Identification of post‐translational modifications that occur during sperm maturation using difference in two‐dimensional gel electrophoresis

Baker, Mark A.; Witherdin, R.; Hetherington, Louise; Cunningham-Smith, K.; Aitken, R. John

doi:10.1002/pmic.200401100

Cited by 110 publications

(117 citation statements)

References 36 publications

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“…In other studies, calcium-binding proteins and proteins undergoing tyrosine phosphorylation during capacitation were identified using the proteomic approach [24,25]. A modified proteomic method, difference 2D gel electrophoresis, was developed to analyze post-translational modification of proteins during sperm maturation [26]. Using this powerful approach, several studies, including ours, have revealed a growing list of proteins that may be involved in regulating sperm motility [27][28][29][30][31][32].…”

Section: Introductionmentioning

confidence: 99%

Characterization of 3-hydroxyisobutyrate dehydrogenase, HIBADH, as a sperm-motility marker

Tasi

Chao

Chung

et al. 2013

J Assist Reprod Genet

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Purpose Asthenozoospermia is a major cause of male infertility. However, the molecular mechanisms underlying spermmotility defects remain largely unknown in the majority of cases. In our previous study, we applied a proteomic approach to identify unknown proteins that were downregulated in spermatozoa with low motility compared to spermatozoa with good motility. Several sperm motility-related proteins have been identified. In this study, 3-hydroxyisobutyrate dehydrogenase (HIBADH), one of the proteins identified using the proteomic tools, is further characterized.Methods Reverse-transcription polymerase chain reactions (RT-PCR), western blotting, and immunofluorescence assays (IFA) were preformed to investigate the expression pattern. The enzymatic activity of HIBADH was evaluated in sperm with good (>50 %), moderate (< 50 %) and lower motility (< 20 %). Results Using RT-PCR, we found that transcripts of HIBADH are enriched in the cerebellum, heart, skeletal muscle, uterus, placenta, and testes of male humans. In western blotting, it is expressed in the placenta, testes, and spermatozoa. During spermiogenesis, HIBADH is located at Capsule HIBADH is involved in the mitochondrial function of spermatozoa and maintains sperm motility. It may serve as a spermmotility marker.Pao-Lin Kuo and Ying-Hung Lin contributed equally to co-corresponding author. Assist Reprod Genet (2013) 30:505-512 DOI 10.1007 the mid-piece (a specialized development from the mitochondria) of elongating, elongated, and mature sperm. The enzymatic activity of HIBADH in sperm with moderate and lower motility were significantly reduced compared with good motility (P<0.0001 and P<0.05, respectively). Conclusions Our study indicated that HIBADH is involved in the mitochondrial function of spermatozoa, and maintains sperm motility. It may serve as a sperm-motility marker. Electronic supplementary material

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Section: Introductionmentioning

confidence: 99%

Characterization of 3-hydroxyisobutyrate dehydrogenase, HIBADH, as a sperm-motility marker

Tasi

Chao

Chung

et al. 2013

J Assist Reprod Genet

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“…A previous case report has already provided some evidence of the potential of the present proteomic tools to identify proteins involved in infertility [3]. More recently, the PTMs during sperm maturation in the rat have also been studied using fluorescent 2-DE [31]. We initiated the present work to characterize the most abundant proteins extracted from human sperm cells fractionated on Percoll gradients, through 2-DE and subsequent MS (MALDI-TOF and nano ESI IT) identification.…”

Section: Introductionmentioning

confidence: 99%

Proteomic identification of human sperm proteins

Martínez-Heredia¹,

Estanyol²,

Ballescà³

et al. 2006

Proteomics

230

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Conventional 1-DE has in the past provided a wealth of information concerning the major sperm proteins. However, so far there are relatively few reports exploiting the potential of the present proteomic tools to identify and to study additional yet-unidentified important proteins present in human spermatozoa. In the present work, 2-DE of proteins extracted from human normozoospermic spermatozoa led to the resolution of over 1000 spots. Subsequent excision from the gels of 145 spots and MALDI-TOF MS analysis allowed the identification of 98 different proteins. The function of these proteins turned out to be energy production (23%), transcription, protein synthesis, transport, folding and turnover (23%), cell cycle, apoptosis and oxidative stress (10%), signal transduction (8%), cytoskeleton, flagella and cell movement (10%), cell recognition (7%), metabolism (6%) and unknown function (11%). As many as 23% of the proteins identified have not been previously described as being expressed in human spermatozoa. The present data provide an important clue towards determining the function of these proteins and opens up the possibility to perform additional experiments.

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“…2,12 In addition, the distribution and post-translational modification of numerous proteins are also altered. [13][14][15] Through the process, the sperm gain the ability to swim forward and the capacity to undergo acrosome reaction and fertilize an ovum. 16 The mature sperm are then stored in the cauda epididymidis.…”

Section: Introductionmentioning

confidence: 99%

Mouse sperm acquire a new structure on the apical hook during epididymal maturation

Lin¹,

Hsu²,

Yen³

2013

Asian J Androl

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Spermatozoa emerging from the testis undergo a maturation process in the epididymis during which they change morphologically, biochemically and physiologically to gain motility and the ability to fertilize ova. We examined mouse epididymal sperm with immunostaining and transmission electron microscopy (EM) and identified a previously unknown structure on the apical hook. The structure has a coiled configuration around 11 nm in thickness and is present at the tip of each corner of the triangular-rod shaped perforatorium. Surveying sperm isolated from various regions of the epididymis indicated that mouse sperm acquire the hook rim (HR) structure during its passage through the proximal two-thirds of the caput epididymidis. The structure withstands vigorous sonication and harsh chemical treatments and remains intact after the acrosome reaction. Its location and sturdiness suggest a function in protecting the apical hook from mechanical wear during fertilization. Our EM images of epididymal sperm also revealed additional novel structures as well as lateral asymmetry of the sperm head, indicating that mouse sperm head has a structure more complex than previously recognized.

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Identification of post‐translational modifications that occur during sperm maturation using difference in two‐dimensional gel electrophoresis

Cited by 110 publications

References 36 publications

Characterization of 3-hydroxyisobutyrate dehydrogenase, HIBADH, as a sperm-motility marker

Characterization of 3-hydroxyisobutyrate dehydrogenase, HIBADH, as a sperm-motility marker

Proteomic identification of human sperm proteins

Mouse sperm acquire a new structure on the apical hook during epididymal maturation

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