The three-dimensional morphology and distribution of human chromosomes 3 were studied in nuclei of spermatogonia and spermatocytes I from formaldehyde-fixed human testis sections. Chromosome arms, pericentromeres and telomeric regions were painted by a three-color, five-probe fluorescence in situ hybridization protocol. Light optical serial sections of premeiotic and meiotic nuclei obtained by confocal laser scanning microscopy revealed that premeiotic chromosomes 3 are separate from each other and occupy variably shaped territories, which are sectored in distinct 3 p- and q-arm domains. Three-dimensional reconstructions of the painted chromosome domains by a Voronoi tessellation approach showed that mean chromosome volumes did not differ significantly among the premeiotic and meiotic stages investigated. A significant increase in surface area and reduction of dimensionless ‘roundness factor’ estimates of arm domains indicated that the restructuring of spatially separate chromosome territories initiates during preleptotene. Telomeric regions, which in meiotic stem cells located predominantly in arm-domain chromatin, showed a redistribution towards the domain surface during this stage. At leptotene homologues were generally misaligned and displayed intimate intermingling of non-homologous chromatin. Pairing initiated at the ends of bent zygotene chromosomes, which displayed a complex surface structure with discernible sister chromatids. The results indicate that, in mammals, homology search is executed during leptotene, after remodeling of chromosome territories.

REFERENCES

Adler
I. D.
(
1996
).
Comparison of the duration of spermatogenesis between male rodents and humans.
Mutat. Res
352
,
169
172
Ambros
P. F.
,
Sumner
A. T.
(
1987
).
Correlation of pachytene chromomeres and metaphase bands of human chromosomes and distinctive properties of telomeric regions.
Cytogenet. Cell Genet
44
,
223
228
Anderson
L. K.
,
Offenberg
H. H.
,
Verkuilen
W. M. H. C.
,
Heyting
C.
(
1997
).
RecA-like proteins are components of early meiotic nodules in Lilly.
Proc. Nat. Acad. Sci. USA
94
,
6868
6873
Aragon-Alcaide
L.
,
Reader
S.
,
Beven
A.
,
Shaw
P.
,
Miller
T.
,
Moore
G.
(
1997
).
Association of homologous chromosomes during floral development.
Curr. Biol
7
,
905
908
Armstrong
S. J.
,
Kirkham
A. J.
,
Hulten
M. A. J.
(
1994
).
XY chromosome behavior in the germ-line of the human male: a FISH analysisof spatial orientation, chromatin condensation and pairing.
Chromosome Res
2
,
445
452
Ashley
T.
,
Plug
A. W.
,
Xu
J.
,
Solari
A. J.
,
Reddy
G.
,
Golub
E. I.
,
Ward
D. C.
(
1995
).
Dynamic changes in Rad51 distribution on chromatin during meiosis in male and female vertebrates.
Chromosoma
104
,
19
28
Baker
S. M.
,
Bronne
C. E.
,
Zhang
L.
,
Plug
A. W.
,
Robatzek
M.
,
Warren
G.
,
Elliott
E. A.
,
Yu
J.
,
Ashley
T.
,
Arnheim
N.
,
Flavell
R. A.
,
Liskay
R. M.
(
1995
).
Male mice defective in the DNA mismatch repair gene PMS2 exhibit abnormal chromosome synapsis in meiosis.
Cell
82
,
309
319
Barlow
A. L.
,
Hulten
M. A.
(
1996
).
Combined immunocytogenetic and molecular cytogenetic analysis of meiosis I human spermatocytes.
Chromosome Res.
4
562
573
Barlow
A. L.
,
Benson
F. E.
,
West
S. C.
,
Hulten
M. A.
(
1997
).
Distribution of the Rad51 recombinase in human and mouse spermatocytes.
EMBO J.
16
,
5207
5215
Bascom-Slack
C. A.
,
Dawson
D. S.
(
1997
).
The yeast motor protein, Kar3p, is essential for meiosis I.
J. Cell Biol
139
,
459
467
Bass
H. W.
,
Marshall
W. F.
,
Sedat
J. W.
,
Agard
D. A.
,
Cande
W. Z.
(
1997
).
Telomeres cluster de novo before the initiation of synapsis: athree-dimensional spatial analysis of telomere positions before and duringmeiotic prophase.
J. Cell Biol
137
,
5
18
Baumann
P.
,
Benson
F. E.
,
West
S. C.
(
1996
).
Human Rad51 protein promotes ATP-dependent homologous pairing and strand transfer reactions in vitro.
Cell
87
,
757
766
Bishop
D. K.
(
1994
).
RecA homologs Dmc1 and Rad51 interact to form multiple nuclear complexes prior to meiotic chromosome synapsis.
Cell
79
,
1081
1092
Boiko
M.
(
1983
).
Human meiosis VIII. Chromosome pairing and formation of the synaptonemal complex in oocytes.
Carlsberg Res. Commun
48
,
457
483
Carpenter
A. T.
(
1987
).
Gene conversion, recombination nodules and initiation of meiotic synapsis.
BioEssays
6
,
232
236
Cheng
E. Y.
,
Gartler
S. M.
(
1994
).
A fluorescent in situ hybridization analysis of X chromosome pairing in human female meiosis.
Hum. Genet
94
,
389
394
Chikashige
Y.
,
Ding
D. Q.
,
Funabiki
H.
,
Haraguchi
T.
,
Mashiko
S.
,
Yanagida
M.
,
Hiraoka
Y.
(
1994
).
Telomere-led premeiotic chromosome movement in fission yeast.
Science
264
,
270
273
Chikashige
Y.
,
Ding
D. Q.
,
Imai
Y.
,
Yamamoto
M.
,
Haraguchi
T.
,
Hiraoka
Y.
(
1997
).
Meiotic nuclear reorganization: switching the position of centromeres and telomeres in the fission yeast Schizosaccharomyces pombe.
EMBO J.
16
193
202
Comings
D. E.
(
1980
).
Arrangement of chromatin in the nucleus.
Hum. Genet
53
,
131
143
Cook
P. R.
(
1997
).
The transcriptional basis of chromosome pairing.
J. Cell Sci
110
,
1033
1040
Craig
J. M.
,
Bickmore
W. A.
(
1994
).
The distribution of CpG islands in mammalian chromosomes.
Nat. Genet
7
,
376
382
Cremer
T.
,
Kurz
A.
,
Zirbel
R.
,
Rinke
B.
,
Dietzel
S.
,
Schröck
E.
,
Speicher
M. R.
,
Ried
T.
,
Matthieu
U.
,
Jauch
A.
,
Emmerich
P.
,
Scherthan
H.
,
Cremer
C.
,
Lichter
P.
(
1993
).
Role of chromosome territories in the functional compartmentalization of the cell nucleus.
Cold Spring Harbor Symp. Quant. Biol
58
,
777
792
Dawe
K. R.
,
Sedat
J. W.
,
Agard
D. A.
,
Cande
Z. W.
(
1994
).
Meiotic chromosome pairing in maize is associated with a novel chromatin organization.
Cell
76
,
901
912
Dernburg
A. F.
,
Sedat
J. W.
,
Hawley
R. S.
(
1996
).
Direct evidence of a role for heterochromatin in meiotic chromosome segregation.
Cell
86
,
135
146
Dietrich
A. J. J.
,
de Boer
P.
(
1983
).
A sequential analysis of the development of the synaptonemal complex in spermatocytes of the mouse by electron microscopy using hydroxyurea and agar filtration.
Genetica
61
,
119
129
Dietzel
S.
,
Jauch
A.
,
Kienle
D.
,
Qu
G.
,
Holtgreve-Grez
H.
,
Eils
R.
,
Munkel
C.
,
Bittner
M.
,
Meltzer
P. S.
,
Trend
J. M.
,
Cremer
T.
(
1998
).
Separate and variably shaped chromosome arm domains are disclosed by chromosome arm painting in human cell nuclei.
Chromosome Res
6
,
25
33
Ding
D. Q.
,
Chikashige
Y.
,
Haraguchi
T.
,
Hiraoka
Y.
(
1998
).
Oscillatory nuclear movement in fission yeast meiotic prophase is driven byastral microtubules, as revealed by continuous observation of chromosomes and microtubules in living cells.
J. Cell Sci
111
,
701
712
Eils
R.
,
Bertin
E.
,
Saracoglu
K.
,
Rinke
B.
,
Schröck
E.
,
Parazza
F.
,
Usson
Y.
,
Robert-Nicoud
M.
,
Stelzer
E. H. K.
,
Chassery
J.-M.
,
Cremer
T.
,
Cremer
C.
(
1995
).
Application of laser confocal microscopy and 3D-Voronoi diagrams for volume and surface estimates of interphase chromosomes.
J. Microscopy
177
,
150
161
Eils
R.
,
Dietzel
S.
,
Bertin
E.
,
Schröck
E.
,
Speicher
M. R.
,
Ried
T.
,
Robert-Nicoud
M.
,
Cremer
C.
,
Cremer
T.
(
1996
).
Three-dimensional reconstruction of painted human interphase chromosomes: active and inactive X-chromosome territories have similar volumes but differ in shape and surface structure.
J. Cell Biol
135
,
1427
1440
Ferguson
M.
,
Ward
D. C.
(
1992
).
Cell cycle dependent chromosomal movement in pre-mitotic human T-lymphocyte nuclei.
Chromosoma
101
,
557
565
Ferreira
J.
,
Paolella
G.
,
Ramos
C.
,
Lamond
A. I.
(
1997
).
Spatial organization of large-scale chromatin domains in the nucleus: a magnified view of single chromosome territories.
J. Cell Biol
139
,
1597
1610
Funabiki
H.
,
Hagan
I.
,
Uzawa
S.
,
Yanagida
M.
(
1993
).
Cell cycle-dependent specific positioning and clustering of centromeres and telomeres in fission yeast.
J. Cell Biol
121
,
961
976
Goldman
A. S.
,
Lichten
M.
(
1996
).
The efficiency of meiotic recombination between dispersed sequences in Saccharomyces cerevisiae depends upon their chromosomal location.
Genetics
144
,
43
55
Guan
X. Y.
,
Zhang
H.
,
Bittner
M.
,
Jiang
Y.
,
Meltzer
P.
,
Trent
J.
(
1996
).
Chromosome arm painting probes.
Nat. Genet
12
,
10
11
Haber
J. E.
,
Leung
W. Y.
,
Borts
R. H.
,
Lichten
M.
(
1991
).
The frequency of meiotic recombination in yeast is independent of the number and position of homologous donor sequences: implications for chromosome pairing.
Proc. Nat. Acad. Sci. USA
88
,
1120
1124
Hawley
R. S.
,
Arbel
T.
(
1993
).
Yeast genetics and the fall of the classical view of meiosis.
Cell
72
,
301
303
Heng
H. H.
,
Chamberlain
J. W.
,
Shi
X. M.
,
Spyropoulos
B.
,
Tsui
L. C.
,
Moens
P. B.
(
1996
).
Regulation of meiotic chromatin loop size by chromosomal position.
Proc. Nat. Acad. Sci. USA
93
,
2795
2800
Heyting
C.
(
1996
).
Synaptonemal complexes: structure and function.
Curr. Opin. Cell Biol
8
,
389
396
Hiraoka
Y.
,
Dernburg
A. F.
,
Parmerlee
S. J.
,
Rykowski
M. C.
,
Agard
D. A.
,
Sedat
J. W.
(
1993
).
The onset of homologous chromosome pairing during Drosophila melanogaster embryogenesis.
J. Cell Biol
120
,
591
600
Jackson
D. A.
,
Cook
P. R.
(
1995
).
The structural basis of nuclear function.
Int. Rev. Cytol
162
,
125
149
Jin
Q.-W.
,
Trelles-Sticken
E.
,
Scherthan
H.
,
Loidl
J.
(
1998
).
Yeast nuclei display prominent centromere clustering which is reduced in non-dividing cells and in meiotic prophase.
J. Cell Biol
141
,
21
29
Jinks-Robertson
S.
,
Petes
T. D.
(
1985
).
High-frequency meiotic gene conversion between repeated genes on non-homologous chromosomes.
Proc. Nat. Acad. Sci. USA
82
,
3350
3354
Karpen
G. H.
,
Le
M. H.
,
Le
H.
(
1996
).
Centric heterochromatin and the efficiency of achiasmate disjunction in Drosophila female meiosis.
Science
273
,
118
122
Kleckner
N.
(
1996
).
Meiosis: how could it work?.
Proc. Nat. Acad. Sci. USA
93
,
8167
8174
Kleckner
N.
,
Padmore
R.
,
Bishop
D. K.
(
1991
).
Meiotic chromosome metabolism: one view.
Cold Spring Harbor Symp. Quant. Biol
56
,
729
743
Kleckner
N.
,
Weiner
B.
(
1993
).
Potential advantages of unstable interactions for pairing in meiotic, somatic, and premeiotic cells.
Cold Spring Harbor Symp. Quant. Biol
58
,
553
565
Kohli
J.
(
1994
).
Telomeres lead chromosome movement.
Curr. Biol
4
,
724
727
Kurz
A.
,
Lampel
S.
,
Nickolenko
J. E.
,
Bradl
J.
,
Benner
A.
,
Zirbel
R. M.
,
Cremer
T.
,
Lichter
P.
(
1996
).
Active and inactive genes localizepreferentially in the periphery of chromosome territories.
J. Cell Biol
135
,
1195
1205
Loidl
J.
(
1990
).
The initiation of meiotic chromosome pairing: the cytological view.
Genome
33
,
759
778
Loidl
J.
,
Länger
H.
(
1993
).
Evaluation of models of homologue search with respect to their efficiency on meiotic pairing.
Heredity
71
,
342
351
Loidl
J.
,
Klein
F.
,
Scherthan
H.
(
1994
).
Homologous pairing is reduced but not abolished in asynaptic mutants of yeast.
J. Cell Biol
125
,
1191
1200
Loidl
J. H.
,
Scherthan
J. T.
,
Dunnen
D.
,
Klein
F.
(
1995
).
Morphology of a human-derived YAC in Yeast meiosis.
Chromosoma
104
,
183
188
Luciani
J. M.
,
Guichaoua
M. R.
,
Cau
P.
,
Devictor
B.
,
Salagnon
N.
(
1988
).
Differential elongation of autosomal pachytene bivalents related to their DNA content in human spermatocytes.
Chromosoma
97
,
19
25
Maguire
M. P.
(
1972
).
Role of heterochromatin in homologous chromosome pairing: evaluation of evidence.
Science
176
,
543
544
Marshall
W. F.
,
Straight
A.
,
Marko
J. F.
,
Swedlow
J.
,
Dernburg
A.
,
Belmont
A.
,
Murray
A. W.
,
Agard
D. A.
,
Sedat
J. W.
(
1997
).
Interphase chromosomes undergo constrained diffusional motion in living cells.
Curr. Biol
7
,
930
939
Moens
P. B.
(
1974
).
Quantitative electron microscopy of chromosome organization at meiotic prophase.
Cold Spring Harbor Symp. Quant. Biol.
,
38
,
99
107
Moens
P. B.
,
Pearlman
R. E.
(
1988
).
Chromatin organization at meiosis.
BioEssays
9
,
151
153
Moses
M. J.
(
1968
).
Synaptinemal Complex.
Ann. Rev. Genetics
2
,
363
412
Murti
J. R.
,
Bumbulis
M.
,
Schimenti
J. C.
(
1994
).
Gene conversion between unlinked sequences in the germline of mice.
Genetics
137
,
837
843
Nagele
R.
,
Freeman
T.
,
McMorrow
L.
,
Lee
H.-Y.
(
1995
).
Precise spatial positioning of chromosomes during prometaphase: evidence for chromosomal order.
Science
270
,
1831
1835
Oud
J. L.
,
Reutlinger
A. H. H.
(
1981
).
Chromosome behaviour during early meiotic prophase of mouse spermatocytes.
Chromosoma
83
,
395
407
Parvinnen
M.
,
Söderström
K.-O.
(
1976
).
Chromosome rotation and formation of synapsis.
Nature
260
,
534
535
Plug
A. W.
,
Peters
A. H.
,
Keegan
K. S.
,
Hoekstra
M. F.
,
De Boer
P.
,
Ashley
T.
(
1998
).
Changes in protein composition of meiotic nodules during mammalian meiosis.
J. Cell Sci
111
,
413
423
Radman
M.
,
Wagner
R.
(
1993
).
Mismatch recognition in chromosomal interactions and speciation.
Chromosoma
102
,
369
373
Rasmussen
S. W.
,
Holm
P. B.
(
1978
).
Human Meiosis II. Chromosome pairing and recombination nodules in human spermatocytes.
Carlsberg Res. Com
43
,
275
327
Rasmussen
S. W.
,
Holm
P. B.
(
1980
).
Mechanics of Meiosis.
Hereditas
93
,
187
216
Robinett
C. C.
,
Straight
A.
,
Li
G.
,
Willhelm
C.
,
Sudlow
G.
,
Murray
A.
,
Belmont
A. S.
(
1996
).
In vivo localization of DNA sequences and visualization of large-scale chromatin organization using lac operator/repressor recognition.
J. Cell Biol
135
,
1685
1700
Rockmill
B.
,
Sym
M.
,
Scherthan
H.
,
Roeder
G. S.
(
1995
).
Two RecA homologs facilitate meiotic chromosome synapsis by establishing connections between homologous chromosomes.
Genes Dev
9
,
2684
2695
Roeder
G. S.
(
1995
).
Sex and the single cell: meiosis in yeast.
Proc. Nat. Acad. Sci. USA
92
,
10450
10456
Roeder
G. S.
(
1997
).
Meiotic chromosomes: it takes two to tango.
Genes Dev
11
,
2600
2621
Sachs
R. K.
,
van den Engh
G.
,
Trask
B.
,
Yokota
H.
,
Hearst
J. E.
(
1995
).
A random-walk/giant-loop model for interphase chromosomes.
Proc. Nat. Acad. Sci. USA
92
,
2710
2714
Salonen
K.
,
Paranko
J.
,
Parvinen
M.
(
1982
).
A colcemid sensitive mechanism involved in regulation of chromosome movements during meiotic pairing.
Chromosoma
85
,
611
618
Scherthan
H.
,
Loidl
J.
,
Schuster
T.
,
Schweizer
D.
(
1992
).
Meiotic chromosome condensation and pairing in Saccharomyces cerevisiae studied by chromosome painting.
Chromosoma
101
,
590
595
Scherthan
H.
,
Bähler
J.
,
Kohli
J.
(
1994
).
Dynamics of chromosomeorganization and pairing during meiotic prophase of fission yest.
J. Cell Biol
127
,
273
285
Scherthan
H.
,
Weich
S.
,
Schwegler
H.
,
Heyting
C.
,
Härle
M.
,
Cremer
T.
(
1996
).
Centromere and telomere movements during early meiotic prophase of mouse and man are associated with the onset of chromosome pairing.
J. Cell Biol
134
,
1109
1125
Schulze
W.
,
Rehder
U.
(
1984
).
Organization and morphogenesis of the human seminiferous epithelium.
Cell Tissue Res
237
,
395
407
Schwarzacher
T.
(
1997
).
Three stages of meiotic homologous chromosome pairing in wheat:cognition, alignment and synapsis.
Sexual Plant Reprod
10
,
324
331
Selig
S.
,
Okumura
K.
,
Ward
D. C.
,
Cedar
H.
(
1992
).
Delineation of replication time zones by fluorescence in situ hybridization.
EMBO J
11
,
1217
1225
Smith
A.
,
Benavente
R.
(
1995
).
An M r51,000 protein of mammalian spermatogenic cells that is common to the whole XY body and centromeric heterochromatin of autosomes.
Chromosoma
103
,
591
596
Smithies
O.
,
Powers
P. A.
(
1986
).
Gene conversions and their relation to to homologous chromosome pairing.
Phil. Trans. R. Soc. Lond. B
312
,
291
302
Stack
S. M.
(
1984
).
Heterochromatin, the synaptonemal complex and crossing over.
J. Cell Sci
71
,
159
176
Svoboda
A.
,
Bähler
J.
,
Kohli
J.
(
1995
).
Microtubule-driven nuclear movements and linear elements as meiosis-specific characteristics of the fission yeasts Schizosaccharomyces versatilis and Schizosaccharomyces pombe.
Chromosoma
104
,
203
214
Terasawa
M.
,
Shinohara
A.
,
Hotta
Y.
,
Ogawa
H.
,
Ogawa
T.
(
1995
).
Localization of RecA-like recombination proteins on chromosomes of the lily at various meiotic stages.
Genes Dev
9
,
925
934
Vocero-Akbani
A.
,
Helms
C.
,
Wang
J. C.
,
Sanjurjo
F. J.
,
Korte-Sarfaty
J.
,
Veile
R. A.
,
Liu
L.
,
Jauch
A.
,
Burgess
A. K.
,
Hing
A. V.
,
Holt
M. S.
,
Ramachandra
S.
,
Whelan
A. J.
,
Anker
R.
,
Ahrent
L.
,
Chen
M.
,
Gavin
M. R.
,
Iannantuoni
K.
,
Morton
S. M.
,
Pandit
S. D.
,
Read
C. M.
,
Steinbrueck
T.
,
Warlick
C.
,
Smoller
D. A.
,
Donis-Keller
H.
(
1996
).
Mapping human telomere regions with YAC and P1 clones: Chromosome specific markers for 27 telomeres including 149 STSs and 24 polymorphisms for 14 proterminal regions.
Genomics
36
,
492
506
von Wettstein
D.
,
Rasmussen
S. W.
,
Holm
P. B.
(
1984
).
The synaptonemal complex in genetic segregation.
Annu. Rev. Genet
18
,
331
413
Vourc'h
C.
,
Taruscio
D.
,
Boyle
A. L.
,
Ward
D. C.
(
1993
).
Cell cycle-dependent distribution of telomeres, centromeres and chromosome-specific subsatellite domains in the interphase nucleus of mouse lymphocytes.
Exp. Cell Res
205
,
142
151
Wandall
A.
,
Svendsen
A.
(
1985
).
Transition from somatic to meiotic pairing and progressional changes of the synaptonemal complex.
Chromosoma
92
,
254
264
Waye
J. S.
,
Willard
H. F.
(
1989
).
Chromosome specificity of satellite DNAs: short-and long-range organization of a diverged dimeric subset of human alpha satellite from chromosome 3.
Chromosoma
97
,
475
480
Weiner
B.
,
Kleckner
N.
(
1994
).
Chromosome pairing via multiple interstitial interactions before and during meiosis in yeast.
Cell
77
,
977
991
Wu
T. C.
,
Lichten
M.
(
1994
).
Meiosis-induced double-strand break sites determined by yeast chromatin structure.
Science
263
,
515
518
Yokota
H.
,
van den Engh
G.
,
Hearst
J. E.
,
Sachs
R. K.
,
Trask
B. J.
(
1995
).
Evidence for the organization of chromatin in megabase pair-sized loops arranged along a random walk path in the human G0/G1interphase nucleus.
J. Cell Biol
130
,
1239
1249
Young
I. T.
(
1977
).
Proof without prejudice: Use of the Kolmogorov-Smirnov test for the analysis of histograms from flow systems and other sources.
J. Histochem. Cytochem
25
,
935
941
Zalensky
A. O.
,
Tomilin
N. V.
,
Zalenskaya
I. A.
,
Teplitz
R. L.
,
Bradbury
E. M.
(
1997
).
Telomere-telomere interactions and candidate telomere binding protein(s) in mammalian sperm cells.
Exp. Cell Res
232
,
29
41
Zink
D.
,
Cremer
T.
,
Saffrich
R.
,
Fischer
R.
,
Trendelenburg
M. F.
,
Ansorge
W.
,
Stelzer
E. H.
(
1998
).
Structure and dynamics of human interphase chromosome territories in vivo.
Hum. Genet
102
,
241
251
This content is only available via PDF.