Identification and molecular analysis of cashmere goat lncRNAs reveal their integrated regulatory network and potential roles in secondary hair follicle.

Long non-coding RNAs (lncRNAs) is a class of eukaryotic transcripts with length of more than 200 bp. They contribute to the regulation of gene expressions involved in multiple processes including the skin cell proliferation, differentiation, and reconstruction of the secondary hair follicles (SHFs). In this study, firstly, we identified 16 putative lncRNAs from SHFs of cashmere goat based on the EST sequences from NCBI database. Secondly, we investigated their transcriptional pattern in SHFs of cashmere goat, and constructed their ceRNA regulatory networks. The RT-qPCR results showed four lncRNAs (lncRNA-475074, -052149, -052140, and -051789) were significantly up-regulated, and nine lncRNAs (lncRNA-711032, -475083, -475070, -052139, -052127, -052037, -051903, -051847, and -051804) were significantly down-regulatd in anagen SHFs of cashmere goat. CeRNA networks analysis revealed complex interactional relationship among lncRNAs, miRNAs and mRNAs. Further, the KEGG pathway enrichment was performed for the potential target genes of the identified lncRNAs based on bioinformatics technique, and the results indicated that differentially expressed lncRNAs directly or indirectly might regulate potential target genes. Our results from this study will provide a significant information for further exploring the functions and possible mechanisms of the identified lncRNAs in SHFs of cashmere goat.

LncRNA-599554 sponges miR-15a-5p to contribute inductive ability of dermal papilla cells through positive regulation of the expression of Wnt3a in cashmere goat

BACKGROUND: Long non-coding RNAs (lncRNAs), as post-transcriptional regulators, were thought to function in the inductive property of dermal papilla cells (DPCs) in cashmere goat. Previously, lncRNA-599554 was identified in secondary hair follicle (SHF) of cashmere goat, but its functional significance is unknown. RESULTS: In the present investigation, we verified that lncRNA-599554 had significantly higher expression at the anagen dermal papilla of cashmere goat SHF than that at telogen. Based on overexpression and knockdown techniques, we found that lncRNA-599554 contributes the inductive property of DPCs of cashmere goat, which was assessed by detecting the changes in the expression of several typical indictor genes in DPCs including ET-1, SCF, Versican, ALP, Lef1 and Ptc-1. Based on RNA pull-down assay, we verified that lncRNA-599554 directly interacted with chi-miR-15a-5p. Also, we showed that lncRNA-599554 positively regulated the Wnt3a expression in DPCs but which did not appear to involve its modulating of promoter methylation. Based on the use of Dual-luciferase reporter assays, our data indicated that lncRNA-599554 regulated the Wnt3a expression through chi-miR-15a-5p-mediated post-transcriptional level. CONCLUSIONS: We showed that lncRNA-599554 contributes the inductive property of DPCs in cashmere goat which might be achieved through sponging chi-miR-15b-5p to promote the Wnt3a expression. The results from the present investigation provided a novel insight into the functional mechanism of lncRNA-599554 in the SHF regeneration of cashmere goat along with the formation and growth of cashmere fiber.

The Seed Semipermeable Layer and Its Relation to Seed Quality Assessment in Four Grass Species.

The existence of a semipermeable layer in grass seeds has been extensively reported, yet knowledge of its influence on tests for seed viability and vigor that depend upon measurement of electrical conductivity (EC) is limited. This study determined the presence and location of the semipermeable layer, and its relation to seed viability and vigor assessment, in seeds of four important grass species-Elymus nutans Griseb., Lolium perenne L., Leymus chinensis (Trin.) Tzvel., and Avena sativa L. Intact seeds of E. nutans, Lolium perenne, and Leymus chinensis exhibited little staining with triphenyl tetrazolium chloride (TTC), and there were no differences in EC between seeds with different germination percentage (GP) (P > 0.05). After piercing the seed coat, however, all three species displayed positive staining with TTC, along with a significant negative correlation between EC and GP (E. nutans: R2 = 0.7708; Lolium perenne: R2= 0.8414; Leymus chinensis: R2 = 0.859; P < 0.01). In contrast, both intact and pierced seeds of A. sativa possessed a permeable seed coat that showed positive staining with TTC and EC values that were significantly negatively correlated with GP [R2 = 0.9071 (intact) and 0.9597 (pierced); P < 0.01]. In commercial seed lots of A. sativa, a field emergence test indicated that EC showed a significant negative correlation with field emergence at two sowing dates (R2= 0.6069, P < 0.01 and 0.5316, P < 0.05). Analysis of seed coat permeability revealed the presence of a semipermeable layer located in the seed coat adjacent to the endosperm in E. nutans, Lolium perenne, and Leymus chinensis; however, no semipermeable layer was observed in A. sativa. This is the first report of the absence of a semipermeable layer in a grass species. The existence of a semipermeable layer is one of the most important factors affecting seed viability and vigor testing (based on EC measurement) in E. nutans, Lolium perenne, and Leymus chinensis. Increasing the permeability of the semipermeable layer, e.g., by piercing the seed coat, may permit the use of EC measurement to assess seed vigor in species that possess such a layer.