The Effects of Negative Pressure on the Gene Expression of Motility Related Proteins in Boar Spermatozoa during Liquid Storage at 17oC

2021 ◽  
Author(s):  
Yanbing Li ◽  
Jingchun Li ◽  
Qun Zhang ◽  
Qian Wang ◽  
Minghui Guo ◽  
...  
Keyword(s):  
Gene Expression ◽  
Negative Pressure ◽  
Protein Gene ◽  
Adhesion Protein ◽  
Spermatozoa Motility ◽  
Liquid Storage ◽  
Boar Sperm ◽  
Boar Semen ◽  
Related Proteins ◽  
Boar Spermatozoa

Background: The present study was aimed to investigate the effects of negative pressure applied before storage on the gene expression of motility related proteins in boar spermatozoa.Methods: Boar semen samples were collected and pooled and diluted with Modena solution containing 0.4% (w/v) of bovine serum albumin. Negative pressure was applied for 2-5 min using a vacuum pump with a barometer. The pressure applied was 0 (Control), -0.02 MPa (P2), -0.04 MPa (P4) and -0.08 MPa (P8). The expression of AQN-1, AQN-3, AWN, PSP-I, PSP-II gene in boar spermatozoa was evaluated.Result: Application of -0.04 MPa improved the sperm motility compared with the other groups. In conclusion, our results confirmed thatnegative pressure preservation at 17oC had an effect on the expression of boar sperm adhesion protein gene. The relative expression of AWN0PSP-I and PSP-II genes in boar sperm were lower under -0.04 MPa and -0.08 MPa pressure, which was beneficial to protect spermatozoa motility. 

scholarly journals Implication of Polyhistidine, a Novel Apoptosis Inhibitor, in Inhibiting Lipopolysaccharide-Induced Apoptosis in Boar Sperm

Animals ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 719 ◽  
Author(s):  
Tianzeng Song ◽  
Yi Shi ◽  
Yangang Wang ◽  
Izhar Hyder Qazi ◽  
Christiana Angel ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Storage Conditions ◽  
Potential Implication ◽  
Liquid Storage ◽  
Boar Sperm ◽  
Tlr4 Agonist ◽  
Reasonable Evidence ◽  
Boar Semen ◽  
Induced Apoptosis ◽  
Apoptosis Inhibitor

Lipopolysaccharide (LPS) released from Gram-negative bacteria binds to toll-like receptor 4 (TLR4) and induces boar sperm apoptosis. Similarly, polyhistidine (pHis), a TLR4 agonist, can also bind to TLR4. We hypothesized that pHis could inhibit LPS-induced sperm apoptosis by competitively binding to TLR4 to then improve sperm quality. Therefore, the objective of this study was to examine whether pHis can inhibit LPS-induced sperm apoptosis and affect sperm quality. The results showed that the concentrations of bacterial colonies were significantly increased from 36 to 120 h under liquid storage conditions (p < 0.05); however, concentrations of LPS in boar semen showed a relatively constant trend (4.98 ± 1.55 EU/mL) following 120 h storage. The addition of 100 μg/mL pHis in the BTS extender significantly improved boar sperm motility and viability at 37 °C, and it significantly (p < 0.05) inhibited boar sperm apoptosis under liquid storage (17 °C) and at 37 °C incubation conditions. The co-treatment of LPS and pHis further confirmed that pHis played its role in inhibiting LPS-induced sperm apoptosis. In conclusion, our preliminary findings provide reasonable evidence that pHis could act as an inhibitor of LPS-induced apoptosis in boar sperm stored for longer periods of time. pHis might inhibit LPS-induced sperm apoptosis by competitively binding to TLR4. Nevertheless, further mechanistic studies are awaited to fully elucidate its potential implication in inhibiting LSP-induced apoptosis.

Effects of negative pressure applied before storage (liquid storage, 17°C) on boar semen quality and fertilization ability

2019 ◽  
Author(s):  
LI. Jingchun ◽  
LI. Qi ◽  
LI. Yanbug ◽  
WEI Guosheng ◽  
SUN Dongbo
Keyword(s):  
Sperm Motility ◽  
Negative Pressure ◽  
Semen Quality ◽  
The Other ◽  
Blastocyst Development ◽  
Cleavage Rate ◽  
Liquid Storage ◽  
Fertilizing Ability ◽  
Boar Semen ◽  
Acrosome Integrity

The present study was aimed to investigate the effects of negative pressure applied before storage on the quality and fertilization ability of boar semen. Boar semen samples were collected and pooled, and diluted with Modena solution containing 0.4% (w/v) of bovine serum albumin. Negative pressure was applied for 2–5 min using a vacuum pump with a barometer. The pressure applied were 0 (Control), -0.02 MPa (P200), -0.04 MPa (P400), and -0.08 MPa (P800). The sperm motility, acrosome integrity and sperm fertilizing ability were evaluated. Application of –0.04 MPa improved the sperm motility, acrosome integrity and fertilizing ability, compared with the other groups. The sperm motility and acrosome integrity decreased with increasing storage time in vitro. After 5 days, the sperm motility and acrosome integrity of the P400 group were all higher than those of the other groups (P less than 0.05). The cleavage rate (64.5% ± 2.4%) and blastocyst development rate (33.9% ± 2.8%) for semen stored for 7 days were similar to those of fresh semen. In conclusion, application of –0.04 MPa before liquid storage at 17°C can improve the quality and fertilization ability of boar semen.

scholarly journals Elimination of apoptotic boar spermatozoa using magnetic activated cell sorting

2014 ◽  
Vol 83 (1) ◽  
pp. 13-18 ◽  
Author(s):  
Janko Mrkun ◽  
Tamara Dolenšek ◽  
Tanja Knific ◽  
Anja Pišlar ◽  
Marjan Kosec ◽  
...  
Keyword(s):  
Cell Sorting ◽  
Semen Quality ◽  
Annexin V ◽  
Alexa Fluor ◽  
Liquid Storage ◽  
Boar Semen ◽  
And Control ◽  
Boar Spermatozoa ◽  
First Time ◽  
Magnetic Activated Cell Sorting

One of the features of apoptosis is the externalization of phosphatidylserine which could be used to remove apoptotic cells from semen preparations. Magnetic-activated cell sorting using annexin V-conjugated microbeads which bind to phosphatidylserine could be used to enhance semen quality. Twelve boar semen samples after 3 days of liquid storage at 16­­–17 °C were subjected to magnetic-activated cell sorting. Bound and unbound fractions and control samples were subjected to flow cytometry following the staining of spermatozoa with Annexin V conjugated with Alexa Fluor 488 and propidium iodide. Four subpopulations were obtained: live, early apoptotic live, late apoptotic, early necrotic dead and late necrotic dead. The frequency of early apoptotic and late necrotic spermatozoa was significantly higher (P< 0.05) in bound (14.1 ± 10.6% and 24.1 ± 10.2%, respectively) than in unbound fractions (3.4 ± 2.1% and 12.7 ± 3.1%) and control (3.5 ± 1.6% and 12.0 ± 5.0%). The lowest concentration of live spermatozoa was found in the bound fraction (10.6 ± 8.0 %), which differed significantly (P< 0.05) from the control. In unbound fractions there was a significantly higher concentration (P< 0.05) of morphologically normal spermatozoa (31.8 ± 12.6%) compared to bound ones (5.9 ± 7.3%). A significantly (P< 0.05) lower proportion of morphologically normal spermatozoa was observed in both fractions compared to control (67.2 ± 17.0%). Boar spermatozoa were separated by the above method for the first time, however, the results showed this method to be inappropriate for boar semen separation under the tested conditions.

Effect of commercial long-term extenders on metabolic activity and membrane integrity of boar spermatozoa stored at 17°C

2013 ◽  
Vol 16 (3) ◽  
pp. 517-525 ◽  
Author(s):  
A. Dziekońska ◽  
L. Fraser ◽  
A. Majewska ◽  
M. Lecewicz ◽  
Ł. Zasiadczyk ◽  
...  
Keyword(s):  
Metabolic Activity ◽  
Storage Time ◽  
Membrane Integrity ◽  
Prolonged Storage ◽  
Atp Content ◽  
Liquid Storage ◽  
Semen Storage ◽  
Boar Semen ◽  
Boar Spermatozoa

Abstract This study was aimed to analyze the metabolic activity and membrane integrity of boar spermatozoa following storage in long-term semen extenders. Boar semen was diluted with AndrohepR EnduraGuardTM (AeG), DILU-Cell (DC), SafeCell PlusTM (SCP) and Vitasem LD (VLD) extenders and stored for 10 days at 17oC. Parameters of the analyzed sperm metabolic activity included total motility (TMOT), progressive motility (PMOT), high mitochondrial membrane potential (MMP) and ATP content, whereas those of the membrane integrity included plasma membrane integrity (PMI) and normal apical ridge (NAR) acrosome. Extender type was a significant (P < 0.05) source of variation in all the analyzed sperm parameters, except for ATP content. Furthermore, the storage time had a significant effect (P < 0.05) on the sperm metabolic activity and membrane integrity during semen storage. In all extenders the metabolic activity and membrane integrity of the stored spermatozoa decreased continuously over time. Among the four analyzed extenders, AeG and SCP showed the best performance in terms of TMOT and PMI on Days 5, 7 and 10 of storage. Marked differences in the proportions of spermatozoa with high MMP were observed between the extenders, particularly on Day 10 of storage. There were not any marked differences in sperm ATP content between the extenders, regardless of the storage time. Furthermore, the percentage of spermatozoa with NAR acrosomes decreased during prolonged storage, being markedly lower in DC-diluted semen compared with semen diluted with either AeG or SCP extender. The results of this study indicated that components of the long-term extenders have different effects on the sperm functionality and prolonged semen longevity by delaying the processes associated with sperm ageing during liquid storage.

scholarly journals The in Vitro Effect of Taurine on Boar Spermatozoa Quality

2018 ◽  
Vol 66 (1) ◽  
pp. 131-137
Author(s):  
Dušan Paál ◽  
František Strejček ◽  
Eva Tvrdá ◽  
Grzegorz Formicki ◽  
Sabine Klein ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Fluorescent Dyes ◽  
Semen Quality ◽  
Sperm Analysis ◽  
Spermatozoa Motility ◽  
Boar Semen ◽  
Acrosome Integrity ◽  
Boar Spermatozoa ◽  
Vitro Effect

The aim of this in vitro study was to evaluate the effects of taurine (TAU) supplementation on boar spermatozoa motility, viability, acrosome integrity and morphology. Eighteen boar semen samples were diluted with the Androhep PlusTM extender containing no TAU (control) or supplemented with 1.5 mM, 7 mM, 12.5 mM TAU and cultured at 4 °C for 18 days. Sperm motility was evaluated using the computer-aided sperm analysis (CASA) system. Furthermore, the samples were fixed and assessed for the occurrence of morphological abnormalities using phase contrast microscopy. Fluorescent dyes SYBR – 14 and propidium iodide were used to determine the sperm viability. Acrosome integrity was examined using PNA – Alexa Fluor 647 and flow cytometry. A gradual decrease of the semen quality was detected in all experimental groups over the course of the study. CASA revealed no selective advantage of TAU supplementation on the spermatozoa motility (p > 0.05). TAU administration showed to be ineffective in preserving spermatozoa viability as well as acrosome integrity as measured by flow cytometry (p > 0.05). Under the conditions of this study, no significant positive effect of TAU was recorded following its administration to the Androhep PlusTM boar semen extender with respect to spermatozoa quality.

Seminal plasma affects sperm sex sorting in boars

2016 ◽  
Vol 28 (5) ◽  
pp. 556 ◽  
Author(s):  
Diego V. Alkmin ◽  
Inmaculada Parrilla ◽  
Tatiana Tarantini ◽  
David del Olmo ◽  
Juan M. Vazquez ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Holding Time ◽  
Oxygen Species ◽  
Plasma Membrane Fluidity ◽  
Progressive Motility ◽  
Liquid Storage ◽  
Boar Semen ◽  
Sorting Efficiency ◽  
Boar Spermatozoa

Two experiments were conducted in boar semen samples to evaluate how both holding time (24 h) and the presence of seminal plasma (SP) before sorting affect sperm sortability and the ability of sex-sorted spermatozoa to tolerate liquid storage. Whole ejaculate samples were divided into three aliquots immediately after collection: one was diluted (1 : 1, v/v) in Beltsville thawing solution (BTS; 50% SP); the SP of the other two aliquots was removed and the sperm pellets were diluted with BTS + 10% of their own SP (10% SP) or BTS alone (0% SP). The three aliquots of each ejaculate were divided into two portions, one that was processed immediately for sorting and a second that was sorted after 24 h storage at 15–17°C. In the first experiment, the ability to exhibit well-defined X- and Y-chromosome-bearing sperm peaks (split) in the cytometry histogram and the subsequent sorting efficiency were assessed (20 ejaculates). In contrast with holding time, the SP proportion influenced the parameters examined, as evidenced by the higher number of ejaculates exhibiting split and better sorting efficiency (P < 0.05) in semen samples with 0–10% SP compared with those with 50% SP. In a second experiment, the quality (viability, total and progressive motility) and functionality (plasma membrane fluidity and intracellular generation of reactive oxygen species) of sex-sorted spermatozoa were evaluated after 0, 72 and 120 h storage at 15–17°C (10 ejaculates). Holding time and SP proportion did not influence the quality or functionality of stored sex-sorted spermatozoa. In conclusion, a holding time as long as 24 h before sorting did not negatively affect sex sorting efficiency or the ability of sorted boar spermatozoa to tolerate long-term liquid storage. A high proportion of SP (50%) in the semen samples before sorting reduced the number of ejaculates to be sorted and negatively influenced the sorting efficiency, but did not affect the ability of sex-sorted spermatozoa to tolerate liquid storage.

Interplay between Pur α and Egr-1 in the transcriptional regulation of amyloid precursor protein gene expression

2010 ◽  
Vol 34 (8) ◽  
pp. S27-S27
Author(s):  
Jianqi Cui ◽  
Xiuying Pei ◽  
Qian Zhang ◽  
Bassel E. Sawaya ◽  
Xiaohong Lu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcriptional Regulation ◽  
Amyloid Precursor Protein ◽  
Protein Gene ◽  
Precursor Protein ◽  
Amyloid Precursor Protein Gene

scholarly journals Actin-Related Protein 4 Interacts with PIE1 and Regulates Gene Expression in Arabidopsis

Genes ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 520
Author(s):  
Wenfeng Nie ◽  
Jinyu Wang
Keyword(s):  
Gene Expression ◽  
Plant Growth ◽  
Chromatin Remodeling ◽  
Leaf Size ◽  
Early Flowering ◽  
Physical Interaction ◽  
Loss Of Function ◽  
Single Nucleotide Change ◽  
Chromatin Remodeling Complex ◽  
Related Proteins

As essential structural components of ATP-dependent chromatin-remodeling complex, the nucleolus-localized actin-related proteins (ARPs) play critical roles in many biological processes. Among them, ARP4 is identified as an integral subunit of chromatin remodeling complex SWR1, which is conserved in yeast, humans and plants. It was shown that RNAi mediated knock-down of Arabidopsis thaliana ARP4 (AtARP4) could affect plant development, specifically, leading to early flowering. However, so far, little is known about how ARP4 functions in the SWR1 complex in plant. Here, we identified a loss-of-function mutant of AtARP4 with a single nucleotide change from glycine to arginine, which had significantly smaller leaf size. The results from the split luciferase complementation imaging (LCI) and yeast two hybrid (Y2H) assays confirmed its physical interaction with the scaffold and catalytic subunit of SWR1 complex, photoperiod-independent early flowering 1 (PIE1). Furthermore, mutation of AtARP4 caused altered transcription response of hundreds of genes, in which the number of up-regulated differentially expressed genes (DEGs) was much larger than those down-regulated. Although most DEGs in atarp4 are related to plant defense and response to hormones such as salicylic acid, overall, it has less overlapping with other swr1 mutants and the hta9 hta11 double-mutant. In conclusion, our results reveal that AtARP4 is important for plant growth and such an effect is likely attributed to its repression on gene expression, typically at defense-related loci, thus providing some evidence for the coordination of plant growth and defense, while the regulatory patterns and mechanisms are distinctive from other SWR1 complex components.

scholarly journals Serine-rich ultra high sulfur protein gene expression in murine hair and skin during the hair cycle.

1990 ◽  
Vol 265 (34) ◽  
pp. 21375-21380
Author(s):  
L Wood ◽  
M Mills ◽  
N Hatzenbuhler ◽  
G Vogeli
Keyword(s):  
Gene Expression ◽  
Protein Gene ◽  
Hair Cycle ◽  
Sulfur Protein
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