Molecular Cytogenetic and Y Copy Number Analysis of a Reciprocal ECAY-ECA13 Translocation in a Stallion with Complete Meiotic Arrest
Abstract
:Simple Summary
Abstract
1. Introduction
2. Material and Methods
2.1. Animal and Samples
2.2. Chromosome Preparations for Molecular Cytogenetic Analysis
2.3. Selection of Probes for Fluorescence In Situ Hybridization (FISH)
2.4. Genomic and BAC DNA Isolation
2.5. Fluorescence In Situ Hybridization (FISH)
2.6. Digital Droplet PCR Analysis (ddPCR)
3. Results
3.1. Molecular Cytogenetic Analysis of ECAY and ECA13 Reciprocal Translocation
3.2. Copy Number Analysis of Horse MSY Multicopy Genes
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Marker ID | CH241 BAC | Cytogenetic Location | Location in EquCab3 or ECAY BAC Contig Map | Reference Marker | Reference |
---|---|---|---|---|---|
Y-1 | 069E11 | Yq proximal 2/3 | Y and Xq heterochromatin | ETSTY7 ampliconic array | [9] |
Y-2 | 022P7 | Yqdistal | Y Contig Ia, single copy | KDM5D | [9] |
Y-3 | 140M23 | Yqdistal | Y Contig Ib, multicopy (MC) | SRY | [9] |
Y-4 | 017D15 | Yqdistal | Y Contig Ib, multicopy (MC) | TSPY | [9] |
Y-5 | 090G18 | Yqdistal | Y Contig Ic, single copy | n/a | [9] |
Y-6 | 112E12 | Yqdistal | Y Contig II, single copy | NLGN4Y | [9] |
Y-7 | 011B8 | Yqdistal | Y Contig II, single copy | n/a | [9] |
Y-8 | 125H6 | Yqdistal | Y Contig III, single copy | TMSB4Y | [9] |
Y-9 | 102J15 | Yqdistal | Y Contig III, single copy | TMSB4Y | [9] |
Y-10 | 106F1 | Yqdistal | Y Contig IV single copy | ZFY | [9] |
PAR | 194E12 | Xpter/Yqter | chrX:3945-246,703 | PLCXD1 | [20] |
13-1 | 078E13 | 13p15 | chr13:5,913,678-6,105,097 | GPER1 | This study |
13-2 | 060D24 | 13p13 | chr13:11,481,169-11,662,804 | ELN | This study |
13-3 | 158P20 | 13q12 | chr13:18,059,033-18,254,381 | LEX041 | [19] |
Gene | Forward Primer 5′–3′ | Reverse Primer 5′–3′ | Probe Sequence 5′–3′ |
---|---|---|---|
TSPY | CATAGTGGAGGAAGAGGATGAAA | GGCAATGGTTTAACCCTGAAA | CTCTTTCTGGGAGACCTGCCCTTT |
SRY | TTCTGTGATCTATGCTGGCG | TTACCCTCCGGACTTTCTCA | AACAGGGACTCTGCCGCCACCA |
RBMY | GAAGCTCCACAACTTGAGGT | CTCTGACCTATGATGGAAGCA | TGTCTGCCACCATGCTCACGACCA |
ETSTY1 | GACGGACGACCTTGTGTT | ACGCTCACAGATGACAGTAG | TGTCCCGGCCACCTCAGGGC |
ETSTY2 | TTGTTGTTAGGCTACCTGGC | AAGGGCAAACCATAACCTCC | TGGGCAAGCTTCTCCATGGTTGCTGCA |
ETSTY5 | GAGGCAGGTACTTCGTTACC | TCACTCACAAAGTCAACGCT | TGCCGTGAGCTTGAGGGCGAA |
UBA1Y | TTTCTGTTGTCTGGACGGAG | CTCCACGGATGTAGTCAGAG | AGCAGAGGCCTCCTGTGTCTGAGCT |
HSFY | AGGCTTTCTCCACTGGTTTC | GAGGCTGTCCCGAACTTTTA | CCCCTGCTCTAAAGTGCTTCCTGTCG |
MYOZ1 | GACTTTCCAGATGCCCAAGT | ACCAGAACCTCTCCAACAGGCCTTCT | GCTCCTCTGTTTCTCCATCC |
Sample ID | TSPY | ETSTY1 | ETSTY2 | ETSTY5 | SRY | RBMY | HSFY | UBA1Y |
---|---|---|---|---|---|---|---|---|
H787 | 5.6 | 1.9 | 3.8 | 2.6 | 0.8 | 1.9 | 0.9 | 3.1 |
23346 | 9.1 | 3.6 | 5.7 | 5.3 | 1 | 1.5 | 0.9 | 3.8 |
23348 | 8.2 | 7.4 | 5.7 | 4.8 | 1 | 2.3 | 0.9 | 3.5 |
70858 | 9.7 | 4.1 | 4.2 | 4.1 | 0.8 | 2 | 0.9 | 3.2 |
70980 | 10.8 | 3.9 | 5.1 | 4.4 | 0.9 | 1.7 | 0.9 | 3.8 |
70981 | 11.3 | 3.8 | 4.6 | 4.4 | 0.8 | 1.7 | 1.1 | 3 |
73901 | 8.6 | 3.9 | 4.1 | 3.9 | 1 | 2 | 1 | 3 |
74413 | 8.2 | 3.9 | 4.3 | 4.3 | 0.9 | 1.9 | 1 | 3.6 |
74836 | 8.2 | 3.7 | 4.6 | 4 | 0.8 | 1.7 | 1.2 | 3.3 |
74837 | 8.6 | 3.9 | 4.5 | 3.7 | 0.7 | 1.9 | 1 | 3.8 |
75052 | 9.2 | 3.6 | 4.6 | 4.2 | 0.9 | 2 | 1 | 3.7 |
TR007 | 9 | 4.2 | 3.8 | 4.1 | 1 | 1.9 | 1 | 3.9 |
TR008 | 9.5 | 4 | 3.7 | 4.1 | 0.9 | 2.1 | 1 | 4.3 |
TR009 | 8.2 | 3.9 | 3.8 | 3.8 | 0.9 | 2 | 1 | 4 |
H061 | 8 | 4.1 | 4.5 | 3.6 | 0.8 | 1.9 | 1.1 | 1.8 |
H294 | 13.3 | 3.8 | 5.4 | 4.4 | 0.8 | 1.7 | 1.1 | 3.3 |
Bravo | 8.5 | 4.9 | 5.6 | 3.6 | 0.9 | 2 | 1.1 | 1.5 |
p-value | 0.0249 * | 0.0293 * | 0.2494 | 0.0038 ** | 0.4003 | 0.9756 | 0.2365 | 0.7583 |
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Castaneda, C.; Ruiz, A.J.; Tibary, A.; Raudsepp, T. Molecular Cytogenetic and Y Copy Number Analysis of a Reciprocal ECAY-ECA13 Translocation in a Stallion with Complete Meiotic Arrest. Genes 2021, 12, 1892. https://doi.org/10.3390/genes12121892
Castaneda C, Ruiz AJ, Tibary A, Raudsepp T. Molecular Cytogenetic and Y Copy Number Analysis of a Reciprocal ECAY-ECA13 Translocation in a Stallion with Complete Meiotic Arrest. Genes. 2021; 12(12):1892. https://doi.org/10.3390/genes12121892
Chicago/Turabian StyleCastaneda, Caitlin, Agustin J. Ruiz, Ahmed Tibary, and Terje Raudsepp. 2021. "Molecular Cytogenetic and Y Copy Number Analysis of a Reciprocal ECAY-ECA13 Translocation in a Stallion with Complete Meiotic Arrest" Genes 12, no. 12: 1892. https://doi.org/10.3390/genes12121892