evenly and ubiquitously throughout the body.Figure 3.

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