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1.
BMC Genomics ; 25(1): 386, 2024 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-38641604

RESUMO

BACKGROUND: The growth and development of organism were dependent on the effect of genetic, environment, and their interaction. In recent decades, lots of candidate additive genetic markers and genes had been detected by using genome-widely association study (GWAS). However, restricted to computing power and practical tool, the interactive effect of markers and genes were not revealed clearly. And utilization of these interactive markers is difficult in the breeding and prediction, such as genome selection (GS). RESULTS: Through the Power-FDR curve, the GbyE algorithm can detect more significant genetic loci at different levels of genetic correlation and heritability, especially at low heritability levels. The additive effect of GbyE exhibits high significance on certain chromosomes, while the interactive effect detects more significant sites on other chromosomes, which were not detected in the first two parts. In prediction accuracy testing, in most cases of heritability and genetic correlation, the majority of prediction accuracy of GbyE is significantly higher than that of the mean method, regardless of whether the rrBLUP model or BGLR model is used for statistics. The GbyE algorithm improves the prediction accuracy of the three Bayesian models BRR, BayesA, and BayesLASSO using information from genetic by environmental interaction (G × E) and increases the prediction accuracy by 9.4%, 9.1%, and 11%, respectively, relative to the Mean value method. The GbyE algorithm is significantly superior to the mean method in the absence of a single environment, regardless of the combination of heritability and genetic correlation, especially in the case of high genetic correlation and heritability. CONCLUSIONS: Therefore, this study constructed a new genotype design model program (GbyE) for GWAS and GS using Kronecker product. which was able to clearly estimate the additive and interactive effects separately. The results showed that GbyE can provide higher statistical power for the GWAS and more prediction accuracy of the GS models. In addition, GbyE gives varying degrees of improvement of prediction accuracy in three Bayesian models (BRR, BayesA, and BayesCpi). Whatever the phenotype were missed in the single environment or multiple environments, the GbyE also makes better prediction for inference population set. This study helps us understand the interactive relationship between genomic and environment in the complex traits. The GbyE source code is available at the GitHub website ( https://github.com/liu-xinrui/GbyE ).


Assuntos
Locos de Características Quantitativas , Seleção Genética , Teorema de Bayes , Modelos Genéticos , Fenótipo , Genótipo , Estudo de Associação Genômica Ampla/métodos , Polimorfismo de Nucleotídeo Único
2.
J Biophotonics ; 17(4): e202300422, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38211977

RESUMO

Multiphoton microscopy (MPM) enables deep brain imaging. Three optical windows: NIR-I, NIR-II, and NIR-III are widely used. Recently, NIR-IV (the 2200 nm window) has been demonstrated to be the last and longest window for deep tissue MPM. However, so far MPM covers only two optical windows labeled by single fluorescent probe, one for emission and one for excitation. Here we demonstrate in vivo deep brain MPM covering three optical windows, with emission at NIR-I, NIR-II, and excitation at NIR-IV, labeled by ICG. The innovations include: (1) characterizing both 3-photon excitation and emission properties of ICG emitting at both NIR-I and NIR-II, in water, plasma, and circulating blood; (2) a home-built multiphoton microscope with simultaneous dual channel detection, with which we demonstrate deep brain MPM 950 µm (NIR-I) and 850 µm (NIR-II) into the mouse brain in vivo, verifying that multi-optical window MPM is promising for deep brain imaging.


Assuntos
Encéfalo , Microscopia de Fluorescência por Excitação Multifotônica , Camundongos , Animais , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Encéfalo/diagnóstico por imagem , Corantes Fluorescentes , Imagem Óptica/métodos , Espectroscopia de Luz Próxima ao Infravermelho/métodos
3.
Nanoscale Adv ; 6(2): 511-515, 2024 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-38235073

RESUMO

3-Photon microscopy (3PM) excited at the 1700 nm window features a smaller tissue attenuation and hence a larger penetration depth in brain imaging compared with other excitation wavelengths in vivo. While the comparison of the penetration depth quantified by effective attenuation length le with other excitation wavelengths have been extensively investigated, comparison within the 1700 nm window has never been demonstrated. This is mainly due to the lack of a proper excitation laser source and characterization of the in vivo emission properties of fluorescent labels within this window. Herein, we demonstrate detailed measurements and comparison of le through the 3-photon imaging of the mouse brain in vivo, at different excitation wavelengths (1600 nm, 1700 nm, and 1800 nm). 3PF imaging and in vivo spectrum measurements were performed using AIE nanoparticle labeling. Our results show that le derived from both 3PF imaging and THG imaging is the largest at 1700 nm, indicating that it enables the deepest penetration in brain imaging in vivo.

4.
Int J Mol Sci ; 25(1)2024 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-38203803

RESUMO

miR-129 plays a crucial role in regulating various cellular processes, including adipogenesis; however, its downstream molecular mechanisms remain unclear. In this study, we demonstrated that miR-129 promotes yak adipogenesis in vitro via the PI3K/AKT pathway. Overexpression and interference of miR-129 in yak intramuscular preadipocytes (YIMAs) enhanced and inhibited cell differentiation, respectively, with corresponding changes in cell proliferation. Further investigation revealed that miR-129 enhances AKT and p-AKT activity in the AKT pathway without affecting cell apoptosis, and a specific inhibitor (LY294002) was used to confirm that miR-129 regulates YIMAs proliferation and differentiation through the PI3K/AKT pathway. Our findings suggest that miR-129 promotes yak adipogenesis by enhancing PI3K/AKT pathway activity. This study provides the foundation to precisely elucidate the molecular mechanism of miR-129 in YIMAs adipogenesis and develop advanced miRNA-based strategies to improve meat nutrition and obesity-related ailments in beef production.


Assuntos
MicroRNAs , Fosfatidilinositol 3-Quinases , Animais , Bovinos , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Diferenciação Celular/genética , Proliferação de Células/genética , MicroRNAs/genética
5.
ACS Omega ; 9(1): 2000-2011, 2024 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-38222569

RESUMO

The increase in the styrene content in styrene-butadiene rubber (SBR) can improve the abrasion performance and cutting resistance of rubber, which has received attention in the tire industry. The fatigue performance is the main evaluation index of rubber materials applied to tires. In this study, the effect of the styrene content and its interaction with carbon black (CB) on the dynamic fatigue performance and mechanism of SBR were investigated. The results indicated that the dynamic fatigue life of the rubber composite materials was prolonged with increasing styrene content; furthermore, it showed a trend of increasing and then decreasing with increasing CB content. At a certain CB content, styrene and CB displayed a synergistic effect on improving the dynamic fatigue life of the composite materials. The dynamic fatigue performance of SBR40/CB20 was the best. The expansion of the fatigue cracks followed the secondary cracking mechanism, which consumed a large amount of strain energy and slowed the development of the main crack. However, when the CB content exceeded 40 phr, the mechanism transformed to main crack self-propagation and the fatigue life decreased.

6.
BMC Genomics ; 25(1): 69, 2024 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-38233755

RESUMO

BACKGROUND: The yak is a symbol of the Qinghai-Tibet Plateau and provides important basic resources for human life on the plateau. Domestic yaks have been subjected to strong artificial selection and environmental pressures over the long-term. Understanding the molecular mechanisms of phenotypic differences in yak populations can reveal key functional genes involved in the domestication process and improve genetic breeding. MATERIAL AND METHOD: Here, we re-sequenced 80 yaks (Maiwa, Yushu, and Huanhu populations) to identify single-nucleotide polymorphisms (SNPs) as genetic variants. After filtering and quality control, remaining SNPs were kept to identify the genome-wide regions of selective sweeps associated with domestic traits. The four methods (π, XPEHH, iHS, and XP-nSL) were used to detect the population genetic separation. RESULTS: By comparing the differences in the population stratification, linkage disequilibrium decay rate, and characteristic selective sweep signals, we identified 203 putative selective regions of domestic traits, 45 of which were mapped to 27 known genes. They were clustered into 4 major GO biological process terms. All known genes were associated with seven major domestication traits, such as dwarfism (ANKRD28), milk (HECW1, HECW2, and OSBPL2), meat (SPATA5 and GRHL2), fertility (BTBD11 and ARFIP1), adaptation (NCKAP5, ANTXR1, LAMA5, OSBPL2, AOC2, and RYR2), growth (GRHL2, GRID2, SMARCAL1, and EPHB2), and the immune system (INPP5D and ADCYAP1R1). CONCLUSIONS: We provided there is an obvious genetic different among domestic progress in these three yak populations. Our findings improve the understanding of the major genetic switches and domestic processes among yak populations.


Assuntos
ATPases Associadas a Diversas Atividades Celulares , Domesticação , Receptores de Esteroides , Animais , Humanos , Bovinos/genética , Genoma , Análise de Sequência de DNA , Tibet , Genética Populacional , Proteínas dos Microfilamentos , Receptores de Superfície Celular , DNA Helicases , Proteínas do Tecido Nervoso , Ubiquitina-Proteína Ligases
7.
Int J Biol Macromol ; 257(Pt 1): 128531, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38042314

RESUMO

The regulatory mechanisms and functions of circular RNAs (circRNAs) in yak intramuscular fat (IMF) deposition remain unclear. This study aimed to investigate yak circRNAs with high and low IMF content using high-throughput sequencing. A total of 270 differentially expressed circRNAs were identified, of which 129 were upregulated and 141 were downregulated. Among these circRNAs, circCWC22, derived from the yak CWC22 gene, was further studied to understand its functions and regulatory mechanisms. Sequencing and RNase R processing confirmed the circular nature of circCWC22. By constructing a circRNA-miRNA-mRNA co-expression network, the potential regulatory pathway of circCWC22/miR-3059-x/HMGCL was identified. To investigate the roles of circCWC22, miR-3059-x, and HMGCL in the deposition of yak intramuscular preadipocytes (YIMAs), CCK-8, EdU, BODIPY, triglyceride content, and qRT-PCR analyses were performed. The results demonstrated that circCWC22, miR-3059-x, and HMGCL promoted the differentiation and inhibited the proliferation of YIMAs. Using the dual-luciferase reporter system and qRT-PCR, we confirmed that circCWC22 adsorbed miR-3059-x, and HMGCL was identified as a target gene of miR-3059-x. In conclusion, this study uncovered a large number of potential circRNAs involved in IMF deposition and highlighted the significant role of circCWC22 in yak IMF deposition via the circCWC22/miR-3059-x/HMGCL axis.


Assuntos
MicroRNAs , RNA Circular , Animais , Bovinos , RNA Circular/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Ontologia Genética , Redes Reguladoras de Genes
8.
Front Vet Sci ; 10: 1272238, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37915947

RESUMO

The concentration of intramuscular fat (IMF) is a crucial determinant of yak meat quality. However, the molecular mechanisms that regulate IMF in yak remain largely elusive. In our study, we conducted transcriptome sequencing on the longissimus dorsi muscle tissues of yaks with varying IMF contents. We then filtered differentially expressed genes (DEGs), microRNAs (DEMs), and long non-coding RNAs (DELs) to elucidate potential regulatory pathways of adipogenesis in yaks. Overall, our research sheds light on an array of potential mRNAs and noncoding RNAs implicated in IMF deposition and elaborates on the role of HIF1α in yaks. These findings contribute valuable insights that can serve as a guide for further research into the molecular mechanisms governing IMF deposition.

9.
Anim Biotechnol ; 34(8): 3978-3987, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37593948

RESUMO

Meat tenderness is an important sensory index when consumers choose meat products, which determines the value of meat products and consumers' buying intentions. Yak meat is rich in nutrition and unique in flavor, which is favored by consumers. However, its meat has the deficiencies of low tenderness and poor taste, which has a negative impact on the value of its meat products and customer satisfaction. To identify the genes affecting the yak meat tenderness, we used RNA-seq to analyze the longissimus dorsi muscle of yaks with different tenderness, screened a total of 1120 differentially expressed genes (DEGs). Meanwhile, 23 pathways were significantly enriched. By further analysis, we identified eight genes related to yak meat tenderness (WNT5A, ARID5B, SERPINE1 KLHL40, RUNX1, MAFF, RFX7 and ARID5A). Notably, SERPINE1 was involved in the significant enrichment pathways of 'complement and coagulation cascade pathway', 'HIF-1 signaling pathway' and 'AGE-RAGE signaling pathway in diabetic complications' which can regulate meat tenderness. This implies that SERPINE1 may play an important regulatory role among them. The DEGs associated with yak meat quality screened in this work will be helpful to identify potential biomarkers related to meat tenderness.


Assuntos
Perfilação da Expressão Gênica , Carne , Bovinos/genética , Animais , Carne/análise , RNA-Seq , Músculo Esquelético/metabolismo , Transcriptoma/genética
10.
Int J Mol Sci ; 24(14)2023 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-37511041

RESUMO

Sirtuin 1 (SIRT1) overexpression significantly inhibits lipid deposition during yak intramuscular preadipocyte (YIMA) differentiation; however, the regulatory mechanism remains unknown. We elucidated the role of SIRT1 in YIMA differentiation using lentivirus-mediated downregulation technology and conducted mRNA-seq and ChIP-seq assays using H3K9ac antibodies after SIRT1 overexpression in order to reveal SIRT1 targets during YIMA adipogenesis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed in order to identify the functional annotation of common genes. In addition, a potential target of SIRT1 was selected to verify its effects on the differentiation and proliferation of YIMAs. SIRT1 interfered with lipid deposition and promoted YIMA differentiation. In total, 143,518 specific peaks were identified after SIRT1 overexpression, where genes associated with downregulation peaks were enriched in transcription, gene expression, lipid-related processes, and classical lipid-related pathways. The H3K9ac signal in the whole genome promoter region (2 kb upstream and downstream of the transcription start site (TSS)) was weakened, and the peaks were distributed across all gene functional regions. Genes that lost signals in their TSS region or gene body region were enriched in both biological processes and pathways associated with lipogenesis. The ChIP-seq results revealed 714 common differential genes in mRNA-seq, which were enriched in "MAPK signaling", "lipid and atherosclerosis", "mTOR signaling", and "FoxO signaling" pathways. A total of 445 genes were downregulated in both their H3K9ac signals and mRNA expression, and one of their most significantly enriched pathways was FoxO signaling. Nine genes (FBP2, FPGT, HSD17B11, KCNJ15, MAP3K20, SLC5A3, TRIM23, ZCCHC10, and ZMYM1) lost the H3K9ac signal in their TSS regions and had low mRNA expression, and three genes (KCNJ15, TGM3, and TRIM54) had low expression but lost their H3K9ac signal in the gene body region. The interference of TRIM23 significantly inhibited fat deposition during preadipocyte differentiation and promoted cell proliferation by increasing S-phase cell numbers. The present study provides new insights into the molecular mechanism of intramuscular fat content deposition and the epigenetic role of SIRT1 in adipocyte differentiation.


Assuntos
Adipogenia , Epigenômica , Sirtuína 1 , Adipócitos/metabolismo , Diferenciação Celular/genética , Lipídeos/farmacologia , RNA Mensageiro/metabolismo , Sirtuína 1/genética , Sirtuína 1/metabolismo , Adipogenia/genética
11.
Gene ; 878: 147574, 2023 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-37336271

RESUMO

Ubiquitination is a widespread post-transcriptional modification (PTM) that occurs during protein degradation in eukaryotes and participates in almost all physiological and pathological processes, including animal adipogenesis. Ubiquitination is a cascade reaction regulated by the activating enzyme E1, conjugating enzyme E2, and ligase E3. Several recent studies have reported that E3 ligases play important regulatory roles in adipogenesis. However, as a key influencing factor for the recognition and connection between the substrate and ubiquitin during ubiquitination, its regulatory role in adipogenesis has not received adequate attention. In this review, we summarize the E3s' regulation and modification targets in animal adipogenesis, explain the regulatory mechanisms in lipogenic-related pathways, and further analyze the existing positive results to provide research directions of guiding significance for further studies on the regulatory mechanisms of E3s in animal adipogenesis.


Assuntos
Adipogenia , Ubiquitina-Proteína Ligases , Animais , Ubiquitinação , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina/metabolismo , Proteólise , Enzimas de Conjugação de Ubiquitina/metabolismo
12.
Animals (Basel) ; 13(9)2023 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-37174506

RESUMO

Yaks have evolved several breeds or genetic resources owing to their geographical and ecological environment, and investigating the genetic construction of body size among breeds is key for breeding. Here, a genome-wide association study (GWAS) was performed for five body size traits in 31 yak breeds and genetic resources. The information from clustering individuals according to their habitats was used for kinship grouping in the compressed mixed linear model (CMLM). We named this approach the pCMLM method. A total of 3,584,464 high-quality single nucleotide polymorphisms (SNPs) were obtained, and six markers were found to be significantly associated with height by pCMLM. Four candidate genes, including FXYD6, SOHLH2, ADGRB2, and OSBPL6, were identified. Our results show that when CMLM cannot identify optimal clustering groups, pCMLM can provide sufficient associated results based on population information. Moreover, this study provides basic information on the gene localization of quantitative traits of body size among yak breeds.

13.
Cell Prolif ; 56(9): e13430, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36855961

RESUMO

Skeletal muscle is a complex heterogeneous tissue and characterizing its cellular heterogeneity and transcriptional and epigenetic signatures are important for understanding the details of its ontogeny. In our study, we applied scRNA-seq and scATAC-seq to investigate the cell types, molecular features, transcriptional and epigenetic regulation, and patterns of developing bovine skeletal muscle from gestational, lactational and adult stages. Detailed molecular analyses were used to dissect cellular heterogeneity, and we deduced the differentiation trajectory of myogenic cells and uncovered their dynamic gene expression profiles. SCENIC analysis was performed to demonstrate key regulons during cell fate decisions. We explored the future expression states of these heterogeneous cells by RNA velocity analysis and found extensive networks of intercellular communication using the toolkit CellChat. Moreover, the transcriptomic and chromatin accessibility modalities were confirmed to be highly concordant, and integrative analysis of chromatin accessibility and gene expression revealed key transcriptional regulators acting during myogenesis. In bovine skeletal muscle, by scRNA-seq and scATAC-seq analysis, different cell types such as adipocytes, endothelial cells, fibroblasts, lymphocytes, monocytes, pericyte cells and eight skeletal myogenic subpopulations were identified at the three developmental stages. The pseudotime trajectory exhibited a distinct sequential ordering for these myogenic subpopulations and eight distinct gene clusters were observed according to their expression pattern. Moreover, specifically expressed TFs (such as MSC, MYF5, MYOD1, FOXP3, ESRRA, BACH1, SIX2 and ATF4) associated with muscle development were predicted, and likely future transcriptional states of individual cells and the developmental dynamics of differentiation among neighbouring cells were predicted. CellChat analysis on the scRNA-seq data set then classified many ligand-receptor pairs among these cell clusters, which were further categorized into significant signalling pathways, including BMP, IGF, WNT, MSTN, ANGPTL, TGFB, TNF, VEGF and FGF. Finally, scRNA-seq and scATAC-seq results were successfully integrated to reveal a series of specifically expressed TFs that are likely to be candidates for the promotion of cell fate transition during bovine skeletal muscle development. Overall, our results outline a single-cell dynamic chromatin/transcriptional landscape for normal bovine skeletal muscle development; these provide an important resource for understanding the structure and function of mammalian skeletal muscle, which will promote research into its biology.


Assuntos
Cromatina , Epigênese Genética , Bovinos , Animais , Cromatina/genética , Células Endoteliais/metabolismo , Fatores de Transcrição/metabolismo , Desenvolvimento Muscular/genética , Mamíferos/metabolismo
14.
BMC Genomics ; 24(1): 149, 2023 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-36973659

RESUMO

BACKGROUND: Interspecific hybridization plays vital roles in enriching animal diversity, while male hybrid sterility (MHS) of the offspring commonly suffered from spermatogenic arrest constitutes the postzygotic reproductive isolation. Cattle-yak, the hybrid offspring of cattle (Bos taurus) and yak (Bos grunniens) can serve as an ideal MHS animal model. Although meiotic arrest was found to contribute to MHS of cattle-yak, yet the cellular characteristics and developmental potentials of male germline cell in pubertal cattle-yak remain to be systematically investigated. RESULTS: Single-cell RNA-seq analysis of germline and niche cell types in pubertal testis of cattle-yak and yak indicated that dynamic gene expression of developmental germ cells was terminated at late primary spermatocyte (meiotic arrest) and abnormal components of niche cell in pubertal cattle-yak. Further in vitro proliferation and differentially expressed gene (DEG) analysis of specific type of cells revealed that undifferentiated spermatogonia of cattle-yak exhibited defects in viability and proliferation/differentiation potentials. CONCLUSION: Comparative scRNA-seq and in vitro proliferation analysis of testicular cells indicated that not only meiotic arrest contributed to MHS of cattle-yak. Spermatogenic arrest of cattle-yak may originate from the differentiation stage of undifferentiated spermatogonia and niche cells of cattle-yak may provide an adverse microenvironment for spermatogenesis.


Assuntos
Infertilidade Masculina , Testículo , Animais , Masculino , Humanos , Bovinos , Testículo/metabolismo , Análise da Expressão Gênica de Célula Única , Infertilidade Masculina/genética , Infertilidade Masculina/veterinária , Espermatogênese/genética , Espermatogônias
15.
ACS Nano ; 17(4): 3686-3695, 2023 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-36799427

RESUMO

Multiphoton microscopy (MPM) is an enabling technology for visualizing deep-brain structures at high spatial resolution in vivo. Within the low tissue absorption window, shifting to longer excitation wavelengths reduces tissue scattering and boosts penetration depth. Recently, the 2200 nm excitation window has emerged as the last and longest window suitable for deep-brain MPM. However, multiphoton fluorescence imaging at this window has not been demonstrated, due to the lack of characterization of multiphoton properties of fluorescent labels. Here we demonstrate technologies for measuring both the multiphoton excitation and emission properties of fluorescent labels at the 2200 nm window, using (1) 3-photon (ησ3) and 4-photon action cross sections (ησ4) and (2) 3-photon and 4-photon emission spectra both ex vivo and in vivo of quantum dots. Our results show that quantum dots have exceptionally large ησ3 and ησ4 for efficient generation of multiphoton fluorescence. Besides, the 3-photon and 4-photon emission spectra of quantum dots are essentially identical to those of one-photon emission, which change negligibly subject to the local environment of circulating blood. Based on these characterization results, we further demonstrate deep-brain vasculature imaging in vivo. Due to the superb multiphoton properties of quantum dots, 3-photon and 4-photon fluorescence imaging reaches a maximum brain imaging depth of 1060 and 940 µm below the surface of a mouse brain, respectively, which enables the imaging of subcortical structures. We thus fill the last gap in multiphoton fluorescence imaging in terms of wavelength selection.


Assuntos
Pontos Quânticos , Animais , Camundongos , Pontos Quânticos/química , Encéfalo/diagnóstico por imagem , Encéfalo/irrigação sanguínea , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Corantes Fluorescentes/química , Imagem Óptica
16.
J Biophotonics ; 16(5): e202200365, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36633161

RESUMO

The brain arteriolar wall is a multilayered structure, whose integrity is of key significance to the brain function. However, resolving these different layers in anmial models in vivo is hampered by the lack of either labeling or imaging technology. Here, we demonstrate that three-photon microscopy (3PM) is an ideal solution. In mouse brain in vivo, excited at the 1700-nm window, label-free third-harmonic generation imaging and three-photon fluorescence (3PF) imaging with Alexa 633 labeling colocalize and resolve the internal elastic lamina. Furthermore, Alexa Fluor 594-conjugated Wheat Germ Agglutinin (WGA-594) shows time-dependent labeling behavior. As time lapses, WGA-594 first labels endothelium, and then vascular smooth muscle cells, which are readily captured and resolved with 3PF imaging. Our results show that 3PM, in combination with proper labeling, is a promising technology for investigating the structures of brain arteriolar wall in vivo.


Assuntos
Encéfalo , Microscopia de Fluorescência por Excitação Multifotônica , Camundongos , Animais , Encéfalo/diagnóstico por imagem , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Endotélio
17.
Epigenetics ; 18(1): 2135194, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-36264146

RESUMO

The SIRT1 epigenetic regulator is involved in hepatic lipid homoeostasis. However, the role of SIRT1 in regulating intramuscular fat deposition as well as the pathways and potential epigenetic targets involved remain unknown. Herein, we investigate SIRT1 function, its genome-wide epigenetic target profile, and transcriptomic changes under SIRT1 overexpression during yak intramuscular preadipocytes differentiation. To this end, we analysed the relationship between SIRT1 and intramuscular fat content as well as lipid metabolism-related genes in longissimus dorsi tissue. We found that SIRT1 expression negatively correlates with intramuscular fat content as well as with the expression of genes related to lipid synthesis, while positively correlating with that of fatty acid oxidation-involved genes. SIRT1 overexpression in intramuscular preadipocytes significantly reduced adipose differentiation marker expression, intracellular triacylglycerol content, and lipid deposition. Chromatin immunoprecipitation coupled with high-throughput sequencing of H3K4ac (a known direct target of SIRT1) and high-throughput mRNA sequencing results revealed that SIRT1 may regulate intramuscular fat deposition via three potential new transcription factors (NRF1, NKX3.1, and EGR1) and four genes (MAPK1, RXRA, AGPAT1, and HADH) implicated in protein processing within the endoplasmic reticulum pathway and the MAPK signalling pathway in yaks. Our study provides novel insights into the role of SIRT1 in regulating yak intramuscular fat deposition and may help clarify the mechanistic determinants of yak meat characteristics.


Assuntos
Tecido Adiposo , Sequenciamento de Cromatina por Imunoprecipitação , Tecido Adiposo/metabolismo , Sirtuína 1/metabolismo , Metilação de DNA , Lipídeos , Epigênese Genética , RNA Mensageiro/metabolismo
18.
Animals (Basel) ; 12(22)2022 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-36428402

RESUMO

This experiment was conducted to investigate the effects of methionine analogues 2-hydroxy-4-methylthio butanoic acid isopropyl ester (HBMi) on growth performance, nutrient apparent digestibility, serum metabolite, serum free amino acids, and rumen fermentation parameters of yaks. Twenty-four male Maiwa yaks (252.79 ± 15.95 kg) were randomly allocated to four dietary treatments: basic diet (CON), or three HBMi (MetaSmart (MS); Adisseo Inc., Antony, France) supplementation treatments: MS1 (5 g), MS2 (10 g), and MS3 (15 g). The results showed that the increase in the supplemented MS levels linearly increased the average daily gain (p < 0.05), while the serum alkaline phosphatase activity and malondialdehyde content were increased when yaks were fed with 15 g/d MS (p < 0.05). The diet supplemented with MS linearly increased the percentages of glutamic acid and proline, and linearly or quadratically decreased the percentages of isoleucine, phenylalanine, and valine (p < 0.05). Furthermore, supplementation of 10 g/d and 15 g/d MS increased ruminal microbial crude protein (p < 0.05). The ratio of acetate to propionate in the MS2 group was lower than those in CON and MS1 groups (p < 0.05). In summary, a diet supplemented with 10 g/d MS could be an effective way to improve the growth performance of fattening yaks without negative effects.

19.
Mol Reprod Dev ; 89(12): 632-645, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36409004

RESUMO

Hybrid male sterility (HMS) is a reproductive isolation mechanism limiting the formation of fertile offspring through interspecific fertilization. Cattleyak is the interspecific hybrid presenting significant heterosis in several economic traits, but HMS restricted its wide reproduction in cettleyak breeding. In this study, we detected the specifically expressed genes of a variety of cells (undifferentiated spermatogonia, primary spermatocytes, secondary spermatocytes, haploid spermatids, sperm, Sertoli cells, Leydig cells, and macrophages) in the testis of yak and cattleyak, and found that the spermatogenesis of cattleyak might be blocked at meiosis I, and the expression of niche factors (NR5A1, GATA4, STAR, CYP11A1, CD68, TNF, and CX3CR1) in undifferentiated spermatogonia niche was abnormal. Then we isolated the undifferentiated spermatogonia and Sertoli cells from yak and cattleyak by enzyme digestion, and detected the specific genes in the two bovid testicular cells as well as the proliferation capacity of the undifferentiated spermatogonia. These results indicated that weak proliferation ability and scarce number of undifferentiated spermatogonia and abnormal gene expressions in Sertoli cells may contribute to male sterility of cattleyak.


Assuntos
Infertilidade Masculina , Espermatogônias , Humanos , Masculino , Espermatogônias/metabolismo , Células de Sertoli/metabolismo , Sêmen , Testículo/metabolismo , Espermatogênese/genética , Infertilidade Masculina/metabolismo
20.
Int J Mol Sci ; 23(20)2022 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-36293266

RESUMO

Due to its prominent secretory activity, adipose tissue (AT) is now considered a major player in the crosstalk between organs, especially with skeletal muscle. In which, exosomes are effective carriers for the intercellular material transfer of a wide range of molecules that can influence a series of physiological and pathological processes in recipient cells. Considering their underlying roles, the regulatory mechanisms of adipose-secreted exosomes and their cellular crosstalk with skeletal muscle have received great attention in the field. In this review, we describe what is currently known of adipose-secreted exosomes, as well as their applications in skeletal muscle pathophysiology.


Assuntos
Exossomos , Exossomos/metabolismo , Tecido Adiposo/metabolismo , Músculo Esquelético/metabolismo , Transporte Biológico
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