Quantification of HSP70 gene expression and determination of capacitation status of magnetically separated cryopreserved bovine spermatozoa at different thawing temperature and time

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

Research output: Contribution to journalArticle

Abstract

The role of heat shock protein in reproduction is widely known as a molecular chaperone in aiding and repairing protein formation when stress occurred. The present objectives were to evaluate the effect of different thawing temperature and time on the expression of HSP70 gene expression and the capacitation status in cryopreserved bovine spermatozoa. Briefly, fresh ejaculates were obtained from three different adult bulls. The semen then underwent a sperm washing technique known as Magnetic Activated Cell Sorting System (MACS) and later on, cryopreserved. The sperm-containing straws were then thawed at five different thawing temperatures and time post-cryostorage; 20°C for 13 s, 37°C for 30 s, 40°C for 7 s, 60°C for 6 s and 80°C for 5 s. The RNA was extracted from each of the sperm’s pellets and converted to cDNA prior to the qPCR process. Capacitation status was then determined by means of CTC assay. The results showed that after the process of amplification, there is a significant different of HSP70 gene expression in MACS process samples when the thawing process was performed at 37°C for 30 s, with p<0.05. Furthermore, the CTC assay also showed that thawing at the same temperature gave less capacitated spermatozoa with p<0.05. As a conclusion, MACS yield spermatozoa with a better expression of HSP70 gene and less capacitated spermatozoa when thawing was done at 37°C for 30 s.

Original languageEnglish
Pages (from-to)1101-1108
Number of pages8
JournalSains Malaysiana
Volume47
Issue number6
DOIs
Publication statusPublished - 1 Jun 2018

Fingerprint

thawing
spermatozoa
gene expression
cattle
temperature
sorting
molecular chaperones
assays
cells
heat shock proteins
washing
pellets
semen
straw
bulls
RNA
genes
proteins

Keywords

  • CTC
  • Heat-shock protein
  • MAC
  • Sperm cryopreservation
  • Thawing

ASJC Scopus subject areas

  • General

Cite this

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; Nang, Chew Fang; Zuraina, Farida; Yusof, Mohd.

In: Sains Malaysiana, Vol. 47, No. 6, 01.06.2018, p. 1101-1108.

Research output: Contribution to journalArticle

Mohamad, Syarifah Faezah Syed ; Ibrahim, Siti Fatimah ; Ismail, Nur Hilwan ; Osman, Khairul ; Jaafar, Farah Hanan Fathihah ; Nang, Chew Fang ; Zuraina, Farida ; Yusof, Mohd. / Quantification of HSP70 gene expression and determination of capacitation status of magnetically separated cryopreserved bovine spermatozoa at different thawing temperature and time. In: Sains Malaysiana. 2018 ; Vol. 47, No. 6. pp. 1101-1108.
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AU - Ibrahim, Siti Fatimah

AU - Ismail, Nur Hilwan

AU - Osman, Khairul

AU - Jaafar, Farah Hanan Fathihah

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AU - Zuraina, Farida

AU - Yusof, Mohd

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AB - The role of heat shock protein in reproduction is widely known as a molecular chaperone in aiding and repairing protein formation when stress occurred. The present objectives were to evaluate the effect of different thawing temperature and time on the expression of HSP70 gene expression and the capacitation status in cryopreserved bovine spermatozoa. Briefly, fresh ejaculates were obtained from three different adult bulls. The semen then underwent a sperm washing technique known as Magnetic Activated Cell Sorting System (MACS) and later on, cryopreserved. The sperm-containing straws were then thawed at five different thawing temperatures and time post-cryostorage; 20°C for 13 s, 37°C for 30 s, 40°C for 7 s, 60°C for 6 s and 80°C for 5 s. The RNA was extracted from each of the sperm’s pellets and converted to cDNA prior to the qPCR process. Capacitation status was then determined by means of CTC assay. The results showed that after the process of amplification, there is a significant different of HSP70 gene expression in MACS process samples when the thawing process was performed at 37°C for 30 s, with p<0.05. Furthermore, the CTC assay also showed that thawing at the same temperature gave less capacitated spermatozoa with p<0.05. As a conclusion, MACS yield spermatozoa with a better expression of HSP70 gene and less capacitated spermatozoa when thawing was done at 37°C for 30 s.

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