Project description:Diapause and diapause-like states constitute natural, inducible, and evolutionarily conserved examples of lifespan plasticity that are well-suited to represent physiologically accurate models for longevity. We integrated ATAC-seq and RNA-seq technologies to probe the transcriptional regulatory pathways controlling lifespan plasticity in diapause.
Project description:Diapause and diapause-like states constitute natural, inducible, and evolutionarily conserved examples of lifespan plasticity that are well-suited to represent physiologically accurate models for longevity. We integrated ATAC-seq and RNA-seq technologies to probe the transcriptional regulatory pathways controlling lifespan plasticity in diapause.
Project description:The most common ladybird beetle, Coccinella septempunctata L., is an excellent predator of crop pests such as aphids and white flies, and it shows a wide range of adaptability, a large appetite and a high reproductive ability. In this study, we collected female adults in three different states, i.e., non-diapause, diapause and diapause termination, for transcriptome sequencing. The experimental insects consisted of three different states as follows: Non-diapause female insects were reared at 24±1°C, with a RH of 70±10% and a 16:8 h light: dark (L: D) photoperiod and collected after their first oviposition. Female adults in diapause were reared at 18±1°C at an RH of 70±10% and a 10:14-h (L:D) photoperiod. The experimental diapause insects were collected after 30 days. Diapause-terminated adults were transferred to another climatic cabinet with the 30-day diapause insects and reared under the same conditions as the non-diapause insects. After their first oviposition, the female insects were collected and stored at -80°C. Three biological replicates per treatment (non-diapause, diapause, diapause-terminated) were sequenced using Illumina HiSeq 2500.
Project description:Investigating essential physiological processes in diapausing mites by analyzing genome wide gene expression changes using custom-built microarray. We investigated the molecular biology of facultative reproductive diapause in the chelicerate Tetranychus urticae (Acari: Tetranychidae) by analyzing genome-wide gene expression differences in diapausing and non diapausing T. urticae, using an Agilent custom-built two color gene expression microarray. Analysis of this dataset showed that a remarkable number, 11% of the total number of predicted T. urticae genes, were differentially expressed. Gene Ontology analysis revealed that many metabolic pathways were affected in diapausing females. Genes related to digestion and detoxification, cryo-protection, carotenoid synthesis and the organization of the cytoskeleton were profoundly influenced by the state of diapause. We also further confirmed the importance of horizontally transferred carotenoid synthesis genes in diapause and different color morphs of T. urticae. We made one comparison: diapausing mites (DIA) vs non-diapausing mites (NON-DIA), in 4 replicates. Both types of mites belonged to the T. urticae LS-VL strain. In this strain, approximately 30% of mites enter diapause under the experimental conditions applied. Hence, we were able to sample RNA of mites with similar genetic background that were reared under identical environmental conditions. The labeled cRNA samples were pooled and hybdrized to a custom Sureprint genome wide G3 Gene Expression 8x60K microarray. Data was normalized by Agilent Feature Extraction software (using protocol GE2_107_SEP09). Genespring software (Agilent technologies) was used for the statistical analysis of the data.
Project description:Most northern insect species experience a period of developmental arrest, diapause, which enables them to survive over the winter and postpone reproduction until favorable conditions. We studied the timing of reproductive diapause and its long-term effects on the cold tolerance of Drosophila montana, D. littoralis and D. ezoana females in seasonally varying environmental conditions. At the same time we traced expression levels of 219 genes in D. montana using a custom-made microarray. We show that the seasonal switch to reproductive diapause occurs over a short time period, and that overwintering in reproductive diapause has long-lasting effects on cold tolerance. Some genes, such as Hsc70, Jon25bi and period, were upregulated throughout the diapause, while others, including regucalcin, couch potato and Thor, were upregulated only at its specific phases. Some of the expression patterns induced during the sensitive stage, when the females either enter diapause or not, remained induced regardless of the later conditions. qPCR analyses confirmed the findings of the microarray analysis in D. montana and revealed similar gene expression changes in D. littoralis and D. ezoana. The present study helps to achieve a better understanding of the genetic regulation of diapause and of the plasticity of seasonal responses in general. Custom made DNA microarray for Drosophila montana and D. virilis. Current experiment includes 8 samples (7 to 250 days old diapausing or non-diapausing D. montana females) with two or three biological replicates
Project description:The most common ladybird beetle, Coccinella septempunctata L., is an excellent predator of crop pests such as aphids and white flies, and it shows a wide range of adaptability, a large appetite and a high reproductive ability. In this study, we collected female adults in three different states, i.e., non-diapause, diapause and diapause termination, for transcriptome sequencing.
Project description:Regulation of embryonic diapause, dormancy that interrupts the tight connection between develop- mental stage and time, is still poorly understood. Here, we characterize the transcriptional and metabolite profiles of mouse diapause embryos and identify unique gene expression and metabolic signatures with activated lipolysis, glycolysis, and metabolic pathways regulated by AMPK. Lipolysis is increased due to mTORC2 repression, increasing fatty acids to support cell survival. We further show that starvation in pre-implantation ICM-derived mouse ESCs induces a reversible dormant state, transcriptionally mimicking the in vivo diapause stage. During starvation, Lkb1, an upstream kinase of AMPK, represses mTOR, which induces a revers- ible glycolytic and epigenetically H4K16Ac-negative, diapause-like state. Diapause furthermore activates expression of glutamine transporters SLC38A1/2. We show by genetic and small molecule inhibitors that glutamine transporters are essential for the H4K16Ac-negative, diapause state. These data sug- gest that mTORC1/2 inhibition, regulated by amino acid levels, is causal for diapause metabolism and epigenetic state.
Project description:Gene expression microarray exprement comparing differential expression between: 1) Early Diapause (ED), 2) Late Diapause (LD), 3) Non-Diapause (ND), 4) Hexane-induced diapause break (HEX). Four phenotypes (ED,ND,LD,HEX), four replicate pools of four individuals (four individuals in each replicate) in each phenotype, four competitve hybs comparing each phenotype to every other phenotype.
Project description:Embryonic diapause is a widely occurring evolutionary adaptation phenomenon in animals. Artemia is one of the classic animal models for diapause research. The current studies of Artemia diapause mainly focus on the induction and maintenance of the embryonic diapause, but there is little research on the molecular regulatory mechanism of Artemia embryonic diapause termination (EDT) and embryonic reactivation. Here the gene expression of Artemia cyst in diapause stage and 5 hours after embryonic diapause termination (EDT), which is in post-diapause stage were tested by ATAC-seq to analyze the mechanism of signal regulation involved in Artemia EDT at the molecular level.
Project description:Embryonic diapause is a widely occurring evolutionary adaptation phenomenon in animals. Artemia is one of the classic animal models for diapause research. The current studies of Artemia diapause mainly focus on the induction and maintenance of the embryonic diapause, but there is little research on the molecular regulatory mechanism of Artemia embryonic diapause termination (EDT) and embryonic reactivation. Here the gene expression of Artemia cyst in diapause stage and 5 hours after embryonic diapause termination (EDT), which is in post-diapause stage were tested by RNA-seq to analyze the mechanism of signal regulation involved in Artemia EDT at the molecular level.