A parthenogenetic maternal and double paternal contribution to an ovotesticular disorder of sex development
- Xin-Yi Xia†1,
- Wei-Ping Wang†1,
- Tian-Fu Li†1,
- Wei-Wei Li†1,
- Qiu-Yue Wu1,
- Na Li1,
- Cui Zhang1,
- Hong-Liu Gao1,
- Xiao-Jun Li1 and
- Ying-Xia Cui1Email author
© Xia et al.; licensee BioMed Central Ltd. 2014
Received: 31 December 2013
Accepted: 12 February 2014
Published: 28 February 2014
An ovotesticular disorder of sex development (OT-DSD) was rarely found in human. The mechanism causing such condition is poorly understood. We hereby reported a 11-year-old child with OT-DSD and a karyotype 46,XX/46,XY, a single maternal and double paternal genetic contribution to the patient.
Fluorescence in situ hybridization (FISH), blood grouping, HLA (human leukocyte antigen) haplotyping and a genome-wide scanning of lymphocytes with 398 short tandem repeat microsatellite markers were performed to investigate the origin of the cell lines concerned. ABO typing revealed that two populations of red cells were in the patient, which were group A and group B, both from paternal alleles. HLA haplotyping showed the patient had three haplotypes. Haplotype 1 was inherited from maternity, haplotype 2 and 3 were from paternity. The STR microsatellite analysis showed 25 of the 74 fully informative markers in both parents, three alleles were inherited: one of them was from mother, another two were from father. Seventeen of the thirty-eight paternal markers, the patient inherited two paternal alleles. For 121 informative maternal markers, the patient had a single maternal allele. There were two distinct alleles in locus DXS6810 and DXS1073 on X-chromosome, in which one was from the mother and the other from the father.
The patient was a single maternal and double paternal genetic, which was a type of a parthenogenetic division of a maternal haploid nucleus into two identical nuclei, followed by fertilization by two spermatozoa and fusion of the two zygotes into a single individual at the early embryonic stage. To the best of our knowledge, this is the oldest OT-DSD case of parthenogenetic chimerism. These data provide additional evidence that a parthenogenetic maternal and double paternal contribution causes 46,XX/46,XY OT-DSD.
KeywordsOvotesticular disorder of sex development Parthenogenetic chimera Molecular genetics
Ovotesticular disorder of sex development (OT-DSD), replaced the terminology ‘true hermaphrodite’ in 2006, is a rare condition of sexual differentiation and defined as the presence of ovarian and testicular tissue in the same individual [1, 2]. It constitutes 3%-10% of the total DSD, and presents significant diagnostic and therapeutic challenges . Few cases of OT-DSD with such studies have been reported [4–7]. The mechanism causing such 46,XX/46,XY chimerism is poorly understood . We hereby report an 11-year–old child with an OT-DSD and a 46,XX/46,XY karyotype in the cultures from peripheral lymphocytes, skin fibroblasts and two kinds of gonadal tissues.
Materials and methods
Cytogenetic analysis and fluorescent in situ hybridization
Chromosomes were prepared from phytohemagglutinin-stimulated lymphocytes and cultured fibroblasts, from the patient’s dermatic, ovarian and testicular tissues, respectively. G-banding was according to standard techniques. Fifty G-banding metaphases were analyzed for each sample. Fluorescence in situ hybridization (FISH) using a mixture of probes specific for DXZ1 and DYZ3 (CEPX Spectrum green, CEPY Spectrum orange; Vysis, Downers Grove, IL) to determine X/Y-chromosome. FISH was performed on 500 metaphases for each sample from the patient. Tests were performed according to the manufacturer’s instructed protocols. Signals were visualized under an Olympus BX51 microscope (Center Valley, Pennsylvania) equipped with a cooled, charged coupled device camera and Cytovision 3.0 image analysis software (Applied Imaging, Sunderland, United Kingdom).
Short tandem-repeat (STR) microsatellite markers
DNA was isolated from the peripheral blood of the patient and the parents. Using two commercial kits (PRISM Human Linkage Mapping Set v2.5, ABI, USA and PowerPlex 16, Promega, USA) with a total of 379 short- tandem-repeat (STR) microsatellite markers distributed over all 22 autosomes and 19 markers over X-chromosome. PCR products were analyzed in the ABI 377 DNA Sequencer (Applied Biosystems, USA). The results were analyzed by GeneMapper Software v4.0 (ABI).
Blood grouping and HLA studies
Red cell typing for ABO (DiaMed-ID micro typing system, DiaMed, Switzerland)  and other blood-group antigens for the patient and the parents were assessed. Genomic samples of blood from the patient and the parents were also used for molecular typing of HLA class I and II markers by polymerase chain reaction (ABDR003vl-20020412 kits, Pel-Freez Clinical Systems, USA) sequence-specific primer amplification .
Results of HLA haplotyping on the patient and the parents
HLA-A*02, B*51, DRB1*09
HLA-A*0203, B*38, DRB1*16
HLA-A*24, B*46, DRB1*04
HLA-A*24, B*54, DRB1*09
HLA-A*02, B*51, DRB1*09
HLA-A*24, B*46, DRB1*04
HLA-A*24, B*54, DRB1*09
Although no case of parthenogenesis has been reported in humans, it is possible that some individuals are partly parthenogenetic. For the first time, Strain et al. described a ‘parthenogenetic chimera’ where one of the clones contained cells derived exclusively from a single duplicated maternal genome. Molecular studies using microsatellite markers showed a double paternal and a single maternal allelic contribution. In 1998, Giltay et al. reported another patient with ambiguous genitalia. A parthenogenetic activation of the oocyte taking place before fertilization was suggested. In 2005, Chen et al. presented the prenatal diagnosis, sonographic findings and, molecular genetic analysis of a 46,XX/46,XY true hermaphrodite chimera. Informative sex chromosome and pericentromeric autosome markers demonstrated double paternal and single maternal genetic contributions. In 2007, Souter et al. reported a rare case of 46,XX/46,XY twins, twin A presented with ambiguous genitalia and twin B was a phenotypically normal male. The twins are chimeric and share a single genetic contribution from their mother but have two genetic contributions from their father thus supporting the existence of a third, previously unreported type of twinning. In 2008, Hersmus et al. experienced two late-diagnosed children who presented with proximal hypospadias and bilateral scrotal gonads. One should consider the possibility of ovotesticular DSD when managing patients with proximal hypospadias even if both gonads are palpable in the scrotum.
The present case demonstrated that only a single maternal genetic contributes to the patient’s genome. First, an allele revealed in locus DXS6810 and locus DXS1073, which implied only a maternal X chromosomal contribution to the patient’s genome. Second, HLA haplotyping presented a single maternal allele and third, 146 marker loci on autosomes clearly demonstrated that the patient was inherited one maternal allele. These results are compatible with the involvement of one maternal gamete. With respect to the paternal genetic contribution, there were a number of evidences to identify two different spermatozoa from the father. For example, both a paternal X chromosome and a Y chromosome contributed to the patient karyotype; the ABO typing revealed two populations of red cells, group A and group B, both from paternal alleles; HLA haplotyping and 42 informative marker loci demonstrated the inheritance of the two paternal alleles, so the possibility of the first three mechanisms can be ruled out. It is very likely that in the present case there may be a parthenogenetic division of a haploid nucleus to give two identical nuclei followed by fertilization by two spermatozoa, just as the mode of origin of the chimerism described by Giltay et al. . To the best of our knowledge, this is the oldest OT-DSD case of parthenogenetic chimerism.
We presented the clinical, cytogenetic, and molecular genetic findings of an OT-DSD of parthenogenetic chimera, to possess a single maternal genetic contribution and two paternal genetic contributions. These data provide additional evidence that a parthenogenetic maternal and double paternal contribution causes 46,XX/46,XY OT-DSD. The mechanism causing such parthenogenetic chimera need to be further investigated.
Written informed consent was obtained from the parents of the patient for publication of this Case report and any accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.
Ovotesticular disorder of sex development
Fluorescence in situ hybridization
Human leukocyte antigen
We thank the patient’s parents for their kind participation and support. We are grateful to Dr. Xiao-Qin Ye, Dr. Yuan-Zhe Wu, Dr. Jian-Feng Luan and Hong-Lin Yin for clinical data and histopathological diagnosis. The authors thank the patient and her parents for cooperation in this work. This work was supported by Natural Science Foundation of Jiangsu Province (BK2011660) and Key Foundation of Jiangsu Science and Technology Bureau (BM2013058), the Foundation of Nanjing Jinling Hospital (2013059).
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