et al., 1971 a and b) and bivalents 10 and m (F!RGUSO:!

G-BANDING OF PACHYTENE BIVALENTS
IN HUMAN SPERMATOCYTES
J.
M. LUCIANI
Marie-Régine
MORAZZANI
A. STAHL
Labovatoive d’Histologie et Enibryologic 11,
U. E. R. de Me’!et’KC,
27, boulevard Jean-Moulin,
4
13385 MarseiLLe Cedex
SUMMARY
G-banding of meiotic pachytene chromosomes is obtained using trypsine procedure. G-bands
sufficiently pronounced to allow identification of several bivalents. Our results show the equivalence of the G-band patterns between mitotic chromosomes and meiotic bivalents. The G-band
patterns of mitotic-meiotic chromosomes also correlate closely with chromomere patterns of
meiotic pachytene bivalents.
are
!!-!
For several years, the identification of pachytene bivalents in human male
meiosis has aroused the interest of cytogeneticists.
The difficulty in obtaining 23 dispersed bivalents explains the paucity of results.
Analysis of chromomere patterns normally visible along pachytene bivalents has
led to the identification of acrocentric bivalents i
, z
3
, 21 and 22 (H
5
, i
4
RFORD
E
UNG
et al., 1971 a and b) and bivalents 10 and m (F!RGUSO:!-SMrTH and PAGE, 1973
).
Another bivalent, n
° 9
, has been identified thanks to its structural (presence of
parameres : Hu:vG!R!oxn et al., 1972
) and staining properties (Giemsa stain at
AGNE et al., z
pH m.6 : G
973
; PAGE, 1973).
Banding techniques have been too little used in studying meiotic chromosomes.
The A
RRIGHI and Hsu technique )
1971 permits one to obtain C-bands, however these
(
are often present in the absence of any denaturing treatment (I,u
rANr et al., 1972
C
and fig. r). Quinacrine mustard staining (C
RSSON et at., 1971
E
ASP
) allows the identification of bivalents during diakinesis, yet this method has not been applied to the
analysis of pachytene bivalents.
The use of trypsin digestion permits one to obtain G-bands along pachytene bivalents. This method, plus a modification of technique permitting bivalent dispersion,
leads to the identification of certain bivalents.
MATERIAL AND METHODS
Meiotic preparations were obtained from two subjects examined for sterility. A testicular
fragment obtained by biopsy, was inunediatly immersed in a o.q
4 P
. ioo hypotonic potassium
chloride (Lucmrr et al., 1971
). The hypotonic treatment was prolonged from 8 to 10 hours. The
3 vol/tI
sample was then transfered and dilacerated in a methanol and glacial acetic acid solution (
00 r.p.m. for minutes. The pellet was
vol.). The resulting cellular suspension was centrifuged at 8
resuspended in 5 mls of 45 p. ioo acetic acid, centrifuged once again and the final suspension was
spread on pre-cooled slides.
Slides thus obtained were placed in 2 x SSC solution at 40
C for 2 hours, then treated with
0
a o.2! p. ioo trypsin solution for io seconds at 1
8°C and finally stained in Giemsa solution.
o
RESULTS
AND DISCUSSION
Under trypsin action G-bands appear along the pachytene bivalents (fig. 2
).
Sequence analysis of the bands on isolated bivalents shows an exact correspondance
with G-bands characteristic of mitotic chromosomes. Thus, it is possible to identify
with certitude certain bivalents (fig.3
and 4
).
Our results show the existance of an equivalence between G-bands of meiotic
and mitotic chromosomes. In addition, the equivalence exists between G-banding
patterns of meiotic or mitotic chromosomes and the chromomere sequence described
UNGERFORD et al., for acrocentric bivalents (
USO
G
R
PE
by H
1971 a, b) and by rr-S:vmTH
and PAGE for bivalents 10 and 11 (r
). Finally, the reproductibility of the method
973
should rapidly lead to the mapping of pachytene bivalents.
Colloque
D. G. 12. S.
7’., Biologic de la Procy,!ation,
Paris, 7-8 inars 1975.
RÉSUMÉ
IDENTIFICATION DES I3IVAI,RNTS DU ST.1D!
PACHYTÉNE
o
DANS
PAR LA
LE SPËRMATOCYTE HUMAIN
TECHNIQUE
DES BANDES G
(TRYPSINE)
Sur les chromosomes du stade pachytène l’utilisation de la trypsine permet d’obtenir des
bandes G suffisamment nettes pour permettre l’identification de plusieurs bivalents. Nos résultats
montrent l’équivalence des bandes G entre les chromosomes mitotiques et les bivalents méiotiques.
RÉFÉRENCES BIBLIOGRAPHIQUES
F. E., Hsu T. C., 1971
. Localization of heterochromatin in human chromosomes. Cytogene8
86.
tics, 10, 1
INOSTEN J., Z
ECH L., 1971
ASPERSSON T., Hur.TEN D
C
, L
L
. Identification of chromosome bivalents
in human male meiosis by quinacrine mustard fluorescence analysis. Hereditas, Litnd, 67, .
149
147
ERGUSON NI. A., PAGE B. NI., 1973
F
MlTH
-S
10 II
(
) trans. Pachytene analysis in human reciprocal ;
.
7
8
location. ,J. Med. Genet., 10, 2
ABERGE C., T
ANGUAV R., r9!3. Aspect cytologique et localisation intranucléaire de l’héGAGNE R., L
térochromatine constitutive des chromosomes C, chez l’Homme. Cl:romosoma, 41, 159
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1
ALABAN t
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r
NI L. R., H
I
ALLER G., àTILLER A.
ABADIE G. LT., B
HUNGERFOIW D. A., L
L., r
97 a. Chromosome structure and function in man. IV. Ilrovisioilal map of the three long acrocentric
autosomes (chromosomes I3
.
0
6
2
, 14
, and 15
) at pachytene in the iiiale. Ann. Genet., 14, 257
ALABAN G. B., 1971 b. Chromosome structure and function
ABADIE G. U., B
HuNGERr·oRD D. A., L
in me.,. Il. Provisional maps of the two smallest autosomes (chromosomes 21 and 22
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i
the n.a
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LER G.,
L
A
UNGERFORD HTO
H
G. B.,
I
)
E
LA G.U.,M
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ESS
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21
5
6
21
Sci., U. S. A., 69, 68.
LucIANI J. M., DsvccTOR-VuILLET !L, STAHL A., 1971
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3
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D
UILLET
APODANO A. :’IL, -V
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AGNER
UCIANI J. M., -V
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973 Identification of chromosome 9 in human male meiosis. Cytogenet. Cell Genet., 12,
r
’ S4-z6
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3
RRIGUI
A