GH Overexpression Alters Spermatic Cells MicroRNAome Profile in Transgenic Zebrafish

Overexpression of growth hormone (GH) in gh-transgenic zebrafish of a highly studied lineage F0104 has earlier been reported to cause increased muscle growth. In addition to this, GH affects a broad range of cellular processes in transgenic fish, such as morphology, physiology, and behavior. Reports show changes such as decreased sperm quality and reduced reproductive performance in transgenic males. It is hypothesized that microRNAs are directly involved in the regulation of fertility potential during spermatogenesis. The primary aim of our study was to verify whether gh overexpression disturbs the sperm miRNA profile and influences the sperm quality in transgenic zebrafish. We report a significant increase in body weight of gh-transgenic males along with associated reduced sperm motility and other kinetic parameters in comparison to the non-transgenic group. MicroRNA transcriptome sequencing of gh-transgenic zebrafish sperms revealed expressions of 186 miRNAs, among which six miRNA were up-regulated (miR-146b, miR-200a-5p, miR-146a, miR-726, miR-184, and miR-738) and sixteen were down-regulated (miR-19d-3p, miR-126a-5p, miR-126b-5p, miR-22a-5p, miR-16c-5p, miR-20a-5p, miR-126b-3p, miR-107a-3p, miR-93, miR-2189, miR-202–5p, miR-221–3p, miR-125a, miR-125b-5p, miR-126a-3p, and miR-30c-5p) in comparison to non-transgenic zebrafish. Some of the dysregulated miRNAs were previously reported to be related to abnormalities in sperm quality and reduced reproduction ability in other species. In this study, an average of 134 differentially expressed miRNAs-targeted genes were predicted using the in silico approach. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis demonstrated that the genes of affected pathways were primarily related to spermatogenesis, sperm motility, and cell apoptosis. Our results suggested that excess GH caused a detrimental effect on sperm microRNAome, consequently reducing the sperm quality and reproductive potential of zebrafish males.

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Transcriptomic analysis of Procambarus clarkii affected by “Black May” disease

Scientific Reports ◽

10.1038/s41598-020-78191-8 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Guoqing Shen ◽

Xiao Zhang ◽

Jie Gong ◽

Yang Wang ◽

Pengdan Huang ◽

...

Keyword(s):

Procambarus Clarkii ◽

Kegg Pathway ◽

Enrichment Analysis ◽

Differentially Expressed ◽

Pathway Enrichment Analysis ◽

Apoptosis Pathway ◽

Pathway Enrichment ◽

Cellular Processes ◽

Expression Of Genes ◽

Muscle Tissues

AbstractEach year from April to May, high mortality rates are reported in red swamp crayfish (Procambarus clarkii) cultured in Jiangsu and other regions, in China, and this phenomenon has come to be known as “Black May” disease (BMD). Therefore, in order to investigate the possible causes of this disease, this study gathered BMD-affected P. clarkii samples and performed transcriptome analysis on hepatopancreas, gill, and muscle tissues. A total of 19,995,164, 149,212,804, and 222,053,848 clean reads were respectively obtained from the gills, muscle, and hepatopancreas of BMD-affected P. clarkii, and 114,024 unigenes were identified. The number of differentially expressed genes (DEGs) in gill, muscle, and hepatopancreas was 1703, 964, and 476, respectively. GO and KEGG enrichment analyses of the DEGs were then conducted. Based on KEGG pathway enrichment analysis, the most significantly differentially expressed pathways were mainly those involved with metabolism, human disease, and cellular processes. Further analysis of the significantly DEGs revealed that they were mainly related to the mitochondrial-mediated apoptosis pathway and that the expression of these DEGs was mostly down-regulated. Moreover, the expression of genes related to immune and metabolism-related pathways was also significantly down-regulated, and these significantly-inhibited pathways were the likely causes of P. clarkii death. Therefore, our results provide a basis for the identification of BMD causes.

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Subordinate male cichlids retain reproductive competence during social suppression

Proceedings of The Royal Society B Biological Sciences ◽

10.1098/rspb.2011.0997 ◽

2011 ◽

Vol 279 (1728) ◽

pp. 434-443 ◽

Cited By ~ 27

Author(s):

Jacqueline M. Kustan ◽

Karen P. Maruska ◽

Russell D. Fernald

Keyword(s):

Social Status ◽

Sperm Motility ◽

Sperm Quality ◽

Reproductive Potential ◽

Testis Size ◽

Reproductive Suppression ◽

Tightly Coupled ◽

Astatotilapia Burtoni ◽

Save Energy ◽

Social Suppression

Subordinate males, which are excluded from reproduction often save energy by reducing their investment in sperm production. However, if their position in a dominance hierarchy changes suddenly they should also rapidly attain fertilization capability. Here, we asked how social suppression and ascension to dominance influences sperm quality, spermatogenesis and reproductive competence in the cichlid Astatotilapia burtoni , where reproduction is tightly coupled to social status. Dominant territorial (T) males are reproductively active while subordinate non-territorial (NT) males are suppressed, but given the opportunity, NT males will perform dominance behaviours within minutes and attain T male testes size within days. Using the thymidine analogue 5-bromo-2-deoxyuridine (BrdU) to label germ cell proliferation, we found that the spermatogenic cycle takes approximately 11–12 days, and social status had no effect on proliferation, suggesting that spermatogenesis continues during reproductive suppression. Although sperm velocity did not differ among social states, NT males had reduced sperm motility. Remarkably, males ascending in status showed sperm motility equivalent to T males within 24 h. Males also successfully reproduced within hours of social opportunity, despite four to five weeks of suppression and reduced testis size. Our data suggest that NT males maintain reproductive potential during suppression possibly as a strategy to rapidly improve reproductive fitness upon social opportunity.

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Sperm Motility, Oxidative Status, and Mitochondrial Activity: Exploring Correlation in Different Species

Antioxidants ◽

10.3390/antiox10071131 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1131

Author(s):

Alessandra Gallo ◽

Maria Consiglia Esposito ◽

Elisabetta Tosti ◽

Raffaele Boni

Keyword(s):

Sperm Motility ◽

Membrane Lipids ◽

Sperm Quality ◽

Marine Invertebrate ◽

Sperm Concentration ◽

Reproductive Potential ◽

Relevant Information ◽

Membrane Lipid ◽

Oxidative Status ◽

Mitochondrial Activity

Sperm quality assessment is the first step for evaluating male fertility and includes the estimation of sperm concentration, motility, and morphology. Nevertheless, other parameters can be assessed providing additional information on the male reproductive potential. This study aimed to evaluate and correlate the oxidative status, mitochondrial functionality, and motility in spermatozoa of two marine invertebrate (Ciona robusta and Mytilus galloprovincialis) and one mammalian (Bos taurus) species. By combining fluorescent staining and spectrofluorometer, sperm oxidative status was evaluated through intracellular reactive oxygen species (ROS) and plasma membrane lipid peroxidation (LPO) analysis. Mitochondrial functionality was assessed through the mitochondrial membrane potential (MMP). In the three examined species, a negative correlation emerged between sperm motility vs ROS levels and LPO. Sperm motility positively correlated with MMP in bovine, whereas these parameters were not related in ascidian or even negatively related in mussel spermatozoa. MMP was negatively related to ROS and LPO levels in ascidians, only to LPO in bovine, and positively related in mussel spermatozoa. These results suggest that energy sources for sperm motility vary between species and that ROS causes a decline in sperm motility via oxidative damage of membrane lipids. Overall, this study validates the use of fluorescent probes in combination with spectrofluorometer as a simple and powerful methodology for supplementary evaluation of sperm quality shedding light on new potential quality markers and provided relevant information on sperm energetic metabolism.

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A conserved role of Parkinson-associated DJ-1 metabolites in sperm motility, mitosis, and embryonic development

10.1101/2021.01.16.426934 ◽

2021 ◽

Cited By ~ 1

Author(s):

Susanne Bour ◽

Yanina Dening ◽

Melanie Balbach ◽

Ina Poser ◽

Inés Ramírez Álvarez ◽

...

Keyword(s):

Sperm Motility ◽

Mitochondrial Function ◽

Sperm Quality ◽

Model Systems ◽

Mitochondrial Activity ◽

Loss Of Function ◽

C Elegans ◽

Cellular Processes ◽

Positive Effects

AbstractFertility rates in the developing world have dramatically dropped in the last decades. This drop is likely due to a decline in sperm quality and women having children at older ages. Loss of function mutations in DJ-1, a Parkinson’s associated gene, are linked to alterations in multiple cellular processes such as mitochondrial activity, ROS production or sperm motility and lead to an early onset of Parkinson’s disease and male infertility in humans and other species. Glycolate (GA) and D-lactate (DL), products of DJ-1 glyoxalase activity, sustain mitochondrial function and protect against environmental aggressions. We, therefore, tested whether these substances could also have a rescue effect on these phenotypes. Here, we show that DJ-1 loss of function not only affects sperm motility but also leads to defects in mitosis and an age-dependent increase in the abortion rate. Remarkably, whereas DL was only able to rescue embryonic lethality in C. elegans, GA rescued these phenotypes in all model systems tested and even increased sperm motility in wild-type sperm. These positive effects seem to be mediated through an increase in NAD(P)H production and the regulation of intracellular calcium. These findings not only strongly suggest GA as a new therapeutic candidate to improve male and female fertility but also show its potential to treat diseases associated with a decline in mitochondrial function or to improve mitochondrial function in aging.

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Identification of miRNA Signature Associated With Erectile Dysfunction in Type 2 Diabetes Mellitus by Support Vector Machine-Recursive Feature Elimination

Frontiers in Genetics ◽

10.3389/fgene.2021.762136 ◽

2021 ◽

Vol 12 ◽

Author(s):

Haibo Xu ◽

Baoyin Zhao ◽

Wei Zhong ◽

Peng Teng ◽

Hong Qiao

Keyword(s):

Diabetes Mellitus ◽

Erectile Dysfunction ◽

Target Genes ◽

Enrichment Analysis ◽

Recursive Feature Elimination ◽

Support Vector ◽

Pathway Enrichment Analysis ◽

Cellular Processes ◽

Mirna Sequencing

Diabetic mellitus erectile dysfunction (DMED) is one of the most common complications of diabetes mellitus (DM), which seriously affects the self-esteem and quality of life of diabetics. MicroRNAs (miRNAs) are endogenous non-coding RNAs whose expression levels can affect multiple cellular processes. Many pieces of studies have demonstrated that miRNA plays a role in the occurrence and development of DMED. However, the exact mechanism of this process is unclear. Hence, we apply miRNA sequencing from blood samples of 10 DMED patients and 10 DM controls to study the mechanisms of miRNA interactions in DMED patients. Firstly, we found four characteristic miRNAs as signature by the SVM-RFE method (hsa-let-7E-5p, hsa-miR-30 days-5p, hsa-miR-199b-5p, and hsa-miR-342–3p), called DMEDSig-4. Subsequently, we correlated DMEDSig-4 with clinical factors and further verified the ability of these miRNAs to classify samples. Finally, we functionally verified the relationship between DMEDSig-4 and DMED by pathway enrichment analysis of miRNA and its target genes. In brief, our study found four key miRNAs, which may be the key influencing factors of DMED. Meanwhile, the DMEDSig-4 could help in the development of new therapies for DMED.

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Transcriptomic Analysis of Early Stages of Intestinal Regeneration in Holothuria glaberrima

10.1101/2020.09.23.310599 ◽

2020 ◽

Author(s):

David J. Quispe-Parra ◽

Joshua G. Medina-Feliciano ◽

Sebastián Cruz-González ◽

Humberto Ortiz-Zuazaga ◽

José E. García-Arrarás

Keyword(s):

De Novo ◽

Focal Point ◽

Expression Patterns ◽

Enrichment Analysis ◽

Regenerative Process ◽

Pathway Enrichment Analysis ◽

Cellular Processes ◽

Early Stages ◽

Intestinal Regeneration ◽

Holothuria Glaberrima

AbstractEchinoderms comprise a group of animals with impressive regenerative capabilities. They can replace complex internal organs following injury or autotomy. In holothurians or sea cucumbers, cellular processes of intestinal regeneration have been extensively studied. The molecular machinery behind this faculty, however, remains to be understood. Here we assembled and annotated a de novo transcriptome using RNA-seq data consisting of regenerating and non-regenerating intestinal tissues from the sea cucumber Holothuria glaberrima. Comparisons of differential expression were made using the mesentery as a reference against 24 hour and 3 days regenerating intestine, revealing a large number of differentially expressed transcripts. Gene ontology and pathway enrichment analysis showed evidence of increasing transcriptional activity. Further analysis of transcripts associated with transcription factors revealed diverse expression patterns with mechanisms involving developmental and cancer-related activity that could be related to the regenerative process. Our study demonstrates the broad and diversified gene expression profile during the early stages of the process using the mesentery as the focal point of intestinal regeneration. It also establishes the genes that are the most important candidates in the cellular processes that underlie regenerative responses.

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Black Sea turbot sperm motility depending on the dilution of seawater

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/937/2/022076 ◽

2021 ◽

Vol 937 (2) ◽

pp. 022076

Author(s):

Iu Baiandina

Keyword(s):

Sperm Motility ◽

Black Sea ◽

Sperm Quality ◽

Abiotic Factors ◽

Reproductive Potential ◽

The Black Sea ◽

Energy Reserves ◽

External Fertilization ◽

Sperm Activation ◽

Fertilizing Ability

Abstract The sperm motility of fish reflects their fertilizing ability. Sperm activation in fish with external fertilization occurs in an isotonic medium (sea or freshwater). The duration of sperm motility is a compromise between the level of energy reserves possessed by the sperm and the process of osmotic damage it experiences in the activating medium. Under natural conditions, various biotic and abiotic factors can affect the characteristics of motility. At the same time, when assessing the reproductive potential of males in laboratory conditions, the dilution and methods of its activation have a significant effect on the characteristics of sperm quality (namely, speed, the proportion of motile spermatozoa, and the time of their activity). In our work, we determined the main characteristics of the motility of the Black Sea turbot sperm at a dilution of 1:10 and 1: 100 with seawater. We have shown that, the average speed and percentage of motile sperm significantly decrease with higher dilution of seawater.

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miR-27b-3p a Negative Regulator of DSB-DNA Repair

Genes ◽

10.3390/genes12091333 ◽

2021 ◽

Vol 12 (9) ◽

pp. 1333

Author(s):

Ricardo I. Peraza-Vega ◽

Mahara Valverde ◽

Emilio Rojas

Keyword(s):

Dna Damage ◽

Dna Repair ◽

Excision Repair ◽

Enrichment Analysis ◽

Dna Lesions ◽

Negative Regulator ◽

Pathway Enrichment Analysis ◽

Cellular Processes ◽

Repair Mechanisms ◽

Base Excision

Understanding the regulation of DNA repair mechanisms is of utmost importance to identify altered cellular processes that lead to diseases such as cancer through genomic instability. In this sense, miRNAs have shown a crucial role. Specifically, miR-27b-3 biogenesis has been shown to be induced in response to DNA damage, suggesting that this microRNA has a role in DNA repair. In this work, we show that the overexpression of miR-27b-3p reduces the ability of cells to repair DNA lesions, mainly double-stranded breaks (DSB), and causes the deregulation of genes involved in homologous recombination repair (HRR), base excision repair (BER), and the cell cycle. DNA damage was induced in BALB/c-3T3 cells, which overexpress miR-27b-3p, using xenobiotic agents with specific mechanisms of action that challenge different repair mechanisms to determine their reparative capacity. In addition, we evaluated the expression of 84 DNA damage signaling and repair genes and performed pathway enrichment analysis to identify altered cellular processes. Taken together, our results indicate that miR-27b-3p acts as a negative regulator of DNA repair when overexpressed.

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