Molecular analysis of polymerase gamma gene and mitochondrial polymorphism in fertile and subfertile men

T P Harris, K P Gomas, F Weir, A J Holyoake, P McHugh, M Wu, Y Sin, I L Sin, F Y T Sin

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

CAG trinucleotide repeat length in the nuclear polymerase gamma gene (POLgamma) has been shown to be associated with men with reduced fertility. The present study investigated the frequency of CAG repeat length genotypes and three exonuclease motifs of the POLgamma in relation to the frequency of mitochondrial nucleotide substitutions. DNA from semen samples of 93 normozoospermic men and 192 non-normozoospermic men was isolated and the specific regions of the genes were amplified by polymerase chain reactions (PCR) and sequenced to identify mutations. The genotypic frequencies of pooled POLgamma CAG repeat lengths, =10/ not equal 10 heterozygotes and not equal 10/ not equal 10 homozygotes, were significantly different between normozoospermic and non-normozoospermic men (p < 0.05), with non-normozoospermic men having a slightly higher frequency of the =10/=10 genotypes. The allelic frequency for =10 is 0.79 and not equal10 is 0.21 for normozoospermic men and 0.85 and 0.15, respectively, for non-normozoospermic men (p < 0.025). There was no mutation detected in the exonuclease motifs in all the samples tested. Eighty normozoospermic and 124 non-normozoospermic semen samples were analysed for nucleotide substitutions in mitochondrial genes by PCR and sequencing. Heteroplasmic mutations were found in one azoospermic man, four asthenozoospermic men and two normozoospermic men. Only one asthenozoospermic man was heterozygous for the POLgamma genotype. Of the 17 men with non-synonymous nucleotide substitutions, 14 were homozygous for the POLgamma genotype. Non-normozoospermic men had twice as many nucleotide substitutions than normozoospermic men. However, there were no significant differences in the frequencies of nucleotide substitution and POLgamma genotypes in the two groups of men.

Original languageEnglish
Pages (from-to)421-433
Number of pages13
JournalInternational Journal of Andrology
Volume29
Issue number3
DOIs
Publication statusPublished - Jun 2006
Externally publishedYes

Fingerprint

Mitochondrial Genes
Nucleotides
Genotype
Genes
Exonucleases
Semen
Mutation
Trinucleotide Repeats
Polymerase Chain Reaction
Homozygote
Heterozygote

Cite this

Harris, T P ; Gomas, K P ; Weir, F ; Holyoake, A J ; McHugh, P ; Wu, M ; Sin, Y ; Sin, I L ; Sin, F Y T. / Molecular analysis of polymerase gamma gene and mitochondrial polymorphism in fertile and subfertile men. In: International Journal of Andrology. 2006 ; Vol. 29, No. 3. pp. 421-433.
@article{8e42880705c149aa965db17d64b0509c,
title = "Molecular analysis of polymerase gamma gene and mitochondrial polymorphism in fertile and subfertile men",
abstract = "CAG trinucleotide repeat length in the nuclear polymerase gamma gene (POLgamma) has been shown to be associated with men with reduced fertility. The present study investigated the frequency of CAG repeat length genotypes and three exonuclease motifs of the POLgamma in relation to the frequency of mitochondrial nucleotide substitutions. DNA from semen samples of 93 normozoospermic men and 192 non-normozoospermic men was isolated and the specific regions of the genes were amplified by polymerase chain reactions (PCR) and sequenced to identify mutations. The genotypic frequencies of pooled POLgamma CAG repeat lengths, =10/ not equal 10 heterozygotes and not equal 10/ not equal 10 homozygotes, were significantly different between normozoospermic and non-normozoospermic men (p < 0.05), with non-normozoospermic men having a slightly higher frequency of the =10/=10 genotypes. The allelic frequency for =10 is 0.79 and not equal10 is 0.21 for normozoospermic men and 0.85 and 0.15, respectively, for non-normozoospermic men (p < 0.025). There was no mutation detected in the exonuclease motifs in all the samples tested. Eighty normozoospermic and 124 non-normozoospermic semen samples were analysed for nucleotide substitutions in mitochondrial genes by PCR and sequencing. Heteroplasmic mutations were found in one azoospermic man, four asthenozoospermic men and two normozoospermic men. Only one asthenozoospermic man was heterozygous for the POLgamma genotype. Of the 17 men with non-synonymous nucleotide substitutions, 14 were homozygous for the POLgamma genotype. Non-normozoospermic men had twice as many nucleotide substitutions than normozoospermic men. However, there were no significant differences in the frequencies of nucleotide substitution and POLgamma genotypes in the two groups of men.",
keywords = "DNA Mutational Analysis, DNA Polymerase gamma, DNA Replication, DNA, Mitochondrial/physiology, DNA-Directed DNA Polymerase/genetics, Exons/genetics, Genotype, Heterozygote, Homozygote, Humans, Infertility, Male/genetics, Male, Mitochondria/enzymology, Polymorphism, Single Nucleotide, Sperm Count, Sperm Motility/genetics, Trinucleotide Repeats",
author = "Harris, {T P} and Gomas, {K P} and F Weir and Holyoake, {A J} and P McHugh and M Wu and Y Sin and Sin, {I L} and Sin, {F Y T}",
year = "2006",
month = "6",
doi = "10.1111/j.1365-2605.2005.00633.x",
language = "English",
volume = "29",
pages = "421--433",
journal = "International Journal of Andrology",
issn = "0105-6263",
publisher = "Wiley",
number = "3",

}

Molecular analysis of polymerase gamma gene and mitochondrial polymorphism in fertile and subfertile men. / Harris, T P; Gomas, K P; Weir, F; Holyoake, A J; McHugh, P; Wu, M; Sin, Y; Sin, I L; Sin, F Y T.

In: International Journal of Andrology, Vol. 29, No. 3, 06.2006, p. 421-433.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Molecular analysis of polymerase gamma gene and mitochondrial polymorphism in fertile and subfertile men

AU - Harris, T P

AU - Gomas, K P

AU - Weir, F

AU - Holyoake, A J

AU - McHugh, P

AU - Wu, M

AU - Sin, Y

AU - Sin, I L

AU - Sin, F Y T

PY - 2006/6

Y1 - 2006/6

N2 - CAG trinucleotide repeat length in the nuclear polymerase gamma gene (POLgamma) has been shown to be associated with men with reduced fertility. The present study investigated the frequency of CAG repeat length genotypes and three exonuclease motifs of the POLgamma in relation to the frequency of mitochondrial nucleotide substitutions. DNA from semen samples of 93 normozoospermic men and 192 non-normozoospermic men was isolated and the specific regions of the genes were amplified by polymerase chain reactions (PCR) and sequenced to identify mutations. The genotypic frequencies of pooled POLgamma CAG repeat lengths, =10/ not equal 10 heterozygotes and not equal 10/ not equal 10 homozygotes, were significantly different between normozoospermic and non-normozoospermic men (p < 0.05), with non-normozoospermic men having a slightly higher frequency of the =10/=10 genotypes. The allelic frequency for =10 is 0.79 and not equal10 is 0.21 for normozoospermic men and 0.85 and 0.15, respectively, for non-normozoospermic men (p < 0.025). There was no mutation detected in the exonuclease motifs in all the samples tested. Eighty normozoospermic and 124 non-normozoospermic semen samples were analysed for nucleotide substitutions in mitochondrial genes by PCR and sequencing. Heteroplasmic mutations were found in one azoospermic man, four asthenozoospermic men and two normozoospermic men. Only one asthenozoospermic man was heterozygous for the POLgamma genotype. Of the 17 men with non-synonymous nucleotide substitutions, 14 were homozygous for the POLgamma genotype. Non-normozoospermic men had twice as many nucleotide substitutions than normozoospermic men. However, there were no significant differences in the frequencies of nucleotide substitution and POLgamma genotypes in the two groups of men.

AB - CAG trinucleotide repeat length in the nuclear polymerase gamma gene (POLgamma) has been shown to be associated with men with reduced fertility. The present study investigated the frequency of CAG repeat length genotypes and three exonuclease motifs of the POLgamma in relation to the frequency of mitochondrial nucleotide substitutions. DNA from semen samples of 93 normozoospermic men and 192 non-normozoospermic men was isolated and the specific regions of the genes were amplified by polymerase chain reactions (PCR) and sequenced to identify mutations. The genotypic frequencies of pooled POLgamma CAG repeat lengths, =10/ not equal 10 heterozygotes and not equal 10/ not equal 10 homozygotes, were significantly different between normozoospermic and non-normozoospermic men (p < 0.05), with non-normozoospermic men having a slightly higher frequency of the =10/=10 genotypes. The allelic frequency for =10 is 0.79 and not equal10 is 0.21 for normozoospermic men and 0.85 and 0.15, respectively, for non-normozoospermic men (p < 0.025). There was no mutation detected in the exonuclease motifs in all the samples tested. Eighty normozoospermic and 124 non-normozoospermic semen samples were analysed for nucleotide substitutions in mitochondrial genes by PCR and sequencing. Heteroplasmic mutations were found in one azoospermic man, four asthenozoospermic men and two normozoospermic men. Only one asthenozoospermic man was heterozygous for the POLgamma genotype. Of the 17 men with non-synonymous nucleotide substitutions, 14 were homozygous for the POLgamma genotype. Non-normozoospermic men had twice as many nucleotide substitutions than normozoospermic men. However, there were no significant differences in the frequencies of nucleotide substitution and POLgamma genotypes in the two groups of men.

KW - DNA Mutational Analysis

KW - DNA Polymerase gamma

KW - DNA Replication

KW - DNA, Mitochondrial/physiology

KW - DNA-Directed DNA Polymerase/genetics

KW - Exons/genetics

KW - Genotype

KW - Heterozygote

KW - Homozygote

KW - Humans

KW - Infertility, Male/genetics

KW - Male

KW - Mitochondria/enzymology

KW - Polymorphism, Single Nucleotide

KW - Sperm Count

KW - Sperm Motility/genetics

KW - Trinucleotide Repeats

U2 - 10.1111/j.1365-2605.2005.00633.x

DO - 10.1111/j.1365-2605.2005.00633.x

M3 - Article

VL - 29

SP - 421

EP - 433

JO - International Journal of Andrology

JF - International Journal of Andrology

SN - 0105-6263

IS - 3

ER -