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Copy number variation in the region harboring SOX9 gene in dogs with testicular/ovotesticular disorder of sex development (78,XX; SRY-negative).

Marcinkowska-Swojak M, Szczerbal I, Pausch H, Nowacka-Woszuk J, Flisikowski K, Dzimira S, Nizanski W, Payan-Carreira R, Fries R, Kozlowski P, Switonski M - Sci Rep (2015)

Bottom Line: No copy variation of SOX9 was observed.Our extensive studies have excluded duplication of SOX9 as the common cause of XX DSD in analyzed samples.However, it remains possible that the causative mutation is hidden in highly polymorphic CNVR1.

View Article: PubMed Central - PubMed

Affiliation: European Centre of Bioinformatics and Genomics, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznan, Poland.

ABSTRACT
Although the disorder of sex development in dogs with female karyotype (XX DSD) is quite common, its molecular basis is still unclear. Among mutations underlying XX DSD in mammals are duplication of a long sequence upstream of the SOX9 gene (RevSex) and duplication of the SOX9 gene (also observed in dogs). We performed a comparative analysis of 16 XX DSD and 30 control female dogs, using FISH and MLPA approaches. Our study was focused on a region harboring SOX9 and a region orthologous to the human RevSex (CanRevSex), which was located by in silico analysis downstream of SOX9. Two highly polymorphic copy number variable regions (CNVRs): CNVR1 upstream of SOX9 and CNVR2 encompassing CanRevSex were identified. Although none of the detected copy number variants were specific to either affected or control animals, we observed that the average number of copies in CNVR1 was higher in XX DSD. No copy variation of SOX9 was observed. Our extensive studies have excluded duplication of SOX9 as the common cause of XX DSD in analyzed samples. However, it remains possible that the causative mutation is hidden in highly polymorphic CNVR1.

No MeSH data available.


The MLPA and FISH analysis in the Pug family.The family consisted of two XX, SRY-negative DSD females (A6 and B4) and their healthy parents: father (C2) and mother (C3). Bar plots represent the relative copy number (y-axis) of each CanSOX9+ probe (x-axis). The red bracket marks probes, where the signal is increased (5′SOX9_01–5′SOX9_07). Corresponding diagrams of the results of FISH analysis are shown next to the MLPA bar plots.
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f4: The MLPA and FISH analysis in the Pug family.The family consisted of two XX, SRY-negative DSD females (A6 and B4) and their healthy parents: father (C2) and mother (C3). Bar plots represent the relative copy number (y-axis) of each CanSOX9+ probe (x-axis). The red bracket marks probes, where the signal is increased (5′SOX9_01–5′SOX9_07). Corresponding diagrams of the results of FISH analysis are shown next to the MLPA bar plots.

Mentions: We then analyzed the segregation of copy number changes in Pug family, which consisted of two affected siblings and their healthy parents (Fig. 4). We did not analyze grandparents of the affected siblings, due to the lack of grandfather DNA (C5) and the repeatedly poor quality of MLPA (non interpretable) results of the grandmother sample (C4). Both XX DSD animals (A6 and B4) showed a clear signal increase for probes 5′SOX9_01-07. A similar signal increase was observed in the father (C2), but not the mother (C3). The increased MLPA signal correlated with results of the FISH analysis. Unfortunately, we could not analyze other littermates, and were thus unable to conclude, whether the increase of copies in XX DSD animals represents an association or is coincidental.


Copy number variation in the region harboring SOX9 gene in dogs with testicular/ovotesticular disorder of sex development (78,XX; SRY-negative).

Marcinkowska-Swojak M, Szczerbal I, Pausch H, Nowacka-Woszuk J, Flisikowski K, Dzimira S, Nizanski W, Payan-Carreira R, Fries R, Kozlowski P, Switonski M - Sci Rep (2015)

The MLPA and FISH analysis in the Pug family.The family consisted of two XX, SRY-negative DSD females (A6 and B4) and their healthy parents: father (C2) and mother (C3). Bar plots represent the relative copy number (y-axis) of each CanSOX9+ probe (x-axis). The red bracket marks probes, where the signal is increased (5′SOX9_01–5′SOX9_07). Corresponding diagrams of the results of FISH analysis are shown next to the MLPA bar plots.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4589768&req=5

f4: The MLPA and FISH analysis in the Pug family.The family consisted of two XX, SRY-negative DSD females (A6 and B4) and their healthy parents: father (C2) and mother (C3). Bar plots represent the relative copy number (y-axis) of each CanSOX9+ probe (x-axis). The red bracket marks probes, where the signal is increased (5′SOX9_01–5′SOX9_07). Corresponding diagrams of the results of FISH analysis are shown next to the MLPA bar plots.
Mentions: We then analyzed the segregation of copy number changes in Pug family, which consisted of two affected siblings and their healthy parents (Fig. 4). We did not analyze grandparents of the affected siblings, due to the lack of grandfather DNA (C5) and the repeatedly poor quality of MLPA (non interpretable) results of the grandmother sample (C4). Both XX DSD animals (A6 and B4) showed a clear signal increase for probes 5′SOX9_01-07. A similar signal increase was observed in the father (C2), but not the mother (C3). The increased MLPA signal correlated with results of the FISH analysis. Unfortunately, we could not analyze other littermates, and were thus unable to conclude, whether the increase of copies in XX DSD animals represents an association or is coincidental.

Bottom Line: No copy variation of SOX9 was observed.Our extensive studies have excluded duplication of SOX9 as the common cause of XX DSD in analyzed samples.However, it remains possible that the causative mutation is hidden in highly polymorphic CNVR1.

View Article: PubMed Central - PubMed

Affiliation: European Centre of Bioinformatics and Genomics, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznan, Poland.

ABSTRACT
Although the disorder of sex development in dogs with female karyotype (XX DSD) is quite common, its molecular basis is still unclear. Among mutations underlying XX DSD in mammals are duplication of a long sequence upstream of the SOX9 gene (RevSex) and duplication of the SOX9 gene (also observed in dogs). We performed a comparative analysis of 16 XX DSD and 30 control female dogs, using FISH and MLPA approaches. Our study was focused on a region harboring SOX9 and a region orthologous to the human RevSex (CanRevSex), which was located by in silico analysis downstream of SOX9. Two highly polymorphic copy number variable regions (CNVRs): CNVR1 upstream of SOX9 and CNVR2 encompassing CanRevSex were identified. Although none of the detected copy number variants were specific to either affected or control animals, we observed that the average number of copies in CNVR1 was higher in XX DSD. No copy variation of SOX9 was observed. Our extensive studies have excluded duplication of SOX9 as the common cause of XX DSD in analyzed samples. However, it remains possible that the causative mutation is hidden in highly polymorphic CNVR1.

No MeSH data available.