Tuesday, May 15, 2007


European Journal of Human Genetics

Linear increase of structural and numerical
chromosome 9 abnormalities in human sperm
regarding age

Merce` Bosch1, Osvaldo Rajmil2, Josep Egozcue3 and Cristina Templado*,1
1Departament de Biologia Cel.lular, Fisiologia i Immunologia, Facultat de Medicina, Universitat Auto`noma de
Barcelona, Bellaterra 08193, Spain; 2Servei d’Andrologia, Fundacio´ Puigvert, Barcelona 08025, Spain; 3Departament
de Biologia Cel.lular, Fisiologia i Immunologia, Facultat de Cie`ncies, Universitat Auto`noma de Barcelona, Bellaterra
08193, Spain
A simultaneous four-colour fluorescence in situ hybridisation (FISH) assay was used in human sperm in
order to search for a paternal age effect on: (1) the incidence of structural aberrations and aneuploidy of
chromosome 9, and (2) the sex ratio in both normal spermatozoa and spermatozoa with a numerical or
structural abnormality of chromosome 9. The sperm samples were collected from 18 healthy donors, aged
24–74 years (mean 48.8 years old). Specific probes for the subtelomeric 9q region (9qter), centromeric
regions of chromosomes 6 and 9, and the satellite III region of the Y chromosome were used for FISH
analysis. A total of 190 117 sperms were evaluated with a minimum of 10 000 sperm scored from each
donor. A significant linear increase in the overall level of duplications and deletions for the centromeric
and subtelomeric regions of chromosome 9 (Pr0.002), chromosome 9 disomy (Po0.0001) as well as
diploidy (Po0.0001) was detected in relation to age. The percentage of increase for each 10-year period
was 29% for chromosome 9 disomy, 18.8% for diploidy, and ranged from 14.6 to 28% for structural
aberrations. Our results indicate a linear increase in structural aberrations and disomy for chromosome 9 in
sperm with respect to age.
European Journal of Human Genetics (2003) 11, 754–759. doi:10.1038/sj.ejhg.5201049
Keywords: paternal age; spermatozoa; structural aberrations; aneuploidy; diploidy; sex ratio; chromosome 9;

In conclusion, a linear relationship between donor age
and increasing frequencies of structural chromosome
aberrations and disomy for chromosome 9, and diploidy
in human spermatozoa have been herein demonstrated.
Thus far, disomy frequencies for 11 of the 24 chromosomes
of the human sperm karyotype have been evaluated
by FISH in relation to age, while structural aberrations
have only been analysed for chromosomes 110 and 9
(present study). It would be necessary to include other
Figure 1 Relationship between age and frequencies of
numerical and structural abnormalities for chromosome 9,
and diploidy.
Age and chromosome abnormalities in human sperm
M Bosch et al
European Journal of Human Genetics
chromosomes in these studies to define the real effect of
advanced paternal age on chromosomal abnormalities in
We thank Dr Pere Puig Casado for performing the statistical analysis
and also Mr Chuck Simmons for the revision and correction of the
English of this manuscript. This work received financial support from
Ministerio de Ciencia y Tecnologı´a (Project BFI2002-01193) and
Generalitat de Catalunya (CIRIT, 2001 SGR-00201), Spain.
1 Hassold T, Hunt PA: To err (meiotically) is human: the genesis of
human aneuploidy. Nat Rev 2001; 2: 280– 291.
2 Jacobs P: The chromosome complement of human gametes.
Oxford Rev Reprod Biol 1992; 14: 48– 72.
3 Olson SD, Magenis RE: Preferential paternal origin of de novo
structural chromosome rearrangements; in Daniel A (ed): The
cytogenetics of mammalian autosomal rearrangements. New York:
Alan R Liss, 1988, pp 583– 599.
4 Estop AM, Ma´rquez C, Munne S et al: An analysis of human sperm
chromosome breakpoints. Am J Hum Genet 1995; 56: 452– 460.
5 Templado C, Ma´rquez C, Munne S et al: An analysis of human
sperm chromosome aneuploidy. Cytogenet Cell Genet 1996; 74:
194– 200.
6 Shi Q, Martin RH: Aneuploidy in human sperm: a review of
frequency and distribution of aneuploidy, effects of donor age
and lifestyle factors. Cytogenet Cell Genet 2000; 90: 219– 226.
7 Martin RH, Rademaker AW: The effect of age on the frequency of
sperm chromosomal abnormalities in normal men. Am J Hum
Genet 1987; 41: 484– 492.
8 Prestes Sartorelli EM, Mazzucatto LF, Monterio de Pina-Neto J:
Effect of paternal age in human sperm chromosomes. Fertil Steril
2001; 79: 1119– 1123.
9 Martin RH, Spriggs E, Ko E, Rademaker AW: The relationship
between paternal age, sex ratios, and aneuploidy frequencies in
human sperm, as assessed by multicolor FISH. Am J Hum Genet
1995; 57: 1395– 1399.
10 McInnes B, Rademaker AW, Martin RH: Donor age and the
frequency of disomy for chromosomes 1, 13, 21 and structural
abnormalities in human spermatozoa using multicolor
fluorescence in-situ hybridization. Hum Reprod 1998; 13:
2489– 2494.
11 Bosch M, Rajmil O, Martı´nez-Pasarell O, Egozcue J, Templado C:
Linear increase of diploidy in human sperm with age: a fourcolour
FISH study. Eur J Hum Genet 2001; 9: 533– 538.
12 Robbins WA, Baulch JE, Moore II D, Weier HU, Blakey D,
Wyrobek AJ: Three-probe fluorescence in situ hybridisation to
assess chromosome X, Y, and aneuploidy in sperm of 14 men
from two healthy groups: evidence for a paternal age effect on
sperm aneuploidy. Reprod Fertil Dev 1995; 7: 799– 809.
13 Luetjens CM, Rolf C, Gassner P, Werny JE, Nieschlag E: Sperm
aneuploidy rates in younger and older men. Hum Reprod 2002; 7:
1826– 1832.
14 Rousseaux S, Hazzouri M, Pelletier R, Monteil M, Usson Y, Sele B:
Disomy rates for chromosomes 14 and 21 studied by fluorescent
in-situ hybridization in spermatozoa from three men over 60
years of age. Mol Hum Reprod 1998; 4: 695– 699.
15 Griffin DK, Abruzzo MA, Millie EA et al: Nondisjunction in
human sperm: evidence for an age effect of increasing paternal
age. Hum Mol Genet 1995; 4: 2227– 2232.
16 Asada H, Sueoka K, Hashiba T, Kuroshima M, Kobayashi N,
Yoshimura Y: The effect of age and abnormal sperm count on the
nondisjunction of spermatozoa. J Assist Reprod Genet 2000; 17:
51– 59.
17 Kinakin B, Rademaker A, Martin R: Paternal age effect of YY
aneuploidy in human sperm, as assessed by fluorescence in situ
hybridization. Cytogenet Cell Genet 1997; 78: 116–119.
18 Gardner RJM, Sutherland GR: Chromosome abnormalities
and genetic counseling. Oxford monograph on medical
genetics. New York: Oxford University Press, 1996, Vol 29:
pp 139– 152.
19 Cohen O, Cans C, CuillelMet al: Cartographic study: breakpoints
in 1574 families carrying human reciprocal translocations. Hum
Genet 1996; 97: 659– 667.
20 Brandriff BF, Gordon LA, Moore II DH, Carrano AV: An analysis of
structural aberrations in human sperm chromosomes. Cytogenet
Cell Genet 1988; 47: 29– 36.
21 Estop AM, Cieply K, Vankirt V, Munne S, Garver K: Cytogenetics
studies in human sperm. Hum Genet 1991; 87: 447– 451.
22 Starke H, Seidel J, HennWet al: Homologous sequences at human
chromosome 9 bands p12 and q13– 21.1 are involved in different
patterns of pericentric rearrangements. Eur J Hum Genet 2002; 10:
790– 800.
23 Sloter ED, Lowe X, Moore II DH, Nath J, Wyrobek AJ: Multicolor
FISH analysis of chromosomal breaks, duplications, deletions,
and numerical abnormalities in the sperm of healthy men. Am J
Hum Genet 2000; 67: 862– 872.
24 Benet J, Genesca` A, Navarro J, Egozcue J, Templado C:
Cytogenetic studies in motile sperm from normal men. Hum
Genet 1992; 89: 176– 180.
25 Yamada K: Population studies of inv(9) chromosomes in 4,300
Japanese: incidence, sex difference and clinical significance. Jpn J
Hum Genet 1992; 37: 293– 301.
26 Pellestor F, Girardet A, Coignet L, Andreo B, Charlieu JP:
Assessment of aneuploidy for chromosomes 8, 9, 13, 16, and
21 in human sperm by using primed in situ labeling technique.
Am J Hum Genet 1996; 58: 797– 802.
27 Rives N, Mazurier S, Bellet D, Joly G, Mace B: Assessment
of autosome and gonosome disomy in human sperm
nuclei by chromosome painting. Hum Genet 1998; 102:
616– 623.
28 Martin RH, Rademaker A: The frequencies of aneuploidy among
individual chromosomes in 6,821 human sperm complements.
Cytogenet Cell Genet 1990; 53: 103– 107.
29 Guttenbach M, Engel W, Schmid M: Analysis of structural and
numerical chromosome abnormalities in sperm of normal men
and carriers of constitutional chromosome aberrations. A review.
Hum Genet 1997; 100: 1– 21.
30 Gagne R, Laberge C, Tanguay R: Cytological aspect and
intranuclear localization of the heterochromatic segments of
C9 chromosomes in man. Chromosoma 1973; 41: 159–166.
31 Renaud H, Gasser SM: Heterochromatin: a meiotic matchmaker?
Trends Cell Biol 1997; 7: 201–205.
32 Amiel A, Sardos-Albertini F, Fejgin MD, Sharony R, Diukman R,
Bartoov B: Interchromosomal effect leading to an increase in
aneuploidy in sperm nuclei in a man heterozygous for pericentric
inversion (inv 9) and C-heterochromatin. J Hum Genet 2001; 46:
245– 250.
33 Serra A, Brahe C, Millington-Ward A et al: Pericentric inversion of
chromosome 9: prevalence in 300 Down syndrome families and
molecular studies of nondisjunction. Am J Med Genet 1990; 7:
162– 168.
34 Mroz K, Hassold TJ, Hunt PA: Meiotic aneuploidy in the XXY
mouse: evidence that a compromised testicular environment
increases the incidence of meiotic errors. Hum Reprod 1999; 14:
1151– 1156.
35 Egozcue S, Blanco J, Vendrell JM et al: Human male infertility:
chromosome anomalies, meiotic disorders, abnormal
spermatozoa and recurrent abortion. Hum Reprod Update 2000;
6: 93– 105.
Age and chromosome abnormalities in human sperm
M Bosch et al
European Journal of Human Genetics



Post a Comment

Subscribe to Post Comments [Atom]

<< Home