Micro-dissection of Rat Brain for RNA or Protein Extraction from Specific Brain Region

Chiu, Kin; Lau, Benson Wui-Man; Lau, Ho Tak; So, Kwok-Fai; Chang, Raymond Chuen‐Chung

doi:10.3791/269

Cited by 102 publications

(82 citation statements)

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“…The rats (N = 3) were exposed to low oxygen (2% O 2 ) for 24 h while the controls (N = 3) remained in a normal environment. The frontal cortex was removed from the brains of the rats by micro-dissection, as previously described (Chiu et al, 2007) The RNAs were then purified with TRIzol ® Reagent (Invitrogen, Carlsbad, CA, USA) according to manufacturer instructions. RNase-free DNase (Promega, Madison, WI, USA) was used to remove DNA from the RNA isolation procedure.…”

Section: Ethics Statementmentioning

confidence: 99%

A microRNA-152 that targets the phosphatase and tensin homolog to inhibit low oxygen induced-apoptosis in human brain microvascular endothelial cells

Cao

et al. 2016

Genet. Mol. Res.

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ABSTRACT. Brain damage caused by perinatal asphyxia is dangerous for neonatal infants, but the mechanism by which it occurs remains elusive. In this study, microRNA-152 (miR-152) expression was induced by low oxygen levels in rat models of hypoxia brain damage, as well as in human brain microvascular endothelial cells (HBMECs) cultured in vitro. Analysis of the sequence of miR-152 revealed that the phosphatase and tensin homolog gene (PTEN) is probably the target of miR-152 both in humans and rats. When HBMECs were transfected with miR-152 mimics, PTEN expression was inhibited at both the mRNA and protein levels. Moreover, transfection with the miR-152 mimic also inhibited apoptosis induced by hypoxia. Furthermore, expression of the pro-apoptotic gene Bax was downregulated while the anti-apoptotic gene Bcl2 was upregulated after miR-152 mimic transfection. Taken together, these results indicate that miR-152 induced by hypoxia suppresses cell apoptosis and acts as a protective factor during hypoxia by repressing PTEN.

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Section: Ethics Statementmentioning

confidence: 99%

A microRNA-152 that targets the phosphatase and tensin homolog to inhibit low oxygen induced-apoptosis in human brain microvascular endothelial cells

Cao

et al. 2016

Genet. Mol. Res.

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“…Animals were sacrificed immediately following completion of anesthesia at 15:00. Within 3 mins after decapitation, the whole brain was rapidly removed and the bilateral hippocampus was directly removed from the brain, as described previously (27,28).…”

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confidence: 99%

Changes in the gene expression levels of microRNAs in the rat hippocampus by sevoflurane and propofol anesthesia

Goto

Hori

Ishikawa

et al. 2014

Molecular Medicine Reports

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Abstract. General anesthesia is commonly used in the surgical arena, but little is known regarding its influence at the genomic and molecular levels. MicroRNAs (miRNAs) belong to a new class of non-coding RNA molecules which influence cell biology. In the present study, it was hypothesized that miRNAs alter gene expression levels under general anesthesia. The aim was to compare the miRNA expression profiles in the rat hippocampus in response to anesthesia with representative volatile (sevoflurane) and intravenous (propofol) anesthetics. Wistar Rats were randomly assigned to either a 2.4% sevoflurane, 600 µg/kg/min propofol or control (without anesthetics) group. Total RNA from hippocampal samples which contained miRNA was subjected to quantitative reverse transcriptionpolymerase chain reaction and Taqman Low-Density Arrays (TLDA). A total of 373 miRNAs are associated with rats and the TLDA analysis revealed that 279 expressed miRNAs (74.8%) were expressed in all three groups. Significant differences in the levels of 33 of the 279 expressed miRNAs (11.8%) were observed among the three groups in response to the anesthetic agents, and when compared with the control group, significant differences were found in 26 of the 279 expressed miRNAs (9.3%). Following sevoflurane anesthesia, the levels of four miRNAs were significantly increased and those of 12 were significantly reduced. By contrast, following propofol anesthesia, the levels of 11 miRNAs were significantly reduced but no miRNAs exhibited significantly elevated levels. Fourteen miRNAs were significantly differentially expressed between the two anesthesia groups. In conclusion, sevoflurane and propofol exerted different effects on miRNA expression in the rat hippocampus.

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“…30 Plasma membranes from these regions were purified through sucrose gradients as described previously. 31 Briefly, previously dissected tissue was homogenized in 20 volumes of isolation buffer (10 mM Tris-HCl, pH 7.4, 0.1 g/L bacitracin, and 100 lM phenyl methyl sulfonyl fluoride) containing 8.5% sucrose using a Potter-Elvehjem homogenizer.…”

Section: Plasma Membrane Preparationmentioning

confidence: 99%

Differential Effect of Caffeine Consumption on Diverse Brain Areas of Pregnant Rats

Ballesteros-Yáñez

Castillo

Amo-Salas

et al. 2012

Journal of Caffeine Research

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Background: It has previously been shown that during gestation, the mother's brain has an increase in glial fibrillary acidic protein (GFAP)-immunoreactivity (-ir) and a decrease in the mRNA level of A1 adenosine receptor. Little is known about the A2A adenosine receptor in the maternal brain, and whether caffeine consumption throughout gestational period modifies GFAP and adenosine receptor density in specific brain areas. This study was undertaken to investigate the protein density of GFAP and adenosine receptors (A1 and A2A subtypes) in different regions of pregnant rat brain and the possible effect of caffeine on these proteins. Methods: For this purpose, we examined the GFAP-, A1-and A2A-ir in the cingulate cortex (Cg2), dentate gyrus (DG), medial preoptic area (mPOA), secondary somatosensory cortex (S2), and striatum (Str) of pregnant Wistar rats (drug-free tap water or water with 1g/L diluted caffeine). Results: We show a consistent and highly significant reduction of GFAP-ir in caffeine-treated pregnant rats in most of the areas analyzed. Our data demonstrate that caffeine consumption induces a significant increase of A2A-ir in Str. Concerning A1 receptor, the observed changes are dependent on the region analyzed; this receptor density is increased in Cg2, DG, and mPOA and decreased in the somatosensory cortex and Str. The results were confirmed by Western blotting. Conclusions: Our results suggest that chronic caffeine exposure could modify the physiolological situation of gestation by a reorganization of the neural circuits and the adenosine neuromodulator system.

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Micro-dissection of Rat Brain for RNA or Protein Extraction from Specific Brain Region

Cited by 102 publications

References 0 publications

A microRNA-152 that targets the phosphatase and tensin homolog to inhibit low oxygen induced-apoptosis in human brain microvascular endothelial cells

A microRNA-152 that targets the phosphatase and tensin homolog to inhibit low oxygen induced-apoptosis in human brain microvascular endothelial cells

Changes in the gene expression levels of microRNAs in the rat hippocampus by sevoflurane and propofol anesthesia

Differential Effect of Caffeine Consumption on Diverse Brain Areas of Pregnant Rats

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