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Human Reproduction vol.15 no.4 pp.853856, 2000

Detection of chromosomal abnormalities by fluorescentin-situ hybridization in immotile viable spermatozoadetermined by hypo-osmotic sperm swelling test*

Hulusi Bulent Zeyneloglu1! "ol#an Baltaci$%evinc &ge1 'li Haberal1 and %(Batioglu1

1Department of Obstetrics/Gnaecolo!, an" 2Department of#e"ical Genetics, $as%ent &niversit, 'n%ara, ()r%e

3(o *+om correspon"ence s+o)l" be a""resse" at #esa -or)itesi #anola $lo% 24, aol), 0630, ()r%e

)f randomly selected immotile spermatozoa are used for

intracytoplasmic sperm inection +),%) pregnancy rates

are significantly decreased( .he hypo-osmotic swelling test

+H/%. is the only method available to detect the viable

but immotile spermatozoa for ),%)( However evidence is

still lac#ing for the chromosomal abnormalities for the

normal-loo#ing but immotile spermatozoa positive for

H/%.( %perm samples from $0 infertile men with normal

chromosomal constitution were obtained( 'fter ercoll

separation morphologically normal but immotile sperma-

tozoa were transported individually into H/%. solution

for 1 min using micropipettes( ,ells that showed tail

curlingwith swelling in H/%. were then transferred

bac# intohuman tubal fluid solution to allow reversal

of swelling(.hese sperm cells were fi2ed and processed

for the multi-colour fluorescence in-situ hybridization

+3)%H for chro-mosomes 4 5 and 16( .he same

3)%H procedure wasapplied for the motile spermatozoafrom the same cohortwhich formed the control group(

.he average aneuploidyrates were 1(70 and 1(89: in

1000 H/%. positive immotileand motile spermatozoa

respectively detected by 3)%H foreach patient( /ur

results indicate that morphologicallynormal immotile

but viable spermatozoa have an aneuplo-idy rate similar

to that of normal motile spermatozoa(Key words:

c+romosome abnormalit/+poosmotic s*ellin! test/male

infertilit/oli!ooospermia/spermatooa

)ntroductionntractoplasmic sperm inection is t+e assiste" repro

")ctive tec+nolo! proce")re of c+oice in cases of severe

malefactor infertilit an" )neplaine" fertiliation fail)re

afterconventional invitro fertiliation 7 9alermo et al.,

1::2.(+e a"vanta!es of are i t+e nee" of onl one

spermatooon per oocte an" ii t+e abilit to )se not onl

t+eeac)late" spermatooa, b)t also spermatooa from testis

an"

immat)re forms of sperm cells from t+e testis ma be

obtaine"*it+ varin! s)ccess.

9oor or absent fertiliation rates are obtaine" in cases of

)sin! immotile eac)late" spermatooa of )n%no*n

viabilit ;a! et al., 1::5. (otal immotilit ma be ca)se"

b necrooospermia, )ltrastr)ct)ral "efects or metabolic

immat)rit of t+e spermatooa.

n cases of total necrooospermia in t+e eac)late, motile

spermatooa can sometimes be recovere" from a testic)lar

biops (o)rnae et al., 1::6.

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@)ropean ociet of

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H(B(Zeyneloglu et al.

3igure 1( (+e spermatooa inc)bate" in +poosmotic sol)tion, t+eapanF if t+e final

*as+o)t "i" not pro")ce s)fficient immotile spermatooa, t+en t+e

spermatooa from t+e interface bet*een t+e !ra"ients of 9ercoll *ere

)se". (+e spermatooa *ere inc)bate" in +poosmotic sol)tion for

3igure $( 9+oto!rap+s of immotile spermatooa after < analsis*it+ centromeric probes for t+e c+romosomes 18 bl)e, !reenan" = oran!e in a t+reecolo)r +bri"iation protocol. 'll probes*ere "irectl labelle" *it+ fl)oroc+romes an" obtaine"commerciall. cale bar = 5 m.

.able )( omparison of c+romosomal stat)s of motile an" immotile sperm cells from 20 infertile men after < analsis

mmotile

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(+e t*o tpes of sperm cell *ere not si!nificantl

"ifferent.

Resul

ts

(+e mean sperm concentration of t+e patients *as C.3

2.C106/ml an" me"ian *as C.5106/ml ran!e 2.312.

#eanpercenta!e total motile tpe ' + tpe $ + tpe

spermatooa *as 18.8 C.6B me"ian 1:BF ran!e C

35. (+e meanpercenta!e normal morp+olo! accor"in! to(!erber! strict criteria *as 1.8 1.0B me"ian 2.0F

ran!e 03. 'vera!e c+romosome, = c+romosome an"

18 c+romosome anomalrates in"ivi")all *ere 0.46, 0.33

an" 0.2CB respectivel. Overall, t+e avera!e inci"ence of

spermatooa *it+ abnormal c+romosomal stat)s *as 1.C0

4.48B 0.:2.6B in immotile