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DNA microarray-based analysis of voluntary resistance wheel running reveals novel transcriptome leading robust hippocampal plasticity.

Lee MC, Rakwal R, Shibato J, Inoue K, Chang H, Soya H - Physiol Rep (2014)

Bottom Line: RWR rats showed a significant decrease in average running distances although average work levels increased immensely, by about 11-fold compared to WR, resulting in muscular adaptation for the fast-twitch plantaris muscle.Interestingly, RWR down-regulated multiple inflammatory cytokines (IL1B, IL2RA, and TNF) and chemokines (CXCL1, CXCL10, CCL2, and CCR4) with the SYCP3, PRL genes, which are potentially involved in regulating hippocampal neuroplastic changes.These results provide understanding of the voluntary-RWR-related hippocampal transcriptome, which will open a window to the underlying mechanisms of the positive effects of exercise, with therapeutic value for enhancing hippocampal functions.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Exercise Biochemistry and Neuroendocrinology, Faculty of Health and Sport Sciences, University of Tsukuba, TsukubaIbaraki, Japan International Research Fellow of the Japan Society for the Promotion of Science, Tokyo, Japan.

No MeSH data available.


Related in: MedlinePlus

Workflow from sampling and grinding of the hippocampus, total RNA extraction, and DNA microarray analysis of the hippocampus. (A) The rats hippocampi were rapidly dissected and ground to a fine powder in liquid nitrogen and stored at –80°C. (B) Total RNA extraction from the finely powdered hippocampus. Total RNA quality was confirmed by both spectrophotometry and agarose‐gel electrophoresis. (C) DNA microarray chip showing the hybridized sample combinations (Sed × WR and Sed × RWR) and dye‐swap (Cy3 vs. Cy5).
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fig01: Workflow from sampling and grinding of the hippocampus, total RNA extraction, and DNA microarray analysis of the hippocampus. (A) The rats hippocampi were rapidly dissected and ground to a fine powder in liquid nitrogen and stored at –80°C. (B) Total RNA extraction from the finely powdered hippocampus. Total RNA quality was confirmed by both spectrophotometry and agarose‐gel electrophoresis. (C) DNA microarray chip showing the hybridized sample combinations (Sed × WR and Sed × RWR) and dye‐swap (Cy3 vs. Cy5).

Mentions: Between 08:00 and 12:00 on the day after the final day of exercise, the animals were decapitated using a guillotine. To measure the physiological states of the rats, adrenal glands, thymus, soleus, and plantaris muscles were sampled and weighed (wet weight) immediately after decapitation. The hippocampi were also rapidly dissected, snap frozen in liquid nitrogen, and stored at −80°C prior to further analysis. The deep‐frozen hippocampi were transferred to a precooled (in liquid nitrogen) mortar and ground with a pestle to a very fine powder with liquid nitrogen. The workflow for preparation of fine tissue powders for hippocampal gene analysis is given in Fig. 1A. The powdered samples were transferred to 2 mL Eppendorf microtubes and stored in aliquots at −80°C until used for extraction of total RNA. Total RNA was extracted from ~50 mg sample powder using the QIAGEN RNeasy Mini Kit (QIAGEN, Germantown, MD). To verify the quality of this RNA, the yield and purity were determined spectrophotometrically (IMPLEN, Germany) and visually confirmed using formaldehyde‐agarose gel electrophoresis (Fig. 1B).


DNA microarray-based analysis of voluntary resistance wheel running reveals novel transcriptome leading robust hippocampal plasticity.

Lee MC, Rakwal R, Shibato J, Inoue K, Chang H, Soya H - Physiol Rep (2014)

Workflow from sampling and grinding of the hippocampus, total RNA extraction, and DNA microarray analysis of the hippocampus. (A) The rats hippocampi were rapidly dissected and ground to a fine powder in liquid nitrogen and stored at –80°C. (B) Total RNA extraction from the finely powdered hippocampus. Total RNA quality was confirmed by both spectrophotometry and agarose‐gel electrophoresis. (C) DNA microarray chip showing the hybridized sample combinations (Sed × WR and Sed × RWR) and dye‐swap (Cy3 vs. Cy5).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4255813&req=5

fig01: Workflow from sampling and grinding of the hippocampus, total RNA extraction, and DNA microarray analysis of the hippocampus. (A) The rats hippocampi were rapidly dissected and ground to a fine powder in liquid nitrogen and stored at –80°C. (B) Total RNA extraction from the finely powdered hippocampus. Total RNA quality was confirmed by both spectrophotometry and agarose‐gel electrophoresis. (C) DNA microarray chip showing the hybridized sample combinations (Sed × WR and Sed × RWR) and dye‐swap (Cy3 vs. Cy5).
Mentions: Between 08:00 and 12:00 on the day after the final day of exercise, the animals were decapitated using a guillotine. To measure the physiological states of the rats, adrenal glands, thymus, soleus, and plantaris muscles were sampled and weighed (wet weight) immediately after decapitation. The hippocampi were also rapidly dissected, snap frozen in liquid nitrogen, and stored at −80°C prior to further analysis. The deep‐frozen hippocampi were transferred to a precooled (in liquid nitrogen) mortar and ground with a pestle to a very fine powder with liquid nitrogen. The workflow for preparation of fine tissue powders for hippocampal gene analysis is given in Fig. 1A. The powdered samples were transferred to 2 mL Eppendorf microtubes and stored in aliquots at −80°C until used for extraction of total RNA. Total RNA was extracted from ~50 mg sample powder using the QIAGEN RNeasy Mini Kit (QIAGEN, Germantown, MD). To verify the quality of this RNA, the yield and purity were determined spectrophotometrically (IMPLEN, Germany) and visually confirmed using formaldehyde‐agarose gel electrophoresis (Fig. 1B).

Bottom Line: RWR rats showed a significant decrease in average running distances although average work levels increased immensely, by about 11-fold compared to WR, resulting in muscular adaptation for the fast-twitch plantaris muscle.Interestingly, RWR down-regulated multiple inflammatory cytokines (IL1B, IL2RA, and TNF) and chemokines (CXCL1, CXCL10, CCL2, and CCR4) with the SYCP3, PRL genes, which are potentially involved in regulating hippocampal neuroplastic changes.These results provide understanding of the voluntary-RWR-related hippocampal transcriptome, which will open a window to the underlying mechanisms of the positive effects of exercise, with therapeutic value for enhancing hippocampal functions.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Exercise Biochemistry and Neuroendocrinology, Faculty of Health and Sport Sciences, University of Tsukuba, TsukubaIbaraki, Japan International Research Fellow of the Japan Society for the Promotion of Science, Tokyo, Japan.

No MeSH data available.


Related in: MedlinePlus