Slc7a11 Modulated by POU2F1 is Involved in Pigmentation in Rabbit

Chen, Yang; Hu, Shuaishuai; Mu, Lin; Zhao, Bohao; Wang, Manman; Yang, Neng; Bao, Guolian; Zhu, Cigen; Wu, Xinsheng

doi:10.3390/ijms20102493

Cited by 19 publications

(22 citation statements)

References 37 publications

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“…Recently, SLC7A11 was found to be modulated by POU class 2 homeobox 1 gene (POU2F1) in rabbit pigmentation [14]. POU2F1, also known as OCT1, OTF1, or oct-1B [15], is a potent regulator of stress responses, metabolism, and tumorigenicity and is itself regulated by phosphorylation, ubiquitination, O-GlcNAcylation, and other mechanisms [16].…”

Section: Introductionmentioning

confidence: 99%

“…Recent studies into POU2F1 have focused on its impacts on cancers and tumors [17][18][19][20], especially hepatocellular carcinoma [21,22]. Furthermore, POU2F1 was identified as a transcription factor that binds to the promoter region of SLC7A11 via two binding sites in the Rex rabbit [14]. However, little is known about the role of POU2F1 in fur color formation in mammals, and the effects of the gene on the fur color of the Rex rabbit remain unclear.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of POU2F1 Gene and Its Potential Impact on the Expression of Genes Involved in Fur Color Formation in Rex Rabbit

Yang

Zhao

et al. 2020

Genes

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The naturally colorful fur of the Rex rabbit is becoming increasingly popular in the modern textile market. Our previous study found that POU class 2 homeobox 1 gene (POU2F1) potentially affects the expression of genes involved in fur color formation in the Rex rabbit, but the function and regulation of POU2F1 has not been reported. In this study, the expression patterns of POU2F1 in Rex rabbits of various colors, as well as in different organs, were analyzed by RT-qPCR. Interference and overexpression of POU2F1 were used to identify the potential effects of POU2F1 on other genes related to fur color formation. The results show that the levels of POU2F1 expression were significantly higher in the dorsal skin of the brown and protein yellow Rex rabbits, compared with that of the black one. POU2F1 mRNAs were widespread in the tissues examined in this study and showed the highest level in the lungs. By transfecting rabbit melanocytes with an POU2F1-overexpression plasmid, we found that the POU2F1 protein was located at the nucleus, and the protein showed the classic characteristics of a transcription factor. In addition, abnormal expression of POU2F1 significantly affected the expression of pigmentation-related genes, including SLC7A11, MITF, SLC24A5, MC1R, and ASIP, revealing the regulatory roles of POU2F1 on pigmentation. The results provide the basis for further exploration of the role of POU2F1 in fur color formation of the Rex rabbit.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Characterization of POU2F1 Gene and Its Potential Impact on the Expression of Genes Involved in Fur Color Formation in Rex Rabbit

Yang

Zhao

et al. 2020

Genes

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“…Rabbit melanocytes were isolated from the dorsal skin tissue of Rex rabbits as described by Chen et al (2019). The cells were cultured in M254 medium (Gibco, Carlsbad, CA, USA) supplemented with 1% human melanocyte growth supplement-2 (HMGS-2, Gibco, Carlsbad, CA, USA) and maintained in an incubator at 37 • C in an atmosphere containing 5% CO 2 .…”

Section: Rabbit Melanocyte Culture and Overexpression Transfection Ofmentioning

confidence: 99%

“…Cell proliferation was evaluated using a Cell Counting Kit-8 assay (Vazyme, Nanjing, China), in accordance with the manufacturer's instructions, as described previously by Zhao et al (2019). Cells were harvested and seeded in 96-well plates after 16 h transfection, then the OD at 450 nm of each well was measured at 0, 24, 48 and 72 h using an Infinite M200 Pro spectrophotometer (Tecan, Männedorf, Switzerland).…”

Section: Cell Proliferation Assaymentioning

confidence: 99%

KIT is involved in melanocyte proliferation, apoptosis and melanogenesis in the Rex Rabbit

Chen

Zhao

et al. 2020

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Background Melanocytes play an extremely important role in the process of skin and coat colors in mammals which is regulated by melanin-related genes. Previous studies have demonstrated that KIT is implicated in the process of determining the color of the coat in Rex rabbits. However, the effect of KIT on the proliferation and apoptosis of melanocytes and melanogenesis has not been clarified. Methods The mRNA and protein expression levels of KIT were quantified in different coat colored rabbits by qRT-PCR and a Wes assay. To identify whether KIT functions by regulating of melanogenesis, KIT overexpression and knockdown was conducted in melanocytes, and KIT mRNA expression and melanin-related genes TYR, MITF, PMEL and DCT were quantified by qRT-PCR. To further confirm whether KIT influences melanogenesis in melanocytes, melanin content was quantified using NaOH lysis after overexpression and knockdown of KIT. Melanocyte proliferation was estimated using a CCK-8 assay at 0, 24, 48 and 72 h after transfection, and the rate of apoptosis of melanocytes was measured by fluorescence-activated cell sorting. Results KITmRNA and protein expression levels were significantly different in the skin of Rex rabbits with different color coats (P < 0.05), the greatest levels observed in those with black skin. The mRNA expression levels of KIT significantly affected the mRNA expression of the pigmentation-related genes TYR, MITF, PMEL and DCT (P < 0.01). Melanin content was evidently regulated by the change in expression patterns of KIT (P < 0.01). In addition, KIT clearly promoted melanocyte proliferation, but inhibited apoptosis. Conclusions Our results reveal that KIT is a critical gene in the regulation of melanogenesis, controlling proliferation and apoptosis in melanocytes, providing additional evidence for the mechanism of pigmentation of animal fur.

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“…Rabbit melanocytes were isolated from the dorsal skin tissue of Rex rabbits as described by Chen et al (Chen et al, 2019). The cells were cultured in M254 medium (Gibco, Carlsbad, CA, USA) supplemented with 1% human melanocyte growth supplement-2 (HMGS-2, Gibco, Carlsbad, CA, USA) and maintained in an incubator at 37℃ in an atmosphere containing 5% CO 2 .…”

Section: Rabbit Melanocyte Culture and Overexpression Transfection Ofmentioning

confidence: 99%

Peer Review #3 of "KIT is involved in melanocyte proliferation, apoptosis and melanogenesis in the Rex Rabbit (v0.3)"

Bourneuf¹

2020

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Background: Melanocytes play an extremely important role in the process of skin and coat colors in mammals which is regulated by melanin-related genes. Previous studies have demonstrated that KIT is implicated in the process of determining the color of the coat in Rex rabbits. However, the effect of KIT on the proliferation and apoptosis of melanocytes and melanogenesis has not been clarified. Methods: The mRNA and protein expression levels of KIT were quantified in different coat colored rabbits by qRT-PCR and a Wes assay. To identify whether KIT functions by the regulation of melanogenesis, KIT overexpression and knockdown was conducted in melanocytes, and KIT mRNA expression and that of the melanin-related genes TYR, MITF, PMEL and DCT were quantified by qRT-PCR. To further confirm whether KIT influences melanogenesis in melanocytes, the levels of melanin were quantified using NaOH lysis after overexpression and knockdown of KIT. Melanocyte proliferation was estimated using a CCK-8 assay at 0, 24, 48 and 72 h after transfection, and the rate of apoptosis of melanocytes wasmeasured by fluorescenceactivated cell sorting. Results: KIT mRNA and protein expression levels were significantly different in the skin of Rex rabbits with different color coats (P < 0.05), the greatest levels observed in those with black skin. The mRNA expression levels of KIT significantly affected the mRNA expression of the pigmentation-related genes TYR, MITF, PMEL and DCT (P < 0.01). Melanin content was evidently regulated by the change in expression patterns of KIT (P < 0.01). In addition, KIT clearly promoted melanocyte proliferation, but inhibited apoptosis. Conclusions: Our results reveal that KIT is a critical gene in the regulation of melanogenesis, controlling proliferation and apoptosis in melanocytes, providing additional evidence for the mechanism of pigmentation of animal fur.

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Slc7a11 Modulated by POU2F1 is Involved in Pigmentation in Rabbit

Cited by 19 publications

References 37 publications

Characterization of POU2F1 Gene and Its Potential Impact on the Expression of Genes Involved in Fur Color Formation in Rex Rabbit

Characterization of POU2F1 Gene and Its Potential Impact on the Expression of Genes Involved in Fur Color Formation in Rex Rabbit

KIT is involved in melanocyte proliferation, apoptosis and melanogenesis in the Rex Rabbit

Peer Review #3 of "KIT is involved in melanocyte proliferation, apoptosis and melanogenesis in the Rex Rabbit (v0.3)"

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