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Microarray Analysis of Juvenile Hormone Response in Drosophila melanogaster S2 cells
A microchip array encompassing probes for 14,010 genes of Drosophila melanogaster was used to analyze the effect of juvenile hormone (JH) on genome-wide gene expression. JH is a member of a group of insect hormones involved in regulating larval development and adult reproductive processes. Total RNA...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
University of Wisconsin Library
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3014815/ https://www.ncbi.nlm.nih.gov/pubmed/20672983 http://dx.doi.org/10.1673/031.010.6601 |
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author | Willis, David K. Wang, Jun Lindholm, Joliene R. Orth, Anthony Goodman, Walter G. |
author_facet | Willis, David K. Wang, Jun Lindholm, Joliene R. Orth, Anthony Goodman, Walter G. |
author_sort | Willis, David K. |
collection | PubMed |
description | A microchip array encompassing probes for 14,010 genes of Drosophila melanogaster was used to analyze the effect of juvenile hormone (JH) on genome-wide gene expression. JH is a member of a group of insect hormones involved in regulating larval development and adult reproductive processes. Total RNA was isolated from Drosophila S2 cells after 4 hours treatment with 250 ng/ml (10R) JH III or 250 ng/ml methyl linoleate. A collection of 32 known or putative genes demonstrated a significant change with JH III treatment (r > 2.0, P ≤ 0.005). Of these, the abundance of 13 transcripts was significantly increased and 19 decreased. The expression of a subset of these loci was analyzed by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). Three loci that exhibited constant expression in the presence and absence of JH III (RP49 [FBgn0002626], FBgn0023529, and FBgn0034354) were evaluated and found to be reliable invariant reference transcripts for real-time RT-qPCR analysis using BestKeeper and geNorm software. Increased expression in presence of JH III was confirmed by real-time RTqPCR analysis. However, only one of five loci that exhibited reduced expression on microarrays could be confirmed as significantly reduced (P ≤ 0.05). Among the confirmed JH III up-regulated genes were two loci of unknown function (FBgn0040887 and FBgn0037057) and Epac, an exchange protein directly activated by cyclic AMP, a guanine nucleotide exchange factor for Rap1 small GTPase. |
format | Text |
id | pubmed-3014815 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | University of Wisconsin Library |
record_format | MEDLINE/PubMed |
spelling | pubmed-30148152012-02-09 Microarray Analysis of Juvenile Hormone Response in Drosophila melanogaster S2 cells Willis, David K. Wang, Jun Lindholm, Joliene R. Orth, Anthony Goodman, Walter G. J Insect Sci Article A microchip array encompassing probes for 14,010 genes of Drosophila melanogaster was used to analyze the effect of juvenile hormone (JH) on genome-wide gene expression. JH is a member of a group of insect hormones involved in regulating larval development and adult reproductive processes. Total RNA was isolated from Drosophila S2 cells after 4 hours treatment with 250 ng/ml (10R) JH III or 250 ng/ml methyl linoleate. A collection of 32 known or putative genes demonstrated a significant change with JH III treatment (r > 2.0, P ≤ 0.005). Of these, the abundance of 13 transcripts was significantly increased and 19 decreased. The expression of a subset of these loci was analyzed by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). Three loci that exhibited constant expression in the presence and absence of JH III (RP49 [FBgn0002626], FBgn0023529, and FBgn0034354) were evaluated and found to be reliable invariant reference transcripts for real-time RT-qPCR analysis using BestKeeper and geNorm software. Increased expression in presence of JH III was confirmed by real-time RTqPCR analysis. However, only one of five loci that exhibited reduced expression on microarrays could be confirmed as significantly reduced (P ≤ 0.05). Among the confirmed JH III up-regulated genes were two loci of unknown function (FBgn0040887 and FBgn0037057) and Epac, an exchange protein directly activated by cyclic AMP, a guanine nucleotide exchange factor for Rap1 small GTPase. University of Wisconsin Library 2010-06-15 /pmc/articles/PMC3014815/ /pubmed/20672983 http://dx.doi.org/10.1673/031.010.6601 Text en © 2010 http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Article Willis, David K. Wang, Jun Lindholm, Joliene R. Orth, Anthony Goodman, Walter G. Microarray Analysis of Juvenile Hormone Response in Drosophila melanogaster S2 cells |
title | Microarray Analysis of Juvenile Hormone Response in Drosophila melanogaster S2 cells |
title_full | Microarray Analysis of Juvenile Hormone Response in Drosophila melanogaster S2 cells |
title_fullStr | Microarray Analysis of Juvenile Hormone Response in Drosophila melanogaster S2 cells |
title_full_unstemmed | Microarray Analysis of Juvenile Hormone Response in Drosophila melanogaster S2 cells |
title_short | Microarray Analysis of Juvenile Hormone Response in Drosophila melanogaster S2 cells |
title_sort | microarray analysis of juvenile hormone response in drosophila melanogaster s2 cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3014815/ https://www.ncbi.nlm.nih.gov/pubmed/20672983 http://dx.doi.org/10.1673/031.010.6601 |
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