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Two-dimensional differential in-gel electrophoresis-based proteomics of male gametes in relation to oxidative stress.

Abstract：

OBJECTIVE:To identify the relative abundance of proteins in pooled reactive oxygen species (ROS)-positive (ROS+) and ROS-negative (ROS-) semen samples with the use of two-dimensional differential in-gel electrophoresis (2D-DIGE). DESIGN:Spermatozoa suspensions from ROS+ and ROS- groups by 2D-DIGE analysis. SETTING:Tertiary hospital. PATIENT(S):20 donors and 32 infertile men. INTERVENTION(S):Seminal ejaculates evaluated for semen and proteomic analysis. MAIN OUTCOME MEASURE(S):Semen samples from 20 donors and 32 infertile men were pooled, divided into ROS+ and ROS- groups based on the cutoff value of <20 relative light units/s/10(6) sperm and frozen. From each pooled group, spermatozoa were labeled with Cy3/Cy5 fluorescent dye. Duplicate 2D-DIGE gels were run. Image analysis was performed with the use of Decider software. Protein spots exhibiting ≥1.5-fold difference in intensity were excised from the preparatory gel and identified by liquid chromatography-mass spectrometry. Data were analyzed with the use of Sequest and Blast programs. RESULT(S):A total of 1,343 protein spots in gel 1 (ROS-) and 1,265 spots in gel 2 (ROS+) were detected. The majority of protein spots had similar expression, with 31 spots were differentially expressed. Six spots were significantly decreased and 25 increased in the ROS- sample compared with the ROS+ sample. CONCLUSION(S):Significantly different expression of protective proteins against oxidative stress was found in ROS-compared with ROS+ samples. These differences may explain the role of oxidation species in the pathology of male infertility.

journal_name

Fertil Steril

journal_title

Fertility and sterility

authors

Hamada A,Sharma R,du Plessis SS,Willard B,Yadav SP,Sabanegh E,Agarwal A

doi

10.1016/j.fertnstert.2012.11.046

subject

Has Abstract

pub_date

2013-04-01 00:00:00

pages

1216-1226.e2

issue

5

eissn

0015-0282

issn

1556-5653

pii

S0015-0282(12)02458-2

journal_volume

99

pub_type

杂志文章

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