Quantification of HSP70 Gene Expression and Determination of Capacitation Status of Magnetically Separated Cryopreserved Bovine Spermatozoa at Different Thawing Temperature and Time

Mohamad, Syarifah Faezah Syed; Ibrahim, Siti Fatimah; Ismail, Nur Hilwan; Osman, Khairul; Jaafar, Farah Hanan Fathihah; Chew, Fang Nang; Yusof, Farida Zuraina Mohd

doi:10.17576/jsm-2018-4706-04

Cited by 4 publications

(4 citation statements)

References 24 publications

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“…(32). The quality of bull spermatozoa may also be accessible by the quantification of Hsp70 gene expression rather than motility alone, according to the findings of the (33) study.…”

Section: Resultsmentioning

confidence: 99%

Protective Effects of Autologous Platelet-Rich Plasma (PRP) on the Outcome of Cryopreservation in Rabbit Sperm

Abdulla¹,

Rebaï²,

Al-Delemi³

2022

Cell Mol Biol (Noisy-le-grand)

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Sperm cryopreservation is a cost-effective means of preserving gene resources and transporting sperm across distant locations. However, due to the general disadvantages of using freeze-thawed sperm, to prevent this irreversible damage, cryopreservation techniques must be improved by the addition of additional cryoprotection agents. This study aims to improve the freezability of buck semen using an intratesticular injection of Autologous Platelet-Rich Plasma (PRP) and confirm this theory by Casa laboratory analysis and gene expression detection of three genes (CATSPER1, SPAG5 and Hsp70). Twenty rabbits a New Zealand healthy male were randomly divided into two equal groups; the control and PRP group which enriched with PRP, the semen collection was applied after 10 weeks, then each sample was divided into two fractions; First fractions we apply the laboratory semen analysis directly, and the second fractions were cryopreservation, then thawed after one month to apply the same laboratory analysis. The results of CASA, DNA fragmentation, and real time-PCR analysis had statistically significant differences (P<0.01) when compartment of these values after and before freezing, yet we don't record any statistical differences between the C group and CR group. This study's findings are extremely significant, indicating that intratesticular injection of PRP is a good method for using in enhancing the rabbit sperm procedure after freeze-thawing.

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“…(32). The quality of bull spermatozoa may also be accessible by the quantification of Hsp70 gene expression rather than motility alone, according to the findings of the (33) study.…”

Section: Resultsmentioning

confidence: 99%

Protective Effects of Autologous Platelet-Rich Plasma (PRP) on the Outcome of Cryopreservation in Rabbit Sperm

Abdulla¹,

Rebaï²,

Al-Delemi³

2022

Cell Mol Biol (Noisy-le-grand)

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“…The initial concentration of the mixture was determined using SpermaCueTM (MiniTube, Germany) and adjusted to 25 X 10 6 cells/mL in 0.25mL cryopreservation straws. Cryopreservation was conducted based on the protocol as described by [6]. Thawing protocols for both groups of sperm were conducted in a 37°C water bath for 30secs.…”

Section: Sperm Isolation System Cryopreservation and Thawing Protocolmentioning

confidence: 99%

Analysis of Bovine Spermatozoa CSP-Gene Expression using qPCR and Relative Quantification Method as Biodiagnostic Tool for Fertilizing Capacity

Ismail¹,

Mohamad²,

Jaafar³

et al. 2019

IJET

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Gene expression studies enable for real-time relative quantification of expressed genes. However, the incorporation of a primary discerning tool before amplification for specific sequences of genes of interest is yet to be implemented. The amplification of gene sequences from discriminate sample groups further enhances the results following qPCR and provides an absolute exclusive measurement for the defined sample. Cryopreserved spermatozoa have characteristics of compromised sperm quality as the cells are exposed to a rapid temperature downshift not normally encountered by such cells in vivo. Thus, the use of a separation system before cryopreservation produces a refined outcome in gene expression studies. The proliferation of Cold Shock Protein (CSP) can relatively be quantified by conducting qPCR following sperm selection and cryopreservation. CSPs are indicators of cells adapting to the decrease in temperature and indirectly acts as an intracellular protective mechanism for the cell. The current study compares the effects of sperm selection system in isolating a homogeneous population of spermatozoa for cryopreservation followed by quantification of CSP gene. Method of molecular assessment involved quantifying the amplified sequences via qPCR and gene expression following sperm isolation was significant in increasing spermatozoa viability by 26.2% (P< 0.05). The relative fold expression of CSP gene for treatment group increased from 1.00 fold to 1.38 fold. The Cq values for treatment group had recorded an earlier point of amplification (Cq= 24.87) as compared to control Cq= 25.83. Our findings suggest that the use of a sperm isolation system before cryopreservation would increase the probability of obtaining higher amplifications of CSP genes that would confer protection against extremely low temperatures during cryopreservation. This would increase the likelihood of in vitro fertilization using cryopreserved spermatozoa by implementing qPCR as a potential biomolecular diagnostic tool to ascertain the fertilizing potential of the spermatozoa.

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“…Besides these common sperm transcripts across various livestock species, there were flagellar specific spRNAs and sperm motility proteins such as BSP ( de Souza et al, 2017 ; Pardede et al, 2020 ), AKAP ( Gilbert et al, 2007 ; Chatterjee et al, 2010 ; Kasimanickam and Kastelic, 2016 ; Ing et al, 2020 ), ODF ( Chen et al, 2015 ; Li et al, 2018 ; Pardede et al, 2020 ; Özbek et al, 2021 ), zinc finger nucleases ( Card et al, 2013 ; Kasimanickam and Kastelic, 2016 ; Corral-Vazquez et al, 2021 ) and heat shock proteins. The heat shock proteins are a group of tyrosine regulatory elements that participate in hyperactivation and nitric oxide synthesis during fertilization ( Feugang et al, 2010 ; Mohamad et al, 2018 ; Nicolas et al, 2020 ; Lian et al, 2021 ; Sun et al, 2021 ). The spRNAs such as calmegin ( Guyonnet et al, 2009 ; Card et al, 2013 ), clusterin ( Singh et al, 2019 ; Lian et al, 2021 ; Zhao et al, 2021 ), TSSK ( Bissonnette et al, 2009 ; Li et al, 2020 ; Ablondi et al, 2021 ), and CABYR ( Selvaraju et al, 2017 ) were also reported in cattle and pigs, are known to be essential for male fertility.…”

Section: Introductionmentioning

confidence: 99%

“…The important spRNAs related to sperm capacitation included CABYR ( Bailey, 2010 ) and AKAP ( Maciel et al, 2018 ), which regulate calcium-binding and tyrosine phosphorylation. On the other hand, CatSper, VDAC1, and HSPA2 are involved in calcium signaling whereas , COL11A1, PDE4A can participate in PI3K-Akt and cAMP-PKA signaling pathways ( Li et al, 2018 ), The HSP70 ( Mohamad et al, 2018 ) and HSP90 ( Jin and Yang, 2017 ), involved in calcium signaling, were also reported in spRNA population. Thus, it appears that calcium signaling and tyrosine phosphorylation are important events of capacitation that spRNAs may mediate.…”

Section: Introductionmentioning

confidence: 99%

Significance and Relevance of Spermatozoal RNAs to Male Fertility in Livestock

et al. 2021

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Livestock production contributes to a significant part of the economy in developing countries. Although artificial insemination techniques brought substantial improvements in reproductive efficiency, male infertility remains a leading challenge in livestock. Current strategies for the diagnosis of male infertility largely depend on the evaluation of semen parameters and fail to diagnose idiopathic infertility in most cases. Recent evidences show that spermatozoa contains a suit of RNA population whose profile differs between fertile and infertile males. Studies have also demonstrated the crucial roles of spermatozoal RNA (spRNA) in spermatogenesis, fertilization, and early embryonic development. Thus, the spRNA profile may serve as unique molecular signatures of fertile sperm and may play pivotal roles in the diagnosis and treatment of male fertility. This manuscript provides an update on various spRNA populations, including protein-coding and non-coding RNAs, in livestock species and their potential role in semen quality, particularly sperm motility, freezability, and fertility. The contribution of seminal plasma to the spRNA population is also discussed. Furthermore, we discussed the significance of rare non-coding RNAs (ncRNAs) such as long ncRNAs (lncRNAs) and circular RNAs (circRNAs) in spermatogenic events.

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Quantification of HSP70 Gene Expression and Determination of Capacitation Status of Magnetically Separated Cryopreserved Bovine Spermatozoa at Different Thawing Temperature and Time

Cited by 4 publications

References 24 publications

Protective Effects of Autologous Platelet-Rich Plasma (PRP) on the Outcome of Cryopreservation in Rabbit Sperm

Protective Effects of Autologous Platelet-Rich Plasma (PRP) on the Outcome of Cryopreservation in Rabbit Sperm

Analysis of Bovine Spermatozoa CSP-Gene Expression using qPCR and Relative Quantification Method as Biodiagnostic Tool for Fertilizing Capacity

Significance and Relevance of Spermatozoal RNAs to Male Fertility in Livestock

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