Effects of melatonin implantation on cashmere yield,fibre characteristics,duration of cashmere growth as well as growth and reproductive performance of Inner Mongolian cashmere goats

Background： Exogenous melatonin could induce cashmere growth. However, induced growth of cashmere fleece by melatonin implants cannot be combined with the typical growth, resulting in earlier shedding followed by another cycle of cashmere growth. To address this issue, we examine the effects on the cashmere yield, fibre characteristics, and the growth and reproductive performance of cashmere goats of planned administration of melatonin. Methods： Eighteen half-sib, female goats were assigned to two treatments （n = 9） including a control and a treatment where melatonin （2 mg/kg BW） was implanted at the end of April and end of June. Cashmere growth and shedding were observed for approximately ] year following implantation. Fibre samples were collected monthly to determine cumulative cashmere length. Initiation and cessation dates for cashmere growth as well as the rate of cashmere growth were calculated. Cashmere yield, weight gain of dam, kidding date, litter size, and birth weight were also recorded. Results： Melatonin implantation increased cashmere yield by 34.5 % （control 553.7 g vs. melatonin 745.0 g; P〈 0.01）, cashmere length by 21.3 % （control 95.2 mm vs. melatonin 115.4 mm; P 〈 0.01）, and decreased fibre diameter by 4.4 % （control 14.6 pm vs. melatonin 14.0 IJm; P 〈 0.03）. In melatonin-treated goats, the average initiation date was earlier than in control goats （May 18, 2013 vs. July 2, 20]3; P〈 0.01） but there was a similar cessation date （March 22, 2014 vs. March 27, 2014）. Consequently, the duration of cashmere growth was longer in melatonin-treated goats than in control goats （307 vs.270 days; P 〈 0.01）. The final BW, average daily gain, kidding date, litter size, and birth weight were not influenced by melatonin implantation. Conclusions： These data indicate that melatonin implantation （2 mg/kg BW） on two occasions （late April and June） increased cashmere yield by combining the induced growth of cashmere fleece with the typical growth and decreased the fibre diameter without changing dam growth rate or reproductive performance.

Hoxc13/β-catenin Correlation with Hair Follicle Activity in Cashmere Goat

Seasonal hair follicle activity and fibre growth in some Cashmere-bearing goats （Caprus hircus） is a cyclic process that is well characterized morphologically but understood incompletely at the molecular level. As an initial step in discovering regulators in hair-follicle activity and cycling, we used qPCR to investigate 19 genes expression in Cashmere goat side skin from 12 mon. Many of these genes may be associated with the hair follicle development-relevant genes （HFDRGs） in the literature. Here we show that Hoxc13/β-catenin gene associated with the follicle activity. In addition, Hoxc13 was found to be expressed with an drastic increase between July and November for melatonin treatments. To further investigate the role of Hoxcl3 on HFDRGs, fibroblasts and keratinocytes from Cashmere goat skin were transfected with p-ECFP- Hoxc13. The result suggested that overexpression ofHoxcl3 gene decreased HFDRGs with negative role for hair follicle development and increase HFDRGs with positive role for hair follicle development in vitro. These findings provide data on the Hoxc13 expression profile of normal Cashmere goat skin and Cashmere goat skin with melatonin treatment, and demonstrate hair-follicle-activity dependent regulation of Hoxc13 expression.

Hoxc13 Expression Pattern in Cashmere Goat Skin During Hair Follicle Development

Hoxc13 has an important role in controlling hair formation. In this study, we examine the Hoxc13 RNA expression pattern of skin during embryo development. The result indicated that changes of the Hoxe13 gene expression and thickness of skin have a similar trend during hair follicle morphogenesis. In interpreting these results, we investigated whether the regulation motifs is in Hoxc13 intron, which is a 5.4 kb fragment. To blast with other mammals, we found a very conservative region in all mammal animals and two regions in livestock, such as cow, sheep, horse, dog, and so on, which are not in other Hox genes. We have examined putative pre-miRNA in this region, providing an entry point for elucidating currently unknown mechanisms that are required for regulating quantitative levels of Hoxc13 gene expression.

Prokaryotic Expression, Ascitic Polyclonal Antibody Preparation and Identification of Cashmere Goat Izumo1

Izumol is a novel member of the immunoglobulin superfamily locating on sperm, and is indispensable for sperm-egg fusion. According to its immunoglobulin-like domain in the extracellular region, Izumol was fractionated into 6 fragments （F0-F5） which were ligated with pGEX-4T1 to construct the prokaryotic expression vectors pGEX-Fn. The recombinant plasmids were transformed into Escherichia coli BL21 （DE3） and the GST-Fn fusion proteins were expressed successfully by induction with IPTG. GST-F0, a recombinant fusion protein of GST with the full length of extracellular region of mature cashmere goat Izumol, was purified by polyacrylamide gel slicing method and was used as an antigen to immunize the Kunming mouse to generate anti-GST-Izumol ascetic polyclonal antibody with intraperitoneal injection of S 180 cells. Subsequently, the anti-GST-Izumol polyclonal antibody was purified with miscellaneous antigen by glutaraldehyde cross-linking method. Western blotting analysis showed that the purified ascetic polyclonal antibody had high affinity to all 6 GST-Izumol fragment fusion proteins. Immunohistochemical analysis with this antibody displayed that the cashmere goat Izumol proteins were at the equatorial segment of sperm head surface. These results indicate that this polyclonal antibody has high specificity and lays the foundations for further study on the expression pattern of Izumol in cashmere goat testis and binding abilities of each extra-membrane fragment of Izumol to the egg surface.

Molecular Characterization and Expression Pattern of Rheb Gene in Inner Mongolia Cashmere Goat(Capra hircus)

As one member of the Ras super family, Rheb is an upstream regulator of mTOR signaling pathway, which regulates the process of cell-growth, proliferation and differentiation. In order to study the relationship between Rheb and mTOR in Inner Mongolian Cashmere goat （Capra hircus） cells, Ras homolog enriched in brain （Rheb） gene eDNA was amplified by RT-PCR. It is 555 bp in length and includes the complete ORF encoding 184 amino acids （GenBank accession no. HM569224）. The full eDNA nucleotide sequence has a 99% identity with that of sheep, 98% with cattle and 93% with human while their amino acids sequence shares identity with 98, 97 and 97% of them, correspondingly. The bioinformatics analysis showed that Rheb has a Ras family domain, two casein kinase II phosphorylation sites, two ATP/GTP-binding sites motifA （P-loop）, a prenyl group binding site （CAAX box）. Tissue-specific expression analysis performed by semi- quantitative RT-PCR. The Rheb gene was expressed in all the tested tissues and the highest level ofmRNA accumulation was detected in brain, suggesting that Rheb played an important role in goat cells.

Performances Analysis and Developmental Strategy of Hexi Cashmere Goats

The productive performances of Hexi cashmere goats for breeding nucleus groups and producing groups in different production areas were reviewed. At the same time, the productive performances were analyzed. Moreover, the existing problems and solving methods were put forward. In addition, the Hexi cashmere goats was an ancient and native breed, and it was necessary to further improve its productive performance to protect and utilize the breed resource effectively.

A Single-cell Transcriptome Atlas of Cashmere Goat Hair Follicle Morphogenesis

Cashmere,also known as soft gold,is produced from the secondary hair follicles(SHFs)of cashmere goats.The number of SHFs determines the yield and quality of cashmere;therefore,it is of interest to investigate the transcriptional profiles present during cashmere goat hair follicle development.However,mechanisms underlying this development process remain largely unexplored,and studies regarding hair follicle development mostly use a murine research model.In this study,to provide a comprehensive understanding of cellular heterogeneity and cell fate decisions,single-cell RNA sequencing was performed on 19,705 single cells of the dorsal skin from cashmere goat fetuses at induction(embryonic day 60;E60),organogenesis(E90),and cytodifferentiation(E120)stages.For the first time,unsupervised clustering analysis identified 16 cell clusters,and their corresponding cell types were also characterized.Based on lineage inference,a detailed molecular landscape was revealed along the dermal and epidermal cell lineage developmental pathways.Notably,our current data also confirmed the heterogeneity of dermal papillae from different hair follicle types,which was further validated by immunofluorescence analysis.The current study identifies different biomarkers during cashmere goat hair follicle development and has implications for cashmere goat breeding in the future.

河北本地绒山羊的杂交选育研究

[Objective] To improve the performance of Hebei cashmere goats and thus promote the development of cashmere industry. [Method] Through grading hybridization, the F2 generation was obtained using Liaoning cashmere goats as male parents and local cashmere goats as female parents. Then fixed cross was performed. After selection, the body size indexes, weight and cashmere yield were determined. The quality of cash mere, lambing rate, and net cashmere rate were also analyzed. [ Result] Compared with the local cashmere goats, the hybrid cashmere goats had robust physique and good body size indexes; the weight and cashmere yield increased significantly; cashmere fiber was longer significantly; the lambing rate and net cashmere rate were improved. [ Conclusion] After breeding, the cashmere goats have good quality and high yield of cashmere, and their performance is significantly improved.