Single-cell transcriptomic atlas of goat ovarian aging

Background The ovaries are one of the first organs that undergo degenerative changes earlier in the aging process,and ovarian aging is shown by a decrease in the number and quality of oocytes.However,little is known about the molecular mechanisms of female age-related fertility decline in different types of ovarian cells during aging,especially in goats.Therefore,the aim of this study was to reveal the mechanisms driving ovarian aging in goats at single-cell resolution.Results For the first time,we surveyed the single-cell transcriptomic landscape of over 27,000 ovarian cells from newborn,young and aging goats,and identified nine ovarian cell types with distinct gene-expression signatures.Functional enrichment analysis showed that ovarian cell types were involved in their own unique biological processes,such as Wnt beta-catenin signalling was enriched in germ cells,whereas ovarian steroidogenesis was enriched in granulosa cells(GCs).Further analysis showed that ovarian aging was linked to GCs-specific changes in the antioxidant system,oxidative phosphorylation,and apoptosis.Subsequently,we identified a series of dynamic genes,such as AMH,CRABP2,THBS1 and TIMP1,which determined the fate of GCs.Additionally,FOXO1,SOX4,and HIF1A were identified as significant regulons that instructed the differentiation of GCs in a distinct manner during ovarian aging.Conclusions This study revealed a comprehensive aging-associated transcriptomic atlas characterizing the cell typespecific mechanisms during ovarian aging at the single-cell level and offers new diagnostic biomarkers and potential therapeutic targets for age-related goat ovarian diseases.

Effects of antioxidant‑rich Lactiplantibacillus plantarum inoculated alfalfa silage on rumen fermentation, antioxidant and immunity status, and mammary gland gene expression in dairy goats

Background Milk synthesis in lactating animals demands high energy metabolism,which results in an increased production of reactive oxygen metabolites(ROM)causing an imbalance between oxidants and antioxidants thereby inducing oxidative stress(OS)on the animals.To mitigate OS and postpartum disorders in dairy goats and gain insight into the impact of dietary choices on redox status during lactation,a feeding trial was conducted using alfalfa silage inoculated with a high-antioxidant strain of Lactiplantibacillus plantarum.Methods Twenty-four Guanzhong dairy goats(38.1±1.20 kg)were randomly assigned to two dietary treatments:one containing silage inoculated with L.plantarum MTD/1(RSMTD-1),and the other containing silage inoculated with high antioxidant activity L.plantarum 24-7(ES24-7).Results ES24-7-inoculated silage exhibited better fermentation quality and antioxidant activity compared to RSMTD-1.The ES24-7 diet elevated the total antioxidant capacity(T-AOC),superoxide dismutase(SOD),glutathione peroxi-dase(GSH-Px),and catalase(CAT)activities in milk,serum,and feces of lactating goats(with the exception of T-AOC in milk).Additionally,the diet containing ES24-7 inoculated silage enhanced casein yield,milk free fatty acid(FFA)content,and vitamin A level in the goats’milk.Furthermore,an increase of immunoglobulin(Ig)A,IgG,IgM,inter-leukin(IL)-4,and IL-10 concentrations were observed,coupled with a reduction in IL-1β,IL-2,IL-6,interferon(IFN)-γ,and tumor necrosis factor(TNF)-αconcentrations in the serum of lactating goats fed ES24-7.Higher concentrations of total volatile fatty acid(VFA),acetate,and propionate were observed in the rumen fluid of dairy goats fed ES24-7 inoculated silage.Moreover,the diet containing ES24-7 inoculated silage significantly upregulated the expression of nuclear factor erythroid 2 like 2(NFE2L2),beta-carotene oxygenase 1(BCO1),SOD1,SOD2,SOD3,GPX2,CAT,glu-tathione-disulfide reductase(GSR),and heme oxygenase 1(HMOX1)genes in the mammary gland,while decreased the levels of NADPH oxidase 4(NOX4),TNF,and interferon gamma(IFNG).Conclusions These findings indicated that feeding L.plantarum 24-7 inoculated alfalfa silage not only improved rumen fermentation and milk quality in lactating dairy goats but also boosted their immunity and antioxidant status by modulating the expression of several genes related to antioxidant and inflammation in the mammary gland.

Enhancing milk quality and modulating rectal microbiota of dairy goats in starch‑rich diet:the role of bile acid supplementation

Background Diets rich in starch have been shown to increase a risk of reducing milk fat content in dairy goats.While bile acids(BAs)have been used as a lipid emulsifier in monogastric and aquatic animals,their effect on ruminants is not well understood.This study aimed to investigate the impact of BAs supplementation on various aspects of dairy goat physiology,including milk composition,rumen fermentation,gut microbiota,and BA metabolism.Results We randomly divided eighteen healthy primiparity lactating dairy goats(days in milk=100±6 d)into two groups and supplemented them with 0 or 4 g/d of BAs undergoing 5 weeks of feeding on a starch-rich diet.The results showed that BAs supplementation positively influenced milk yield and improved the quality of fatty acids in goat milk.BAs supplementation led to a reduction in saturated fatty acids(C16:0)and an increase in monounsaturated fatty acids(cis-9 C18:1),resulting in a healthier milk fatty acid profile.We observed a significant increase in plasma total bile acid concentration while the proportion of rumen short-chain fatty acids was not affected.Furthermore,BAs supplementation induced significant changes in the composition of the gut microbiota,favoring the enrichment of specific bacterial groups and altering the balance of microbial populations.Correlation analysis revealed associations between specific bacterial groups(Bacillus and Christensenellaceae R-7 group)and BA types,suggesting a role for the gut microbiota in BA metabolism.Functional prediction analysis revealed notable changes in pathways associated with lipid metabolism,suggesting that BAs supplementation has the potential to modulate lipid-related processes.Conclusion These findings highlight the potential benefits of BAs supplementation in enhancing milk production,improving milk quality,and influencing metabolic pathways in dairy goats.Further research is warranted to elucidate the underlying mechanisms and explore the broader implications of these findings.

Global gene expression profiling of perirenal brown adipose tissue whitening in goat kids reveals novel genes linked to adipose remodeling

Background Brown adipose tissue(BAT)is known to be capable of non-shivering thermogenesis under cold stimulation,which is related to the mortality of animals.In the previous study,we observed that goat BAT is mainly located around the kidney at birth,and changes to white adipose tissue(WAT)in the perirenal adipose tissue of goats within one month after birth.However,the regulatory factors underlying this change is remain unclear.In this study,we systematically studied the perirenal adipose tissue of goat kids in histological,cytological,and accompanying molecular level changes from 0 to 28 d after birth.Results Our study found a higher mortality rate in winter-born goat kids,with goat birthing data statistics.Then we used thermal imaging revealing high temperature in goat hips at postnatal 0 d and gradually decrease during 28 d.This is consistent with the region of perirenal BAT deposition and highlights its critical role in energy expenditure and body temperature regulation in goat kids.Additionally,we found a series of changes of BAT during the first 28 d after birth,such as whitening,larger lipid droplets,decreased mitochondrial numbers,and down-regulation of key thermogenesis-related genes(UCP1,DIO2,UCP2,CIDEA,PPARGC1a,C/EBPb,and C/EBPa).Then,we used RNA-seq found specific marker genes for goat adipose tissue and identified 12 new marker genes for BAT and 10 new marker genes for WAT of goats.Furthermore,12 candidate genes were found to potentially regulate goat BAT thermogenesis.The mechanism of the change of this biological phenomenon does not involve a large-scale death of brown adipocytes and subsequent proliferation of white adipocytes.While apoptosis may play a limited role,it is largely not critical in this transition process.Conclusions We concluded that perirenal BAT plays a crucial role in thermoregulation in newborn goat kids,with notable species differences in the expression of adipose tissue marker genes,and we highlighted some potential marker genes for goat BAT and WAT.Additionally,the change from BAT to WAT does not involve a large-scale death of brown adipocytes and subsequent proliferation of white adipocytes.

Pharmacokinetic Characterization of Xylazine in Goats with Simultaneous Anesthesia Studies

The aim was to study the pharmacokinetics of xylazine as a stable anesthetic in goats.In this study,goats were injected with xylazine at the rate of 0.3 mL·kg-1 intramusculally,and blood samples were collected at 1,3,5,10,20,30,45,60,90,120,180,and 240 min after administration,respectively.Xylazine was extracted by liquid-liquid extraction and separation method,and blood concentration was determined by high performance liquid chromatography(HPLC).The pharmacokinetic characteristics of xylazine in healthy goats were analyzed by pharmacokinetic software.The results showed that the chromatographic peak time of xylazine chromatography was 9-11 min.The specificity of the method was good.The linear correlation coefficient R2 of the standard curve was 0.9982 when the concentration of xylazine was in the range of 10-1×1000 ng.The pharmacokinetic model of xylazine in goats was a one-chamber model with first-order rate absorption,distribution half-life t1/2Ka was(0.49±0.041)min,elimination half-life t1/2Ke was(23.3±2.5)min,and the peak time(Tp)of the highest concentration was(2.8±0.2)min.The total drug clearance CL/F was(0.00016±0.000016)mg·kg-1·min-1(ng·mL-1),and the minimum effective blood concentration was 56.6 ng·mL-1,which was consistent with the clinical anesthetic effect.The results showed that xylazine had the characteristics of rapid absorption,wide distribution,short peak time,slow clearance rate,and long anesthetic time in goats,which could be used as the basic drug for the development of goat complex anesthetic preparation.

Effects of Serum Concentration, Synchronization Time and Confluence on the Cell-Cycle Synchronization Efficiency of Goat Fibroblasts

This study aims to evaluate the effect of serum concentration, synchronization time, and confluence degree on the synchronisation efficiency of goat fibroblast cycle. The results indicated that there was no difference in the percentage of nucleated fibroblasts in the G0/G1 stage between serum concentrations of 0.3% and 0.4% (83.89% and 82.69%, respectively, P > 0.05) as well as between serum concentrations of 0.2% and 0.5% (76.95% and 75.46%, respectively, P > 0.05). The percentage of nucleated fibroblasts in the G0/G1 stage was highest at the concentration of 0.3% and lowest in the control group (83.89% vs. 62.67%, P 0.05). The beneficial effect of high confluence was confirmed by the large percentage of nucleated fibroblasts at the G0/G1 stage. The 60% confluency was significantly lower than the 80% and 100% confluency (73.44%, 86.63%, and 87.17%, respectively, P < 0.05). The results indicate that the goat fibroblast cycle synchronization is the most effective at the serum concentration of 0.3%, 72 hours of synchronization and 100% confluency.

Cysticercus Tenuicollis in Visceral Organs of Goats and Sheep in Uganda: A Case Study in Lira Municipal Abattoir

Cysticercus tenuicollis is a metacestode stage of Taenia hydatigena residing in the visceral organs of small ruminants and its effects cause economic losses due to condemnation of visceral organs in slaughterhouses. This study reports on the prevalence of Cysticercus tenuicollis in small ruminants slaughtered in Lira Municipal Abattoir. A total of 70 sheep and 70 goats were sampled and examined after slaughter for the presence of Cysticercus tenuicollis in the visceral organs using standard meat inspection procedures. An abattoir data collection sheet was used to record data, which was then analysed in SPSS version 20. The study revealed 51.4% of goats and 68.6% of sheep to be infested with Cysticercus tenuicollis. Following the district of origin for the small ruminants, Otuke district had the highest prevalence of the parasite (82.6%), followed by Kole (72.4%), Alebtong (66.7%), Apac (59.4%) and Lira (31.4%). Cysticercus tenuicollis was more frequently seen in the intestines (57.8%) of sheep and goats than in any other visceral organs. The liver had a prevalence of 20.7%, the lungs 17.8%, the kidneys 3.5% and the fetal sac;(15.7%) in goats and (12.9%) in sheep. Cysticercus tenuicollis was detected in 68.8% of females and 58.8% of male small ruminants. It is recommended to routinely deworm goats and sheep by farmers most especially in districts with the highest prevalence of Cysticercus tenuicollis infestation. The sale of the condemned parts of sheep and goats and the feeding of dogs on these contaminated materials should be stopped forthwith. It is also being suggested that a national study be carried out to determine the prevalence and economic impact of Cysticercus tenuicollis in small ruminants.

Effects of dietary Clostridium butyricum and rumen protected fat on meat quality,oxidative stability,and chemical composition of finishing goats

Background Clostridium butyricum(CB)is a probiotic that can regulate intestinal microbial composition and improve meat quality.Rumen protected fat(RPF)has been shown to increase the dietary energy density and provide essential fatty acids.However,it is still unknown whether dietary supplementation with CB and RPF exerts beneficial effects on growth performance and nutritional value of goat meat.This study aimed to investigate the effects of dietary CB and RPF supplementation on growth performance,meat quality,oxidative stability,and meat nutritional value of finishing goats.Thirty-two goats(initial body weight,20.5±0.82 kg)were used in a completely randomized block design with a 2 RPF supplementation(0 vs.30 g/d)×2 CB supplementation(0 vs.1.0 g/d)factorial treatment arrangement.The experiment included a 14-d adaptation and 70-d data and sample collection period.The goats were fed a diet consisted of 400 g/kg peanut seedling and 600 g/kg corn-based concentrate(dry matter basis).Result Interaction between CB and RPF was rarely observed on the variables measured,except that shear force was reduced(P<0.05)by adding CB or RPF alone or their combination;the increased intramuscular fat(IMF)content with adding RPF was more pronounced(P<0.05)with CB than without CB addition.The pH24h(P=0.009),a*values(P=0.007),total antioxidant capacity(P=0.050),glutathione peroxidase activities(P=0.006),concentrations of 18:3(P<0.001),20:5(P=0.003)and total polyunsaturated fatty acids(P=0.048)were increased,whereas the L*values(P<0.001),shear force(P=0.050)and malondialdehyde content(P=0.044)were decreased by adding CB.Furthermore,CB supplementation increased essential amino acid(P=0.027),flavor amino acid(P=0.010)and total amino acid contents(P=0.024)as well as upregulated the expression of lipoprotein lipase(P=0.034)and peroxisome proliferator-activated receptorγ(PPARγ)(P=0.012),and downregulated the expression of stearoyl-CoA desaturase(SCD)(P=0.034).The RPF supplementation increased dry matter intake(P=0.005),averaged daily gain(trend,P=0.058),hot carcass weight(P=0.046),backfat thickness(P=0.006),concentrations of 16:0(P<0.001)and c9-18:1(P=0.002),and decreased the shear force(P<0.001),isoleucine(P=0.049)and lysine content(P=0.003)of meat.In addition,the expressions of acetyl-CoA carboxylase(P=0.003),fatty acid synthase(P=0.038),SCD(P<0.001)and PPARγ(P=0.022)were upregulated due to RPF supplementation,resulting in higher(P<0.001)content of IMF.Conclusions CB and RPF could be fed to goats for improving the growth performance,carcass traits and meat quality,and promote fat deposition by upregulating the expression of lipogenic genes of Longissimus thoracis muscle.

Effect of Moringa oleifera on semen quality profiles in endangered Teressa goat under tropical humid island ecosystem

Objective:To investigate the effect of Moringa oleifera leaf extract on semen quality parameters in Teressa goat buck.Methods:A total of 25 semen samples from six bucks were selected for the study.Spermatozoa of 150×10^(6) were incubated in 0,300,500 and 700μg of leaf extract as groupⅠ,Ⅱ,Ⅲ and Ⅳ,respectively.Liquid stored semen samples were analysed for motility,viability,total sperm abnormalities,plasma membrane,acrosomal and nuclear integrities,seminal plasma intracellular enzymes[aspartate aminotransferase(AST),alanine aminotransferase(ALT)and lactate dehydrogenase(LDH)]and total antioxidant capacity(TAC)and spermatozoa malondialdehyde(MDA)and cholesterol efflux in comparison with the control group(groupⅠ)for up to 72 h.Results:Moringa oleifera leaf extract(500μg/150×10^(6) spermatozoa)-treated semen had significantly higher motility,viability,plasma membrane,acrosomal and nuclear integrities,and TAC(P<0.05),and had significantly lower total sperm abnormalities,AST,ALT,LDH,MDA production and cholesterol efflux compared to those in other Moringa oleifera leaf extract treated(300 or 700μg/150×10^(6) spermatozoa)and control groups at different hours of liquid storage(P<0.05).Semen quality parameters and TAC showed an increasing trend and total sperm abnormalities,MDA production,leakage of intra-cellular enzymes and cholesterol efflux showed a decreasing trend from group Ⅰto group Ⅲand then an opposite trend from group Ⅲ to group Ⅳ at different hours of liquid storage.Conclusions:Moringa oleifera leaf extract 500μg/150×10^(6) spermatozoa can be used for liquid semen preservation in Teressa goat at Andaman and Nicobar Islands.

circRNA3669 promotes goat endometrial epithelial cells proliferation via miR-26a/RCN2 to activate PI3K/AKT-mTOR and MAPK pathways

The development of receptive endometrium(RE) from pre-receptive endometrium(PE) for successful embryo implantation is a complex dynamic process in which the morphology and physiological states of the endometrial epithelium undergo a series of significant changes, including cell proliferation and apoptosis. However, the molecular mechanisms are not yet fully understood. In this study, a higher circRNA3669 level was observed in PE than in RE of goats. Functional assays revealed that this overexpression promoted the proliferation of goat endometrial epithelial cells(GEECs) by activating the expression of genes related to the PI3K/AKT-mTOR and MAPK pathways,thereby inhibiting apoptosis in vitro. Furthermore, circRNA3669 functioned as a competing endogenous RNA(ceRNA) to upregulate Reticulocalbin-2(RCN2) expression at the post-transcriptional level by interacting with and downregulating miR-26a in GEECs. In addition, RCN2, which is highly expressed in the PE of goats, was found to be regulated by β-estradiol(E2) and progesterone(P4). Our results demonstrated that RCN2 also affected the key proteins PI3K, AKT, mTOR, JNK, and P38 in the PI3K/AKT-mTOR and MAPK pathways, thereby facilitating GEECs proliferation and suppressing their apoptosis in vitro. Collectively, we constructed a new circRNA3669-miR-26aRCN2 regulatory network in GEECs, which further provides strong evidence that circRNA could potentially play a crucial regulatory role in the development of RE in goats.

Changes in milk fat globule membrane proteins along lactation stage of Laoshan dairy goat

The milk fat globule membrane(MFGM)is a complex structure with numerous functions,and its composition is affected by many factors.There have been few systematic investigations on goat MFGM proteome profiling during lactation.Individual milk samples from 15 healthy dairy goats were obtained at six lactation time points for investigation of the MFGM proteome using both data-independent acquisition(DIA)and data-dependent acquisition(DDA)proteomics techniques combined with multivariate statistical analysis.Using the DIA method,890 variably abundant MFGM proteins were discovered throughout the lactation cycle.From 1 to 240 d,butyrophilin subfamily 1 member A1,lipoprotein lipase,perilipin-2,and adipose triglyceride lipase were upregulated,while APOE,complement C3,clusterin,and IgG were downregulated.Furthermore,from 1 to 90 d,annexin A1,annexin A2,and antithrombin-ll were downregulated,then upregulated by d 240.Albumin had a high degree of connectedness,indicating that it was a key protein,according to protein-protein interaction research.Overall,our findings gave new insights into the biological features of MFGM protein in goat milk throughout lactation,which may aid in the creation of specialized MFGM products and infant formula.

Goat milk-based infant formula regulates intestinal barrier function and promotes the production of short-chain fatty acids

Infant formula(IF)based on cow milk and goat milk is a substitute food for infants who are underfed with human milk.In our previous study,we reported the composition and physicochemical stability of IF based on milk from cows and goats and a combination of both milks.Here,we investigated the effects of these 3 IFs on intestinal immunity and short-chain fatty acid production(SCFAs)using human microbiota-associated(HMA)mice and selected human milk as a positive control.The results showed that goat milk-based IF is associated with a functional immune advantage,due to the rise in the levels of immune-related cytokines interleukin(IL)-2 and IL-10,decreased levels of intestinal permeability markers D-lactic acid and endotoxin,and increased mRNA levels of intestinal tight junction proteins occludin and claudin.In addition,the intestine of mice fed with goat milk-based IF contained 12.06μmol/g acetate,2.42μmol/g propionate,and 1.72μmol/g butyrate,which reached 69%,79%,and 60%of the levels in human milk,respectively.Our results indicate that goat milk-based IF improves intestinal immune function and promotes the production of intestinal SCFAs.

Slaughter Performance, Muscle Quality and Nutritional Composition of Duoluo Goats in Sichuan Province

[Objectives] This study was conducted to understand the biological characteristics of Duoluo goats. [Methods] With male and female Duoluo goats as the research object, related performance indexes were determined and analyzed after slaughter. [Results] The weights of one-year-old male and female Duoluo goats before slaughter were (25.68±2.37) and (24.51±1.97) kg, respectively;the dressing percentages were (43.55±3.24) % and (43.84±3.42) %, respectively;and the net meat percentages were (33.02±2.32) % and (34.19±2.46) %, respectively. (2) The cooked meat rates of male and female Duoluo goats were (65.49±1.42) % and (65.36±1.55) %, respectively;the shearing force was (5.48±1.24) and (5.65±1.02) N, respectively;and the loin-eye muscle areas were (8.95±1.72) and (8.82±1.15) cm 2, respectively. (3) The protein contents of male and female Duoluo goats were (20.07±0.42) and (19.47±1.22) g/100 g, respectively;and the fat contents were (1.30±0.80) % and (2.92±0.55) %, respectively;and the cholesterol contents were (71.76±11.47) and (74.83±2.68) mg/100 g, respectively;(4) The Ca contents of male and female Duoluo goats were (69.53±10.89) and (63.00±4.56) mg/kg, respectively;the Fe contents were (16.73±1.83) and (14.77±0.58) mg/kg, respectively;and the Zn contents were (38.47±3.68) and (31.83±2.22) mg/kg, respectively. (5) In the muscles of male and female Duoluo goats, the contents of essential amino acids were (7.44±0.24) and (7.26±0.41) g/100 g , respectively;the contents of non-essential amino acids were (11.81±0.33) and (11.42±0.67) g/100 g, respectively;and the contents of flavor amino acids were (8.79±0.27) and (8.42±0.57) g/100 g, respectively. [Conclusions] This study lays a foundation for the excavation and utilization of the resource, Duoluo goats.

Branched-chain fatty acids from goat milk alleviate ulcerative colitis via the TLR4/NF-κB/NLRP3 pathway

Branched-chain fatty acids(BCFAs)are new bioactive fatty acids with anti-inflammatory properties.However,the role of BCFAs in alleviating ulcerative colitis has not been clarified.Herein,we evaluated the protective effect of BCFAs from goat milk in mice with colitis induced using dextran sodium sulfate(DSS)and explored the corresponding mechanism.These results show that BCFAs extracted from goat milk can significantly alleviate weight loss in mice,and reduce the disease activity index and the activity of myeloperoxidase while increasing the content of antioxidant enzymes in colon tissue and reducing the oxidation stress response.These data also show that BCFAs can down-regulate the gene and protein expression of the toll-like receptor 4(TLR4)/nuclear factorκB p65(NF-κB p65)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathway,and at the same time significantly reduce the expression of pro-inflammatory factors tumor necrosis factorα(TNF-α),interleukin 1β(IL-1β),and IL-18 in colon tissue,and significantly increase the expression of the anti-inflammatory factor IL-10.In conclusion,these results demonstrated that BCFAs in goat milk exerted effects on colitis-related inflammatory cytokines and inhibited inflammation by inducing the TLR4/NF-κB/NLRP3 pathway to alleviate DSS-induced ulcerative colitis.This study provides evidence for the potential of BCFAs as bioactive fatty acids in food products and to ameliorate ulcerative colitis development in mice.

鳙goat基因序列特征及其时空表达模式分析

【目的】探究鳙的ghrelin O-acyltransferase(goat)基因序列特征及时空表达模式,解析goat基因的潜在生物学功能。【方法】通过RACE扩增得到goat基因cDNA全长序列,采用实时荧光定量PCR对其在早期发育阶段、各组织和冬夏季4个组织(脑、肠道、肌肉和肝脏)中的表达特征进行检测;结合原位杂交和免疫组化技术分别从基因和蛋白水平解析goat基因在肠组织中的分布特征,并通过实时荧光定量PCR评估goat基因及通路相关基因的共表达模式。【结果】鉴定得到goat基因cDNA序列全长2255 bp,开放阅读框(ORF)长1116 bp,共编码371个氨基酸残基。推导氨基酸序列含7个跨膜结构域,并包含参与催化的天冬酰胺(第251位)和组氨酸(第282位)。同源比对分析发现鳙GOAT氨基酸序列与鲤科鱼类相似度极高。鳙goat基因在早期孵化时和出膜后5 d的表达水平均显著高于其他时期(P<0.05,下同);成体组织差异表达分析发现goat基因在肠道中表达量最高;goat基因在夏季脑、肠道和肌肉组织的相对表达量均显著高于冬季。原位杂交试验结果显示goat基因在肠道中表达信号较强,且肠道切片的免疫组化试验显示肠黏膜中存在GOAT与GHRL共定位的阳性信号。goat基因与生长激素的合成、分泌和作用通路(ko04935)中相关基因存在共表达关系。【结论】goat基因具有MBOAT家族的典型结构特征,可能通过与摄食相关基因(ghrl等)协作来参与鳙摄食调控和生长代谢适应等生物学过程。

A 1.1 Mb duplication CNV on chromosome 17 contributes to skeletal muscle development in Boer goats

The Boer goat is one of the top meat breeds in modern animal husbandry and has attracted widespread attention for its unique growth performance.However,the genetic basis of muscle development in the Boer goat remains obscure.In this study,we identified specific structural variants in the Boer goat based on genome-wide selection signals and analyzed the basis of the molecular heredity of related candidate genes in muscle development.A total of9 959 autosomal copy number variations(CNVs) were identified through selection signal analysis in 127 goat genomes.Specifically,we confirmed that the highest signal CNV(HSV) was a chromosomal arrangement containing an approximately 1.11 Mb(CHIR17:60062304-61171840 bp) duplicated fragment inserted in reverse orientation and a 5 362 bp deleted region(CHIR17:60145940-60151302 bp) with overlapping genes(e.g.,ARHGAP10,NR3C2,EDNRA,PRMT9,and TMEM184C).The homozygous duplicated HSV genotype(+/+) was found in 96% of Boer goats but was not detected in Eurasian goats and was only detected in 4% of indigenous African goats.The expression network of three candidate genes(ARHGAP10,NR3C2,and EDNRA)regulating dose transcription was constructed by RNA sequencing.Results indicated that these genes were involved in the proliferation and differentiation of skeletal muscle satellite cells(SMSCs) and their overexpression significantly increased the expression of SAA3.The HSV of the Boer goat contributed to superior skeletal muscle growth via the dose effects of overlapping genes.

Genomic analysis of indigenous goats in Southwest Asia reveals evidence of ancient adaptive introgression related to desert climate

Understanding how evolutionary pressures related to climate change have shaped the current genetic background of domestic animals is a fundamental pursuit of biology. Here, we generated wholegenome sequencing data from native goat populations in Iraq and Pakistan. Combined with previously published data on modern, ancient(Late Neolithic to Medieval periods), and wild Capra species worldwide, we explored the genetic population structure, ancestry components, and signatures of natural positive selection in native goat populations in Southwest Asia(SWA). Results revealed that the genetic structure of SWA goats was deeply influenced by gene flow from the eastern Mediterranean during the Chalcolithic period, which may reflect adaptation to gradual warming and aridity in the region. Furthermore, comparative genomic analysis revealed adaptive introgression of the KITLG locus from the Nubian ibex(C. nubiana) into African and SWA goats. The frequency of the selected allele at this locus was significantly higher among goat populations located near northeastern Africa. These results provide new insights into the genetic composition and history of goat populations in the SWA region.

Dietary copper supplementation modulates performance and lipid metabolism in meat goat kids

Forty-eight male Lezhi black goat kids with similar body weight((12.09±1.70)kg)and age((60±5)d)were used to determine the effect of dietary copper(Cu),in the form of reagent grade Cu sulfate(CuSO4∙5H2O),on performance,serum lipid profile,and the relative mRNA abundance of genes involved in lipid metabolism.Goat kids were stratified by body weight and randomly assigned to one of 4 treatment groups.Each treatment consisted of 12 replicate pens with each pen containing one goat kid.Treatment groups received the basal diet with no supplemental Cu(control),basal diet plus 10 mg of Cu kg^(-1)of dry matter(DM),basal diet plus 20 mg of Cu kg^(-1)of DM,or basal diet plus 30 mg of Cu kg^(-1)of DM.Goats were housed individually in pens and fed a high-concentrate pelleted diet for 60 d.Average daily gain,average daily feed intake and feed:gain of goats were not affected by dietary Cu supplementation(P>0.10).No differences were detected in serum total cholesterol,triglyceride,and high density lipoprotein cholesterol concentrations of goat kids fed with different Cu concentrations(P>0.05).However,serum low density lipoprotein cholesterol concentrations decreased linearly(P=0.01)as the concentration of dietary Cu increased.Intramuscular fat content of longissimus muscle increased(P=0.002)quadratically and liver Cu concentrations increased(P<0.001)linearly as dietary Cu concentration increased.Compared with the control,dietary supplementation of 20 mg Cu kg^(-1)DM decreased the relative mRNA abundance of fatty acid-binding protein 4(P=0.01)and lipoprotein lipase(P=0.05),and tended to decrease the relative mRNA abundance of carnitine palmitoyltransferase I(P=0.06)in longissimus muscle of goats.The relative mRNA abundance of peroxisome proliferator-activated receptor alpha(P<0.001),carnitine acetyltransferase(P=0.001),and carnitine palmitoyltransferase Ⅰ(P=0.001)were also decreased in liver by Cu supplementation.These results indicate that dietary supplementation of Cu modified lipid metabolism by increasing muscular fat and decreasing serum low density lipoprotein cholesterol,and the modification might be associated with the reduction of relative mRNA abundance of genes for oxidation of long-chain fatty acid in muscle and liver of Lezhi black goat kids.

Sulforaphane prevents LPS‑induced inflammation by regulating the Nrf2‑mediated autophagy pathway in goat mammary epithelial cells and a mouse model of mastitis

Background Mastitis not only deteriorates the composition or quality of milk,but also damages the health and pro-ductivity of dairy goats.Sulforaphane(SFN)is a phytochemical isothiocyanate compound with various pharmacologi-cal effects such as anti-oxidant and anti-inflammatory.However,the effect of SFN on mastitis has yet to be elucidated.This study aimed to explore the anti-oxidant and anti-inflammatory effects and potential molecular mechanisms of SFN in lipopolysaccharide(LPS)-induced primary goat mammary epithelial cells(GMECs)and a mouse model of mastitis.Results In vitro,SFN downregulated the mRNA expression of inflammatory factors(tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6),inhibited the protein expression of inflammatory mediators(cyclooxygenase-2(COX2),and inducible nitric oxide synthase(iNOS))while suppressing nuclear factor kappa-B(NF-κB)activation in LPS-induced GMECs.Additionally,SFN exhibited an antioxidant effect by increasing Nrf2 expression and nuclear translocation,up-regulating antioxidant enzymes expression,and decreasing LPS-induced reactive oxygen species(ROS)produc-tion in GMECs.Furthermore,SFN pretreatment promoted the autophagy pathway,which was dependent on the increased Nrf2 level,and contributed significantly to the improved LPS-induced oxidative stress and inflammatory response.In vivo,SFN effectively alleviated histopathological lesions,suppressed the expression of inflammatory factors,enhanced immunohistochemistry staining of Nrf2,and amplified of LC3 puncta LPS-induced mastitis in mice.Mechanically,the in vitro and in vivo study showed that the anti-inflammatory and anti-oxidative stress effects of SFN were mediated by the Nrf2-mediated autophagy pathway in GMECs and a mouse model of mastitis.Conclusions These results indicate that the natural compound SFN has a preventive effect on LPS-induced inflam-mation through by regulating the Nrf2-mediated autophagy pathway in primary goat mammary epithelial cells and a mouse model of mastitis,which may improve prevention strategies for mastitis in dairy goats.

Whole-genome methylation analysis reveals epigenetic variation between wild-type and nontransgenic cloned,ASMT transgenic cloned dairy goats generated by the somatic cell nuclear transfer

Background:SCNT(somatic cell nuclear transfer)is of great significance to biological research and also to the livestock breeding.However,the survival rate of the SCNT cloned animals is relatively low compared to other transgenic methods.This indicates the potential epigenetic variations between them.DNA methylation is a key marker of mammalian epigenetics and its alterations will lead to phenotypic differences.In this study,ASMT(acetylserotonin-Omethyltransferase)ovarian overexpression transgenic goat was produced by using SCNT.To investigate whether there are epigenetic differences between cloned and WT(wild type)goats,WGBS(whole-genome bisulfite sequencing)was used to measure the whole-genome methylation of these animals.Results:It is observed that the different m Cp G sites are mainly present in the intergenic and intronic regions between cloned and WT animals,and their CG-type methylation sites are strongly correlated.DMR(differentially methylated region)lengths are located around 1000 bp,mainly distributed in the exonic,intergenic and intronic functional domains.A total of 56 and 36 DMGs(differentially methylated genes)were identified by GO and KEGG databases,respectively.Functional annotation showed that DMGs were enriched in biological-process,cellularcomponent,molecular-function and other signaling pathways.A total of 10 identical genes related to growth and development were identified in GO and KEGG databases.Conclusion:The differences in methylation genes among the tested animals have been identified.A total of 10 DMGs associated with growth and development were identified between cloned and WT animals.The results indicate that the differential patterns of DNA methylation between the cloned and WT goats are probably caused by the SCNT.These novel observations will help us to further identify the unveiled mechanisms of somatic cell cloning technology,particularly in goats.