Analysis Tools
reproductive system
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• female mice are fertile with litter sizes comparable to those of wild-type females
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• numbers of pachytene spermatocytes, diplotene spermatocytes, round spermatids and elongated spermatids are all significantly reduced
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• TUNEL assays showed a significant increase in the number of apoptotic germ cells in the testes; apoptotic cells are mainly spermatocytes
• sperm exhibit low mitochondrial membrane potential (MMP) levels; protein expression levels of BAX and CASPASE3 are significantly increased whereas BCL-2 level is significantly reduced, indicating increased sperm apoptosis
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• testes show a significant reduction of spermatogenic cells after the zygotene stage
• however, testis/body weight ratio and testis gross morphology and histology are normal
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• protein level of KLC3 (involved in mitochondrial migration and sperm midpiece formation) is significantly reduced in testes, suggesting impaired mitochondrial development during spermiogenesis
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• TEM analysis of sperm showed blurred or absent inner mitochondrial membrane cristae
• number of sperm with abnormal mitochondrial structure is significantly higher than that in wild-type males
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• cauda epididymal sperm concentration is significantly lower than that in wild-type males
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• immunofluorescence staining of SYCP1 and SYCP3 on spermatocyte spreads showed chromothripsis, no synapsed sex chromosomes, and abnormal SYCP1 signals in the non-pseudoautosomal region (non-PAR) in pachytene spermatocytes
• phosphorylated H2AX (gammaH2AX) signal is not restricted to the sex chromosomes at the pachytene stage, suggesting failure to repair meiotic DSBs
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• percentage of pachytene spermatocytes with abnormal synapses is significantly increased
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• CTC staining showed that the rate of sperm capacitation is significantly decreased
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• male mice are sterile and produce no litters
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• fertilization ability of sperm is impaired: in vivo fertilization rate (%) is zero while in vitro fertilization (IVF) rate is significantly lower than that in wild-type controls
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• sperm exhibit a significantly lower ATP content and oxygen consumption level than wild-type sperm
• mRNA and protein levels of MTATP and MTCYB, key markers in the mitochondrial electron transfer chain, are significantly reduced in sperm
• however, sperm glycolysis levels are normal
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• sperm progressive motility is significantly lower than that in wild-type males
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• cauda epididymal sperm motility is significantly lower than that in wild-type males
• however, sperm morphology is normal
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• CTC staining showed that the percentage of acrosome-reacted sperm is significantly decreased
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cellular
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• protein level of KLC3 (involved in mitochondrial migration and sperm midpiece formation) is significantly reduced in testes, suggesting impaired mitochondrial development during spermiogenesis
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• TEM analysis of sperm showed blurred or absent inner mitochondrial membrane cristae
• number of sperm with abnormal mitochondrial structure is significantly higher than that in wild-type males
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• numbers of pachytene spermatocytes, diplotene spermatocytes, round spermatids and elongated spermatids are all significantly reduced
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• cauda epididymal sperm concentration is significantly lower than that in wild-type males
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• TEM analysis of sperm showed blurred or absent inner mitochondrial membrane cristae
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• immunofluorescence staining of SYCP1 and SYCP3 on spermatocyte spreads showed chromothripsis, no synapsed sex chromosomes, and abnormal SYCP1 signals in the non-pseudoautosomal region (non-PAR) in pachytene spermatocytes
• phosphorylated H2AX (gammaH2AX) signal is not restricted to the sex chromosomes at the pachytene stage, suggesting failure to repair meiotic DSBs
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• percentage of pachytene spermatocytes with abnormal synapses is significantly increased
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• TUNEL assays showed a significant increase in the number of apoptotic germ cells in the testes; apoptotic cells are mainly spermatocytes
• sperm exhibit low mitochondrial membrane potential (MMP) levels; protein expression levels of BAX and CASPASE3 are significantly increased whereas BCL-2 level is significantly reduced, indicating increased sperm apoptosis
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• sperm progressive motility is significantly lower than that in wild-type males
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• cauda epididymal sperm motility is significantly lower than that in wild-type males
• however, sperm morphology is normal
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• mRNA and protein levels of MTATP and MTCYB, key markers in the mitochondrial electron transfer chain, are significantly reduced in sperm
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• percentage of pachytene spermatocytes with failed DSB repair is significantly increased
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homeostasis/metabolism
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• percentage of pachytene spermatocytes with failed DSB repair is significantly increased
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