Unknown,Transcriptomics,Genomics,Proteomics

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Changes in gene expression during cold exposure in Drosophila montana and D. virilis

ABSTRACT: We utilized a candidate gene approach using custom microarray constructed for our study species Drosophila montana and D. virilis to identify genes with modulated expression patterns under low temperature conditions. The flies were exposed to four different treatments (+5°C for 6 days, 0°C for 1 hour, two periods of recovery after cold stress and a control treatment). The aim of the study was to identify potential cold-responsive genes and to investigate differences in gene expression between the species. Microarray analysis revealed altogether 31 out of 219 genes on the array to show expression changes during different stages of cold stress. Among the potential stress tolerance genes detected earlier in D. melanogaster, hsr-omega was upregulated in both species during cold acclimation, expression changes in other genes being treatment- and species specific. Our microarray study clearly showed that different stages of cold response elicit changes at least in genes involved in heat shock response, circadian rhythm and metabolism. Cold- induced gene expression was investigated comparing four different treatments to the control treatment: 1) Cold acclimation: 14 days in control conditions, then 6 days at +5°C 2) Cold hardening: 20 days in control conditions, then1 hour at 0°C 3) 15-min recovery from chill coma: the 20-day-old flies were exposed to -6°C for 16 hours, after which they were let to recover for 15 minutes 4) 1-hour recovery from chill coma: the 20-day-old flies were exposed to -6°C for 16 hours, after which they were let to recover for 1 hour. In control treatment flies were kept for 20 days at 19°C. All the flies were 20-days old at the time of sample collection, and the light:dark cycle was 22 hours of light and 2 hours of dark. Because of the limited space on the array plate, the recovery samples were collected only for D. montana. All the samples were collected 5-6 hours after the lights had been turned on in the chamber and the flies were immediately immersed in liquid nitrogen, after which they were stored at -84ºC. Three pools of ten flies were collected from each treatment group.

ORGANISM(S): Drosophila virilis

SUBMITTER: Laura Vesala

PROVIDER: E-GEOD-31103 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Background All animals must be able to adapt to changes in nutrient quality and supply. One striking aspect of this adaptive response is the extension of lifespan when caloric intake is reduced. These animals are also more resistant to various stresses, including starvation. We have characterized the response of adult Drosophila males and females to starvation and sugar conditions using high density microarrays. Results We have organized the regulated genes into specific metabolic and physiologic pathways. Most of the highly regulated genes have a strong sex bias. In addition to expected sex specific changes, such as egg production in females, we found unexpected biases in sensory systems, amino acid and purine metabolism, and signaling pathways. Comparison of starvation to caloric restriction revealed numerous overlaps. Conclusions Our results support the notion that starvation response shares similarities to caloric restriction. They have relevance for sex specific differences in longevity and in response to nutrient conditions which alter lifespan. Nutrient Conditions and Fly handling Larvae were kept on apple juice agar plates with yeast, and the larvae were raised at 25°C. For the developmental control larvae were collected 39-41 h AEL, for the developmental time-points at 51-53 hours and every 12 hours until 99-101 hours. Adult flies were kept at 25°C in 50 ml PS-tubes flat bottom (Greiner bio-one, Frickenhausen, Germany) on standard fly food (16.5 g yeast, 81,5 g cornmeal, 8 g agar, 100 ml sugar beet sirup and 200 ml 10% Nipagin in EtOH). Flies were collected that had hatched in 24 hours and kept in groups of ~200 flies/tube. After 5 days we separated males and females and put them in groups of 80/28 ml PS-tube. We allowed for one day CO2 recovery on standard fly food before flies were moved to 1) tubes with yeast paste on PBS soaked filter paper (control) 2) tubes with only PBS soaked filter paper (starvation) or 3) tubes with filter paper soaked in PBS with 20% Sucrose (sugar). After 24 or 48 hours flies were shock frozen in liquid nitrogen and stored at - 80°C for further handling. Males and females were assayed separately. For females, we performed 24 and 48 starvation, and 24 and 48 hours sugar. For the males only 24 hour starvation was done, since most died upon 48 hour starvation; and 24 and 48 hour sugar. Briefly, flies were collected within a 24 hour period, allowed to stay together for 4 days, separated into males and females, and allowed one day recovery in normal fly food. Afterwards, the flies were placed into three conditions: fresh yeast plus PBS, PBS or PBS plus 20 % sucrose. These experiments essentially parallel the experimental set up of previous larval experiments (7). The starvation and PBS were then compared with yeast (control). Keywords: ordered

Project description:Background All animals must be able to adapt to changes in nutrient quality and supply. One striking aspect of this adaptive response is the extension of lifespan when caloric intake is reduced. These animals are also more resistant to various stresses, including starvation. We have characterized the response of adult Drosophila males and females to starvation and sugar conditions using high density microarrays. Results We have organized the regulated genes into specific metabolic and physiologic pathways. Most of the highly regulated genes have a strong sex bias. In addition to expected sex specific changes, such as egg production in females, we found unexpected biases in sensory systems, amino acid and purine metabolism, and signaling pathways. Comparison of starvation to caloric restriction revealed numerous overlaps. Conclusions Our results support the notion that starvation response shares similarities to caloric restriction. They have relevance for sex specific differences in longevity and in response to nutrient conditions which alter lifespan. Nutrient Conditions and Fly handling Larvae were kept on apple juice agar plates with yeast, and the larvae were raised at 25Â°C. For the developmental control larvae were collected 39-41 h AEL, for the developmental time-points at 51-53 hours and every 12 hours until 99-101 hours. Adult flies were kept at 25Â°C in 50 ml PS-tubes flat bottom (Greiner bio-one, Frickenhausen, Germany) on standard fly food (16.5 g yeast, 81,5 g cornmeal, 8 g agar, 100 ml sugar beet sirup and 200 ml 10% Nipagin in EtOH). Flies were collected that had hatched in 24 hours and kept in groups of ~200 flies/tube. After 5 days we separated males and females and put them in groups of 80/28 ml PS-tube. We allowed for one day CO2 recovery on standard fly food before flies were moved to 1) tubes with yeast paste on PBS soaked filter paper (control) 2) tubes with only PBS soaked filter paper (starvation) or 3) tubes with filter paper soaked in PBS with 20% Sucrose (sugar). After 24 or 48 hours flies were shock frozen in liquid nitrogen and stored at - 80Â°C for further handling. Males and females were assayed separately. For females, we performed 24 and 48 starvation, and 24 and 48 hours sugar. For the males only 24 hour starvation was done, since most died upon 48 hour starvation; and 24 and 48 hour sugar. Briefly, flies were collected within a 24 hour period, allowed to stay together for 4 days, separated into males and females, and allowed one day recovery in normal fly food. Afterwards, the flies were placed into three conditions: fresh yeast plus PBS, PBS or PBS plus 20 % sucrose. These experiments essentially parallel the experimental set up of previous larval experiments (7). The starvation and PBS were then compared with yeast (control).

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Changes in gene expression during cold exposure in Drosophila montana and D. virilis

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