Analysis Tools
reproductive system
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• cauda epididymal spermatozoa exhibit abnormal head folding (backward sperm head bending)
• TEM analysis of cauda epididymal spermatozoa shows retained residual cytoplasm in the sperm head; moreover, the sperm plasma membrane continuously extends from the bent head to the midpiece instead of invaginating at the bending site
• >80% of spermatozoa with completely bent heads are found in the corpus and cauda epididymides while folded heads are rarely observed in testicular spermatozoa
• % of spermatozoa with backward head folding is gradually increased as they transition from the caput to the corpus and finally to the cauda epididymidis
• however, no major abnormalities are detected in the sperm flagella
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• immunofluorescence staining shows strong accumulation of SYPL1 -- a cytoplasmic droplet (CD) marker -- at the bent sperm heads, unlike in control spermatozoa where SYPL1 localizes to the junction of the midpiece and principal piece, suggesting that CDs are not formed properly and excess residual cytoplasm remains in the sperm head
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• round cells, presumably degenerated early spermatids, are detected in the cauda epididymides
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• spermatids are abnormally retained in the seminiferous epithelium of stage VIII and IX seminiferous tubules
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• % of progressively motile spermatozoa is significantly lower than in controls at 10 and 120 min after incubation in capacitation medium
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• % of motile spermatozoa is significantly lower than in controls at 120 min but not at 10 min after incubation in capacitation medium
• moreover, VAP (average path velocity) and VSL (straight-line velocity) are significantly decreased at 10 and 120 min after incubation
• however, VCL (curvilinear velocity) is not significantly altered
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• testes show a significant decrease in the protein levels of ZDHHC19 (zinc finger, DHHC domain containing 19), a testis-enriched acyltransferase that catalyzes protein S-palmitoylation; co-IP and immunoblotting confirmed the interaction between TEX38 and ZDHHC19 in testis lysates
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• histology of cauda epididymides indicates round cells, presumably degenerated early spermatids
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• despite successful copulation with wild-type females, adult males failed to sire any pups over a 2-month mating period
• however, testis-to-body weight ratio is similar to that in wild-type controls
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• spermatozoa fail to fertilize oocytes under all IVF conditions: IVF rate is completely blunted when using cumulus-intact, cumulus-free, or zona pellucida (ZP)-free oocytes
• however, after intracytoplasmic sperm injection (ICSI), ~43% of oocytes injected with sperm heads reach the 2-cell stage and 2 healthy pups are obtained after embryo transfer, indicating that sperm DNA integrity and embryogenesis are unaffected
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cellular
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• cauda epididymal spermatozoa exhibit abnormal head folding (backward sperm head bending)
• TEM analysis of cauda epididymal spermatozoa shows retained residual cytoplasm in the sperm head; moreover, the sperm plasma membrane continuously extends from the bent head to the midpiece instead of invaginating at the bending site
• >80% of spermatozoa with completely bent heads are found in the corpus and cauda epididymides while folded heads are rarely observed in testicular spermatozoa
• % of spermatozoa with backward head folding is gradually increased as they transition from the caput to the corpus and finally to the cauda epididymidis
• however, no major abnormalities are detected in the sperm flagella
|
|
|
• immunofluorescence staining shows strong accumulation of SYPL1 -- a cytoplasmic droplet (CD) marker -- at the bent sperm heads, unlike in control spermatozoa where SYPL1 localizes to the junction of the midpiece and principal piece, suggesting that CDs are not formed properly and excess residual cytoplasm remains in the sperm head
|
|
|
• round cells, presumably degenerated early spermatids, are detected in the cauda epididymides
|
|
|
• spermatids are abnormally retained in the seminiferous epithelium of stage VIII and IX seminiferous tubules
|
|
|
• % of progressively motile spermatozoa is significantly lower than in controls at 10 and 120 min after incubation in capacitation medium
|
|
|
• % of motile spermatozoa is significantly lower than in controls at 120 min but not at 10 min after incubation in capacitation medium
• moreover, VAP (average path velocity) and VSL (straight-line velocity) are significantly decreased at 10 and 120 min after incubation
• however, VCL (curvilinear velocity) is not significantly altered
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endocrine/exocrine glands
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• testes show a significant decrease in the protein levels of ZDHHC19 (zinc finger, DHHC domain containing 19), a testis-enriched acyltransferase that catalyzes protein S-palmitoylation; co-IP and immunoblotting confirmed the interaction between TEX38 and ZDHHC19 in testis lysates
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