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1.
Aging (Albany NY) ; 12(5): 4445-4462, 2020 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-32155132

RESUMO

Forest musk deer (Moschus berezovskii, FMD) is an endangered artiodactyl species, male FMD produce musk. We have sequenced the whole genome of FMD, completed the genomic assembly and annotation, and performed bioinformatic analyses. Our results showed that microsatellites (SSRs) displayed nonrandomly distribution in genomic regions, and SSR abundances were much higher in the intronic and intergenic regions compared to other genomic regions. Tri- and hexanucleotide perfect (P) SSRs predominated in coding regions (CDSs), whereas, tetra- and pentanucleotide P-SSRs were less abundant. Trifold P-SSRs had more GC-contents in the 5'-untranslated regions (5'UTRs) and CDSs than other genomic regions, whereas mononucleotide P-SSRs had the least GC-contents. The repeat copy numbers (RCN) of the same mono- to hexanucleotide P-SSRs had different distributions in different genomic regions. The RCN of trinucleotide P-SSRs had increased significantly in the CDSs compared to the transposable elements (TEs), intronic and intergenic regions. The analysis of coefficient of variability (CV) of P-SSRs showed that the RCN of mononucleotide P-SSRs had relative higher variation in different genomic regions, followed by the CV pattern of RCN: dinucleotide P-SSRs > trinucleotide P-SSRs > tetranucleotide P-SSRs > pentanucleotide P-SSRs > hexanucleotide P-SSRs. The CV variations of RCN of the same mono- to hexanucleotide P-SSRs were relative higher in the intron and intergenic regions, followed by that in the TEs, and the relative lower was in the 5'UTR, CDSs and 3'UTRs. 58 novel polymorphic SSR loci were detected based on genotyping DNA from 36 captive FMD and 22 SSR markers finally showed polymorphism, stability, and repetition.

2.
Biomed Res Int ; 2019: 9291216, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31886268

RESUMO

Animal gut microbiota begins to colonize after birth and is functionally indispensable for maintaining the health of the host. It has been reported that gender and age influence the composition of the intestinal microbiome. However, the effects of gender and age on the intestinal microorganism of forest musk deer (FMD) remain unclear. The aim of this study was to establish the relationship between the structure and composition of fecal microbiota of male and female forest musk deer with age. Here, Illumina Miseq 300PE sequencing platform targeting 16S rRNA V3-V4 hypervariable region applied to define the fecal microbiota of male and female FMD with two age groups, juvenile (age 1-2 years) and adult (age 4-10 years). Alpha diversity index did not show significant difference in bacterial diversity between the males and females or among age groups. The intestinal microbiota of FMD was dominated by three phyla, the Firmicutes, Proteobacteria and Bacteroidetes regardless of gender and different ages. Higher proportions of Proteobacteria were found in adult male and juvenile female individuals. The composition of Bacteroidetes was stable with the gender and age of FMD. Interestingly, the relative abundance of genera Clostridiales and Bacteroidales were higher in the juvenile FMD. Conversely, proportions of Pseudomonas and Lachnospiraceae were abundant in the adult FMD. Higher proportions of Ruminococcaceae, Dore, and 5-7N15 were found in the juvenile male groups. They may reflect the different immune resistance of male and female individuals at different stages of development. This study explored the fecal microbiota composition of forest musk deer in relation to gender and age, which may provide an effective strategy for developing intestinal microecological preparations and potential musk deer breeding.


Assuntos
Envelhecimento/fisiologia , Cervos/microbiologia , Microbioma Gastrointestinal , Caracteres Sexuais , Animais , Biodiversidade , Ácidos Graxos Monoinsaturados , Feminino , Masculino , Filogenia
3.
Zhongguo Zhong Yao Za Zhi ; 44(20): 4448-4453, 2019 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-31872631

RESUMO

Musk,with unique and intense perfume,was a kind of deep brown precious medicinal material in traditional Chinese medicine. However,the immature musk in musk pot was white and stench. Given the fact that bacterial diversity generated odorous metabolites in animal hosts,in this study,musk samples at three different mature stages,including MJ( the end of June),MA( the end of August) and MO( the end of October) were harvested from three male forest musk deer,and then next-generation sequencing was used to intensively survey the bacterial communities in musk harvested at different mature stages. RESULTS: indicated that the average OTUs per sample at the end of June,August and October were 47 116. 00 ± 1 567. 24( SE),52 009. 00 ± 8 958. 75( SE) and50 004. 67±4 135. 57( SE),respectively. Feature of the musk 16 S rRNA gene showed a total of 418 genera belonging to 52 phyla were observed in all samples. The main microbiota was bacteria,which accounted for 98. 82%,99. 95% and 99. 58% in MJ,MA and MO,respectively. At phylum level,Firmicutes was the most abundant bacterial of MA( 32. 75%) and MO( 39. 19%). While,the major bacterial in MJ was Proteobacteria( 49. 14%). PICRUSt analysis revealed the functions of bacterial in MJ were mainly involved in secretion,while bacterial functions of MA and MO were mainly involved in amino acid or other substance metabolism,which was in accord with the musk secretion physiological process of forest musk deer. This is the first study involved in the bacterial diversity in musk of forest musk deer across the maturation process,while may provide a new insight into the musk generation mechanism.


Assuntos
Cervos/microbiologia , Ácidos Graxos Monoinsaturados , Animais , Florestas , Sequenciamento de Nucleotídeos em Larga Escala , Masculino
4.
Biomed Res Int ; 2019: 6301915, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31781630

RESUMO

Bitterness is an important taste sensation for chickens, which provides useful sensory information for acquisition and selection of diet, and warns them against ingestion of potentially harmful and noxious substances in nature. Bitter taste receptors (T2Rs) mediate the recognition of bitter compounds belonging to a family of proteins known as G-protein coupled receptors. The aim of this study was to identify and evaluate the expression of T2R7 in chicken tongue tissue and construct cT2R7-1 and cT2R7-2-expressing HEK-293T cells to access the expression of PLCß2 and ITPR3 after exposure with different concentrations of the bitter compounds. Using real-time PCR, we show that the relative expression level of T2R7 mRNA in 5, 1, 0.1, and 10-3 mM of camphor and erythromycin solutions and 5 mM of chlorpheniramine maleate solutions was significantly higher than that in 50 mM KCL solutions. We confirmed that the bitter taste receptor T2R7 and downstream signaling effectors are sensitive to different concentrations of bitter compounds. Moreover, T2R7-1 (corresponding to the unique haplotype of the Tibetan chicken) had higher sensitivity to bitter compounds compared with that of T2R7-2 (corresponding to the unique haplotype of the Jiuyuan black-chicken). These results provide great significance of taste response on dietary intake to improve chicken feeding efficiency in poultry production and have certain reference value for future taste research in other bird species.


Assuntos
Proteínas Aviárias/biossíntese , Cânfora/farmacologia , Clorfeniramina/farmacologia , Eritromicina/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Animais , Proteínas Aviárias/genética , Galinhas , Relação Dose-Resposta a Droga , Feminino , Células HEK293 , Humanos , Receptores de Inositol 1,4,5-Trifosfato/biossíntese , Receptores de Inositol 1,4,5-Trifosfato/genética , Masculino , Fosfolipase C beta/biossíntese , Fosfolipase C beta/genética , Receptores Acoplados a Proteínas-G , Transdução de Sinais/genética
5.
Animals (Basel) ; 9(8)2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31344924

RESUMO

The forest musk deer (Moschus berezovskii) is a small-sized artiodactyl species famous for the musk secreted by adult males. In the captive population, this species is under the threat of infection diseases, which greatly limits the increase of individual numbers. In the present study, we computationally analyzed the repertoire of the cathelicidin (CATHL) family from the genome of forest musk deer and investigated their expression pattern by real-time PCR. Our results showed that the entire genome of forest musk deer encodes eight cathelicidins, including six functional genes and two pseudogenes. Phylogenetic analyses further revealed that all forest musk deer cathelicidin members have emerged before the split of the forest musk deer and cattle and that forest musk deer CATHL3L2 and CATHL9 are orthologous with two cattle pseudogenes. In addition, the gene expression results showed that the six functional genes are not only abundantly expressed in the spleen and lung, but are also differently expressed in response to abscesses, which suggests that forest musk deer cathelicidins may be involved in infections. Taken together, identification and characterization of the forest musk deer cathelicidins provide fundamental data for further investigating their evolutionary process and biological functions.

6.
Biomed Res Int ; 2019: 4370704, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31214615

RESUMO

The Chinese forest musk deer (Moschus berezovskii) is an economically important species distributed throughout southwest China and northern Vietnam. Occurrence and development of disease are aggravated by inbreeding and genetic diversity declines in captive musk deer populations. Deep transcriptomics investigation may provide a promising way to improve genetic health of captive and wild FMD population. MicroRNAs (miRNAs), which regulate gene expression by targeting and suppressing of mRNAs, play an important role in physiology and organism development control. In this study, RNA-seq technology was adopted to characterize the miRNA transcriptome signature among six tissues (heart, liver, spleen, lung, kidney, and muscle) in Chinese forest musk deer at two years of age. Deep sequencing generated a total of 103,261,451 (~87.87%) good quality small RNA reads; of them 6,622,520 were unique across all six tissues. A total of 2890 miRNAs were identified, among them 1129 were found to be expressed in all tissues. Moreover, coexpression of 20 miRNAs (>2000RPM) in all six tissues and top five highly expressed miRNAs in each tissue implied the crucial and particular function of them in FMD physiological processes. Our findings of forest musk deer miRNAs supplement the database of transcriptome information for this species and conduce to our understanding of forest musk deer biology.


Assuntos
Cervos , Regulação da Expressão Gênica/fisiologia , MicroRNAs , Animais , China , Cervos/genética , Cervos/metabolismo , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , MicroRNAs/biossíntese , MicroRNAs/genética , Especificidade de Órgãos/fisiologia
7.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 37(3): 242-247, 2019 Jun 01.
Artigo em Chinês | MEDLINE | ID: mdl-31218855

RESUMO

OBJECTIVE: To evaluate the effect of zoledronate acid (ZA) on the proliferation and osteogenic differentiation of rat mesenchymal stem cells (BMSCs). METHODS: The BMSCs isolated from the SD rats were cultured with different concentrations of ZA (1, 5, 10, and 20 µmol·L), and the contro1 group received the same volume of culture medium but without ZA. Cell counting kit-8 was used to detect proliferation activity in each group. Alkaline phosphatase (ALP) staining and alizarin red staining were used to detect the osteogenic differentiation ability in each group. The gene expression levels of ALP, bone morphogenetic protein-2 (BMP-2), typeⅠcollagenase (COL-Ⅰ), runt-related transcription factor-2 (Runx-2), zinc finger structure transcription factor (Osx), osteocalcin (OCN), and osteopontin (OPN) were evaluated by real-time quantitative polymerase chain reaction (qRT-PCR). RESULTS: Zoledronate at 1 µmol·L⁻¹ concentration had no effect on the proliferation and osteogenic differentiation of BMSCs. No significant difference was observed between this group and the control group (P>0.05). When the ZA concentration was more than 1 µmol·L⁻¹, ZA inhibited the proliferation and osteogenic differentiation of BMSCs, and the effect was concentration dependent. The difference between each group and the control group was statistically significant (P<0.05). At ZA concentration of 5 µmol·L⁻¹, ZA enhanced the expression of ALP, BMP-2, COL-Ⅰ, Runx-2, Osx, OCN, and OPN (P<0.05). However, at ZA concentration of more than 5 µmol·L⁻¹, the expression levels of osteogenicrelated genes in each group was lower than those of the control group (P<0.05). CONCLUSIONS: Low ZA concentration has no effect on the proliferation and osteogenic differentiation of BMSCs. ZA at 5 µmol·L⁻¹ concentration inhibits the proliferation but promotes the osteogenic differentiation of BMSCs. High ZA concentration inhibits the proliferation and osteogenic differentiation of BMSCs.


Assuntos
Células-Tronco Mesenquimais , Osteogênese , Animais , Células da Medula Óssea , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Ratos , Ratos Sprague-Dawley
8.
In Vitro Cell Dev Biol Anim ; 54(8): 545-548, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30083840

RESUMO

Research of epithelial cells in musk gland is lacking. There are no good characterized epithelial cell lines that can provide complementary in vitro models for in vivo research. We successfully cultivated epithelial cells of musk gland for the first time. The protocol described here produces epithelial cell lines from the mature secreting musk gland. Based on morphological observation, epithelial cells of musk gland were isolated and cultured in vitro. After the third passage, the musk gland-derived cells were filled with many lipid droplets and proliferated well. We used gas chromatography and mass spectrometry to explore the chemical composition of lipid droplets in the musk gland-derived cells. The main components of secreted lipid droplet were alkanes, esters, amines, alcohols, ketones, organic acids, and aldehydes. Muscone, which is the main active compound of musk, was not found. This is a new attempt in the field of animal musk to obtain naturally secreted animal musk in vitro by cloning specialized cells. In conclusion, this study provides a reference at the cellular level to further analyze the biology and physiology of the musk gland epithelium and secretion mechanism of musk deer.


Assuntos
Estruturas Animais/citologia , Separação Celular/métodos , Forma Celular , Cervos/anatomia & histologia , Células Epiteliais/citologia , Epitélio/metabolismo , Ácidos Graxos Monoinsaturados/química , Estruturas Animais/anatomia & histologia , Animais , Cruzamento , Células Cultivadas , Florestas , Cromatografia Gasosa-Espectrometria de Massas , Estações do Ano
9.
J Genet ; 96(6): 1033-1040, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29321364

RESUMO

The Chinese forest musk deer (Moschus berezovskii Flerov) is an endangered artiodactyl mammal. The musk secreted by sexually mature males is highly valued for alleged pharmaceutical properties and perfume manufacturing. However, the genomic and transcriptomic resources of musk deer remain deficiently represented and poorly understood. Next-generation sequencing technique is an efficient method for generating an enormous amount of sequence data that can represent a large number of genes and their expression levels. In the present study, we used Illumina HiSeq technology to perform de novo assembly of heart and musk gland transcriptomes from the Chinese forest musk deer. A total of 239,383 transcripts and 176,450 unigenes were obtained, of which 37,329 unigenes were matched to known sequences in the NCBI nonredundant protein (Nr) database; 31,039 unigenes were assigned to 61 GO terms, and 11,782 to 332 KEGG pathways. Additionally, 592 and 2282 differentially expressed genes were found to be specifically expressed in the heart and musk gland, respectively. The abundant transcriptomic data generated in the present report will provide a comprehensive sequence resource for Chinese forest musk deer as well as lay down a foundation which will help in accelerating genetic and functional genomics research in this species.


Assuntos
Cervos/genética , Transcriptoma/genética , Animais , China , Florestas , Sequenciamento de Nucleotídeos em Larga Escala , Masculino , Repetições de Microssatélites/genética
10.
Sci Rep ; 6: 18975, 2016 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-26744067

RESUMO

Male musk deer secrete musk from the musk gland located between their naval and genitals. Unmated male forest musk deer generate a greater amount of musk than mated males, potentially allowing them to attract a greater number of females. In this study, we used gas chromatography and mass spectrometry (GC/MS) to explore musk chemical composition of the musk pods of captive mated and unmated sexually mature Chinese forest musk deer and used next-generation sequencing to intensively survey the bacterial communities within them. Analysis of the chemical composition of the musk showed that unmated males have more muscone and cholesterol. Features of the musk16S rRNA gene showed that mated Chinese forest musk deer have both a greater Shannon diversity (p < 0.01) and a greater number of estimated operational taxonomic units than unmated ones; many bacterial genera were overrepresented in unmated Chinese forest musk deer males. Members of these genera might be involved in musk odor fermentation. PICRUSt analysis revealed that metabolic pathways such as aldosterone-regulated sodium reabsorption, metabolism of terpenoids and polyketides, flavone and flavonol biosynthesis, and isoflavonoid biosynthesis were enriched in the musk of unmated Chinese forest musk deer males.


Assuntos
Cervos/fisiologia , Glândulas Exócrinas/química , Microbiota/fisiologia , Odorantes/análise , Comportamento Sexual Animal/fisiologia , Aldosterona/análise , Aldosterona/metabolismo , Animais , Colesterol/análise , Colesterol/metabolismo , Cicloparafinas/análise , Cicloparafinas/metabolismo , Cervos/microbiologia , Glândulas Exócrinas/metabolismo , Glândulas Exócrinas/microbiologia , Ácidos Graxos Monoinsaturados/análise , Ácidos Graxos Monoinsaturados/metabolismo , Feminino , Fermentação , Flavonoides/análise , Flavonoides/metabolismo , Florestas , Masculino , Policetídeos/análise , Policetídeos/metabolismo , Simbiose , Terpenos/análise , Terpenos/metabolismo
11.
Zhongguo Zhong Yao Za Zhi ; 40(22): 4319-23, 2015 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-27097400

RESUMO

Forest musk deer is one of the large-scale farming musk deer animals with the largest population at the same time. The male musk deer can secrete valuable medicines, which has high medicinal and economic value. Due to the loss of habitat and indiscriminate hunting, the numbers of wild population specie and the distribution have been drastically reduced. Therefore, in-depth understanding of the molecular genetics progress of forest musk deer will pave a way for musk deer protection and breeding. In this review, the progress associated with the molecular marker, genetic classification, artificial breeding, musk secretion and disease in past decades were reviewed, in order to provide a theoretical basis for subsequent molecular genetic researches in forest musk deer.


Assuntos
Cervos/genética , Animais , Cruzamento , Cervos/classificação , Cervos/crescimento & desenvolvimento , Cervos/metabolismo , Ecossistema , Ácidos Graxos Monoinsaturados/química , Ácidos Graxos Monoinsaturados/metabolismo , Feminino , Masculino
12.
Zhongguo Zhong Yao Za Zhi ; 39(23): 4522-5, 2014 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-25911794

RESUMO

Forest musk deer (Moschus berezovskii), a rare wild medicinal animal, is listed under the category of the state key protected wildlife list of China. Musk, secreted by the musk glands, is with high economic and medicinal value and used as precious traditional medicine in China. In order to meet the needs of musk in Chinese traditional medicine, forest musk deer farming was conducted in 1950s, but the research progress on musk secretion mechanism was slow. Therefore, by reviewing the histological and anatomical structure of forest musk deer musk gland, the relationship between sex hormones and the musk secretion process, and the molecular mechanism of the musk secretion, the existing problems in investigating the musk secretion mechanism were analyzed and the development trends in this field were also discussed, in order to provide a reference for further studies on the musk secretion mechanism and improve musk production of forest musk deer.


Assuntos
Cervos/metabolismo , Glândulas Exócrinas/química , Ácidos Graxos Monoinsaturados/química , Animais , Glândulas Exócrinas/anatomia & histologia , Glândulas Exócrinas/metabolismo , Ácidos Graxos Monoinsaturados/metabolismo , Masculino
13.
Cancer Biol Ther ; 6(3): 335-42, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17312384

RESUMO

RhoE, a small G protein, is constitutively GTP bound within the cell and can regulate actin cytoskeleton reorganization, leading to the appearance of aggregates of actin filaments. Although emerging evidence suggests that RhoE is causally involved in cancer formation and progression, little is known about its significance in solid cancer, including lung cancer. In the present study, the expression of RhoE was analyzed using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR), real-time RT-PCR, immunohistochemistry and western blot in 30 patients with Non-small Cell Lung Cancer (NSCLC). Then the correlation of RhoE overexpression with clinical parameters was evaluated. Furthermore, the possible reasons contributing to the RhoE expression were examined by real-time genomic PCR and mutation analysis on DNA sequence and cDNA sequence. Our results revealed that RhoE expression was dramatically increased in lung cancer tissues compared with adjacent nontumoral lung tissues (p <0.01). Immunohistochemistry showed a strong cytoplasmic staining in cancer cells compared with positive membrane staining in adjacent nontumoral proliferative alveolar epitheliums. Moreover, the overexpression of RhoE was significantly associated with the patients' smoking history (p <0.05). 72% tumor tissues displayed DNA copy number changes based on the DNA levels in the matched adjacent nontumoral lung tissues and this copy number changes correlated significantly with RhoE expression and smoking history (p <0.05). Three polymorphisms were identified but no correlation was found with the clinicopathological features. To our knowledge, this is the first report demonstrating that overexpression of RhoE correlated with smoking and DNA copy number changes, suggesting that RhoE may serve as a molecular marker to identify high-risk individuals and assist in the identification of additional pathways of carcinogenesis.


Assuntos
Biomarcadores Tumorais/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Neoplasias Pulmonares/genética , Fumar/genética , Proteínas rho de Ligação ao GTP/genética , Regiões 3' não Traduzidas/genética , Biomarcadores Tumorais/análise , Carcinoma Pulmonar de Células não Pequenas/química , DNA de Neoplasias/análise , Amplificação de Genes , Dosagem de Genes , Humanos , Imuno-Histoquímica , Neoplasias Pulmonares/química , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima , Proteínas rho de Ligação ao GTP/análise
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