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Neo-XY body: An analysis of XY1Y2 meiotic

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Phyllostomidae) by chromosome painting

Article in Cytogenetic and Genome Research · February 2009

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 Original Article
Cytogenet Genome Res 2009;124:37–43
DOI: 10.1159/000200086
Neo-XY body: an analysis of XY1Y2
meiotic behavior in Carollia (Chiroptera,
Phyllostomidae) by chromosome painting
R.C.R. Noronha a C.Y. Nagamachi a P.C.M. O’Brien b M.A. Ferguson-Smith b
J.C. Pieczarka a
a
Laboratório de Citogenética, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belem (Brazil)
b
Molecular Cytogenetics Laboratory, Department of Veterinary Medicine, University of Cambridge, Cambridge (UK)
Abstract
Classical and molecular cytogenetic analyses of mitotic and
meiotic cells were performed on two species of Carollia from
the family Phyllostomidae (Chiroptera), which have an XX/
XY1Y2 sex determination system. Our results show that the
species Carollia perspicillata and Carollia brevicauda have the
same Xq-autosome translocation (neo-X). Using multicolor
FISH we observed different levels of condensation of the
original X and Y chromosomes when compared to the trans-
located autosomal segment, a likely consequence of the nu-
cleolar organizer region blocking spreading of inactivation
to the autosomal region of the neo-X. The use of chromo-
some painting showed the behavior of the sex chromosome
trivalent – here called the ‘neo-XY body’ – in meiosis. We
compared the variation between the condensation of the
original X and Y and the autosome-sex chromosome axis
and described the pairing between the original X-Y seg-
ments (pseudoautosomal region) and the XY2 homologous
segments, suggesting genetic activity of the latter during
meiosis. Copyright © 2009 S. Karger AG, Basel
This study was supported by CNPq, CAPES and UFPa.
© 2009 S. Karger AG, Basel
1424–8581/09/1241–0037$26.00/0
Fax +41 61 306 12 34
E-Mail karger@karger.ch Accessible online at:
www.karger.com www.karger.com/cgr
Accepted after revision: October 15, 2008
by M. Schmid
Sex chromosome condensation is a meiotic adaptation
to prevent the initiation of potentially damaging recom-
bination events within nonhomologous regions of the
X and Y chromosome (McKee and Handel, 1993). Re-
cently a new proposal for the process of XY inactivation
called meiotic sex chromosome inactivation (MSCI) has
emerged as a novel paradigm for the study of the epigen-
etic regulation of gene expression (Handel, 2004; Turner,
2007).
The sex chromosomes of mammalian spermatocytes
form a specialized nuclear territory known as the XY
body, where both transcription and homologous recom-
bination are restricted. The array of proteins assembled
into the XY body is typical of heterochromatin. This spe-
cial subnuclear domain is in distinct contrast to the au-
tosomal domain of the spermatocyte nucleus, where both
homologous recombination and transcription occur
(Handel, 2004). The special features of the XY body might
reflect absence of homology between the sex chromo-
somes, rather than any form of dosage compensation.
According to Turner (2007), MSCI is a manifestation
of a general meiotic silencing mechanism called meiotic
silencing of unsynapsed chromatin (MSUC). It is initi-
ated by several DNA repair proteins that are maintained
by histone modifications which are associated with tran-
Dr. Julio Cesar Pieczarka
Instituto de Ciências Biológicas, UFPA, CCB, 3º Andar
Av. Augusto Corrêa SN, Bairro Guamá, Belém, Pará (Brazil) 66.075-900
telephone/fax: +91 3201 7931
e-mail: julio@ufpa.br, juliopieczarka@pesquisador.cnpq.br
scriptional silencing in a wide variety of developmental
contexts. This strongly suggests that the formation of
the transcriptionally repressed XY chromatin domain is
a meiotic phenomenon, unrelated to either X inactiva-
tion for dosage compensation or any possible require-
ment for X- or Y-encoded proteins. Following meiosis, X
and Y chromosome repression is maintained and the X
and Y chromosomes appear as heterochromatic do-
mains called postmeiotic sex chromatin (PMSC; Turner
2007).
In all species with multiple (XY1Y2) chromosomal sex
determination systems the sex chromosome-autosome
complexes form a trivalent. Thus, synapses occur be-
tween the original X and Y segments (at the pseudoauto-
somal region, PAR) and in the translocated autosome
segments (Solari, 1994; Solari and Pigozzi, 1994).
The subfamily Carolliinae (Phyllostomidae, Chirop-
tera) is a group of frugivore neotropical bats widely dis-
tributed in the New World. This subfamily is composed
of the genera Carollia and Rhinophylla and is character-
ized by intrageneric karyotype stability, with chromo-
somal sex determination systems of simple XX/XY and
multiple XX/XY1Y2 types (Baker et al., 1989). The XY1Y2
system could have arisen in the ancestor of all Carollia
species after the fusion of an autosome to the X chromo-
some. Thus, the homologous autosome becomes a sex
chromosome named Y2, while Y1 is the true male sex
chromosome of the system. Stock (1975) compared the
karyotype of C. castanea with that of C. perspicillata and
identified the C. castanea autosome that was translocated
to the X chromosome of C. perspicillata.
There are only conventional studies on the meiotic be-
havior of the sex chromosomes of Carollia. Hsu et al.
(1968) described the diplotene phase in Carollia perspicil-
lata and observed the formation of a sex trivalent (XY1Y2),
pointing out the pairing of the homologous region be-
tween Xq and Y2. Noronha et al. (2004) applied the con-
ventional technique for meiotic analysis to C. perspicil-
lata and suggested the formation of a typical sex vesicle
(SV) in pachytene. From the more advanced phases on,
such as diplotene and diakinesis, the sex trivalent (XY1Y2)
presented differences in the behavior of the original XY
axes as compared to the translocated autosome. Accord-
ing to McKee and Handel (1993), after the zygotene-to-
pachytene transition when meiotic synapsis between au-
tosomes is complete, the X and Y chromosomes are rap-
idly silenced and compartmentalized into a peripheral
nuclear subdomain called the sex or XY body. MSCI then
persists throughout the rest of pachytene and diplotene
(Turner, 2007).
38 Cytogenet Genome Res 2009;124:37–43
The frequent presence of an interstitial heterochro-
matic block (LINES-IHB) between the translocated auto-
some and the sex chromosome is thought to be respon-
sible for the evolutionary stability of such systems in a
variety of mammals (Dobigny et al., 2004). In C. perspi-
cillata, instead of heterochromatin, it seems that the nu-
cleolar organizing region (NOR) blocks the spreading of
X inactivation to the autosomal region (Noronha et al.,
2004). According to Handel (2004), the XY body forma-
tion is under regulation, controlled intrinsically and au-
tonomously in male germ cells.
Unlike the rest of the spermatocyte nucleus, the XY
body is a domain characterized by nonhomologous mei-
otic interactions. The X and Y chromosomes are largely
differentiated and share full sequence homology only in
the distal ‘pseudoautosomal’ pairing region, the PAR. Af-
ter synapsis, the PAR undergoes reciprocal recombina-
tion that is visible as a chiasma in diplotene; this serves to
ensure their disjunction during meiosis I anaphase (Han-
del, 2004). However, there is no published report so far on
a meiotic analysis with chromosome painting showing
the PAR.
The availability of whole chromosome probes for the
sex chromosomes of two phyllostomid bat species (Piec-
zarka et al., 2005) made it possible to analyze the meiotic
chromosomes of Carollia perspicillata. The chromosome
painting technique applied to meiosis can clarify many
persisting questions about the evolution of the sex chro-
mosomes in Carollia and the behavior of autosomes
translocated to sex chromosomes during the different
phases of meiosis.
Materials and methods
A cytogenetic study was performed on five Carollia perspicil-
lata (CPE, 2n = 20, 21) and five C. brevicauda (CBR, 2n = 20, 21)
specimens. The specimens were collected from natural popula-
tions and identified by Dr. Suely Marques-Aguiar from the Mu-
seu Paraense Emílio Goeldi (MPEG) and are deposited there. The
conventional meiosis technique followed Eicher (1966). The mi-
totic cells were obtained from bone marrow according to Ford
and Hamerton (1956) and also from fibroblast cell cultures es-
tablished from skin biopsies. NOR technique followed Howell
and Black (1980), and C-banding followed Sumner (1972). The
bright field images were photographed with a Carl Zeiss III pho-
tomicroscope, lens 100! oil immersion, optovar 1.25, with green
filter, ASA 3.2 (DIN 6) and 6.3 (DIN 9). The films employed were
Agfa Copex Pan and Imagelink HQ, developed and fixed with
Kodak D76 developer and Kodak fixer, respectively. The images
were copied onto Kodabrome Print F3 (Kodak) paper and digi-
tized.
Noronha/Nagamachi/O’Brien/
Ferguson-Smith/Pieczarka
 a b
Fig. 1. Mitotic analysis of the species Ca-
rollia perspicillata and C. brevicauda,
XY1Y2. (a) C-banding pattern of chromo-
somes XY1Y2 of C. perspicillata. (b) Multi-
color FISH of chromosomes X, Y1, and Y2
of C. brevicauda. (c) Ag-NOR labeling in
the interstitial region of the long arm of the
X chromosome of C. brevicauda (arrow).
(d) Metaphase of C. brevicauda hybridized
with PY1CBR, labeling heterochromatic
regions, and counterstained with DAPI
that enhances the secondary constriction
of the X chromosome (arrow).
For multicolor FISH we used the Carollia brevicauda probes
previously described by Pieczarka et al. (2005). The neo-X, Y1, and
Y2 chromosomes were sorted by flow cytometry and the DNA
amplified by DOP-PCR. Here we designate the neo-X probe as
PXCBR (probe for X from Carollia brevicauda) and, in a similar
way, the Y1 probe as PY1CBR and the Y2 probe as PY2CBR. For
tricolor experiments PXCBR was labeled with FITC (green) and
PY2CBR with Cy3 (red), both by direct labeling. The Y1 probe was
directly labeled with Cy5 (pink). Both PXCBR and PY2CBR paint-
ed the homologous regions of the neo-X and Y2, producing a yel-
low signal on each (due to the combination of red and green). For
two-color experiments PXCBR was labeled with FITC (green)
and PY1CBR with Cy3 (red). In situ hybridization of painting
probes was performed as previously described (Wienberg et al.,
1990, 1992; Scherthan et al., 1994; Yang et al., 1995). Briefly, 14 !l
of the hybridization mixture (50% formamide, 1! SSC, 10% dex-
tran sulfate, 5 mg salmon sperm DNA, 2 mg mouse Cot-1 DNA)
and 1 !l of labeled PCR product were denatured at 65 ° C, dropped
onto denatured chromosome preparations, and mounted with 22
! 22 mm cover slips. Slides were denatured in 70% formamide,
2! SSC at 65 ° C for 1 min. In situ hybridization was performed
for 48–72 h at 37 ° C. The hybridization signal was detected as de-
scribed earlier (Yang et al., 1995). Digital images were obtained
using a cooled CCD camera (Photometrics NU200 series equipped
with a Kodak KAF 1400 chip) coupled to a Zeiss Axiophot micro-
scope. The software SmartCaptureVP (Digital Scientific) was
used for camera control, digital image acquisition (8-bit grey
scale), and the merging of DAPI and the fluorochrome images of
the paints.
Results
Classical and molecular cytogenetic analyses were
performed on the sex chromosomes in mitotic and mei-
otic cells of C. perspicillata and C. brevicauda, which have
an XX/XY1Y2 chromosomal sex determination system.
Figure 1a shows the sex chromosomes of C. perspicillata
after C-banding. The constitutive heterochromatin is lo-
calized in the distal short arm and in the pericentromer-
Analysis of XY1Y2 meiotic behavior in
Carollia
d
c
ic region of the X chromosome, while chromosome Y1 is
heterochromatic and Y2 does not present any sign of pos-
itive C-banding. In the species C. brevicauda, silver ni-
trate staining identified a NOR only in the interstitial sec-
ondary constriction of the long arm of the X chromo-
some (Fig. 1c).
Specific probes for the sex chromosomes of C. brevi-
cauda (CBR) were hybridized to mitotic cells of the same
species in order to identify the morphology of the origi-
nal X and Y sex chromosomes and of the autosome trans-
located to the X. As explained in Materials and methods,
on the neo-X chromosome the original X portion re-
mains green while the autosomal portion is yellow, as is
Y2. The Y1 is pink because the PY1CBR probe was labeled
with Cy5 (Fig. 1b). The PY1CBR also labels heterochro-
matic regions of some autosomes, while in the neo-X it
labels only the extremity of the short arm and the centro-
mere (Fig. 1d). DAPI staining was negative at the second-
ary constriction on the long arm of the composite neo-X
chromosome.
Conventional meiotic analysis: XY1Y2 system
The C-banding technique labels several heterochro-
matic blocks in pachytene cells (Fig. 2a), but the location
of the neo-XY body (XY1Y2) cannot be determined. In
diplotene the formation of chiasmata was observed along
the entire extension of the nine autosome bivalents
(Fig. 2b). In the sex trivalent the formation of chiasmata
was seen in the distal half of the long arm of the neo-X
with the Y2, and an end-to-end pairing of the Y1 with the
extremity of the short arm of the X (Fig. 2b). In meta-
phase II, cells with n = 11 were found containing the Y1
and Y2 chromosomes (Fig. 2c) and with n = 10 containing
the neo-X chromosome (Fig. 2d).
Cytogenet Genome Res 2009;124:37–43 39
 a b c d
Y1 Y2
X-Y2

X-Y1

Fig. 2. Classical meiotic analysis of Carollia perspicillata. (a) C-banding pattern of a pachytene cell. (b) Diplo-
tene cell with nine autosomal bivalents presenting chiasmata all along their length and one sex trivalent with
chiasmata between Xq and the distal part of Y2 (arrow) and end-to-end pairing between Xp and Y1 (solid ar-
row). (c) Metaphase II with n = 11 (Y1Y2) and (d) Metaphase II with n = 10 (X).

Fig. 3. Meiotic analysis of species Carollia perspicillata. (a, b) Mul-
ticolor FISH sequential to DAPI in pachytene cells. (c, d) Pachy-
tene cells in which the sex bodies are dislocated to the periph-
ery of the cells, accurately identified after in situ hybridization
with fluorochromes (colors: pink: Y1, green: X, and yellow: Y2).
(e) Two-color FISH in pachytene cell enhancing the original XY
axes (yellow arrow), and the region of the secondary constriction
of the X, where the X-autosome segment binds to it (grey arrow).
(f) Diplotene cell with multicolor FISH on the sex trivalent (ST);
in green a segment of the original X paired end-to-end with Y1
(pink), and at the other extremity the original X is bound to the
autosome, which is paired to Y2 (yellow). (g) Multicolor FISH on
the ST in a diplotene cell; the arrow indicates the region of the
secondary constriction localized on the X chromosome (green).

In situ hybridization using whole sex chromosome
probes (XY1Y2): meiosis
Specific probes for the sex chromosomes of Carollia
brevicauda were hybridized to meiotic cells of the species
Carollia perspicillata. The results reveal the behavior of
the sex trivalent in the meiosis of Carollia. Figure 3a
shows a pachytene cell with DAPI staining, without iden-
tification of the trivalent (MSCI-AX). After labeling with
multicolor FISH (Fig. 3b), it was possible to identify the
sex chromosomes and to accurately describe their behav-
ior in prophase I. Figures 3c and d show pachytene cells
with the neo-XY body located at the cell periphery, drag-
ging the X-autosome bivalent with it. The neo-X chromo-
some pairs at one extremity with the Y1, and their ten-
dency toward self-pairing can be noted (Fig. 3c green and
pink axes); however, at the other end, the neo X-Y2 pair-
ing region shows the same degree of decondensation as
the autosomal bivalents, probably with meiotic activity.

40 Cytogenet Genome Res 2009;124:37–43 Noronha/Nagamachi/O’Brien/

Ferguson-Smith/Pieczarka

Fig. 4. (a) Meiotic analysis of Carollia perspicillata with two-color
FISH using the sex chromosome probes PXCBR (green) and
PY1CBR (red) in diplotene cells. The grey bars limit the regions of
homology between the original X and Y (chiasmata). (b) Tri-color
diagram representing the synaptic behavior of the sex chromo-
some axes in the XY1Y2 multiple system.
Fig. 5. The diagram shows the origin of the multiple XY1Y2 system
in Carolliinae. The Phyllostomus hastatus (PHA) autosomes 2 and
5 fused, forming a mid-sized acrocentric pair in C. benkeithi
(CBE). A translocation of the X chromosome to one of the acro-
centrics produced the XY1Y2 system in C. brevicauda (CBR) and
C. perspicillata (CPE), where the neo-X is called X, the Y is the Y1,
and the remaining acrocentric is the Y2.
Figure 3e shows the labeled sex trivalent (XY1Y2) of a
pachytene cell, where the original sex chromosomes X
and Y1 paired end-to-end to form a ring, with the Y1 chro-
mosome almost touching the secondary constriction of
the X, which is the border between the original X chro-
mosome and the autosomal part.
The diplotene cells labeled by multicolor FISH show
the most outstanding of our results, since the probes ac-
curately labeled each one of the paired or unpaired re-
gions which are homologous to the XY1Y2 sex chromo-
some trivalent. The nine autosome bivalents are connect-
ed by chiasmata. In the trivalent the tendency toward
self-pairing of the XY was noted, with thinner axes as
Analysis of XY1Y2 meiotic behavior in
Carollia
compared to the X-autosome and with a thicker axis and
chiasmata points all along (Fig. 3f).
Figure 3g shows a characteristic diplotene cell, high-
lighting the decondensed sex trivalent in which it is pos-
sible to localize the DAPI-negative secondary constric-
tion of C. perspicillata that belongs to the original X chro-
mosome because it is included in the green portion of
PXCBR.
The results obtained for the sex trivalent after apply-
ing the two-color FISH technique showed diplotene cells
with green labeling of the neo-X and Y2 and the Y1 labeled
red. It was possible to localize and outline the X-Y1 PAR,
with the probable formation of chiasmata therein (Fig. 4a).
The diagram in Fig. 4b represents the synaptic behavior
of the neo-XY body as observed using tri-color FISH (see
Fig. 1b).
Discussion
According to Solari and Baker (2006), the taxon previ-
ously known as Carollia castanea now is accepted as two
species, where the Central American taxon preserved the
name and the South American one is now Carollia ben-
keithi. This last species has a karyotype with 2n = 22 and
is close to the ancestral karyotype of the genus, since it
does not have an X-autosome translocation (Solari and
Baker, 2006). It has an additional pair of mid-sized acro-
centric autosomes when compared to C. brevicauda and
C. perspicillata (Solari and Baker, 2006). The NOR of C.
benkeithi is located at the tip of the short arm of the orig-
inal X (Goodpasture and Bloom, 1975). However, accord-
ing to Hsu et al. (1968), it is possible that the ancestral
Phyllostomidae X chromosome had a terminal NOR in
the long arm, which resulted in the interstitial NOR after
fusion with the autosome.
Pieczarka et al. (2005) demonstrated that the Y2 chro-
mosome paint probe of C. brevicauda hybridized to two
chromosomes (2 and 5) of Phyllostomus hastatus (PHA).
We concluded that the two autosomes are rearranged in
a karyotype similar to C. benkeithi. After fusion of the
autosomes 2 and 5, forming a large acrocentric chromo-
some, a translocation to the X of C. perspicillata and C.
brevicauda occurred, producing the XY1Y2 system. This
suggests an Xq-autosome rearrangement with an inter-
stitial NOR, indicating a synapomorphy among those
Carollia with an XY1Y2 system. Figure 5 shows the evolu-
tion of the sex chromosomes with the pairing of the Y2
with the Xq-A (yellow), suggesting the fusion of the auto-
some (A) to the Xq in C. perspicillata. Previously we pos-
Cytogenet Genome Res 2009;124:37–43 41
tulated an Xq-A translocation in C. perspicillata (No-
ronha et al., 2004), but our new data do not corroborate
that.
Solari and Pigozzi (1994) analyzed the meiotic behav-
ior of the sex chromosome trivalent in Artibeus lituratus.
However they only used the synaptonemal complex tech-
nique, while in our study we were able to observe the
chromosome structure in different colors, allowing the
visualization of different levels of chromosome conden-
sation.
XY1Y2 meiosis: conventional analysis
In the diplotene stage of C. perspicillata, the sex triva-
lent (XY1Y2) presents differences in the condensation of
the original XY axes as compared to the translocated au-
tosome. The visualization of the chiasmata in the X-au-
tosome pairing region suggests the existence of gene ac-
tivity in this region of the sex trivalent, just as in the
autosomal bivalents. According to Ashley (2000), in mice
and humans errors in autosomal synapsis are usually as-
sociated with meiotic arrest and impaired fertility, with
the severity of the impairment increasing in proportion
with the degree of asynapsis. We did not detect any de-
generation of the spermatocytes of C. perspicillata, but
rather the presence of metaphases II with differences in
the haploid number (10 and 11), justified by alternate
segregation of the trivalent, with the two Y chromosomes
moving to one pole and the X to the other, ensuring the
production of balanced gametes. In the C. perspicillata
specimens studied here, the autosome translocated to
the X presumably maintains its original gene activity
without interfering with sex determination.
XY1Y2 meiosis: FISH analysis
The FISH analyses revealed a characteristic behavior
of the XY1Y2 in pachytene, previously named the sex ves-
icle or sex body (Solari and Pigozzi, 1994; Noronha et al.,
2004). In this work we define it as the neo-XY body, since
the autosomal segment is not compacted as it should be
in a sex body. According to Kasahara and Dutrillaux
(1983), the autosomal segment of the phyllostomid bat
species A. lituratus shows early replication, even when
linked to the late-replicating X. The chromosome paint-
ing results in pachytene cells in the present study cor-
roborate this hypothesis by demonstrating the visible dif-
ference in condensation between the original X segment
and the X-autosome (Fig. 3).
The autosomal part of the neo-X of C. perspicillata is
separated from the original X chromosome by the pres-
ence of rDNA sequences of the NOR. Painting with
42 Cytogenet Genome Res 2009;124:37–43
PXCBR shows that the secondary constriction belongs to
the original X segment rather than to the translocated
autosome (Fig. 3g). Studies in some mammals that have
an X-autosome translocation show that the original X is
separated from the autosomal sequence by a heterochro-
matic block (Kasahara and Dutrillaux, 1983; Noronha et
al., 2001; Dobigny et al., 2004). This block probably does
not allow the spreading of X inactivation to the autoso-
mal segment of the X during meiosis (Solari and Pigozzi,
1994; Noronha et al., 2001). In Carollia the NOR could
have the same effect as the heterochromatic block in
those species (Noronha et al., 2004).
Recently Turner et al. (2004) proposed that the pro-
teins involved in MSCI are different from those involved
in Xist gene inactivation. MSCI may be a consequence of
synaptic failure (Turner, 2007). In brief, the unsynapsed
sequences attract BRCA1, a tumor suppressor protein. In
turn, BRCA1 attracts the DNA repair protein ATR (atax-
ia telangiectasia and Rad3 related) which is responsible
for H2AX (a variant of the H2 histone abundant in mam-
malian testis) phosphorylation. Phosphorylated H2AX is
postulated to have a role in MSCI. In females the dosage
compensation due to X inactivation is caused by the gene
Xist. The autosomal portion of the neo-X remains active
because the NOR (in Carollia) does not allow the spread-
ing of the X inactivation. In males MSCI results from the
failure of the differential X and Y segments to pair. The
pairing of the autosomal portion of the neo-X with Y2
prevents the inactivation of this region. However, if this
occurred in all cases, all males with X-autosome or Y-au-
tosome translocations would have normal fertility since
the autosomal portion linked to the sex chromosome
would always pair with its homologue region. But there
are many examples of males with sex chromosome-auto-
some translocations being infertile. Delobel et al. (1998)
analyzed one of these situations, a Y-autosome transloca-
tion, and observed that the male was infertile and the
autosome portion that was linked to the Y was inactivat-
ed. Probably the XY body condensation spreads over the
whole chromosome causing MSCI, suggesting that the
NOR in Carollia is important in the male to avoid inacti-
vation of the autosomal portion of the neo-X and thus the
possibility of infertility.
According to Delobel et al. (1998), moving the auto-
somes to the inside of the sex body causes them to become
hypercondensed like the sex chromosomes. This could
explain the degeneration of the spermatocytes through
autosomal inactivation by a spreading effect. X-autosome
translocations are highly deleterious due to meiotic
breakage, the XY inactivation effect on the autosomes,
Noronha/Nagamachi/O’Brien/
Ferguson-Smith/Pieczarka
and the need to maintain a replication time pattern be-
tween the two segments that is different from normal.
The X-linked autosome segment stays highly decon-
densed, just like the autosomal bivalents. The original sex
chromosomes keep a characteristic behavior only when
paired end-to-end (Fig. 3b, c). According to Solari (1994),
the XY body usually condenses early compared to the au-
tosomal bivalents, and its chromatin undergoes self-pair-
ing. The results of our work also show an early condensa-
tion of the original sex chromosomes with a tendency to
self-pairing and movement of the translocated autosomes
to the periphery of the sex body.
According to Graves et al. (1998), the Y chromosome
of mammals became progressively degraded, keeping
only a small region of homology (PAR). The present work
is the first to detect the PAR of C. perspicillata and C.
brevicauda (Fig. 4a) by multicolor FISH. Our results con-
firm the existence of the PAR at the end of the chromo-
some opposite to the X-autosome rearrangement in Car-
ollia.
This meiotic analysis highlights the importance of
multicolor FISH in studies on meiotic mechanisms.
Acknowledgement
The sample collect was authorized by IBAMA (Instituto
Brasileiro do Meio Ambiente) permission 020/2005 (IBAMA reg-
istration: 207419).

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Analysis of XY1Y2 meiotic behavior in Cytogenet Genome Res 2009;124:37–43 43

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