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The Esg Gene Is Involved in Nicotine Sensitivity in Drosophila melanogaster.

Sanchez-Díaz I, Rosales-Bravo F, Reyes-Taboada JL, Covarrubias AA, Narvaez-Padilla V, Reynaud E - PLoS ONE (2015)

Bottom Line: Two fly lines, L4 and L70, whose HRT was significantly longer than control´s were identified.In this work, we demonstrate that esg loss of function induces nicotine sensitivity possibly by altering development of sensory organs and neurons in the medial section of the thoracoabdominal ganglion.The ectopic expression of the miR-310c also induces nicotine sensitivity by lowering Esg levels thus disrupting sensory organs and possibly to the modulation of other miR-310c targets.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Avenida Universidad, 2001, Apartado Postal, 510-3, Cuernavaca 62210, México.

ABSTRACT
In humans, there is a strong correlation between sensitivity to substances of abuse and addiction risk. This differential tolerance to drugs has a strong genetic component. The identification of human genetic factors that alter drug tolerance has been a difficult task. For this reason and taking advantage of the fact that Drosophila responds similarly to humans to many drugs, and that genetically it has a high degree of homology (sharing at least 70% of genes known to be involved in human genetic diseases), we looked for genes in Drosophila that altered their nicotine sensitivity. We developed an instantaneous nicotine vaporization technique that exposed flies in a reproducible way. The amount of nicotine sufficient to "knock out" half of control flies for 30 minutes was determined and this parameter was defined as Half Recovery Time (HRT). Two fly lines, L4 and L70, whose HRT was significantly longer than control´s were identified. The L4 insertion is a loss of function allele of the transcriptional factor escargot (esg), whereas L70 insertion causes miss-expression of the microRNA cluster miR-310-311-312-313 (miR-310c). In this work, we demonstrate that esg loss of function induces nicotine sensitivity possibly by altering development of sensory organs and neurons in the medial section of the thoracoabdominal ganglion. The ectopic expression of the miR-310c also induces nicotine sensitivity by lowering Esg levels thus disrupting sensory organs and possibly to the modulation of other miR-310c targets.

No MeSH data available.


Related in: MedlinePlus

L4 P-element insertion causes Esg loss of function.(A) Western blot of L4 embryos showing a reduced expression of the Esg protein compared to w1118. (B) Quantitative densitometry of western blots. n = 3 (C) Half recuperation time (HRT) of the different esg alleles used. EP = P{EP}esgEU143, esg35ce-3 (loss of function allele) ** = P≤ 0.001, **** = P<< 0.001.
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pone.0133956.g002: L4 P-element insertion causes Esg loss of function.(A) Western blot of L4 embryos showing a reduced expression of the Esg protein compared to w1118. (B) Quantitative densitometry of western blots. n = 3 (C) Half recuperation time (HRT) of the different esg alleles used. EP = P{EP}esgEU143, esg35ce-3 (loss of function allele) ** = P≤ 0.001, **** = P<< 0.001.

Mentions: To evaluate how the P-element insertion was affecting Esg levels in the L4/+ line, we performed western blot analysis using embryos because esg expression levels are reported to be highest at this developmental stage [6]. We found that in embryos, L4 has approximately half of Esg protein compared to the control line w1118 (Fig 2A and 2B). We attempted to perform western blots of adult extracts but Esg could not be detected in these conditions. This result is consistent with reports showing a very low esg expression at this stage [7,8]. Additionally, we found that in approximately 8% of the population, L4 flies have the characteristic cuticle defects reported in other esg loss of function allelles, such as loss of bristles in the tergites and sternites, and loss of pigmentation in the dorsal abdomen [7]. These data strongly suggest that L4 leads to a loss of function of esg.


The Esg Gene Is Involved in Nicotine Sensitivity in Drosophila melanogaster.

Sanchez-Díaz I, Rosales-Bravo F, Reyes-Taboada JL, Covarrubias AA, Narvaez-Padilla V, Reynaud E - PLoS ONE (2015)

L4 P-element insertion causes Esg loss of function.(A) Western blot of L4 embryos showing a reduced expression of the Esg protein compared to w1118. (B) Quantitative densitometry of western blots. n = 3 (C) Half recuperation time (HRT) of the different esg alleles used. EP = P{EP}esgEU143, esg35ce-3 (loss of function allele) ** = P≤ 0.001, **** = P<< 0.001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4519288&req=5

pone.0133956.g002: L4 P-element insertion causes Esg loss of function.(A) Western blot of L4 embryos showing a reduced expression of the Esg protein compared to w1118. (B) Quantitative densitometry of western blots. n = 3 (C) Half recuperation time (HRT) of the different esg alleles used. EP = P{EP}esgEU143, esg35ce-3 (loss of function allele) ** = P≤ 0.001, **** = P<< 0.001.
Mentions: To evaluate how the P-element insertion was affecting Esg levels in the L4/+ line, we performed western blot analysis using embryos because esg expression levels are reported to be highest at this developmental stage [6]. We found that in embryos, L4 has approximately half of Esg protein compared to the control line w1118 (Fig 2A and 2B). We attempted to perform western blots of adult extracts but Esg could not be detected in these conditions. This result is consistent with reports showing a very low esg expression at this stage [7,8]. Additionally, we found that in approximately 8% of the population, L4 flies have the characteristic cuticle defects reported in other esg loss of function allelles, such as loss of bristles in the tergites and sternites, and loss of pigmentation in the dorsal abdomen [7]. These data strongly suggest that L4 leads to a loss of function of esg.

Bottom Line: Two fly lines, L4 and L70, whose HRT was significantly longer than control´s were identified.In this work, we demonstrate that esg loss of function induces nicotine sensitivity possibly by altering development of sensory organs and neurons in the medial section of the thoracoabdominal ganglion.The ectopic expression of the miR-310c also induces nicotine sensitivity by lowering Esg levels thus disrupting sensory organs and possibly to the modulation of other miR-310c targets.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Avenida Universidad, 2001, Apartado Postal, 510-3, Cuernavaca 62210, México.

ABSTRACT
In humans, there is a strong correlation between sensitivity to substances of abuse and addiction risk. This differential tolerance to drugs has a strong genetic component. The identification of human genetic factors that alter drug tolerance has been a difficult task. For this reason and taking advantage of the fact that Drosophila responds similarly to humans to many drugs, and that genetically it has a high degree of homology (sharing at least 70% of genes known to be involved in human genetic diseases), we looked for genes in Drosophila that altered their nicotine sensitivity. We developed an instantaneous nicotine vaporization technique that exposed flies in a reproducible way. The amount of nicotine sufficient to "knock out" half of control flies for 30 minutes was determined and this parameter was defined as Half Recovery Time (HRT). Two fly lines, L4 and L70, whose HRT was significantly longer than control´s were identified. The L4 insertion is a loss of function allele of the transcriptional factor escargot (esg), whereas L70 insertion causes miss-expression of the microRNA cluster miR-310-311-312-313 (miR-310c). In this work, we demonstrate that esg loss of function induces nicotine sensitivity possibly by altering development of sensory organs and neurons in the medial section of the thoracoabdominal ganglion. The ectopic expression of the miR-310c also induces nicotine sensitivity by lowering Esg levels thus disrupting sensory organs and possibly to the modulation of other miR-310c targets.

No MeSH data available.


Related in: MedlinePlus