Unknown,Transcriptomics,Genomics,Proteomics

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Gene Expression Changes Associated with Respiratory Muscle Hypertrophy after Intermittent Transient Tracheal Occlusions

ABSTRACT: The effect of intermittent transient tracheal occlusion (ITTO) on gene expression was compared in the diaphragm and parasternal intercostals. We hypothesized that genes responsible for protein metabolism, myogenic activation, neuromuscular transmission, and inflammatory modulation would be altered in animals exposed to this rodent model of respiratory load compensation rehabilitation. Eight adult male Sprague-Dawley rats underwent placement of a tracheal occluder cuff and were divided into ITTO (n=4) or SHAM (n=4) treatment groups. Animals received 10 days of their assigned treatments. The day following the last treatment session, the animals were euthanized and the diaphragm and parasternals extracted. Messenger RNA was profiled with microarray analysis (Amadid: 028279, Agilent Technologies, Palo Alto, CA). Results were further analyzed with immunofluorescent examination of cross-sectional area and with quantitative reverse transcriptase polymerase chain reaction on a subset of genes exhibiting differential expression. The cross-sectional area of type IIx/b fibers was larger in the muscles of ITTO-trained animals. Microarray analysis identified 650 genes in the diaphragm and 1,181 genes in the intercostals that were differentially expressed between the groups (p<.01). In both muscles, biological processes for cell adhesion and chemical stimulus for the detection of olfaction biological processes were over-expressed. Additionally, we found significant differential expression of the g-protein signaling pathway and protein-amino acid phosphorylation in the intercostals. The results provide insight into neuromuscular plasticity associated with respiratory load compensation rehabilitation. Four experimental animals that received tracheal occlusions and four surgical control animals were used. Intercostal and diaphragm muscles were excised and analyzed separately.

ORGANISM(S): Rattus norvegicus

SUBMITTER: Vipa Bernhardt

PROVIDER: E-GEOD-29574 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The thalamus may be the critical brain area involved in sensory gating and the relay of respiratory mechanical information to the cerebral cortex for the conscious awareness of breathing. We hypothesized that respiratory mechanical stimuli in the form of tracheal occlusions would modulate the gene expression profile of the thalamus. Specifically, it was reasoned that conditioning to the respiratory loading would induce a state change in the medial thalamus consistent with a change in sensory gating and the activation of molecular pathways associated with learning and memory. In addition, respiratory loading is stressful and thus should elicit changes in gene expressions related to stress, anxiety, and depression. Rats were instrumented with inflatable tracheal cuffs. Following surgical recovery, they underwent ten days (5 days/week) of transient tracheal occlusion conditioning. On day 10, the animals were sacrificed and the brains removed. The medial thalamus was dissected and microarray analysis of gene expression performed. Tracheal obstruction conditioning modulated a total of 661 genes (p < 0.05, log2 fold change ≥ 0.58), 250 genes were down-regulated and 411 up-regulated. There was a significant down-regulation of GAD1, GAD2 and HTR1A, HTR2A genes. CCK, PRKCG, mGluR4, and KCJN9 genes were significantly up-regulated. Some of these genes have been associated with anxiety and depression, while others have been shown to play a role in switching between tonic and burst firing modes in the thalamus and thus may be involved in gating of the respiratory stimuli. Furthermore, gene ontology and pathway analysis showed a significant modulation of learning and memory pathways. These results support the hypothesis that the medial thalamus is involved in the respiratory sensory neural pathway due to the state change of its gene expression profile following repeated tracheal occlusions. Four experimental animals that received tracheal occlusions and four surgical control animals were used. A reference design was used to compare all samples.

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Gene Expression Changes Associated with Respiratory Muscle Hypertrophy after Intermittent Transient Tracheal Occlusions

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