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. 2017 Oct 18;7(1):13502.
doi: 10.1038/s41598-017-13986-w.

Skin Transcriptome Reveals the Intrinsic Molecular Mechanisms Underlying Hair Follicle Cycling in Cashmere Goats Under Natural and Shortened Photoperiod Conditions

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Free PMC article

Skin Transcriptome Reveals the Intrinsic Molecular Mechanisms Underlying Hair Follicle Cycling in Cashmere Goats Under Natural and Shortened Photoperiod Conditions

Min Yang et al. Sci Rep. .
Free PMC article

Abstract

The growth of cashmere exhibits a seasonal pattern arising from photoperiod change. However, the underlying molecular mechanism remains unclear. We profiled the skin transcriptome of six goats at seven time points during hair follicle cycling via RNA-seq. The six goats comprised three goats exposed to a natural photoperiod and three exposed to a shortened photoperiod. During hair cycle transition, 1713 genes showed differential expression, and 332 genes showed a pattern of periodic expression. Moreover, a short photoperiod induced the hair follicle to enter anagen early, and 246 genes overlapped with the periodic genes. Among these key genes, cold-shock domain containing C2 (CSDC2) was highly expressed in the epidermis and dermis of Cashmere goat skin, although its function in hair-follicle development remains unknown. CSDC2 silencing in mouse fibroblasts resulted in the decreased mRNA expression of two key hair-follicle factors, leading to reduced cell numbers and a lower cell density. Cashmere growth or molting might be controlled by a set of periodic regulatory genes. The appropriate management of short light exposure can induce hair follicles to enter full anagen early through the activation of these regulators. The CSDC2 gene is a potentially important transcription factor in the hair growth cycle.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Cashmere growth cycle. (A) Schematic diagram showing the cashmere growth cycle, duration of daylight and sampling regimes. The HF from the Inner Mongolian Cashmere goat undergoes circannual changes, including transitions among the anagen phase (May–December), catagen phase (January) and telogen phase (February–April). The SD group was subjected to seven hours of daylight from 9:30–16:30 daily (17 h darkness: 7 h light; 17D: 7 L) from May 1st to October 1st (five months). The colored fonts indicate the sampling time points. At each time point, the skin from 12 goats was sampled, and RNA from six goats was isolated for RNA-seq. (BF) Comparison of the HF structure among stages of the hair growth cycle and between the test and control groups in June. (B) June (early anagen): The HF activity is very low, and the natural photoperiod group is still in early anagen, with an incomplete HF structure. (C) The skin sections from the short-photoperiod goats indicate entry into anagen in early June (Jun_SD), with visible inner root sheaths, dense HFs and developed sebaceous glands. (D) August (anagen): Skin sections show visible inner root sheaths, dense HFs and developed sebaceous glands. (E) January (catagen): Sections show dermal papilla condensation. (F) April (telogen): HF structure is incomplete and undergoes apoptosis. SG, sebaceous gland; IRS, inner root sheath; HS, hair shaft; DP, dermal papilla. Scale bars indicate 100 μm.
Figure 2
Figure 2
Different phase transitions. (A) PCA of the expressed genes in the different HF development stages. (B) The number of DEGs during each transition. The distributions of DEGs with a ≥ 2-fold FPKM difference for the four transitions (Ι, II, Ш and IV). (C) Venn diagrams of the DEGs in the four transitions. (D) Venn diagrams of differentially expressed TFs from transitions Ι and Ш. The number of genes is shown in the individual specific and overlapping areas in the Venn diagrams. (E) Heatmap of the overlapping TFs from transitions Ι and Ш across the different SHF cycling phases, with the HF- higher or specific genes marked in red.
Figure 3
Figure 3
Three major gene clusters with similar expression trends and functional gene annotations. Left: The genes were clustered in eight groups based on the relative expression with the indicated trends. Each expression trend is shown in the red curves. Middle: The GO terms associated with the genes in each cluster are listed. Enrichment significance scores for each GO term are shown as histograms (orange). Right: Previously reported TFs in each cluster are listed; the colorful gene symbols represent the overlapping genes between transitions Ι and Ш, with red denoting the up-regulated genes and, green denoting the down-regulated genes.
Figure 4
Figure 4
The effects of a short photoperiod on the HF. (A) PCA for the expressed genes from all 40 samples. (B) Venn diagram of the overlapping DEGs genes between the periodic and SD responses; 246 genes and 17 TFs overlapped. The expression trends for 238 genes are the same in the pairs from June (treated vs control) and from the transition Ι, and opposite from the transition Ш, including 228 up-regulated and 10 down-regulated genes. (C) The heatmap for the 14 common TFs, with the HF- higher or specific genes marked in red.
Figure 5
Figure 5
RNAi-mediated CSDC2 silencing in NIH/3T3 cells. (A) qPCR analysis of CSDC2 in NIH/3T3 cells that were transfected with control (scrambled) or si-CSDC2. N = 3; P values were calculated using Student’s t test. Error bars indicate SEM. (B) Quantitation of gene expression related to hair-follicle development by qPCR in NIH/3T3 cells at 72 h after transfection with control (scrambled) or si-CSDC2. N = 3; P values were calculated using Student’s t test. Error bars indicate SEM. (C) Cell proliferation at 48 h and 72 h after silencing. (D) Growth of NIH/3T3 cells at 48 h and 72 h after transfection; the images were taken at 40x magnification.

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