CRISPR/Cas9-mediated VDR knockout plays an essential role in the growth of dermal papilla cells through enhanced relative genes

Background Hair follicles in cashmere goats are divided into primary and secondary hair follicles (HFs). HF development, which determines the morphological structure, is regulated by a large number of vital genes; however, the key functional genes and their interaction networks are still unclear. Although the vitamin D receptor (VDR) is related to cashmere goat HF formation, its precise effects are largely unknown. In the present study, we verified the functions of key genes identified in previous studies using hair dermal papilla (DP) cells as an experimental model. Furthermore, we used CRISPR/Cas9 technology to modify the VDR in DP cells to dissect the molecular mechanism underlying HF formation in cashmere goats. Results The VDR expression levels in nine tissues of Shaanbei white cashmere goats differed significantly between embryonic day 60 (E60) and embryonic day 120 (E120). At E120, VDR expression was highest in the skin. At the newborn and E120 stages, the VDR protein was highly expressed in the root sheath and hair ball region of Shaanbei cashmere goats. We cloned the complete CDS of VDR in the Shaanbei white cashmere goat and constructed a VDR-deficient DP cell model by CRISPR/Cas9. Heterozygous and homozygous mutant DP cells were produced. The growth rate of mutant DP cells was significantly lower than that of wild-type DP cells (P < 0.05) and VDR mRNA levels in DP cells decreased significantly after VDR knockdown (P < 0.05). Further, the expression levels of VGF, Noggin, Lef1, and β-catenin were significantly downregulated (P < 0.05). Conclusions Our results indicated that VDR has a vital role in DP cells, and that its effects are mediated by Wnt and BMP4 signaling.

Download Full-text

Regulatory role of LEF-1 in the proliferation of Arbas White Cashmere goat dermal papilla cells

Canadian Journal of Animal Science ◽

10.1139/cjas-2017-0130 ◽

2018 ◽

Vol 98 (4) ◽

pp. 667-674

Author(s):

Fei Hao ◽

Wei Yan ◽

Xiaodong Guo ◽

Bing Zhu ◽

Dongjun Liu

Keyword(s):

Wnt Signaling ◽

Cyclin D1 ◽

Economic Value ◽

Dermal Papilla ◽

Hair Follicles ◽

Cashmere Goat ◽

Follicle Growth ◽

Dermal Papilla Cells ◽

Hair Follicle Growth

Cashmere, which has high economic value, is made from the secondary hair follicles of cashmere goat skin. Dermal papilla cells (DPCs) are considered the center for regulation of hair growth, which is closely related to hair follicle growth. We constructed LEF-1 overexpression and interference experimental groups of goat DPCs to investigate LEF-1 regulation of DPCs proliferation by Wnt signaling, and provide a theoretical basis for improving cashmere yield. In primary DPCs, LEF-1, β-catenin, C-myc, and cyclin D1 expression in the LEF-1 overexpression group was 9.25-, 1.27-, 1.74-, and 1.63-fold, respectively, that of the control. LEF-1, β-catenin, C-myc, and cyclin D1 expression in the LEF-1 interference group was 0.20-, 0.75-, 0.38-, and 0.39-fold, respectively, that of the control. In secondary DPCs, LEF-1, β-catenin, C-myc, and cyclin D1 expression in the LEF-1 overexpression group was 10.53-, 1.48-, 1.64-, and 1.39-fold, respectively, that of the control. LEF-1, β-catenin, C-myc, and cyclin D1 expression in the LEF-1 interference group was 0.21-, 0.71-, 0.40-, and 0.36-fold, respectively, that of the control. Primary and secondary DPCs proliferation rates changed with LEF-1 expression. Therefore, the LEF-1 regulation pattern of cell proliferation through Wnt signaling is similar in both DPCs.

Download Full-text

Single-cell sequencing reveals the intermediate cell state and function of dermal papilla cells in the hair follicle cycle of cashmere goats

10.21203/rs.3.rs-25379/v1 ◽

2020 ◽

Author(s):

Feng Yang ◽

Zhihong Liu ◽

Tianyu Che ◽

Juntao Guo ◽

Yuchun Xie ◽

...

Keyword(s):

Hair Follicle ◽

Progenitor Cells ◽

Dermal Papilla ◽

Hair Follicles ◽

Intermediate Cell ◽

Dermal Papilla Cells ◽

Cashmere Goats ◽

Hair Follicle Cycle ◽

Intermediate Cells ◽

And Function

Abstract Background: Human hair loss and regeneration has stimulated interest in the natural hair cycle worldwide; however, such research is difficult because the periodicity of human or mouse hair is not visually obvious. Dermal papilla cells (DP cells) play an important role in the development of hair follicles, but knowledge of the differentiation and mechanisms of DP stem cells during transition through the hair follicle cycle are still limited, although some studies have reported that DP cells may have an intermediate cell state during differentiation, the classification and function of specific cell states are not clear. Results: Here, we used cashmere goats, that have obvious periodicity of hair follicles, as model animals and, based on unbiased single cell RNA sequencing, we identified and isolated DP cell data. Pseudotime ordering analysis was used to successfully construct a DP cell lineage differentiation trajectory and revealed the sequential activation of key genes, signaling pathways, and functions involved in cell fate decisions. At the same time, we analyzed the mechanisms of different cell fates and revealed the function of four different intermediate cells: Intermediate cells 10 showed important functions in the growth of cashmere and maintenance of cashmere attachment to the skin; intermediate cells 1 revealed important functions in the process of apoptosis and cashmere shedding of secondary hair follicles; intermediate cells 0 initiated new follicular cycles and completed the migration of hair follicles and the occurrence of cashmere; and intermediate cells 15 are suggested to be DP progenitor cells. Conclusions: In development and apoptosis, inner bulge cells not only earlier than outer bulge cells, but occurred faster and was more thorough,this helps a deeper understanding of the role of bulge cells. Pseudogenes play another important role in function which promoted the competitive endogenous RNA (ceRNA) hybridization of pseudogenes.In different hair follicle cycles, DP cells will differentiate into different intermediate state cells and perform different functions, and the marker genes of the cells also changed. Intermediate cells 10 showed important functions in the growth of cashmere and maintenance of cashmere attachment to the skin; intermediate cells 1 revealed important functions in the process of apoptosis and cashmere shedding of secondary hair follicles; intermediate cells 0 initiated new follicular cycles and completed the migration of hair follicles and the occurrence of cashmere; and intermediate cells 15 are DP progenitor cells, this conclusion provides an unprecedented deeper understanding of the function of DP cells.

Download Full-text

Hormones and Hair Growth: Variations in Androgen Receptor Content of Dermal papilla Cells Cultured from Human and Red Deer (Cervus Elaphus) Hair Follicles.

Journal of Investigative Dermatology ◽

10.1111/1523-1747.ep12363039 ◽

1993 ◽

Vol 101 (s1) ◽

pp. 114S-120S ◽

Cited By ~ 24

Author(s):

Valerie Anne Randall ◽

Margaret Julie Thornton ◽

Andrew Guy Messenger ◽

Nigel Andrew Hibberts ◽

Andrew Stewart Irving Loudon ◽

...

Keyword(s):

Androgen Receptor ◽

Cervus Elaphus ◽

Hair Growth ◽

Red Deer ◽

Dermal Papilla ◽

Hair Follicles ◽

Dermal Papilla Cells ◽

Cells Cultured

Download Full-text

Smooth muscle alpha-actin is a marker for hair follicle dermis in vivo and in vitro

Journal of Cell Science ◽

10.1242/jcs.99.3.627 ◽

1991 ◽

Vol 99 (3) ◽

pp. 627-636 ◽

Cited By ~ 2

Author(s):

C.A. Jahoda ◽

A.J. Reynolds ◽

C. Chaponnier ◽

J.C. Forester ◽

G. Gabbiani

Keyword(s):

Smooth Muscle ◽

Hair Follicle ◽

Dermal Papilla ◽

Hair Follicles ◽

Dermal Papilla Cells ◽

Smooth Muscle Alpha Actin ◽

Actin Expression ◽

Alpha Actin ◽

Sheath Cells

We have examined the expression of smooth muscle alpha-actin in hair follicles in situ, and in hair follicle dermal cells in culture by means of immunohistochemistry. Smooth muscle alpha-actin was present in the dermal sheath component of rat vibrissa, rat pelage and human follicles. Dermal papilla cells within all types of follicles did not express the antigen. However, in culture a large percentage of both hair dermal papilla and dermal sheath cells were stained by this antibody. The same cells were negative when tested with an antibody to desmin. Overall, explant-derived skin fibroblasts had relatively low numbers of positively marked cells, but those from skin regions of high hair-follicle density displayed more smooth muscle alpha-actin expression than fibroblasts from areas with fewer follicles. 2-D SDS-PAGE confirmed that, unlike fibroblasts, cultured papilla cells contained significant quantities of the alpha-actin isoform. The rapid switching on of smooth muscle alpha-actin expression by dermal papilla cells in early culture, contrasts with the behaviour of smooth muscle cells in vitro, and has implications for control of expression of the antigen in normal adult systems. The very high percentage of positively marked cultured papilla and sheath cells also provides a novel marker of cells from follicle dermis, and reinforces the idea that they represent a specialized cell population, contributing to the heterogeneity of fibroblast cell types in the skin dermis, and possibly acting as a source of myofibroblasts during wound healing.

Download Full-text

Inhibition of Rab27a and Rab27b Has Opposite Effects on the Regulation of Hair Cycle and Hair Growth

International Journal of Molecular Sciences ◽

10.3390/ijms21165672 ◽

2020 ◽

Vol 21 (16) ◽

pp. 5672

Author(s):

Kyung-Eun Ku ◽

Nahyun Choi ◽

Jong-Hyuk Sung

Keyword(s):

Hair Growth ◽

Expression Patterns ◽

Dermal Papilla ◽

Hair Follicles ◽

Hair Cycle ◽

Outer Root Sheath ◽

Dermal Papilla Cells ◽

Root Sheath ◽

Culture Models

Rab27a/b are known to play an important role in the transport of melanosomes, with their knockout causing silvery gray hair. However, the relationship between Rab27a/b and hair growth is not well known. To evaluate the role of Rab27a/b in hair cycle, we investigated the expression of Rab27a/b during hair cycling and human outer root sheath (hORS) cells. The expression of Rab27a in ORS cells was mainly detected at the anagen, whereas expression of Rab27b in ORS, and epidermal cells was strongly expressed at the telogen. Additionally, Rab27a/b were expressed in the Golgi of hORS cells. To evaluate the role of Rab27a/b in hair growth, telogen-to-anagen transition animal and vibrissae hair follicles (HFs) organ culture models were assayed using Rab27a/b siRNAs. The knockdown of Rab27a or Rab27b suppressed or promoted hair growth, respectively. These results were also confirmed in human dermal papilla cells (hDPCs) and hORS cells, showing the opposite mitogenic effects. Moreover, Rab27b knockdown increased the expression levels of various growth factors in the hDPCs and hORS cells. Overall, the opposite temporal expression patterns during hair cycling and roles for hair growth of Rab27a/b suggested that Rab27a/b might regulate the hair cycle. Therefore, our study may provide a novel solution for the development of hair loss treatment by regulating Rab27a/b levels.

Download Full-text

The local hypothalamic–pituitary–adrenal axis in cultured human dermal papilla cells

BMC Molecular and Cell Biology ◽

10.1186/s12860-020-00287-w ◽

2020 ◽

Vol 21 (1) ◽

Author(s):

Eun Young Lee ◽

You Jin Nam ◽

Sangjin Kang ◽

Eun Ju Choi ◽

Inbo Han ◽

...

Keyword(s):

Hpa Axis ◽

Hair Loss ◽

Human Hair ◽

Stress Hormones ◽

Dermal Papilla ◽

Hair Shaft ◽

Hair Follicles ◽

Follicle Cells ◽

Dermal Papilla Cells ◽

Underlying Mechanisms

Abstract Background Stress is an important cause of skin disease, including hair loss. The hormonal response to stress is due to the HPA axis, which comprises hormones such as corticotropin releasing factor (CRF), adrenocorticotropic hormone (ACTH), and cortisol. Many reports have shown that CRF, a crucial stress hormone, inhibits hair growth and induces hair loss. However, the underlying mechanisms are still unclear. The aim of this study was to examine the effect of CRF on human dermal papilla cells (DPCs) as well as hair follicles and to investigate whether the HPA axis was established in cultured human DPCs. Results CRF inhibited hair shaft elongation and induced early catagen transition in human hair follicles. Hair follicle cells, both human DPCs and human ORSCs, expressed CRF and its receptors and responded to CRF. CRF inhibited the proliferation of human DPCs through cell cycle arrest at G2/M phase and induced the accumulation of reactive oxygen species (ROS). Anagen-related cytokine levels were downregulated in CRF-treated human DPCs. Interestingly, increases in proopiomelanocortin (POMC), ACTH, and cortisol were induced by CRF in human DPCs, and antagonists for the CRF receptor blocked the effects of this hormone. Conclusion The results of this study showed that stress can cause hair loss by acting through stress hormones. Additionally, these results suggested that a fully functional HPA axis exists in human DPCs and that CRF directly affects human DPCs as well as human hair follicles under stress conditions.

Download Full-text