Analysis of microRNA expression profiles during the differentiation of chicken embryonic stem cells into male germ cells.

The differentiation of embryonic stem cells (ESCs) into germ cells in vitro could have very promising applications for infertility treatment and could provide an excellent model for uncovering the molecular mechanisms of germline generation. This study aimed to investigate the differentially expressed miRNAs (DEMs) during the differentiation of chicken ESCs (cESCs) into male germ cells and to establish a profile of the DEMs. Cells before and after induction were subjected to miRNA sequencing (miRNA-seq). A total of 113 DEMs were obtained, including 61 upregulated and 52 downregulated DEMs. GO and KEGG enrichment analyses showed that the target genes were enriched mainly in the MAPK signaling pathway, HTLV infection signaling pathway, cell adhesion molecule (CAM)-related pathways, viral myocarditis, Wnt signaling pathway, ABC transporters, TGF-ß signaling pathways, Notch signaling pathways and insulin signaling pathway. The target genes of the miRNAs were related to cell binding, cell parts and biological regulatory processes. Six DEMs, let-7k-5p, miR-132c-5p, miR-193a-5p, miR-202-5p, miR-383-5p and miR-6553-3p, were assessed by qRT-PCR, and the results were consistent with the results of miRNA-seq. Based on qRT-PCR and western blot verification, miR-383-5p and its putative target gene STRN3 were selected to construct an STRN3 3'-UTR dual-luciferase gene reporter vector and its mutant vector. The double luciferase reporter activity of the cotransfected STRN3-WT + miR-383-5p mimics group was significantly lower (by approximately 46%) than that of the other five groups (p < 0.01). There was no significant difference in luciferase activity among the other 5 groups. This study establishes a DEM profile during the process of cESC differentiation into male germ cells; illustrates the mechanisms by which miRNAs regulate target genes; provides a theoretical basis for further research on the mechanisms of the formation and regulation of male germ cells; and provides an important strategy for gene editing, animal genetic resource protection and transgenic animal production.

Detection of selection signatures in South African Mutton Merino sheep using whole-genome sequencing data.

The South African Mutton Merino (SAMM), a dual-purpose (meat and wool) sheep breed, is characterized by its excellent performance on growth, carcass traits and meat quality compared to other fine-wool Merino breeds. Nowadays, the SAMM breed has been widely used to cross with commercial and indigenous fine-wool or coarse-wool breeds to improve the growth and meat performance in many countries. To date, however, little is known about the genetic basis for its prominent characteristics. In this study, whole-genome sequences of 10 SAMM were sequenced and the selection signatures were analyzed together with those of 39 Australian Merino and Chinese Merino (wool-type Merino) by FST , iHS, and XP-EHH methods. In total, 313 genes in 277 regions were identified by at least 2 methods with the signal of selection and 21 of them were identified by all three methods. We highlighted a list of interesting genes, including GHR, LCORL, SMO, NCAPG, DCC, IBSP, PPARGC1A, PACRGL, PRDM5, XYLB, AHCYL2, TEFM, AFG1L, and FAM184B, which have been shown to be involved in growth, carcass traits, and meat quality by previous studies. Herein, GHR, encoding a transmembrane receptor for growth hormone, is the most notable one. We report the first study on selection signatures analysis of SAMM at whole-genome sequence level. These results provide new insights into the genetic mechanisms underlying the growth and carcass traits in SAMM.

Dual-Functional Eu2+/3+-Complex@ZIF-67 Nanocatalyst Derived from a Green Reduction of Eu3+ Compound.

The Eu2+/3+ mixed-valent complexes have aroused attention because of their potential application in the catalytic field endowed by the variable valence. Herein, we develop an ingenious green strategy to achieve the partial reduction of Eu3+ to Eu2+ ions in the complex of pyrenol-containing cyclen (H4(Pyr)4cyclen, H4[PC]) via a ligand-to-metal charge transfer (LMCT) effect in air at room temperature. To reveal the inherent regulated capacity of the Eu2+/3+ complex in catalysis, we prepared the nanocomposites assembled by the lanthanide complexes encapsulated into ZIF-67 to successively realize the decomposition of H2O2, including europium, gadolinium, and terbium complexes. Among the nanocomposites, Eu2+/3+[PC]H@ZIF-67 exhibits the best catalytic performance due to the achievement of dual regulation of Co3+/Co2+ ratio by the mixed-valent complex. Because of extremely abundant Co2+ active sites, the Eu2+/3+[PC]H@ZIF-67 after catalyzing H2O2 sample was further utilized to degrade organic dye rhodamine B (RhB). The rather outstanding catalytic degradation efficiency was still present in Eu2+/3+[PC]H@ZIF-67. This research presents a facile strategy of Eu3+ reduction based on the ligand design and a new direction for the future development of the composite catalytic materials with mixed-valent complexes.

Effect of HDAC9 on the differentiation of chicken embryonic stem cells into male germ cells.

Histone deacetylase 9 (HDAC9) is a histone deacetylase (HDAC) subtype IIa protein that deacetylates histone 3 (H3), histone 4 (H4), and nonhistone proteins in vivo to alter chromosomal shape and regulate gene transcription. There have been few studies on the regulatory influence of the HDAC9 gene on the differentiation of chicken embryonic stem cells (cESCs) into male germ cells, and the significance of HDAC9 is still unknown. Therefore, we explored the specific role of HDAC9 during differentiation of the cESCs of Jilin Luhua chickens through inhibition or overexpression. In medium supplemented with 10-5 mol/L retinoic acid (RA), cESCs were stimulated to develop into germ cells. HDAC9 and germline marker gene mRNA and protein levels were measured using qRT‒PCR and western blotting. During the differentiation of cESCs into male germ cells, overexpression of the HDAC9 gene greatly increased the mRNA and protein expression levels of the germline marker genes Stra8, Dazl, c-kit, and integrin ɑ6. The HDAC9 inhibitor TMP195 significantly decreased the mRNA and protein expression levels of the above markers. In summary, HDAC9 positively regulates the differentiation of cESCs.

Coordination-induced and tunable layered rare-earth hydroxide-complex intercalated nanohybrid phosphorescent photosensitizer and therapy.

Hydrotalcite intercalated nanohybrid has served as a vital phosphorescent photosensitizer owing to remarkable 1O2 quantum yield and high cell mortality performance. However, it is rather difficult for potential large or complex guest phosphors to directly intercalate into the hydrotalcite gallery. Hence, it is necessary to regulate the interlayer microenvironment of hydrotalcites firstly for outstanding photosensitive properties. Herein, two isomers, 5,5'BDA and 4,4'BDA, with distinctive dual coordinative features were selected to modify the layer microenvironment of the LGdH gallery and induce the introduction of prospective Gd(HPhN)3 phosphorescent complexes into hydrotalcite through two different coordination effects successively. A LGdH-BDA-Gd(HPhN)3 intercalated nanohybrid phosphorescent photosensitizer was successfully obtained. The results indicated that the more efficient improvement was observed from 5,5'BDA due to offering a more spacious and stable space. Specifically, LGdH-5,5'BDA-Gd(HPhN)3 showed significantly better room temperature phosphorescence properties than LGdH-4,4'BDA-Gd(HPhN)3, whose lifetime was nearly 15 times longer than the latter. Additionally, the LGdH-5,5'BDA-Gd(HPhN)3 system displayed superior singlet oxygen generation in vitro under 460 nm irradiation (the quantum yield Φ = 0.48) and outstanding photodynamic therapy performance in tumor cells. LGdH presented more remarkable enhancement performance on the RTP properties of the luminescent molecules. This work provides a novel platform for designing a high-performance hydrotalcite intercalated nanohybrid phosphorescent photosensitizer through coordination induction to regulate the layer microenvironment.

Whole genome sequencing identified genomic diversity and candidated genes associated with economic traits in Northeasern Merino in China.

Introduction: Northeast Merino (NMS) is a breed developed in Northeast China during the 1960s for wool and meat production. It exhibits excellent traits such as high wool yield, superior meat quality, rapid growth rate, robust disease resistance, and adaptability to cold climates. However, no studies have used whole-genome sequencing data to investigate the superior traits of NMS. Methods: In this study, we investigated the population structure, genetic diversity, and selection signals of NMS using whole-genome sequencing data from 20 individuals. Two methods (integrated haplotype score and composite likelihood ratio) were used for selection signal analysis, and the Fixation Index was used to explore the selection signals of NMS and the other two breeds, Mongolian sheep and South African meat Merino. Results: The results showed that NMS had low inbreeding levels, high genomic diversity, and a pedigree of both Merino breeds and Chinese local breeds. A total length of 14.09 Mb genomic region containing 287 genes was detected using the two methods. Further exploration of the functions of these genes revealed that they are mainly concentrated in wool production performance (IRF2BP2, MAP3K7, and WNT3), meat production performance (NDUFA9, SETBP1, ZBTB38, and FTO), cold resistance (DNAJC13, LPGAT1, and PRDM16), and immune response (PRDM2, GALNT8, and HCAR2). The selection signals of NMS and the other two breeds annotated 87 and 23 genes, respectively. These genes were also mainly focused on wool and meat production performance. Conclusion: These results provide a basis for further breeding improvement, comprehensive use of this breed, and a reference for research on other breeds.

The comparison of meat yield, quality, and flavor between small-tailed Han sheep and two crossbred sheep and the verification of related candidate genes.

Introduction: In Northeast China, Dorper and Australian White rams are commonly crossbred with small-tailed Han (STH) ewes to improve the offspring's meat yield and quality. However, the differences in traits and the flavor between the crossbred sheep and STH sheep remain unclear. In addition, the candidate genes potentially influencing the meat quality in the three sheep breeds require further verification. Methods: A total of 18 2-month-old healthy rams were raised over a period of 5 months, which included 6 STH, 6 Dorper and small-tailed Han crossbred (Do × STH), and 6 Australian white and small-tailed Han crossbred (Au × STH) offspring. The differences in slaughter, meat quality traits, fatty acid and amino acid composition in the muscular longissimus dorsi (MLD), and volatile compounds in the semitendinosus muscle were compared between the sheep breeds. The candidate genes related to intramuscular fat (IMF) content and fatty acids were validated. Results: The results of this study revealed that the crossbred sheep had higher body weight, carcass weight, bone weight, net meat weight, and IMF content than the STH sheep (p < 0.05). The Do × STH offspring had a higher pH value (24 h), moisture content, and cooking percentage; they also had redder and brighter meat color. The content of myristate, palmitic, and margaric acids in the crossbred sheep was higher than that in the STH sheep (p < 0.05). The Do × STH offspring had the highest saturated fatty acid content (p < 0.05). The Au × STH offspring had the highest protein content (p < 0.05). The arachidonic acid and amino acid (Asp, Ala, Ile, Leu, Lys, Thr, and essential amino acid) contents were higher in the STH sheep than in the crossbred sheep (p < 0.05). The odor activity value (OAV) analysis showed that most of the aldehydes in the Au × STH offspring had higher values. The PDK4 gene expression was positively associated with the IMF content and was negatively correlated with the linoleic acid content in the Do × STH sheep (p < 0.05). The TMEM273 gene expression was positively associated with linoleic and arachidonic acid contents and was negatively correlated with oleic and palmitic acid contents in the Do × STH sheep (p < 0.05). Discussion: The results showed the differences between the crossbred sheep and STH sheep and provided the candidate genes related to meat quality in sheep.

Melatonin Protects the Apoptosis of Sheep Granulosa Cells by Suppressing Oxidative Stress via MAP3K8 and FOS Pathway.

Melatonin is not only a highly effective active oxygen scavenger but also an important reproductive hormone. Melatonin has a regulatory effect on animal reproduction, especially on the ovaries. It can affect the proliferation and apoptosis of cells in follicles. However, the mechanisms of the dual antioxidation and anti-apoptosis effects of melatonin on granulosa cells are still not clear, especially in sheep. Therefore, we investigated the mechanisms of the protective effect of melatonin against oxidative damage in granulosa cells. At a concentration of 250 µmol/L, H2O2 promoted granulosa cell apoptosis; however, 10 ng/mL melatonin effectively alleviated the pro-apoptotic effect of H2O2. Furthermore, through the application of high-throughput sequencing technology, we identified 109 significantly differentially expressed genes (35 upregulated and 74 downregulated genes) involved in the protective effect of melatonin against apoptosis. The expression levels of nine related genes, i.e., ATF3, FIBIN, FOS, HSPA6, MAP3K8, FOSB, PET117, DLX2, and TRIB1, changed significantly. MAP3K8 and FOS gene overexpression impacted the protective effect of melatonin in granulosa cells; the two genes exhibited an upstream and downstream regulatory relationship. Our findings indicated that melatonin alleviated H2O2-induced apoptosis in sheep granulosa cells through the MAP3K8-FOS pathway.

Extraction of honokiol from Artemisia argyi and in vitro and in vivo investigation of its antifungal activity.

Extracts from plants used in Chinese medicine can be good sources of fungicides for agricultural applications. In this study, we separated and identified antifungal compounds from four traditional Chinese medicine extracts and evaluated their antifungal activities in vitro and in vivo. In vitro, honokiol extracted from Artemisia argyi showed broad-spectrum antimicrobial and mycelial inhibitory activity with EC50 in the range 3.56 - 33.85 µg/mL against eight plant pathogens. q-PCR indicated that honokiol might induce cell cancerisation and inhibit cellular respiration, which provided significant insights into honokiol function in tobacco resistance to molecular mechanisms of the phytopathogenic fungus Phytophthora nicotianae. In vivo, honokiol significantly decreased the rate of fungal infection in eggplants, potatoes, grapes, cherry tomatoes, and cucumbers, and enhanced disease resistance in tobacco. Overall, our results indicate that honokiol has the potential to control a variety of fungal and oomycete diseases, and A. argyi could be a source of honokiol.

Transcriptome Analysis of the Liver and Muscle Tissues of Dorper and Small-Tailed Han Sheep.

It is well known that Dorper (DP) is a full-bodied, fast-growing and high dressing percentage breed, while the production performance of Small-tailed Han sheep (STH) is not so excellent, in contrast to DP. Therefore, in this study, a comparative transcriptomic analysis of liver and muscle tissues from DP and STH breeds was carried out to find differentially expressed genes (DEGs) that affect their growth and meat quality traits. The results showed that the total number of DEGs was 2,188 in the two tissues. There were 950, 160 up-regulated and 1,007, 71 down-regulated genes in the liver and muscle, respectively. Several DEGs such as TGFB1, TGFB3, FABP3, LPL may be associated with growth and development in DP. Also, several GO terms were found to be associated with muscle growth and development, such as developmental growth (GO:0048589), and myofibril (GO:0030016). Further validation of eight genes (6 up-regulated, and 2 down-regulated) was performed using quantitative RT-PCR. These findings will provide valuable information for studying growth and development as well as meat quality traits in sheep.

Cloning and characterization of class 1 and class 2 insulin-like growth factor-I mRNA in Songliao black pig.

Insulin-like growth factor-I (IGF-I) plays a fundamental role in postnatal mammalian growth, development, and metabolism. The mammalian IGF-I gene contains at least six exons from which several different mRNAs are transcribed. In this study, IGF-I mRNA in Songliao black pig liver was investigated using RLM-RACE. Using a 3'-RACE technique, we determined that all mRNA transcripts lacked exon 5 sequence and contained only exon 6 sequence. Using a 5'-RACE technique, we investigated the presence of class 1 and class 2 IGF-I mRNAs. In several other species, the class 1 and class 2 IGF-I polypeptides are generated from mRNAs containing exon 1 or exon 2, respectively. Both class 1 and class 2 IGF-I mRNAs were identified in Songliao black pig liver. Transcription is initiatated upstream of exons 1 and 2 at multiple dispersed start sites to yield two distinct IGF-I mRNA transcript classes which differ in the precursor peptides predicted from their individual leader sequences. Tissue distribution of Songliao black pig class 1 and class 2 IGF-I mRNA was investigated by real-time RT-PCR. Both classes of IGF-I mRNA were expressed in a variety of tissues, however, class 1 IGF-I mRNA was more abundant than class 2 in all tissues.

A smart nanoprobe based on a gadolinium complex encapsulated by ZIF-8 with enhanced room temperature phosphorescence for synchronous oxygen sensing and photodynamic therapy.

The phosphorescence lifetime approach based on the room temperature phosphorescence (RTP) property has received considerable attention in recent years due to its excellent performance in the precise measurement of oxygen. Herein, a smart nanoprobe, Gd[PC]@ZIF-8, was designed and assembled by homogenously encapsulating a rare-earth complex phosphor Gd[(Pyr)4cyclen] (Pyr = pyrenol) into a zeolitic imidazolate framework (ZIF-8). Because of the restriction of the metal-organic framework (MOF) matrix and host-guest interactions, the nanoprobe Gd[PC]@ZIF-8 exhibited highly enhanced RTP properties, including intensity, quantum yield, and elongated decay lifetime. It displayed an outstanding linear relationship between the phosphorescence decay lifetime, intensity and oxygen concentration, which can be applied in the field of oxygen sensing. Moreover, the complex Gd[(Pyr)4cyclen] in the nanoprobe Gd[PC]@ZIF-8 served as a favorable photosensitizer that resulted in the simultaneous conversion of sufficient oxygen molecules into single state oxygen (1O2) under irradiation during the phosphorescence quenching process, which is conducive to photodynamic therapy (PDT). Thus, the design of the smart nanoprobe Gd[PC]@ZIF-8 in this study provides an ingenious strategy of utilizing a MOF as a matrix to enhance the RTP properties of phosphors for synchronous oxygen sensing and PDT.