U6 snRNA is a Suitable Endogenous Control for microRNA-124 and -134 in Cultured Rat Hippocampal Neurons

Ching, Ai Sze; Ahmad‐Annuar, Azlina

doi:10.17576/jsm-2015-4410-14

Cited by 2 publications

(1 citation statement)

References 29 publications

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“…Complementary DNAs (cDNAs) were obtained according to the manufacturer’s instructions using the commercial kit (qScriptTM microRNA synthesis kit-VWR) and used in the BIORAD-CFX connect real-time PCR system with SYBR Green probe technology (Perfecta SYBR Green FastMix Reaction Mixes-VWR) for miRNA expression analysis. Cel-miR-39-3p was used as a spike-in control for normalization (Faraldi et al ., 2019), and U6 snRNA (Ching and Ahmad-Annuar, 2015) was used as an endogenous control for miRNA quantification. According to the results of the microarray analysis, the validation of the genes with altered activities was performed by real-time PCR (BIORAD-CFX connect) method.…”

Section: Methodsmentioning

confidence: 99%

Thymoquinone ameliorates symptoms of Parkinson’s disease in a 6-OHDA rat model by downregulation of miR-204-3p

Pala,

Meral,

Pala Acikgoz

et al. 2024

Behavioural Pharmacology

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microRNAs (miRNAs) play a significant role in the pathophysiology of Parkinson’s disease. In this study, we evaluated the neuroprotective effect of thymoquinone on the expression profiles of miRNA and cognitive functions in the 6-hydroxydopamine (6-OHDA)-induced Parkinson’s model. Male adult Wistar albino rats (200–230 g, n = 36) were randomly assigned to six groups: Sham, thymoquinone (10 mg/kg, p.o.), 6-OHDA, 6-OHDA + thymoquinone (10 mg/kg), 6-OHDA + thymoquinone (20 mg/kg), and 6-OHDA + thymoquinone (50 mg/kg). Behavioral changes were detected using the open field and the elevated plus maze tests. The mature 728 miRNA expressions were evaluated by miRNA microarray (GeneChip miRNA 4.0). Ten miRNAs were selected (rno-miR-212-5p, rno-miR-146b-5p, rno-miR-150-5p, rno-miR-29b-2-5p, rno-miR-126a-3p, rno-miR-187-3p, rno-miR-34a-5p, rno-miR-181d-5p, rno-miR-204-3p, and rno-miR-30c-2-3p) and confirmed by real-time PCR. Striatum samples were stained with hematoxylin-eosin to determine the effect of dopaminergic lesions. One-way ANOVA test and independent sample t-test were used for statistical analyses. rno-miR-204-3p was upregulated at 6-OHDA and downregulated at the 50 mg/kg dose of thymoquinone. In conclusion, thymoquinone at a dose of 50 mg/kg ameliorates symptoms of Parkinson’s disease in a 6-OHDA rat model by downregulation of miR-204-3p. Also, the results showed that thymoquinone can improve locomotor activity and willing exploration and decreased anxiety. Therefore, thymoquinone can be used as a therapeutic agent.

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Section: Methodsmentioning

confidence: 99%

Thymoquinone ameliorates symptoms of Parkinson’s disease in a 6-OHDA rat model by downregulation of miR-204-3p

Pala,

Meral,

Pala Acikgoz

et al. 2024

Behavioural Pharmacology

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Identification of endogenous microRNA references in porcine serum for quantitative real-time PCR normalization

2018

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MicroRNAs (miRNAs) are evolutionarily conserved small non-coding RNAs that regulate the expression of genes, and they affect important biological and physiological states. Circulating miRNAs in blood are useful markers of metabolism and economic traits. Expression levels of circulating miRNAs have been estimated using quantitative real-time PCR (qPCR). Proper normalization is critical for accurate miRNA expression analysis. However, there is no study which systematically presented endogenous reference genes for evaluating circulating miRNA expression in pigs. In this study, ten porcine miRNAs (let-7a, miR-16, miR-17, miR-23a, miR-26a, miR-93, miR-103, miR-107, miR-127 and miR-191), based on the literature, were chosen as candidate reference miRNAs in serum. We evaluated the expression stability value of these miRNAs in Berkshire, Duroc, Landrace and Yorkshire pigs using geNorm and NormFinder. We determined the optimal combination of reference miRNAs for qPCR experiments: miR-127 and miR-17 in Berkshire pigs; miR-127 and miR-93 in Duroc and Landrace pigs; miR-127 and miR-16 in Yorkshire pigs. miR-127 was the best reference gene in pigs, regardless of the breed. Our study is crucial for the discovery of novel biomarkers in pigs. The reference miRNAs presented in this study could be used as appropriate reference genes for the measurement of circulating miRNA levels in studies of physiological blood metabolites.

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U6 snRNA is a Suitable Endogenous Control for microRNA-124 and -134 in Cultured Rat Hippocampal Neurons

Abstract: As researchers seek to determine the cellular mechanisms underlying biological processes

Cited by 2 publications

References 29 publications

Thymoquinone ameliorates symptoms of Parkinson’s disease in a 6-OHDA rat model by downregulation of miR-204-3p

Thymoquinone ameliorates symptoms of Parkinson’s disease in a 6-OHDA rat model by downregulation of miR-204-3p

Identification of endogenous microRNA references in porcine serum for quantitative real-time PCR normalization

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