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1.
Anim Biotechnol ; 34(9): 4559-4568, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36752211

RESUMO

The Yangtze River Delta white goats are the sole goat breed producing brush hair of high quality. Owing to the particularities of its wool production, a higher demand is placed on breeding efforts for this animal. Studies on the developmental mechanisms of the aligned hair follicle stem cells (HFSCs) provide a theoretical basis for molecular breeding. In the present study, HFSCs were isolated using the technique of immunohistochemistry from the cervical spinal skin tissue samples from the fetal sheep, and the miR-133a-3p expression was confirmed using quantitative reverse-transcription PCR (RT-qPCR) and western blotting experiments from the isolated HFSCs. Additionally, the effects on the proliferation and apoptosis of HFSCs were detected using flow cytometry and 5-ethynyl-2'-deoxyuridine assays, along with other methods, following the overexpression of miR-133a-3p or its inhibition. The experimental results revealed that miR-133a-3p overexpressed could inhibit the proliferation of HFSCs and promote apoptosis by specifically targeting DUSP6. While the miR-133a-3p knockdown could promote the proliferation but inhibit the apoptosis of the HFSCs. Meanwhile, the miR-133a-3p knockdown experiments showed opposite outcomes. These results illustrate the presence of a relevant network between DUSP6 and miR-133a-3p, which regulates the production of superior-quality brush hair.


Assuntos
Folículo Piloso , MicroRNAs , Animais , Ovinos , Folículo Piloso/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Cabras/genética , Cabras/metabolismo , Proliferação de Células/genética , Células-Tronco/metabolismo
2.
Anim Biotechnol ; 34(3): 686-697, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37014133

RESUMO

MAP3K1 is a significant member of the MAPK family, and its expressed MEKK1 protein has a wide range of biological activities and is an essential node in the MAPK signaling pathway. A significant number of studies have revealed that MAP3K1 plays a complicated function in the control of cell proliferation, apoptosis, invasion and movement, participates in the regulation of the immune system, and plays an important role in wound healing, tumorigenesis and other processes. In this study, we looked at the involvement of MAP3K1 in the control of hair follicle stem cells (HFSCs). Overexpression of MAP3K1 significantly promoted the proliferation of HFSCs by inhibiting apoptosis and promoting the transition from S phase to G2 phase. The transcriptome identified 189 (MAP3K1_OE) and 414 (MAP3K1_sh) differential genes. The two pathways with the most significant enrichment of differentially expressed genes were the IL-17 signaling pathway and TNF signaling pathway, and the significantly enriched terms in the GO enrichment analysis involved regulation of response of external stimulus, inflammatory and cytokine. Indicate that MAP3K1 can function as a promoting factor in HFSCs through the induction of cell cycle transition from S phase to G2 phase can inhibition apoptosis by mediating crosstalk among several pathways and cytokines.HIGHLIGHTSAbnormal MAP3K1 expression in hair follicle stem cells (HFSCs) can impair HFSC proliferation and apoptosis.MAP3K1 controls hair follicle stem cell proliferation via modulating cell apoptosis and the ratio of cells in S phase/G2 phase.The differential genes shared by MAP3K1_sh and MAP3K1_OE are enriched in GO terms such as inflammation, adipocyte differentiation, acute inflammation, and so on.


Assuntos
Folículo Piloso , MAP Quinase Quinase Quinase 1 , Animais , Folículo Piloso/metabolismo , MAP Quinase Quinase Quinase 1/metabolismo , Células-Tronco/metabolismo , Perfilação da Expressão Gênica , Citocinas/genética , Citocinas/metabolismo , Inflamação/metabolismo
3.
Anim Biotechnol ; 34(4): 1223-1231, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37524308

RESUMO

RNA-seq has shown that the DUSP6 and MAPK signaling pathways are associated with the production of high-quality brush hair (type III hair) in Yangtze River Delta white goats. However, there are few reports on the regulatory effects of DUSP6 expression on hair follicle stem cells (HFSCs) and cellular processes, as well as the underlying mechanism. Here, we investigated the effect of DUSP6 level in HFSCs and the molecular mechanism underlying the functional regulation of HFSCs by DUSP6. Overexpression of DUSP6 significantly suppressed the proliferation of HFSCs by inducing cell cycle arrest in the G1 phase and by promoting apoptosis. Transcriptome analysis revealed a total of 217 differentially expressed genes between DUSP6-overexpressing and control HFSCs, of which 33 (15.2%) were upregulated in DUSP6-overexpressing cells. The two pathways with the most significant enrichment of differentially expressed genes were the TNF signaling pathway and cytokine-cytokine receptor interaction pathway, and the significantly enriched terms in the GO enrichment analysis involved cell attachment and cytokines. These results indicate that DUSP6 can function as an inhibitory factor in HFSCs through the induction of cell cycle arrest in the G1 phase and can promote apoptosis by mediating crosstalk among several pathways and cytokines.HighlightsWe constructed DUSP6 overexpression vectors to detect mRNA and protein expression levels related to high-quality brush hair in MAPK signaling pathway.We found that high expression level of DUSP6 can inhibit the proliferation of hair follicle stem cells (HFSCs) and promote cell apoptosis of HFSCs.DUSP6 may be involved in the growth regulation of HFSCs like Other studies in cancer, tumors by regulating the expression of cytokines, changing the transmission of signals between cells, activating or suppressing immune-related pathways.


Assuntos
Folículo Piloso , Cabelo , Animais , Folículo Piloso/metabolismo , Citocinas/metabolismo , Células-Tronco/metabolismo , Proliferação de Células/genética
4.
Anim Genet ; 53(3): 405-415, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35383992

RESUMO

The Yangtze River Delta white goat is a unique goat species that can produce superior-quality brush hair. The formation of this brush hair is controlled by a series of critical genes and related signaling pathways. Circular RNAs (circRNAs), are ubiquitous endogenous non-coding RNAs that regulate many biological and physiological processes in mammals. However, little is known about the potential regulatory role of circRNAs on superior-quality brush hair formation in Yangtze River Delta white goat. In this study, high-throughput sequencing technology was used to only detect circRNAs in the neck skin tissue of normal-quality goats (NHQs) and superior-quality goats (HQs). A total of 61 803 circRNAs were identified and 32 of them were differentially expressed in the NHQ group vs. the HQ group. Functional enrichment analysis showed that the source gene of differentially expressed circRNAs (DE-circRNAs) was enriched mostly in platelet activation and the focal adhesion signal pathway. Action mechanism analysis revealed that DE-circRNAs could sponge to many identified miRNAs, including miR-31, miR-125b, miR-let-7a and miR-149-5p, which have important roles in goat hair follicle stem cell growth, hair follicle development and morphogenesis. Altogether, our findings provide a valuable basis for studying circRNAs involved in superior-quality brush hair traits and meanwhile advance our understanding of circRNA complex regulation mechanisms in Yangtze River Delta white goat skin hair follicle development.


Assuntos
Cabras , MicroRNAs , Animais , Folículo Piloso , Sequenciamento de Nucleotídeos em Larga Escala , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Circular/genética
5.
Ecotoxicol Environ Saf ; 244: 114029, 2022 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-36055045

RESUMO

Cadmium (Cd) is ubiquitous in our environment and can easily bioaccumulate into the organism after passage through the respiratory and digestive tracts. Long-term exposure to Cd can result in the significant bioaccumulation in organism because of its long biological high-life (10-30 years), which exerts irreversible damages on the health of animals and humans. Although there are increased evidence of impeding the normal function of female reproduction resulted from Cd exposure, the mechanism of the negative action of Cd on the growth and development of ovarian follicle remains enigmatic. Thus, the purpose of the presented study is to summarize available literature which describing Cd-related toxicity involved in the adverse effects on the growth and development of the ovarian follicle. In conclusion, it is suggested that Cd causes damage to the folliculogenesis of mammalians, which results in the decline in the number and quality of ovulated oocytes and the failure in the fertilization. The mechanism behinds that may be linked to the interference to the production of reproductive hormones and the augment of reactive oxygen species (ROS). Furthermore, the enhanced ROS, in turn, impairs various molecules including proteins, lipids and DNA, as well as the balance of the antioxidant defense system, mitochondrial homeostasis, endoplasmic reticulum, autophagy and epigenetic modification. This review is expected to elaborate the toxic mechanism of Cd exposure to the growth and development of ovarian follicles and provide essential remediation strategies to alleviate the damage of Cd to female reproductive health.


Assuntos
Intoxicação por Cádmio , Cádmio , Animais , Antioxidantes , Cádmio/toxicidade , Feminino , Crescimento e Desenvolvimento , Hormônios , Humanos , Lipídeos , Mamíferos/metabolismo , Folículo Ovariano , Espécies Reativas de Oxigênio/metabolismo , Reprodução
6.
Ecotoxicol Environ Saf ; 248: 114311, 2022 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-36410142

RESUMO

Cadmium (Cd) is one major environmental pollutant that can cause detrimental impacts on human as well as animal reproductive systems as a result of oxidative stress. It is widely acknowledged that melatonin secreted principally by the pineal gland is not only a natural potent antioxidant but also a free radical scavenger, whereas concerning how to alleviate the toxic effects of Cd on oocyte maturation remains elusive. In this investigation, it was the first time to explore the protective effects and potential mechanism of melatonin on meiotic maturation of mouse oocytes exposed to Cd in vitro medium. We found that Cd exerts adverse effects on meiotic maturation progression by disrupting the normal function of mitochondrion combined with the aberrant mitochondrial distribution and decreased membrane potential and altering epigenetic modification, including H3K9me2 and H3K4me2. Additionally, it was observed that Cd exposure disrupted the morphology of spindle organization and caused chromosome misalignment, which might be through changing the level of acetylated tubulin, whereas melatonin administration alleviated the toxic impacts of Cd on oocytes. Furthermore, the mitochondrial morphology-related genes mRNA expression and protein expression of autophagy-related genes was also investigated. The results suggested that melatonin supplementation significantly altered the mRNA expression of mitochondrial dynamics-related genes, rather than the expression of mitophagy-related proteins. Taken together, our results validated that melatonin administration has a certain protective impact against oocytes meiosis maturation defects induced by cadmium through changing epigenetic modification and enhancing mitochondrial morphology rather than mitophagy.


Assuntos
Melatonina , Humanos , Camundongos , Animais , Melatonina/farmacologia , Cádmio/toxicidade , Meiose , Oócitos , Mitocôndrias , Epigênese Genética , RNA Mensageiro
7.
Genomics ; 113(2): 463-473, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33358944

RESUMO

In Yangtze River Delta white goat, hypermethylation of CMTM3 leads to a decreased expression level in high quality brush hair. However, the regulation of CMTM3 expression and its function in hair follicle stem cells (HFSCs) remains largely unknown. In this study, we investigated the regulation of CMTM3 expression, function, and molecular mechanism in HFSCs. The re-expression of CMTM3 significantly suppressed the proliferation of HFSCs by inducing G1 cell cycle arrest and promoting apoptosis. Moreover, the downregulation of CMTM3 promoted HFSC proliferation. Treatment with sh_CMTM3 and incubation in a DHT culture medium had the most significant growth-promoting effect. It was hypothesized that transcriptome analysis using RNA sequencing (RNA-seq) in samples would enable the identification of unique protein-coding and non-coding genes that may help uncover the role of CMTM3. Multiple genes and pathways were involved in this process, including 168 common DEGs, such as CXCL8 and E-selectin, which is reportedly involved in multiple regulatory pathways. These results indicated that CMTM3 can function as HFSCs through the induction of a G1 cell cycle arrest and promoted apoptosis by mediating crosstalk between several pathways and transcription factors. Our data is available in the National Center for Biotechnology Information (NCBI) database with the accession number PRJNA657430.


Assuntos
Androgênios/farmacologia , Proliferação de Células , Quimiocinas/genética , Di-Hidrotestosterona/farmacologia , Folículo Piloso/citologia , Proteínas com Domínio MARVEL/genética , Células-Tronco/metabolismo , Adulto , Animais , Apoptose , Células Cultivadas , Cabras , Folículo Piloso/efeitos dos fármacos , Folículo Piloso/metabolismo , Humanos , Células-Tronco/efeitos dos fármacos , Transcriptoma
8.
J Environ Manage ; 151: 22-32, 2015 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-25528270

RESUMO

We study the effects of environmental policy commitments in a futuristic world in which solar radiation management (SRM) can be utilized to reduce climate change damages. Carbon and sulfur dioxide emissions (correlated pollutants) can be reduced through tradable permits. We show that if nations simultaneously commit to carbon permit policies, national SRM levels rise with carbon quotas. Alternatively, if they simultaneously commit to SRM policies, the global temperature falls with each unit increase in the global SRM level. A nation always wishes to be a leader in policymaking, but prefers carbon to SRM policymaking. The globe prefers SRM policy commitments.


Assuntos
Poluentes Atmosféricos/química , Mudança Climática , Política Ambiental , Carbono/química , Conservação dos Recursos Naturais , Previsões , Humanos , Cooperação Internacional , Modelos Teóricos , Formulação de Políticas , Energia Solar , Dióxido de Enxofre/química
9.
Theriogenology ; 210: 110-118, 2023 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-37490796

RESUMO

With increase in women's age, the reproductive capability of female mammals decreases dramatically caused by age-related oxidative stress, coinciding with the decline in the ovarian reserve, and the quality and quantity of oocytes, which is the main determinant of female fertility. Melatonin, as an effective antioxidant and antiaging substance, is secreted by the pineal gland and been found in the follicular fluid as well, which has been turned out to enable to protect oocytes from oxidative stress during ovulation. However, the beneficial effects of melatonin on meiotic maturation in vitro and early embryo development of aged oocytes are still not fully understood. Thus, the aim of this study is to explore the potential mechanism of melatonin to improve the oocytes maturation and early embryonic development. The results suggested that oocyte quality decreased with age, whereas 10-6 M melatonin supplementation can significantly prompt the maturation quality of oocytes, the rate of fertilization and the formation rate of blastocyst. Mechanistic investigation indicated that melatonin supplementation not only restored the function of mitochondria by reducing reactive oxygen species (ROS) generation and early apoptosis, but also increased the level of ATP and total GSH through enhancing the mRNA expression levels of SIRT1, SIRT3, GPX4, SOD1 and SOD2. In conclusion, melatonin could alleviate the impairment of age-related oxidative stress to meiotic maturation and early embryonic development of oocytes. This study may provide a potential remediation strategy to improve the quality of oocytes from aged women and the efficiency of assisted reproductive technologies.


Assuntos
Melatonina , Gravidez , Feminino , Animais , Camundongos , Melatonina/farmacologia , Melatonina/metabolismo , Oócitos , Oogênese , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Desenvolvimento Embrionário , Técnicas de Maturação in Vitro de Oócitos/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Mamíferos
10.
Genes (Basel) ; 13(6)2022 06 09.
Artigo em Inglês | MEDLINE | ID: mdl-35741797

RESUMO

The Yangtze River Delta white goat is a rare goat species capable of producing high-quality brush hair. Dual specificity protein phosphatase 1 (DUSP1) may play a role in the formation of high-quality brush hair, as evidenced by our previous research. We investigated the potential mechanisms that regulate the proliferation and apoptosis of goat hair follicle stem cells. We particularly focused on the relationship between DUSP1 and miR-101, which directly targets DUSP1, predicted and screened through bioinformatics websites. Then, fluorescence assays, flow cytometry, RT-qPCR, and Western blotting were used to investigate the effects of miR-101 on the proliferation and apoptosis of hair follicle stem cells. We found that miR-101 overexpression significantly decreased (p < 0.01) apoptosis and promoted the proliferation of hair follicle stem cells. Furthermore, the overexpression of miR-101 increased (p < 0.05) the mRNA and protein expression levels of the proliferation-related gene (PCNA) and anti-apoptotic gene (Bcl-2), and it decreased (p < 0.05) the mRNA and protein expression levels of the apoptotic gene (Bax). In conclusion, miR-101 can promote the proliferation of and inhibit the apoptosis of hair follicle stem cells by targeting DUSP1, which provides a theoretical basis for further elucidating the molecular mechanism that regulates the production of high-quality brush hair of Yangtze River Delta white goats.


Assuntos
Folículo Piloso , MicroRNAs , Animais , Apoptose/genética , Proliferação de Células/genética , Cabras/genética , Cabras/metabolismo , Folículo Piloso/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Mensageiro/metabolismo , Células-Tronco/metabolismo
11.
Front Cell Dev Biol ; 10: 760466, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35186916

RESUMO

Superior-quality brush hair, also called Type III hair, can be obtained only from the cervical spine region of skin tissues of Yangtze River Delta white goats. The formation of superior-quality brush hair is controlled by a series of critical genes and related signaling pathways. Circular RNAs (circRNAs) are ubiquitous endogenous noncoding RNAs that regulate many biological and physiological processes in mammals. However, little is known about the potential regulatory role of circRNAs in superior-quality brush hair formation. Here, we analyzed circRNA sequencing data from cervical spine region skin tissues of normal-quality brush hair goats and superior-quality brush hair goats and then selected and identified the differentially expressed circRNA circCOL1A1. To investigate the regulatory role and mechanism of action of circCOL1A1, goat hair follicle stem cells (gHFSCs) were cultured and treated with a circCOL1A1 overexpression plasmid and small-interfering RNAs (siRNAs). Functional assays showed that circCOL1A1 knockdown promoted the proliferation and differentiation of gHFSCs cultured in vitro but inhibited stem cell apoptosis, whereas overexpression of circCOL1A1 suppressed stem cell proliferation and differentiation and induced apoptosis. Bioinformatics analysis combined with dual-luciferase reporter assays and RNA binding protein immunoprecipitation (RIP) verified that, mechanistically, circCOL1A1 could bind miR-149-5p directly and then relieve its inhibitory effect on CMTM3 to further control the CMTM3/AR axis. Collectively, our results reveal a novel regulatory pathway for the formation of superior-quality brush hair and indicate that circCOL1A1 plays a role in gHFSC growth and superior-quality brush hair formation by targeting the miR-149-5p/CMTM3/AR axis.

12.
In Vitro Cell Dev Biol Anim ; 58(4): 325-334, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35426064

RESUMO

The Yangtze River Delta white goat is a unique goat species that can produce superior-quality brush hair. The formation of superior-quality brush hair cannot occur without goat hair follicle stem cell differentiation. However, knowledge regarding the regulatory role of miR-149-5p in hair follicle stem cell differentiation is limited. Here, we found that miR-149-5p is widely expressed in the tissues of Yangtze River Delta white goats, but its expression in the skin tissue of superior-quality brush hair goats is high compared to normal- quality goats. The functional studies showed that miR-149-5p overexpression markedly facilitated hair follicle stem cell differentiation, whereas inhibiting miR-149-5p inhibited hair follicle stem cell differentiation. These results more clearly elucidate the regulatory role of miR-149-5p in hair follicle stem cell growth.


Assuntos
Folículo Piloso , MicroRNAs , Animais , Cateninas/metabolismo , Diferenciação Celular/genética , Cabras/genética , Cabras/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , beta Catenina/genética , beta Catenina/metabolismo
13.
Genes (Basel) ; 13(11)2022 10 28.
Artigo em Inglês | MEDLINE | ID: mdl-36360210

RESUMO

Yangtze River Delta white goats are the sole goat breed producing brush hair of high quality. The gene DUSP6 has been extensively studied in tumor cells but rarely in hair follicle stem cells (HFSCs). Per the previous sequencing data, it was determined that DUSP6 expression was up-regulated in superior-quality brush hair tissues, confirming it as a candidate gene associated with this trait. The targeting relationship of miR-145-5p with DUSP6 was determined based on online database prediction and was authenticated using a dual-luciferase gene reporter assay and quantitative reverse-transcription PCR (RT-qPCR). The regulatory effect of miR-145-5p on the growth of HFSCs was determined by targeting DUSP6 with RT-qPCR, 5-ethynyl-2'-deoxyuridine assays, Western blotting, and flow cytometry. The proliferation of HFSCs was inhibited and their apoptosis capacity was enhanced due to the presence of miR-145-5p. Therefore, it was proposed that this may have occurred through a repression effect of DUSP6 on the MAPK signaling pathway. The regulatory network of the HFSCs can be further understood using the theoretical basis established by the findings derived from this study.


Assuntos
Folículo Piloso , MicroRNAs , Animais , Cabras/genética , Cabras/metabolismo , Apoptose/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Proliferação de Células/genética , Células-Tronco/metabolismo
14.
Toxicology ; 457: 152810, 2021 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-33984407

RESUMO

With the rapid development of economic globalization and industrialization, lead (Pb), one of the most important heavy metals, has been used widely since antiquity for several purposes. In fact, its impact on the health of animals and humans is a significant public health risk all the time. Pb could be accumulated in the body for a long time, causing irreversible damage to the health of animals and humans, including hostile reproductive health. Up to now, although there are some published studies on impeding the normal development of ovarian folliculogenesis of female resulted from Pb exposure, with the damage of structure in uterine tissue, the imbalance of female menstrual status, and the change of hormone levels. The potential mechanism of Pb exposure on female reproduction system, however, remains enigmatic. How to alleviate the damage of Pb toxicity to reproductive function of female has become an urgent problem. Therefore, the aim of the present review is to discuss the information on the growth and development of ovarian follicle of mammalians and the potential toxic mechanism when exposed to Pb. The literatures were collected via various websites and consulting books, reports, etc. In summary, Pb impair folliculogenesis of mammalians, which may be related to the interference to the hypothalamic-pituitary-gonadal (HPG) axis and the production of reactive oxygen species (ROS), in turn impairs various molecules including proteins, lipids and DNA, as well as the disruption of the antioxidant defense system, ionic equilibrium and endoplasmic reticulum homeostasis.


Assuntos
Intoxicação por Chumbo/metabolismo , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/crescimento & desenvolvimento , Estresse Oxidativo/efeitos dos fármacos , Animais , Feminino , Humanos , Folículo Ovariano/metabolismo , Estresse Oxidativo/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Reprodução/efeitos dos fármacos , Reprodução/fisiologia
15.
In Vitro Cell Dev Biol Anim ; 57(4): 428-437, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33748907

RESUMO

The Yangtze River Delta White Goat is the only goat breed in the world that can produce superior-quality brush hair. Previous studies have shown that some genes are expressed differentially in the skin tissues between the goats produced superior-quality and normal-quality brush hair. Studies also have shown that different gene play varied roles in regulating the proliferation and apoptosis of hair follicle stem cells. However, the biological function of MAP3K1 (mitogen-activated protein kinase kinase kinase 1) gene in hair follicle stem cells is not fully understood. This study aims to investigate the role of MAP3K1 knockdown during the proliferation and apoptosis of hair follicle stem cells. RT-qPCR and Western blot were used to detect mRNA gene and protein expression level, CCK-8 and EdU assays were used to detect cell proliferation, and cell cycle and apoptosis were detected by flow cytometry. The results showed that the MAP3K1 expression level was significantly higher in the skin tissue of produced superior-quality brush hair than that in produced normal-quality brush hair. Moreover, functional studies indicated that si-MAP3K1 significantly inhibits the proliferation of hair follicle stem cells that came from a superior goat and promotes its apoptosis. Based on aforementioned assays, we speculated that MAP3K1 might play a regulatory effect in superior-quality brush hair traits.


Assuntos
Cabras/genética , Folículo Piloso/crescimento & desenvolvimento , MAP Quinase Quinase Quinase 1/genética , Células-Tronco/citologia , Animais , Apoptose/genética , Proliferação de Células/genética , Inativação Gênica , Cabras/crescimento & desenvolvimento
16.
Front Genet ; 11: 529757, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33262781

RESUMO

The Yangtze River Delta white goat is a unique goat species that can produce superior quality brush hair. CKLF-like MARVEL transmembrane domain-containing 3 (CMTM3), which influences the transcriptional activity of androgen receptor (AR), was identified as a candidate gene related to superior-quality brush hair formation. CMTM3 is generally expressed at low levels, but miR-149-5p is highly expressed in the skin tissues of these goats. The mechanism by which CMTM3 regulates the proliferation and apoptosis of goat hair follicle stem cells has not been elucidated. Here, RT-qPCR, western blotting, 5-ethynyl-2'-deoxyuridine (EdU), cell cycle, apoptosis, and dual-luciferase assays were used to investigate the role and regulatory mechanism of CMTM3 and miR-149-5p. Functional studies showed that CMTM3 overexpression inhibited proliferation and induced apoptosis in cultured hair follicle stem cells, whereas silencing CMTM3 markedly facilitated cell proliferation and deterred apoptosis in cultured hair follicle stem cells. Then, using bioinformatic predictions and the aforementioned assays, including dual-luciferase assays, RT-qPCR, and western blotting, we confirmed that miR-149-5p targets CMTM3 and preliminarily investigated the interaction between CMTM3 and AR in goat hair follicle stem cells. Furthermore, miR-149-5p overexpression significantly accelerated the proliferation and attenuated the apoptosis of hair follicle stem cells. Conversely, miR-149-5p inhibition suppressed the proliferation and induced the apoptosis of hair follicle stem cells. These results reveal a miR-149-5p-related regulatory framework for the miR-149-5p/CMTM3/AR axis during superior quality brush hair formation, in which CMTM3 plays a negative role.

17.
Expert Opin Ther Pat ; 29(2): 127-135, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30632414

RESUMO

INTRODUCTION: By 2020, nearly one million people will live with Parkinson's disease (PD) in the U.S. This disorder has a significant impact on patients' quality of life and is a burden on families and society. Protracted efforts have been made to treat the disease. Cumulative technological innovations are encapsulated by patents, and patent citations have been used to analyze technology diffusion processes in R&D, which is essential to identifying technology evolution trends and providing a review of PD treatment from the perspective of technology flows. AREAS COVERED: A patent citation network was utilized to analyze technology flows. Patents related to anti-PD drugs were collected from the U.S. Patent and Trademark Office (U.S. PTO) database. A total of 1,231 patents and 2,995 internal citations granted between 1988 and 2017 were included and analyzed. EXPERT OPINION: To launch drugs with greater efficiency and safety, approaches such as long-acting sustained release, controlled osmotic release, and other novel drug delivery systems should be emphasized. Multi-target agents could effectively reduce side effects in mono-drug therapy and are worth further exploration. Investors should keep an eye on alpha-synuclein-related therapy, gene therapy, and other experimental therapies that might trigger a historic revolution in the treatment domain.


Assuntos
Antiparkinsonianos/farmacologia , Desenvolvimento de Medicamentos/métodos , Doença de Parkinson/tratamento farmacológico , Antiparkinsonianos/administração & dosagem , Sistemas de Liberação de Medicamentos , Desenho de Fármacos , Humanos , Doença de Parkinson/fisiopatologia , Patentes como Assunto , Qualidade de Vida , Tecnologia Farmacêutica/métodos , Estados Unidos
18.
Gene ; 698: 19-26, 2019 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-30825596

RESUMO

Adult stem cells are self-renewing populations that originate from embryonic progenitor cells during organogenesis and retain multipotency to support tissue and organ regeneration throughout the lifetime of an organism. The hair follicle (HF) is a small organ that is ideal for studying the biology and regulation of adult stem cells. A distinct, permanent pool of adult stem cells is located in the HF bulge region. Most methods used to isolate hair follicle stem cells (HFSCs) begin with mouse or human follicles. Here, we describe two methods of isolating HFSCs from newborn Yangtze River Delta White Goats. A suitable method was found. The cell viability and expression of HFSC marker proteins differed in the two methods. CD49f-positive (integrin alpha 6) HFSCs were sorted by fluorescence activated flow cytometry. Sorted HFSCs can be used in various in vivo grafting models and are useful as an in vitro model to study multipotency, quiescence and activation.


Assuntos
Células-Tronco Adultas/citologia , Técnicas de Cultura de Células/métodos , Folículo Piloso/metabolismo , Células-Tronco Adultas/fisiologia , Animais , Proliferação de Células , Sobrevivência Celular , Cabras/genética , Folículo Piloso/crescimento & desenvolvimento , Organogênese/genética , Cultura Primária de Células/métodos , Regeneração/genética
19.
Gene ; 668: 166-173, 2018 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-29783073

RESUMO

The Yangtze River delta white goat is the only goat breed that produces high-quality brush hair, which is specifically used in top-grade writing brushes. Previous studies have indicated that the CMTM3 and DUSP1 genes are involved in the growth and cycle of high-quality brush hair, and these genes are thought to be involved in the formation of high-quality brush hair traits. In this study, we investigated the relationship between methylation of CMTM3 and DUSP1 and such traits. The results indicated that the relative expression levels of the CMTM3 and DUSP1 genes were higher in non-high-quality brush hair than in high-quality brush hair. Furthermore, the CpG sites of the DUSP1 gene were not methylated, and the methylation level of CMTM3 was negatively correlated with the gene expression level. We believe that the DUSP1 gene regulates the formation of high-quality brush hair by non-methylated, and that methylation of the CMTM3 gene results in a decrease in its expression, causing an increase in the activity of the androgen receptor and the level of androgen. This high androgen level promotes the growth of high-quality brush hair. These study results provide a theoretical basis for further elucidating the molecular mechanism of the formation of high-quality brush hair characteristics, and provide scientific reference for the molecular breeding of high-quality brush hair.


Assuntos
Metilação de DNA , Fosfatase 1 de Especificidade Dupla/genética , Cabras/genética , Cabelo/metabolismo , Proteínas com Domínio MARVEL/genética , Regiões Promotoras Genéticas , Androgênios/sangue , Animais , Sítios de Ligação , Ilhas de CpG , Expressão Gênica , Cabras/sangue , Cabras/metabolismo , Masculino , Fatores de Transcrição/metabolismo
20.
Electron. j. biotechnol ; 39: 74-81, may. 2019. tab, ilus, graf
Artigo em Inglês | LILACS | ID: biblio-1052041

RESUMO

Background: CPEB is considered as an RNA-binding protein first identified in Xenopus oocytes. Although CPEB1 was involved in the growth of oocyte, its role in goat follicular granulosa cell has not been fully elucidated. To clarify the functions of this gene in goat follicular granulosa cells, CPEB1-overexpressing vector and interference vector were structured and transfected into follicular granulosa cells from Jiangsu native white goats of Nantong city, Jiangsu Province, China. The expression levels of differentiation-related genes including CDK1, Cyclin B1, and C-mos were determined 24 h after administration of CPEB1 by quantitative real-time polymerase chain reaction and Western blotting methods. Results: The expression levels of CDK1, Cyclin B1, and C-mos were significantly upregulated after overexpression and significantly downregulated after interference with CPEB1. Conclusions: The CPEB1 gene expression could affect the transcription of genes related to early cleavage divisions, which provided a reference for further research on its role in the growth and maturation of oocytes.


Assuntos
Animais , Feminino , Oócitos , Fatores de Transcrição/genética , Cabras/genética , Transfecção , Fertilização in vitro , Expressão Gênica , Western Blotting , Reação em Cadeia da Polimerase/métodos , Proteínas de Ligação a RNA , Transferência Embrionária , Gado , Fluorescência , Células da Granulosa
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