GENOMICS 30, 287–292 (1995) Genomic Reorganization in the Concolor Gibbon (Hylobates concolor) Revealed by Chromosome Painting U. KOEHLER ,* F. BIGONI,² J. W IENBERG,* ,1 AND R. S TANYON² * Institute of Anthropology and Human Genetics, Richard-Wagner-Strasse 10/I, 80333 Munich, Germany; and ² Institute of Physical Anthropology, University of Genoa, 16126 Genoa, Italy Received May 12, 1995; accepted August 30, 1995 sequence had highly derived karyotypes (Stanyon and We employed fluorescence in situ hybridization Chiarelli, 1983; Van Tuinen and Ledbetter, 1983; (FISH) of specific DNA libraries of all human chromo- Marks, 1983). somes to establish homologies between the human However, with fluorescence in situ hybridization karyotype and the karyotype of Hylobates concolor (2n (FISH) it is now possible to establish unequivocally Å 52). Numerous intra- and interchromosomal re- chromosomal homology between any two primate spe- arrangements have led to a massive reorganization in cies. In the past few years this method has been used the karyotype of H. concolor. There have been a mini- to assign homologies between all human chromosomes mum of 31 translocations. The 25 H. concolor au- and those of great apes, macaques, and two species tosomes are composed of 63 – 67 recognizable segments of lesser apes (H. lar species group (2n Å 44) and H. that show DNA homology to regions of the 22 human syndactylus (2n Å 50); Wienberg et al., 1990, 1992; autosomes. Only 1 autosome, homologous to human Jauch et al., 1992; Stanyon et al., 1992; Koehler et al., chromosome 21, has not been involved in transloca- 1995). Here, we report on the homology between all tions. We compared the gene mapping data for H. con- human chromosomes and those of Hylobates concolor color with our in situ hybridization and found that in (2n Å 52) using in situ hybridization of human chromo- most cases these data are confirmed by our results. H. some-specific DNA libraries. concolor presents a case in a primate closely related to humans, in which chromosome morphology and synteny are highly disturbed in a manner similar to MATERIALS AND METHODS that encountered in rodents.  1995 Academic Press, Inc. Chromosomes of Hylobates concolor were prepared from whole- blood and fibroblast cultures according to standard procedures. The whole-blood cultures were established from two females housed at INTRODUCTION the Budapest Zoo. Low-passage fibroblast cultures of one male and one female of H. concolor were a gift of Dr. O. Ryder, San Diego Although the lesser apes (genus Hylobates) are Zoological Society. All four individuals studied were reported to be- closely related to humans and are classified with great long to the subspecies H. concolor gabriellae. apes and human in the same primate superfamily, Chromosome G-banding previous to in situ hybridization was as described (Arnold et al., 1992b; Wienberg et al., 1992). Briefly, the Hominoidea, they have been systematically left out of G-banded spreads were photographed, slides were destained and chromosomal phylogenies of the primates. A prerequi- postfixed with formaldehyde. In addition to the G-banding protocol, site for reconstructing chromosomal phylogenies and the identification of chromosomes was also facilitated by DAPI-band- understanding genome evolution in higher primates is ing concurrently with in situ hybridization. This double banding sys- establishing between-species chromosomal homologies. tem allowed us to assign easily the in situ hybridization signal to the correct H. concolor chromosomes or, more typically, chromosome Chromosome banding was helpful in establishing chro- segments. Chromosome numbering followed that of Turleau et al. mosomal homologies between human and great apes (1983) to facilitate comparisons with previous gene mapping results and even many lower primates (Dutrillaux, 1979; Seua ´- based on somatic cell hybrids. nez, 1979). However, chromosomal homologies between In situ hybridization. In situ hybridization of DNA library probes lesser apes and other primates including human were to human and primate chromosomes was performed as previously described (Jauch et al., 1992; Wienberg et al., 1992). DNA prepared very difficult to propose solely on the basis of banding from chromosome-specific human bacteriophage libraries (American patterns. It became clear that gibbons had experienced Type Culture Collection) or plasmid libraries (Collins et al., 1991, rapid and massive chromosome evolution and as a con- kindly provided by J. Gray) were used as probes. Library DNA was labeled with biotin or digoxigenin (Arnold et al., 1992a) by standard nick-translation assays. 1 To whom correspondence should be addressed at the Department of Pathology, Tennis Court Road, Cambridge, England. Telephone: Next, 10 ml of the hybridization mixture (50% formamide, 11 SSC, 10% dextran sulfate) containing 1 – 2 mg labeled phage library DNA (/44) 1223 333725. Fax: (/44) 1223 333346. E-mail: jw@mole.bio. cam.ac.uk. or 300 – 500 ng labeled plasmid library DNA, 4 – 8 mg human genomic 287 0888-7543/95 $12.00 Copyright  1995 by Academic Press, Inc. All rights of reproduction in any form reserved.