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Anesthesia for euthanasia influences mRNA expression in healthy mice and after traumatic brain injury.

ABSTRACT: Tissue sampling for gene expression analysis is usually performed under general anesthesia. Anesthetics are known to modulate hemodynamics, receptor-mediated signaling cascades, and outcome parameters. The present study determined the influence of anesthetic paradigms typically used for euthanization and tissue sampling on cerebral mRNA expression in mice. Naïve mice and animals with acute traumatic brain injury induced by controlled cortical impact (CCI) were randomized to the following euthanasia protocols (n=10-11/group): no anesthesia (NA), 1?min of 4?vol% isoflurane in room air (ISO), 3?min of a combination of 5?mg/kg midazolam, 0.05?mg/kg fentanyl, and 0.5?mg/kg medetomidine intraperitoneally (COMB), or 3?min of 360?mg/kg chloral hydrate intraperitoneally (CH). mRNA expression of actin-1-related gene (Act1), FBJ murine osteosarcoma viral oncogene homolog B (FosB), tumor necrosis factor alpha (TNF?), heat shock protein beta-1 (HspB1), interleukin (IL)-6, tight junction protein 1 (ZO-1), IL-1ß, cyclophilin A, micro RNA 497 (miR497), and small cajal body-specific RNA 17 were determined by real-time polymerase chain reaction (PCR) in hippocampus samples. In naïve animals, Act1 expression was downregulated in the CH group compared with NA. FosB expression was downregulated in COMB and CH groups compared with NA. CCI reduced Act1 and FosB expression, whereas HspB1 and TNF? expression increased. After CCI, HspB1 expression was significantly higher in ISO, COMB, and CH groups, and TNF? expression was elevated in ISO and COMB groups. MiR497, IL-6, and IL-1ß were upregulated after CCI but not affected by anesthetics. Effects were independent of absolute mRNA copy numbers. The data demonstrate that a few minutes of anesthesia before tissue sampling are sufficient to induce immediate mRNA changes, which seem to predominate in the early-regulated gene cluster. Anesthesia-related effects on gene expression might explain limited reproduciblity of real-time PCR data between studies or research groups and should therefore be considered for quantitative PCR data.

SUBMITTER: Staib-Lasarzik I

PROVIDER: S-EPMC4170812 | biostudies-literature | 2014 Oct

REPOSITORIES: biostudies-literature

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Anesthesia for euthanasia influences mRNA expression in healthy mice and after traumatic brain injury.

Staib-Lasarzik Irina I Kriege Oliver O Timaru-Kast Ralph R Pieter Dana D Werner Christian C Engelhard Kristin K Thal Serge C SC

Journal of neurotrauma 20140812 19

Tissue sampling for gene expression analysis is usually performed under general anesthesia. Anesthetics are known to modulate hemodynamics, receptor-mediated signaling cascades, and outcome parameters. The present study determined the influence of anesthetic paradigms typically used for euthanization and tissue sampling on cerebral mRNA expression in mice. Naïve mice and animals with acute traumatic brain injury induced by controlled cortical impact (CCI) were randomized to the following euthana ...[more]

PMID: 24945082

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Anesthesia for euthanasia influences mRNA expression in healthy mice and after traumatic brain injury.

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Anesthesia for euthanasia influences mRNA expression in healthy mice and after traumatic brain injury.

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