Cisplatin Induces Apoptosis in Mouse Neonatal Testes Organ Culture
Abstract
:1. Introduction
2. Results
2.1. Cisplatin Induced Testicular Damage in Organ Culture
2.2. Effects of Cisplatin on Gene Expression of Germ Cell Markers in MTFs
2.3. Effects of Cisplatin on Sertoli Cells in the MTF Culture
2.4. Effects of Cisplatin on the Expression of Steroidogenic Genes in MTFs
2.5. Cisplatin Induced Apoptosis-Related Protein Expression in MTFs
3. Discussion
4. Materials and Methods
4.1. Animals
4.2. Organ Culture Method and Drug Treatment
4.3. Histological and Immunohistochemical Analyses
4.4. RNA Isolation and qPCR
4.5. Western Blot Analysis
4.6. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Conflicts of Interest
References
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Antibody | Company | Catalog Number | Dilution (Usage) |
---|---|---|---|
SYCP3 | Abcam | ab97672 | 1:200 (IHC) |
VASA | Abcam | ab13840 | 1:200 (IHC) |
SALL4 | Abcam | ab57577 | 1:200 (IHC) |
Vimentin | Santa Cruz Biotech | sc-373717 | 1:100 (IHC) |
SOX9 | Abcam | ab185966 | 1:200 (IHC) |
3βHSD | Santa Cruz Biotech | sc-30820 | 1:100 (IHC) |
β-actin | Santa Cruz Biotech | sc-47778 | 1:2000 (WB) |
Bcl-2 | Cell signaling | #3498 | 1:2000 (WB) |
Cleaved PARP | Cell signaling | #9541 | 1:2000 (WB) |
Cleaved caspase-3 | Cell signaling | #9664 | 1:2000 (WB) |
Cleaved caspase-8 | Cell signaling | #8592 | 1:2000 (WB) |
Caspase-3 | Cell signaling | #9662 | 1:2000 (WB) |
Caspase-8 | Cell signaling | #9746 | 1:2000 (WB) |
PARP | Cell signaling | #9532 | 1:2000 (WB) |
Gene | Forward Primer | Reverse Primer |
---|---|---|
Sycp3 | 5′-CAGATGCTTCGAGGGTGTG-3′ | 5′-AAGGTGGCTTCCCAGATTTC-3′ |
Vasa | 5′-CCGCATGGCTAGAAGAGATT-3 | 5′-TTCCTCGTGTCAACAGATGC-3 |
Sohlh1 | 5′-CATCTGCTGTGTCTCGGGTA-3′ | 5′-GCTGGAAGACTCTGGCTCAC-3′ |
Sohlh2 | 5′-TGAGACGAGAACGCATCAAG-3′ | 5′-CCTCTGTGATGTGGCTGAGA-3′ |
Dazl | 5′-GTCGAAGGGCTATGGATTTG-3′ | 5′-ACGTGGCTGCACATGATAAG-3 |
Gfra 1 | 5′-CCTGGATTTGCTGATGTCG -3′ | 5′-GCTGAAGTTGGTTTCCTTGC-3′ |
Stra-8 | 5′-CTGAGGCTGTTGGACCAGAT-3′ | 5′-GCAACAGAGTGGAGGAGGAG-3′ |
Sall4 | 5′-CCTCGGTGTTAGATGTCAAGG-3′ | 5′-GGGCACACGTAAGGTCTCTC-3′ |
Sox9 | 5′-AGTACCCGCATCTGCACAAC-3′ | 5′-TACTTGTAATCGGGGTGGTCT-3′ |
Wt1 | 5′-ATCCCAGGCAGGAAAGTGTG-3′ | 5′-GTGCTGTCTTGGAAGTCGGA-3′ |
Amh | 5′-CCTGGAGGAAGTGACATGG-3′ | 5′-CAGGGTAGAGCACCAGCAG-3′ |
AR | 5′-GGCGGTCCTTCACTAATGTC-3′ | 5′-GACAGGTGCCTCATCCTCAC -3′ |
Esr1 | 5′-GCACAAGCGTCAGAGAGATG-3′ | 5′-AGGACAAGGCAGGGCTATTC-3′ |
Cyp19α1 | 5′-TTGAGACGATTCCAGGTGAAG-3′ | 5′-ATTTCCACAAGGTGCCTGTC-3′ |
Cyp11α1 | 5′-GACAATGGTTGGCTAAACCTG-3′ | 5′-GGGTCCACGATGTAAACTGAC-3′ |
Lhr | 5′-GGCCATCCTCATCTTCACAG-3′ | 5′-TTGGCACAAGAATTGACAGG-3′ |
Star | 5′-TGGGCATACTCAACAACCAG-3′ | 5′-GTCTACCACCACCTCCAAGC-3ʹ |
Bad | 5′-GCCCTAGGCTTGAGGAAGTC-3′ | 5′-GGCTCAAACTCTGGGATCTG-3′ |
Bax | 5′-GCTGACATGTTTGCTGATGG-3′ | 5′-GATCAGCTCGGGCACTTTAG-3′ |
Fas | 5′-CTGATCCTCATTCCCGTACC-3′ | 5′-ATCATTGGCACCTCTTCAGC -3′ |
Traf3 | 5′-AGAGTGAGTTGAGTGCACACTT-3′ | 5′-TACCGCGGAGCTGGCCTCAT-3′ |
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Park, H.-J.; Kim, J.-S.; Lee, R.; Song, H. Cisplatin Induces Apoptosis in Mouse Neonatal Testes Organ Culture. Int. J. Mol. Sci. 2022, 23, 13360. https://doi.org/10.3390/ijms232113360
Park H-J, Kim J-S, Lee R, Song H. Cisplatin Induces Apoptosis in Mouse Neonatal Testes Organ Culture. International Journal of Molecular Sciences. 2022; 23(21):13360. https://doi.org/10.3390/ijms232113360
Chicago/Turabian StylePark, Hyun-Jung, Ji-Soo Kim, Ran Lee, and Hyuk Song. 2022. "Cisplatin Induces Apoptosis in Mouse Neonatal Testes Organ Culture" International Journal of Molecular Sciences 23, no. 21: 13360. https://doi.org/10.3390/ijms232113360