Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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The heat shock response of killifish (Fundulus heteroclitus): candidate gene and heterologous microarray approaches.

ABSTRACT: Subspecies of the Atlantic killifish, Fundulus heteroclitus, differ in their maximum thermal tolerance. To determine whether there is a link between the heat shock response (HSR) and maximum thermal tolerance, we exposed 20ÂºC acclimated killifish from these subspecies to a 2hr heat shock at 34ÂºC and examined gene expression during heat shock and recovery using real time quantitative PCR and a heterologous cDNA microarray designed for salmonid fishes. Keywords: Expression profiling by array Microarray analyses were performed on four individual fish per subspecies of killifish (northern and southern) prior to heat shock (control) and after 60 minutes of heat shock, hybridized (one slide per individual) against a common reference RNA pool composed of an equal amount of RNA from all samples in the analysis.

ORGANISM(S): Fundulus heteroclitus

SUBMITTER: Edward Osborne

PROVIDER: E-GEOD-18792 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Heat shock response of killifish (Fundulus heteroclitus): candidate gene and heterologous microarray approaches.

Healy Timothy M TM Tymchuk Wendy E WE Osborne Edward J EJ Schulte Patricia M PM

Physiological genomics 20100126 2

Northern and southern subspecies of the Atlantic killifish, Fundulus heteroclitus, differ in maximal thermal tolerance. To determine whether these subspecies also differ in their heat shock response (HSR), we exposed 20°C acclimated killifish to a 2 h heat shock at 34°C and examined gene expression in fish from both subspecies during heat shock and recovery using real-time quantitative PCR and a heterologous cDNA microarray designed for salmonid fishes. The heat shock proteins Hsp70-1, hsp27, an ...[more]

PMID: 20103695

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Project description:Given the overwhelming evidence that symbiont genotypes differentially affect host processes such as growth, bleaching susceptibility, and nutrient acquisition, we set out to measure gene expression differences in fragments of Montastraea faveolata harboring two different clades of Symbiodinium. On the reefs near Puerto Morelos, MÃ©xico, colonies of M. faveolata are known to shift algal symbiont clade with depth, often associating with clade A at the top, clade B in the middle, and clade C near the bottom of the colony. By measuring photosynthetic efficiency and gene expression in control and heat-stressed fragments containing either clade B, clade C, or a mix of both, we found that: 1) the algal response to thermal stress is due to both host and algal factors; 2) fragments of M. faveolata express different genes in response to sub-bleaching thermal stress depending on algal genotype; 3) the overall effect of heat stress on coral gene expression is less significant than the effect of housing different zooxanthellae types. Overall, we present convincing evidence that different Symbiodinium clades may be functionally distinct, which in turn, greatly influences host gene expression. Six fragments (9.5 Â± 3.5cm2) from the top (2.7m), middle (3.7m), and bottom (5.2m) of one massive colony of Montastraea faveolata were collected with a hammer and chisel at âLa Bocanaâ reef near Puerto Morelos, Quintana Roo, Mexico on 31 July 2007. The fragments were divided evenly between two aquaria (50L) that received a constant flow of seawater (~0.64L/min). Each aquarium was fit with a water pump connected to a spray-bar to provide constant water movement and aeration. Both aquaria were placed in a common pond with flowing water to buffer diurnal temperature fluctuations, and both aquaria were exposed to shaded ambient light. All coral fragments were mounted on plasticene and kept at a depth of ~7cm. From 10 to 19 August 2007 (acclimation #1), both aquaria received an average water temperature of 27.9 Â± 0.6oC (as recorded by HOBO Light/Temperature Data Loggers by Onset Corp.). Effective quantum yield (ÎF/Fmâ) measurements were taken at noon, and maximum quantum yield (Fv/Fm) measurements were taken at dusk for all 18 coral fragments using a DIVING-PAM (Walz) beginning on 11 August. Photosynthetically active radiation (PAR) was measured at noon and averaged 318 Â± 129 umol/m2/sec. From 20 to 21 August 2007, all coral fragments were brought inside during the passage of Hurricane Dean. On 22 August, the experiment was reconstituted, and a second acclimation period began on 23 August and lasted until 1 September. During this time, both aquaria received water at an average temperature of 28.5 Â± 0.8oC, and PAR at an average of 371 Â± 169 umol/m2/sec. On the night of 1 September, one 200-Watt aquarium heater was turned on in the treatment aquarium, and a second heater was turned on 3 days later. During the thermal stress experiment, the control aquarium received an average water temperature of 28.8 Â± 1.2oC; the heated aquarium, 31.5 Â± 1.1oC. PAR present during the thermal stress experiment averaged 420 Â± 152 uE. On the night of 7 September, all fragments were frozen in liquid nitrogen. Eighteen total microarray hybridizations were performed. A reference RNA sample was created by pooling RNA from all 18 prepâs. All control and heat-stressed samples were competitively hybridized against the reference RNA to the M. faveolata microarray (1,310 features).

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Dataset Information

The heat shock response of killifish (Fundulus heteroclitus): candidate gene and heterologous microarray approaches.

Publications

Heat shock response of killifish (Fundulus heteroclitus): candidate gene and heterologous microarray approaches.

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