Analysis Tools
reproductive system
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• females exhibit normal fertility relative to wild-type females
• males show normal testicular size and testis/body weight ratio and intact seminiferous tubules and epithelium with apparently normal spermatogenesis relative to control males
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• number of epididymal sperm is reduced by 37.6% relative to that in wild-type males
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• percentage of morphologically abnormal sperm is 97.5% versus only 13.4% in wild-type males
• PNA staining of epididymal sperm shows that the cytoplasmic droplet is missing
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• SEM analysis of epididymal sperm shows a discontinuous accessory structure in the middle piece
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• epididymal sperm show abnormal mitochondrial position
• however, mitochondrial function is normal, as indicated by MitoTracker Red staining
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• mitochondrial sheath is disorganized, with mitochondria scattered in varied locations
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• TEM analysis of aggregated sperm shows double/multiple heads and multiple (two or more) flagella enfolded by residual cytoplasm
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• epididymal sperm show a significantly higher percentage of 3 types of abnormalities: (1) sperm with head bent back onto the midpiece, (2) sperm clumped together with cellular remnants along their heads, and (3) sperm with tails coiled around their heads
• SEM analysis shows reduced curvature at the head tip
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• round spermatids exhibit vacuolated or irregular shaped acrosomal vesicles
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• PNA staining of epididymal sperm shows abnormal acrosome localization
• round spermatids exhibit a morphologically abnormal Golgi apparatus and vacuolated or irregular shaped acrosomal vesicles causing defects in the maturation phase
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• sperm heads exhibit defective chromatin structure
• some sperm have disrupted nuclear membranes and extra nuclear chromatin material
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• TEM analysis of aggregated sperm shows double/multiple heads
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• late spermatids show irregularly shaped nuclei and abundant cytoplasm
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• sperm show impaired ability to migrate from the uterus into the oviduct
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• CASA analysis shows a significant reduction in both total and rapid sperm motility while the percentages of static cell and straightness are significantly increased
• in addition, lateral amplitude (um) and area (um sq) are significantly increased
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• males show failure of excess cytoplasm shedding from the head and neck region of epididymal sperm; an abnormal large residual body and cytoplasmic body are observed in the testes
• sperm bundles slough into the lumen and subsequently appear in the cauda epididymis
• TEM analysis shows multiple defects, including abnormal acrosome and chromatin structure, multiple nuclei, detached acrosome membranes, extra nuclear chromatin material, retained cytoplasmic remnant in the head, and impaired sperm individualization
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• sperm individualization is impaired: many elongating spermatids or spermatozoa are wrapped in a single cell membrane, indicating defective removal of intercellular bridges between individual spermatids
• TEM analysis of aggregated sperm shows double/multiple heads and multiple (two or more) flagella enfolded by residual cytoplasm
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• spermiation defects include failure of excess cytoplasm shedding and abnormal sperm individualization
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• adult male mice bred with wild-type adult females fail to sire any offspring over a 1-month mating period
• male infertility cannot be rescued by in vitro fertilization
• however, mating behavior is normal with successful ejaculation and vaginal plug formation
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• IVF assays show that sperm bind to the oolemma but still fail to fertilize ZP-free eggs, suggesting that sperm-egg fusion is impaired
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• sperm are unable to fertilize wild-type oocytes by natural mating, possibly due to impaired sperm ability to migrate from the uterus into the oviduct
• impaired fertilization is linked to downregulation of ZPBP, PRSS21, PRSS54, PRSS55, ADAM2, and ADAM3 in cauda epididymal sperm
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• retention of cytoplasmic remnants in the head region results in defective acrosome reaction
• after induction by calcium ionophore A23187, the percentage of non-acrosome-reacted (PNA positive) sperm is significantly higher than that in wild-type controls, suggesting that sperm fail to release their acrosomal contents
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• in vivo fertilization assays show that sperm yielded no (0%) two-cell embryos, whereas the average fertilization rate using sperm from wild-type males is 87.9%
• in vitro fertilization (IVF) assays show that sperm fail to fertilize cumulus-intact (0 of 180), cumulus-free ZP-intact (0 of 86), or ZP-free oocytes (0 of 65)
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cellular
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• number of epididymal sperm is reduced by 37.6% relative to that in wild-type males
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• percentage of morphologically abnormal sperm is 97.5% versus only 13.4% in wild-type males
• PNA staining of epididymal sperm shows that the cytoplasmic droplet is missing
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• SEM analysis of epididymal sperm shows a discontinuous accessory structure in the middle piece
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• epididymal sperm show abnormal mitochondrial position
• however, mitochondrial function is normal, as indicated by MitoTracker Red staining
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• mitochondrial sheath is disorganized, with mitochondria scattered in varied locations
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• TEM analysis of aggregated sperm shows double/multiple heads and multiple (two or more) flagella enfolded by residual cytoplasm
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• epididymal sperm show a significantly higher percentage of 3 types of abnormalities: (1) sperm with head bent back onto the midpiece, (2) sperm clumped together with cellular remnants along their heads, and (3) sperm with tails coiled around their heads
• SEM analysis shows reduced curvature at the head tip
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• round spermatids exhibit vacuolated or irregular shaped acrosomal vesicles
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• PNA staining of epididymal sperm shows abnormal acrosome localization
• round spermatids exhibit a morphologically abnormal Golgi apparatus and vacuolated or irregular shaped acrosomal vesicles causing defects in the maturation phase
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• sperm heads exhibit defective chromatin structure
• some sperm have disrupted nuclear membranes and extra nuclear chromatin material
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• TEM analysis of aggregated sperm shows double/multiple heads
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• late spermatids show irregularly shaped nuclei and abundant cytoplasm
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• sperm show impaired ability to migrate from the uterus into the oviduct
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• CASA analysis shows a significant reduction in both total and rapid sperm motility while the percentages of static cell and straightness are significantly increased
• in addition, lateral amplitude (um) and area (um sq) are significantly increased
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homeostasis/metabolism
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N |
• males exhibit normal serum testosterone levels relative to wild-type males
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