Extraction and Detection of mRNA from Horsehair

Sato, Tetsuo; Sato, Go; Shoji, Youko; Itou, Takuya; Sakai, Takeo

doi:10.1292/jvms.68.503

Cited by 6 publications

(6 citation statements)

References 41 publications

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“…In this experiment, we used the residual mRNA from human stratum corneum to detect the expression of NDUF mRNA. Surprisingly, it has been reported that small amounts of RNA were present in horsehair and human hair 14,26 . Therefore, we tried to detect the residual mRNA in human stratum corneum.…”

Section: Discussionmentioning

confidence: 99%

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Decrease in nicotinamide adenine dinucleotide dehydrogenase is related to skin pigmentation

Nakama¹,

Murakami²,

Tanaka³

et al. 2012

J of Cosmetic Dermatology

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Skin pigmentation is caused by various physical and chemical factors. It might also be influenced by changes in the physiological function of skin with aging. Nicotinamide adenine dinucleotide (NADH) dehydrogenase is an enzyme related to the mitochondrial electron transport system and plays a key role in cellular energy production. It has been reported that the functional decrease in this system causes Parkinson's disease. Another study reports that the amount of NADH dehydrogenase in heart and skeletal muscle decreases with aging. A similar decrease in the skin would probably affect its physiological function. However, no reports have examined the age-related change in levels of NADH dehydrogenase in human skin. In this study, we investigated this change and its effect on skin pigmentation using cultured human epidermal keratinocytes. The mRNA expression of NDUFA1, NDUFB7, and NDUFS2, subunits of NADH dehydrogenase, and its activity were significantly decreased in late passage keratinocytes compared to early passage cells. Conversely, the mRNA expression of melanocyte-stimulating cytokines, interleukin-1 alpha and endothelin 1, was increased in late passage cells. On the other hand, the inhibition of NADH dehydrogenase upregulated the mRNA expression of melanocyte-stimulating cytokines. Moreover, the level of NDUFB7 mRNA was lower in pigmented than in nonpigmented regions of skin in vivo. These results suggest the decrease in NADH dehydrogenase with aging to be involved in skin pigmentation.

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Section: Discussionmentioning

confidence: 99%

“…Surprisingly, it has been reported that small amounts of RNA were present in horsehair and human hair. 14,26 Therefore, we tried to detect the residual mRNA in human stratum corneum. This method is better than invasive methods such as skin biopsy and the suctioning of blisters.…”

Section: Discussionmentioning

confidence: 99%

Decrease in nicotinamide adenine dinucleotide dehydrogenase is related to skin pigmentation

Nakama¹,

Murakami²,

Tanaka³

et al. 2012

J of Cosmetic Dermatology

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“…Genes involved in the differences among the various phenotypes of cells, tissues, organs, and bodies may be elucidated by analyzing their RNA profiles. However, the analysis of RNA profiles using mRNA extracted from hair shafts is considered to be very difficult 14 …”

Section: Discussionmentioning

confidence: 99%

“…However, the analysis of RNA profiles using mRNA extracted from hair shafts is considered to be very difficult. 14 As Fig. 1 shows, we were able to amplify b-actin and GAPDH mRNAs, ubiquitously expressed in various cells, extracted from the scalp hair shafts of three donors by RT-PCR.…”

Section: Discussionmentioning

confidence: 99%

Presence of amplifiable mRNA in acellular hair shafts: utilization to analyze gene expression profiles of black and white hairs

Tochio¹,

Tanaka²,

Nakata³

et al. 2011

Int J Dermatology

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“…Collected fragments were immersed in 800 μl of ISOGEN Reagent (Nippon Gene, Toyama, Japan) in tubes and stirred (15 s × 2 times) using the sonication device, Bioruptor UCD-250 (Cosmo Bio, Tokyo, Japan). Next, RNA was purified from hair lysates with the ISOGEN Kit as described previously [21]. Briefly, tubes were kept at room temperature for 5 min, followed by addition of 200 μl of chloroform.…”

Section: Rna Extractionmentioning

confidence: 99%

Genetic analysis of the human hair roots as a tool for spaceflight experiments

Terada¹,

Seki²,

Higashibata³

et al. 2013

ABB

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The use of hair roots as experimental samples has been a research focus for understanding the effects of spaceflight on astronauts, because it has many advantages, one of which is the fact that hair matrix cells actively divide in a hair follicle and sensitively reflect the physical conditions of the human body. In 2009, a research program focusing on the analysis of astronauts' hairs was initiated to examine the effects of long-term spaceflight on the gene expression and mineral metabolism in the human body. Since the number of samples per astronaut is limited to 5 strands of hairs at each sampling point, due to the ethical viewpoint of astronauts or limited resources in space, it is important to develop an effective method for the molecular analysis of small amounts of hair roots. In this study, mRNA successfully extracted from 1, 5, and 10 hair follicles was amplified and subjected to the DNA microarray analysis to compare the gene expression within subjects. The results indicated that (1) it was possible to perform the genetic analysis on hair samples stored at −80˚C, even without a fixation buffer and (2) the newly modified method of mRNA extraction and analysis was effective in detecting differential gene expression in samples containing only 5 hairs. In conclusion, RNA was efficiently extracted from 5 hair roots, which is the same number of hair roots used in the space experiment; therefore, this method can be applied to genetically analyze astronauts' hair amples. s

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Extraction and Detection of mRNA from Horsehair

Cited by 6 publications

References 41 publications

Decrease in nicotinamide adenine dinucleotide dehydrogenase is related to skin pigmentation

Decrease in nicotinamide adenine dinucleotide dehydrogenase is related to skin pigmentation

Presence of amplifiable mRNA in acellular hair shafts: utilization to analyze gene expression profiles of black and white hairs

Genetic analysis of the human hair roots as a tool for spaceflight experiments

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