Conserved and breed-specific differences in the cervical transcriptome of sheep with divergent fertility at the follicular phase of a natural oestrus cycle

Abstract Background The outcome of cervical artificial insemination (AI) with frozen-thawed semen in sheep is limited by the inability of sperm to traverse the cervix of some ewe breeds. Previous research has demonstrated that cervical sperm transport is dependent on ewe breed, as sperm can traverse the cervix in greater numbers in some higher fertility ewe breeds. However, the molecular mechanisms underlying ewe breed differences in sperm transport through the cervix remain unknown. In this study, we aimed to characterise the cervical transcriptome of four European ewe breeds with known differences in pregnancy rates following cervical AI using frozen-thawed semen at the follicular phase of a natural oestrous cycle. Cervical post mortem tissue samples were collected from two Irish ewe breeds (Belclare and Suffolk; medium and low fertility, respectively) and from two Norwegian ewe breeds (Norwegian White Sheep (NWS) and Fur; high fertility compared to both Irish breeds) at the follicular phase of a natural oestrous cycle (n = 8 to 10 ewes per breed). Results High-quality RNA extracted from biopsies of the mid-region of the cervix was analysed by RNA-sequencing and Gene Ontology (GO). After stringent filtering (P < 0.05 and FC > 1.5), a total of 11, 1539 and 748 differentially expressed genes (DEGs) were identified in Belclare, Fur and NWS compared to the low fertility Suffolk breed, respectively. Gene ontology analysis identified significantly enriched biological processes involved in muscle contraction, extracellular matrix (ECM) development and the immune response. Gene co-expression analysis revealed similar patterns in muscle contraction and ECM development modules in both Norwegian ewe breeds, which differed to the Irish ewe breeds. Conclusions These breed-specific biological processes may account for impaired cervical sperm transport through the cervix in sheep during the follicular phase of the reproductive cycle. This novel and comprehensive dataset provides a rich foundation for future targeted initiatives to improve cervical AI in sheep.

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93 MICRORNA SEQUENCES OF BULL SPERMATOZOA

Reproduction Fertility and Development ◽

10.1071/rdv21n1ab93 ◽

2009 ◽

Vol 21 (1) ◽

pp. 147 ◽

Cited By ~ 3

Author(s):

L. R. Robertson ◽

J. M. Feugang ◽

N. Rodriguez-Osorio ◽

A. Kaya ◽

E. Memili

Keyword(s):

Embryo Development ◽

Molecular Mechanisms ◽

Somatic Cells ◽

Low Fertility ◽

High Fertility ◽

Mammalian Development ◽

Protein Coding ◽

Mirna Microarray ◽

Male Gametes ◽

Enrichment Process

MicroRNA sequences (miRNA) are small RNAs 19 to 22 nucleotides in length that play a crucial role during mammalian development and disease. They are found in plants and animals and regulate the expression of protein-coding genes. Mechanisms regulating gene expression during gamete and embryo development are critical for setting the stage in later development; however, very little is known about the roles of miRNAs in male gametes (spermatozoa). Therefore, the objective of this study was to profile miRNA populations in spermatozoa collected from high and low fertility bulls. Frozen–thawed straws containing spermatozoa from one low and one high fertility bulls were purified using a Percoll gradient (which removes somatic cells) followed by 3 washing steps in PBS. Total RNA were isolated from pelleted spermatozoa using Trizol (3 independent isolations for each bull, total of 6 samples). RNA samples were quantified (NanoDrop Spectrophotometer, Thermo Scientific, Wilmington, DE) and verified for potential somatic cells (Agilent BioAnalyzer, Agilent, Foster City, CA) and DNA (No-RT-PCR) contaminations. Samples devoid of any contaminations went through an enrichment process of miRNA, consisting of dephosphorylation (CIP) and labeling (pCp-Biotin) of the 3′-end of miRNA, followed by their purification (BioSpin6, Bio-Rad, Hercules, CA). The purified miRNAs were analyzed by a miRNA microarray containing approximately 30 000 probes from human, rat, mouse, and other organisms (Ambion/Affymetrix DiscovArray, Asuragen, Austin, TX). The results showed that roughly 48% of total miRNA probe sets (14 215) were successfully analyzed (P < 0.05); of which only 0.5% (7) were significantly differentially expressed between the 2 bulls (fold change >2 and t-test P < 0.001). The roles of these miRNAs are not yet identified because the annotations are unavailable. Nevertheless, our preliminary results suggest that bull spermatozoa are rich in miRNAs, which could play important roles during fertilization and embryo development. Future studies will be aimed at identifying potential molecular mechanisms by which these 7 miRNAs might regulate bovine embryonic development. This study was funded by the Mississippi Agricultural and Forestry Experiment Station and Alta Genetics Inc.

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ON-FARM FORESTRY DEVELOPMENT OPTIONS FOR NORTHLAND

Proceedings of the New Zealand Grassland Association ◽

10.33584/jnzg.1986.47.1762 ◽

1986 ◽

pp. 27-32

Author(s):

J.S. Clark

Keyword(s):

Joint Ventures ◽

Low Fertility ◽

High Fertility ◽

Financial Returns ◽

Farm Forestry ◽

Hill Country ◽

Financial Position ◽

On Farm

Agroforests and woodlots offer Northland hill country farmers investment and diversification opportunities. Agroforests have less effect on the "whole farm" financial position than woodlots, especially where a progressive planting regime is adopted and where no further borrowing is required. Establishment and tending costs for agro-forests are lower, and returns come much sooner. The proven opportunity for continued grazing under trees established in this manner, apart from a short post-planting period, further enhances the agroforesty option. Even where there is reluctance on a farmer's part to plant trees on high fertility land, the expected financial returns from agroforests on low and medium fertility land will increase the overall long-term profitability and flexibility of the whole farming operation. Woodlots may be more appropriate on low fertility areas where weed reversion is likely. Joint ventures may be worth considering where farm finances are a limited factor. Keywords: On-farm forestry development, Northland hill country, agroforestry, woodlots, diversification, joint ventures, progressive planting regimes, grazing availability.

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Effect of cortisol on neurophysin I/oxytocin and peptidyl glycine-α-amidating mono-oxygenase mRNA expression in bovine luteal and granulosa cells

Polish Journal of Veterinary Sciences ◽

10.2478/pjvs-2013-0033 ◽

2013 ◽

Vol 16 (2) ◽

pp. 231-239

Author(s):

A. Ziolkowska ◽

J. Mlynarczuk ◽

J. Kotwica

Keyword(s):

Mrna Expression ◽

Granulosa Cells ◽

Molecular Mechanisms ◽

Hormone Secretion ◽

Oestrous Cycle ◽

Luteal Cells ◽

Ru 486 ◽

Ovarian Cells ◽

Key Genes ◽

Ovary Function

Abstract Cortisol stimulates the synthesis and secretion of oxytocin (OT) from bovine granulosa and luteal cells, but the molecular mechanisms of cortisol action remain unknown. In this study, granulosa cells or luteal cells from days 1-5 and 11-15 of the oestrous cycle were incubated for 4 or 8 h with cortisol (1x10-5, 1x10-7 M). After testing cell viability and hormone secretion (OT, progesterone, estradiol), we studied the effect of cortisol on mRNA expression for precursor of OT (NP-I/OT) and peptidyl glycine-α-amidating mono-oxygenase (PGA). The influence of RU 486 (1x10-5 M), a progesterone receptor blocker and inhibitor of the glucocorticosteroid receptor (GR), on the expression for both genes was tested. Cortisol increased the mRNA expression for NP-I/OT and PGA in granulosa cells and stimulated the expression for NP-I/OT mRNA in luteal cells obtained from days 1-5 and days 11-15 of the oestrous cycle. Expression for PGA mRNA was increased only in luteal cells from days 11-15 of the oestrous cycle. In addition, RU 486 blocked the cortisol-stimulated mRNA expression for NP-I/OT and PGA in both types of cells. These data suggest that cortisol affects OT synthesis and secretion in bovine ovarian cells, by acting on the expression of key genes, that may impair ovary function.

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Molecular regulation of Snai2 in development and disease

Journal of Cell Science ◽

10.1242/jcs.235127 ◽

2019 ◽

Vol 132 (23) ◽

Author(s):

Wenhui Zhou ◽

Kayla M. Gross ◽

Charlotte Kuperwasser

Keyword(s):

Transcription Factor ◽

Cell Biology ◽

Molecular Mechanisms ◽

Epithelial To Mesenchymal Transition ◽

Zinc Finger Protein ◽

Main Role ◽

Biological Processes ◽

Developmental Defects ◽

Mesenchymal Transition ◽

Damage Repair

ABSTRACT The transcription factor Snai2, encoded by the SNAI2 gene, is an evolutionarily conserved C2H2 zinc finger protein that orchestrates biological processes critical to tissue development and tumorigenesis. Initially characterized as a prototypical epithelial-to-mesenchymal transition (EMT) transcription factor, Snai2 has been shown more recently to participate in a wider variety of biological processes, including tumor metastasis, stem and/or progenitor cell biology, cellular differentiation, vascular remodeling and DNA damage repair. The main role of Snai2 in controlling such processes involves facilitating the epigenetic regulation of transcriptional programs, and, as such, its dysregulation manifests in developmental defects, disruption of tissue homeostasis, and other disease conditions. Here, we discuss our current understanding of the molecular mechanisms regulating Snai2 expression, abundance and activity. In addition, we outline how these mechanisms contribute to disease phenotypes or how they may impact rational therapeutic targeting of Snai2 dysregulation in human disease.

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Insights into how environment shapes post-mortem RNA transcription in mouse brain

Scientific Reports ◽

10.1038/s41598-021-92268-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Raphael Severino Bonadio ◽

Larissa Barbosa Nunes ◽

Patricia Natália S. Moretti ◽

Juliana Forte Mazzeu ◽

Stefano Cagnin ◽

...

Keyword(s):

Lipid Metabolism ◽

Mouse Brain ◽

Molecular Mechanisms ◽

Cell Communication ◽

Rna Degradation ◽

The Body ◽

Biological Processes ◽

Post Mortem ◽

Gene Expression Alterations ◽

Upregulated Genes

AbstractMost biological features that occur on the body after death were already deciphered by traditional medicine. However, the molecular mechanisms triggered in the cellular microenvironment are not fully comprehended yet. Previous studies reported gene expression alterations in the post-mortem condition, but little is known about how the environment could influence RNA degradation and transcriptional regulation. In this work, we analysed the transcriptome of mouse brain after death under three concealment simulations (air exposed, buried, and submerged). Our analyses identified 2,103 genes differentially expressed in all tested groups 48 h after death. Moreover, we identified 111 commonly upregulated and 497 commonly downregulated genes in mice from the concealment simulations. The gene functions shared by the individuals from the tested environments were associated with RNA homeostasis, inflammation, developmental processes, cell communication, cell proliferation, and lipid metabolism. Regarding the altered biological processes, we identified that the macroautophagy process was enriched in the upregulated genes and lipid metabolism was enriched in the downregulated genes. On the other hand, we also described a list of biomarkers associated with the submerged and buried groups, indicating that these environments can influence the post-mortem RNA abundance in its particular way.

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Network pharmacology and molecular docking study on the active ingredients of qidengmingmu capsule for the treatment of diabetic retinopathy

Scientific Reports ◽

10.1038/s41598-021-86914-8 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Mingxu Zhang ◽

Jiawei Yang ◽

Xiulan Zhao ◽

Ying Zhao ◽

Siquan Zhu

Keyword(s):

Diabetic Retinopathy ◽

Molecular Docking ◽

Molecular Mechanisms ◽

Hypoxia Inducible Factor ◽

Enrichment Analysis ◽

Docking Study ◽

Network Pharmacology ◽

Biological Processes ◽

Active Ingredients ◽

Molecular Docking Study

AbstractDiabetic retinopathy (DR) is a leading cause of irreversible blindness globally. Qidengmingmu Capsule (QC) is a Chinese patent medicine used to treat DR, but the molecular mechanism of the treatment remains unknown. In this study, we identified and validated potential molecular mechanisms involved in the treatment of DR with QC via network pharmacology and molecular docking methods. The results of Ingredient-DR Target Network showed that 134 common targets and 20 active ingredients of QC were involved. According to the results of enrichment analysis, 2307 biological processes and 40 pathways were related to the treatment effects. Most of these processes and pathways were important for cell survival and were associated with many key factors in DR, such as vascular endothelial growth factor-A (VEGFA), hypoxia-inducible factor-1A (HIF-1Α), and tumor necrosis factor-α (TNFα). Based on the results of the PPI network and KEGG enrichment analyses, we selected AKT1, HIF-1α, VEGFA, TNFα and their corresponding active ingredients for molecular docking. According to the molecular docking results, several key targets of DR (including AKT1, HIF-1α, VEGFA, and TNFα) can form stable bonds with the corresponding active ingredients of QC. In conclusion, through network pharmacology methods, we found that potential biological mechanisms involved in the alleviation of DR by QC are related to multiple biological processes and signaling pathways. The molecular docking results also provide us with sound directions for further experiments.

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Comparing infertility-related stress in high fertility and low fertility countries

Sexual & Reproductive Healthcare ◽

10.1016/j.srhc.2021.100653 ◽

2021 ◽

pp. 100653

Author(s):

Emily A. Groene ◽

Cyrialis Mutabuzi ◽

Dickson Chinunje ◽

Ester Matson Shango ◽

Shalini Kulasingam ◽

...

Keyword(s):

Low Fertility ◽

High Fertility ◽

Related Stress

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Gene expression profiling of bovine endometrium during the oestrous cycle: detection of molecular pathways involved in functional changes

Journal of Molecular Endocrinology ◽

10.1677/jme.1.01799 ◽

2005 ◽

Vol 34 (3) ◽

pp. 889-908 ◽

Cited By ~ 92

Author(s):

S Bauersachs ◽

S E Ulbrich ◽

K Gross ◽

S E M Schmidt ◽

H H D Meyer ◽

...

Keyword(s):

Molecular Mechanisms ◽

Transforming Growth Factor ◽

Cdna Array ◽

Transforming Growth Factor Β ◽

Early Embryo ◽

Oestrous Cycle ◽

Cdna Libraries ◽

Functional Changes ◽

Retinoic Acid Signalling ◽

Subtracted Cdna Libraries

The endometrium plays a central role among the reproductive tissues in the context of early embryo–maternal communication and pregnancy. It undergoes typical changes during the sexual/oestrous cycle, which are regulated by the ovarian hormones progesterone and oestrogen. To identify the underlying molecular mechanisms we have performed the first holistic screen of transcriptome changes in bovine intercaruncular endometrium at two stages of the cycle – end of day 0 (late oestrus, low progesterone) and day 12 (dioestrus, high progesterone). A combination of subtracted cDNA libraries and cDNA array hybridisation revealed 133 genes showing at least a 2-fold change of their mRNA abundance, 65 with higher levels at oestrus and 68 at dioestrus. Interestingly, genes were identified which showed differential expression between different uterine sections as well. The most prominent example was the UTMP (uterine milk protein) mRNA, which was markedly upregulated in the cranial part of the ipsilateral uterine horn at oestrus. A Gene Ontology classification of the genes with known function characterised the oestrus time by elevated expression of genes, for example related to cell adhesion, cell motility and extracellular matrix and the dioestrus time by higher expression of mRNAs encoding for a variety of enzymes and transport proteins, in particular ion channels. Searching in pathway databases and literature data-mining revealed physiological processes and signalling cascades, e.g. the transforming growth factor-β signalling pathway and retinoic acid signalling, which are potentially involved in the regulation of changes of the endometrium during the oestrous cycle.

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Seasonal and steroid-dependent effects on the modulation of LH secretion in the ewe by intracerebroventricularly administered β-endorphin or naloxone

Journal of Endocrinology ◽

10.1677/joe.0.1220509 ◽

1989 ◽

Vol 122 (2) ◽

pp. 509-517 ◽

Cited By ~ 15

Author(s):

R. J. E. Horton ◽

H. Francis ◽

I. J. Clarke

Keyword(s):

Breeding Season ◽

Luteal Phase ◽

Pulse Frequency ◽

Opioid Peptide ◽

Follicular Phase ◽

Oestrous Cycle ◽

Opioid Antagonist ◽

Endogenous Opioid ◽

Endogenous Opioid Peptide ◽

Lh Secretion

ABSTRACT The natural opioid ligand, β-endorphin, and the opioid antagonist, naloxone, were administered intracerebroventricularly (i.c.v.) to evaluate effects on LH secretion in ovariectomized ewes and in ovariectomized ewes treated with oestradiol-17β plus progesterone either during the breeding season or the anoestrous season. Ovary-intact ewes were also studied during the follicular phase of the oestrous cycle. Jugular blood samples were taken at 10-min intervals for 8 h and either saline (20–50 μl), 100 μg naloxone or 10 μg β-endorphin were injected i.c.v. after 4 h. In addition, luteal phase ewes were injected i.c.v. with 25 μg β-endorphin(1–27), a purported endogenous opioid antagonist. In ovariectomized ewes, irrespective of season, saline and naloxone did not affect LH secretion, but β-endorphin decreased the plasma LH concentrations, by reducing LH pulse frequency. The effect of β-endorphin was blocked by administering naloxone 30 min beforehand. Treating ovariectomized ewes with oestradiol-17β plus progesterone during the breeding season reduced plasma LH concentrations from 6–8 μg/l to less than 1 μg/l. In these ewes, saline did not alter LH secretion, but naloxone increased LH pulse frequency and the plasma concentrations of LH within 15–20 min. During anoestrus, the combination of oestradiol-17β plus progesterone to ovariectomized ewes reduced the plasma LH concentrations from 3–5 μg/l to undetectable levels, and neither saline nor naloxone affected LH secretion. During the follicular phase of the oestrous cycle, naloxone enhanced LH pulse frequency, which resulted in increased plasma LH concentrations; saline had no effect. In these sheep, β-endorphin decreased LH pulse frequency and the mean concentrations of LH, and this effect was prevented by the previous administration of naloxone. The i.c.v. administration of β-endorphin(1–27) to luteal phase ewes did not affect LH secretion. These data demonstrate the ability of a naturally occurring opioid peptide to inhibit LH secretion in ewes during the breeding and non-breeding seasons, irrespective of the gonadal steroid background. In contrast, whilst the gonadal steroids suppress LH secretion in ovariectomized ewes during both seasons, they only appear to activate endogenous opioid peptide (EOP)-mediated inhibition of LH secretion during the breeding season. Furthermore, these data support the notion that LH secretion in ovariectomized ewes is not normally under the control of EOP, so that naloxone has no effect. Journal of Endocrinology (1989) 122, 509–517

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miR-146a is a significant brake on autoimmunity, myeloproliferation, and cancer in mice

Journal of Experimental Medicine ◽

10.1084/jem.20101823 ◽

2011 ◽

Vol 208 (6) ◽

pp. 1189-1201 ◽

Cited By ~ 528

Author(s):

Mark P. Boldin ◽

Konstantin D. Taganov ◽

Dinesh S. Rao ◽

Lili Yang ◽

Jimmy L. Zhao ◽

...

Keyword(s):

Molecular Mechanisms ◽

Immune Cell ◽

Myeloid Cell ◽

Genetically Engineered ◽

Biological Processes ◽

Targeted Deletion ◽

Genetically Engineered Mice ◽

Immune Defects ◽

Molecular Brake

Excessive or inappropriate activation of the immune system can be deleterious to the organism, warranting multiple molecular mechanisms to control and properly terminate immune responses. MicroRNAs (miRNAs), ∼22-nt-long noncoding RNAs, have recently emerged as key posttranscriptional regulators, controlling diverse biological processes, including responses to non-self. In this study, we examine the biological role of miR-146a using genetically engineered mice and show that targeted deletion of this gene, whose expression is strongly up-regulated after immune cell maturation and/or activation, results in several immune defects. Collectively, our findings suggest that miR-146a plays a key role as a molecular brake on inflammation, myeloid cell proliferation, and oncogenic transformation.

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