scholarly journals Characterization of GLOD4 in Leydig Cells of Tibetan Sheep During Different Stages of Maturity

Genes ◽  
2019 ◽  
Vol 10 (10) ◽  
pp. 796 ◽  
Author(s):  
Wang ◽  
Li ◽  
Liu ◽  
Zhang ◽  
Zhao ◽  
...  
Keyword(s):  
Leydig Cells ◽  
Structural Characteristics ◽  
Sequence Similarity ◽  
Expression Patterns ◽  
Proteome Analysis ◽  
Testicular Development ◽  
Protein Levels ◽  
Tibetan Sheep ◽  
Pcr Method ◽  
One Year

We have previously reported that glyoxalase domain-containing protein 4 (GLOD4) is expressed in sheep testes by proteome analysis, but its roles during testicular development remain unclear. The aim of this study was to understand the expression characteristics and biological functions of the GLOD4 gene in developmental Tibetan sheep testes. The cDNA sequence of the Tibetan sheep GLOD4 gene was cloned by the RT-PCR method, and the structural characteristics of the GLOD4 protein were analyzed using relevant bioinformatics software, including ProtParam, TMHMM, Signal P 4.1, SOPMA, and phyre2. The expression patterns and immunolocalization of GLOD4 were examined in developmental testes derived from three-month-old (3M), one-year-old (1Y), and three-year-old (3Y) Tibetan sheep using quantitative real-time PCR (qRT-PCR), Western blot, immunohistochemistry, and immunofluorescence staining. The sequence analysis showed that the coding sequence (CDS) region of the GLOD4 gene was 729 bp in length and encoded 242 amino acids. Bioinformatics analysis found that the nucleotide and amino acid sequence of Tibetan sheep GLOD4 exhibited the highest sequence similarity with goat and chiru, and the least with zig-zag eel, of the species compared. GLOD4 expressions at both the mRNA and protein levels were significantly higher in the testes of the 1Y and 3Y groups than those in the 3M group (p < 0.01). Immunohistochemistry and immunofluorescence results indicated that the GLOD4 protein was mainly localized in the cytoplasm of Leydig cells from Tibetan sheep testes throughout the development stages. These results taken together suggest that the GLOD4 gene may be implicated in the development of the Leydig cells of Tibetan sheep during different stages of maturity.

scholarly journals Unraveling Stage-Dependent Expression Patterns of Circular RNAs and Their Related ceRNA Modulation in Ovine Postnatal Testis Development

2021 ◽  
Vol 9 ◽  
Author(s):  
Taotao Li ◽  
Ruirui Luo ◽  
Xia Wang ◽  
Huihui Wang ◽  
Xingxu Zhao ◽  
...  
Keyword(s):  
Germ Cells ◽  
Sertoli Cells ◽  
Leydig Cells ◽  
Expression Patterns ◽  
Interaction Network ◽  
Functional Enrichment ◽  
Circular Rnas ◽  
Testicular Development ◽  
Tibetan Sheep ◽  
Igf1 Expression

Circular RNAs (circRNAs) have been shown to function in the reproductive systems including testis. However, their expression, as well as function in testicular development of sheep remain undefined. Herein, we performed RNA sequencing to reveal circRNA temporal expression patterns in testes of Tibetan sheep from different stages of maturation (3M, 3-month-old; 1Y, 1-year-old; 3Y, 3-year-old). A total of 3,982, 414, and 4,060 differentially expressed (DE) circRNAs were uncovered from 3M vs 1Y, 1Y vs 3Y, and 3M vs 3Y, respectively. Functional enrichment assessment indicated that the source genes of DE circRNAs were primarily engaged in spermatogenesis and testicular immune privilege including blood–testis barrier (BTB). We subsequently constructed the core circRNA–miRNA–mRNA interaction network for genes related to testicular function, such as spermatogenesis, germ cell development, BTB, and cell cycle/meiosis. Furthermore, we validated the target associations between either circ_024949, circ_026259 or IGF1, and oar-miR-29b in this network, and revealed their similar expression signatures in developmental testes that they were extensively expressed in germ cells, Leydig cells, and Sertoli cells, thus suggesting their broad functions in the functional maintenance of Leydig cells and Sertoli cells, as well as the development and maturation of male germ cells. Meanwhile, circ_026259 was shown to promote IGF1 expression through inhibition of oar-miR-29b in sheep Sertoli cells. This work gives the first global view for the expression and regulation of circRNAs in sheep testis, which will be helpful for providing new insights into the molecular mechanism of ovine testis function.

scholarly journals Cloning, Molecular Characterization and Expression Patterns of DMRTC2 Implicated in Germ Cell Development of Male Tibetan Sheep

2020 ◽  
Vol 21 (7) ◽  
pp. 2448 ◽  
Author(s):  
Taotao Li ◽  
Hongyu Zhang ◽  
Xia Wang ◽  
De′en Yin ◽  
Nana Chen ◽  
...  
Keyword(s):  
Amino Acids ◽  
Western Blot ◽  
Molecular Characterization ◽  
Expression Patterns ◽  
Testicular Development ◽  
Reading Frame ◽  
Tibetan Sheep ◽  
Pcr Method ◽  
Conserved Gene ◽  
First Time

The double sex and mab-3-related transcription factors like family C2 (DMRTC2) gene is indispensable for mammalian testicular function and spermatogenesis. Despite its importance, what expression and roles of DMRTC2 possesses and how it regulates the testicular development and spermatogenesis in sheep, especially in Tibetan sheep, remains largely unknown. In this study, DMRTC2 cDNA from testes of Tibetan sheep was firstly cloned by the RT-PCR method, and its molecular characterization was identified. Subsequently, the expression and localization patterns of DMRTC2 were evaluated by quantitative real-time PCR (qPCR), Western blot, and immunofluorescence. The cloning and sequence analysis showed that the Tibetan sheep DMRTC2 cDNA fragment contained 1113 bp open reading frame (ORF) capable of encoding 370 amino acids, and displayed high identities with some other mammals, which shared an identical DM domain sequence of 47 amino acids ranged from residues 38 to 84. qPCR and Western blot results showed that DMRTC2 was expressed in testes throughout the development stages while not in epididymides (caput, corpus, and cauda), with higher mRNA and protein abundance in Tibetan sheep testes of one- and three-year-old (post-puberty) compared with that of three-month-old (pre-puberty). Immunofluorescence results revealed that immune staining for DMRTC2 protein was observed in spermatids and spermatogonia from post-puberty Tibetan sheep testes, and gonocytes from pre-puberty Tibetan sheep testes. Together, these results demonstrated, for the first time, in sheep, that DMRTC2, as a highly conserved gene in mammals, is essential for sheep spermatogenesis by regulating the proliferation or differentiation of gonocytes and development of spermatids in ram testes at different stages of maturity.

scholarly journals Morphometry and histomorphometry of the testis in crossbred pigs fed diets with different protein levels

2013 ◽  
Vol 65 (5) ◽  
pp. 1329-1338
Author(s):  
R.M.B. Valença ◽  
V.A. Silva Junior ◽  
L.P.C. Araújo ◽  
J.C. Reis ◽  
M.M.P. Guerra ◽  
...  
Keyword(s):  
Protein Content ◽  
Sertoli Cell ◽  
Leydig Cells ◽  
Crude Protein ◽  
Seminiferous Tubules ◽  
Testicular Development ◽  
Sperm Production ◽  
Stages Of Development ◽  
Protein Levels ◽  
Crossbred Pigs

Aiming to evaluate the effect of the diet protein content on testicular parameters in pigs, 21 non-gelded male Dalland pigs were used and randomly divided into three groups. Males belonging to groups G2 and G3 received a diet with crude protein levels of 15% below and above, respectively, in relation to G1 (control). At 210 days of age, animals were castrated, and testis and epididymis were collected for morphometric and histomorphometry analyses. No difference was observed in relation to the total length of seminiferous tubules (G1=3239.9±333,3m; G2=2989.4±171,7m and G3=3059.5±254.9m), population of Sertoli cell (G1=4.7±0.5x10(9); G2=4.3±0.3x10(9) and G3=4.7±0.5x10(9)), population (G1=31.6±5.58x10(9); G2=27.3±4.0x10(9) and G3=26.4±3.9x10(9)) and volume of Leydig cells (G1=1289.3±182.6µm³; G2=1179.1±85.4µm³ and G3=1133.3±37.8µm³) and sperm production (G1=5.9±0.9x10(9); G2=5.6±0.6x10(9) and G3=5.1±0.3x10(9)). Protein levels were sufficient to maintain spermatogenesis in different experimental groups. It can be concluded that the magnitude of variation in levels of protein used in different stages of development was not sufficient to promote significant changes in testicular development and spermatogenesis process in adult animals.

scholarly journals Inhibin, activin, follistatin and FSH serum levels and testicular production are highly modulated during the first spermatogenic wave in mice

Reproduction ◽  
2008 ◽  
Vol 136 (3) ◽  
pp. 345-359 ◽  
Author(s):  
Badia Barakat ◽  
Anne E O'Connor ◽  
Elspeth Gold ◽  
David M de Kretser ◽  
Kate L Loveland
Keyword(s):  
Leydig Cells ◽  
Adult Mouse ◽  
Post Partum ◽  
Serum Levels ◽  
Testicular Development ◽  
Myoid Cells ◽  
Protein Levels ◽  
Subunit Mrna ◽  
Spatial And Temporal Distributions

Testicular development is governed by the combined influence of hormones and proteins, including FSH, inhibins, activins and follistatin (FST). This study documents the expression of these proteins and their corresponding mRNAs, in testes and serum from mice aged 0 through 91 dayspost partum(dpp), using real-time PCR,in situhybridisation, immunohistochemistry, ELISA and RIA. Serum immunoactive total inhibin and FSH levels were negatively correlated during development, with FSH levels rising and inhibin levels falling. Activin A production changed significantly during development, with subunit mRNA and protein levels declining rapidly after 4 dpp, while simultaneously levels of the activin antagonists, FST and inhibin/activin βC, increased. Inhibin/activin βAand βBsubunit mRNAs were detected in Sertoli, germ and Leydig cells throughout testis development, with the βAsubunit also detected in peritubular myoid cells. The α, βA, βBand βCsubunit proteins were detected in Sertoli and Leydig cells of developing and adult mouse testes. While βAand βBsubunit proteins were observed in spermatogonia and spermatocytes in immature testes, βCwas localised to leptotene and zygotene spermatocytes in immature and adult testes. Nuclear βAsubunit protein was observed in primary spermatocytes and nuclear βCsubunit in gonocytes and round spermatids. The changing spatial and temporal distributions of inhibins and activins indicate that their modulated synthesis and action are important during onset of murine spermatogenesis. This study provides a foundation for evaluation of these proteins in mice with disturbed testicular development, enabling their role in normal and perturbed spermatogenesis to be more fully understood.

Aromatase and 11β-hydroxysteroid dehydrogenase 2 localisation in the testes of pigs from birth to puberty linked to changes of hormone pattern and testicular morphology

2008 ◽  
Vol 20 (4) ◽  
pp. 505 ◽  
Author(s):  
A. Wagner ◽  
R. Claus
Keyword(s):  
Blood Plasma ◽  
Sertoli Cell ◽  
Germ Cells ◽  
Leydig Cells ◽  
Hydroxysteroid Dehydrogenase ◽  
Blood Collection ◽  
Testicular Development ◽  
Post Mortem ◽  
Cell Numbers ◽  
Testicular Morphology

Oestrogens and glucocorticoids are important for spermatogenesis and are regulated via aromatase for oestradiol synthesis and 11β-hydroxysteroid dehydrogenase 2 (11β-HSD 2) as an inactivator of cortisol. In the present study postnatal changes of these two enzymes were monitored together with testicular development and hormone concentrations. Pigs were assigned to three periods: Weeks 0–5, Weeks 5–11 or Weeks 11–17. In Period 1, groups of four piglets were killed after each week. Blood plasma and testes were sampled immediately post mortem. For Periods 2 and 3, groups of six pigs were fitted with vein catheters for daily blood collection. Testes from all pigs were obtained after killing. Levels of testosterone, oestradiol, LH, FSH and cortisol were determined radioimmunologically. The 11β-HSD 2- and aromatase-expressing cells were stained immunocytochemically. All hormones were maximal 2 weeks after birth. A rise of LH, testosterone and oestradiol occurred again at Week 17. FSH and cortisol remained basal. Parallel to the first postnatal rise, the presence of aromatase and 11β-HSD 2 in Leydig cells increased, together with germ and Sertoli cell numbers. Expression was low from 3 to 5 weeks, was resumed after Week 5 and was maximal at Week 17. The amount of 11β-HSD 2 in germ cells was greatest at birth, decreased thereafter and was absent after Week 3.

scholarly journals Genome-Wide Identification and Expression Analyses of CONSTANS-Like Family Genes in Cucumber (Cucumis sativus L.)

2021 ◽  
Author(s):  
Zhen Tian ◽  
Xiaodong Qin ◽  
Hui Wang ◽  
Ji Li ◽  
Jinfeng Chen
Keyword(s):  
Floral Induction ◽  
Structural Characteristics ◽  
Expression Patterns ◽  
Evolutionary Relationship ◽  
Biological Functions ◽  
Promoter Regions ◽  
Cucumis Sativus L ◽  
Cis Acting ◽  
Genome Wide ◽  
Induction Process

AbstractThe CONSTANS-like (COL) gene family is one of the plant-specific transcription factor families that play important roles in plant growth and development. However, the knowledge of COLs related in cucumber is limited, and their biological functions, especially in the photoperiod-dependent flowering process, are still unclear. In this study, twelve CsaCOL genes were identified in the cucumber genome. Phylogenetic and conserved motif analyses provided insights into the evolutionary relationship between the CsaCOLs. Further, the comparative genome analysis revealed that COL genes are conserved in different plant species, especially collinearity gene pairs related to CsaCOL5. Ten kinds of cis-acting elements were vividly detected in CsaCOLs promoter regions, including five light-responsive elements, which echo the diurnal rhythm expression patterns of seven CsaCOL genes under SD and LD photoperiod regimes. Combined with the expression data of developmental stage, three CsaCOL genes are involved in the flowering network and play pivotal roles for the floral induction process. Our results provide useful information for further elucidating the structural characteristics, expression patterns, and biological functions of COL family genes in many plants

Gene expression patterns and protein cellular localization suggest a novel role for DAZL in developing Tibetan sheep testes

Gene ◽  
2020 ◽  
Vol 731 ◽  
pp. 144335 ◽  
Author(s):  
Taotao Li ◽  
Xia Wang ◽  
Hongyu Zhang ◽  
Haolin Chen ◽  
Ningbo Liu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cellular Localization ◽  
Expression Patterns ◽  
Gene Expression Patterns ◽  
Tibetan Sheep

scholarly journals Birdsong Decreases Protein Levels of FoxP2, a Molecule Required for Human Speech

2008 ◽  
Vol 100 (4) ◽  
pp. 2015-2025 ◽  
Author(s):  
Julie E. Miller ◽  
Elizabeth Spiteri ◽  
Michael C. Condro ◽  
Ryan T. Dosumu-Johnson ◽  
Daniel H. Geschwind ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Target Genes ◽  
Expression Patterns ◽  
Vocal Learning ◽  
Brain Regions ◽  
Motor Deficits ◽  
Mrna Levels ◽  
Adult Males ◽  
Protein Levels ◽  
Area X

Cognitive and motor deficits associated with language and speech are seen in humans harboring FOXP2 mutations. The neural bases for FOXP2 mutation-related deficits are thought to reside in structural abnormalities distributed across systems important for language and motor learning including the cerebral cortex, basal ganglia, and cerebellum. In these brain regions, our prior research showed that FoxP2 mRNA expression patterns are strikingly similar between developing humans and songbirds. Within the songbird brain, this pattern persists throughout life and includes the striatal subregion, Area X, that is dedicated to song development and maintenance. The persistent mRNA expression suggests a role for FoxP2 that extends beyond the formation of vocal learning circuits to their ongoing use. Because FoxP2 is a transcription factor, a role in shaping circuits likely depends on FoxP2 protein levels which might not always parallel mRNA levels. Indeed our current study shows that FoxP2 protein, like its mRNA, is acutely downregulated in mature Area X when adult males sing with some differences. Total corticosterone levels associated with the different behavioral contexts did not vary, indicating that differences in FoxP2 levels are not likely attributable to stress. Our data, together with recent reports on FoxP2's target genes, suggest that lowered FoxP2 levels may allow for expression of genes important for circuit modification and thus vocal variability.

The Expression of Sel1L and Tan-1 in Normal and Neoplastic Cells

2000 ◽  
Vol 15 (1) ◽  
pp. 26-32 ◽  
Author(s):  
M. Cattaneo ◽  
R. Orlandi ◽  
C. Ronchini ◽  
P. Granelli ◽  
G. Malferrari ◽  
...  
Keyword(s):  
Cell Function ◽  
Sequence Similarity ◽  
Expression Patterns ◽  
Negative Regulator ◽  
Chromosomal Mapping ◽  
Normal Adult ◽  
Neoplastic Cells ◽  
C Elegans ◽  
Pancreatic Cancers ◽  
Almost All

We have previously reported on the isolation and chromosomal mapping of a novel human gene (SEL1L), which shows sequence similarity to sel-1, an extragenic suppressor of C. elegans. sel-1 functions as a negative regulator of lin-12 activity, the latter being implicated in the control of diverse cellular differentiation events. In the present study we compare the expression patterns of SEL1L and TAN-1, the human ortholog of lin-12 in normal and neoplastic cells. We found that, whereas both genes are expressed in fetal tissues at similar levels, they are differentially expressed in normal adult and neoplastic cells. In normal adult cells SEL1L is generally present at very low levels; only in the cells of the pancreas does it show maximum expression. By contrast, SEL1L is generally well represented in most neoplastic cells but not in those of pancreatic and gastric carcinomas, where transcription is either downregulated or completely repressed. TAN-1 on the other hand is well represented in almost all normal and neoplastic cells, with very few exceptions. Our observations suggest that SEL1L is presumably implicated in pancreatic and gastric carcinogenesis and that, along with TAN-1, it is very important for normal cell function. Alterations in the expression of SEL1L may be used as a prognostic marker for gastric and pancreatic cancers.

scholarly journals Transcriptome and Proteome Research in Veterinary Science: What Is Possible and What Questions Can Be Asked?

2012 ◽  
Vol 2012 ◽  
pp. 1-14 ◽  
Author(s):  
Robert Klopfleisch ◽  
Achim D. Gruber
Keyword(s):  
Sequential Analysis ◽  
Biomarker Discovery ◽  
Expression Patterns ◽  
Proteome Analysis ◽  
Shotgun Proteomics ◽  
Complete Analysis ◽  
Biological Research ◽  
Veterinary Research ◽  
Gene Sets ◽  
2D Gel

In recent years several technologies for the complete analysis of the transcriptome and proteome have reached a technological level which allows their routine application as scientific tools. The principle of these methods is the identification and quantification of up to ten thousands of RNA and proteins species in a tissue, in contrast to the sequential analysis of conventional methods such as PCR and Western blotting. Due to their technical progress transcriptome and proteome analyses are becoming increasingly relevant in all fields of biological research. They are mainly used for the explorative identification of disease associated complex gene expression patterns and thereby set the stage for hypothesis-driven studies. This review gives an overview on the methods currently available for transcriptome analysis, that is, microarrays, Ref-Seq, quantitative PCR arrays and discusses their potentials and limitations. Second, the most powerful current approaches to proteome analysis are introduced, that is, 2D-gel electrophoresis, shotgun proteomics, MudPIT and the diverse technological concepts are reviewed. Finally, experimental strategies for biomarker discovery, experimental settings for the identification of prognostic gene sets and explorative versus hypothesis driven approaches for the elucidation of diseases associated genes and molecular pathways are described and their potential for studies in veterinary research is highlighted.

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