Analysis Tools
reproductive system
 N |
• males are fertile and show normal testicular histology
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• P0 and P1 ovaries show significantly increased oocyte apoptosis in the absence of efficient DNA damage repair
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• at P0 and P1, ovaries exhibit significantly more TUNEL-positive cells than heterozygous control ovaries
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• absence of primordial follicles
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small ovary
(
J:64677
)
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• 1/20 the size of wild-type
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• P0 ovaries show significant downregulation of genes essential for oogenesis including genes involved in oocyte growth and differentiation (e.g. Nobox, Lhx8, Sohlh1, Sohlh2, Taf4b, Kit, Jag2, Jag1, Gdf9)
• KIT and NOBOX protein expression is totally absent in P0 ovaries; also, only 20.6% of oocytes show weak nuclear localization of LHX8 protein versus 93.9% of oocytes in heterozygous control ovaries
• 96.2% of germ cells exhibit only cytoplasmic localization of SOHLH1 without obvious nuclear localization (3.8%), unlike in heterozygous controls where 24.3% of germ cells show only cytoplasmic localization of SOHLH1
• however, the total number of oocytes is similar to that in heterozygous control ovaries at E18.5 and P0
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• perinatal decrease in the number of oocytes
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• P0 ovaries show significant downregulation of meiosis-related genes (e.g. Syce1, Syce2, Rad51, Sycp3, Cpeb1, Ybx2)
• immunofluorescence staining of PO ovaries with antibodies to DEAD-box helicase 4 (DDX4, a germ cell-specific marker) and Y box protein 2 (YBX2, a marker for the diplotene stage of meiosis that persists in the arrested dictyate stage) shows a significantly lower number of oocytes at the diplotene stage than in heterozygous ovaries (56.8% versus 78.9%), indicating impaired meiotic progression to the diplotene stage of meiotic prophase I
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• immunostaining with antibodies to DDX4 and synaptonemal complex protein 3 (SYCP3) shows that virtually no oocytes develop to the dictyate stage, as indicated by the absence of visible nucleoli
• SYCP3 protein abundance is significantly decreased in P0 ovaries, further suggesting that the few oocytes that reach diplotene fail to progress to the dictyate stage
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• females are infertile but female Mullerian structures are normal
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cellular
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• perinatal decrease in the number of oocytes
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• P0 and P1 ovaries show significantly increased oocyte apoptosis in the absence of efficient DNA damage repair
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• at P0 and P1, ovaries exhibit significantly more TUNEL-positive cells than heterozygous control ovaries
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• at P0, 64.1% of oocytes stain positive for gamma-H2AX (a marker of DNA double stranded breaks) versus 24.8% in heterozygous controls, suggesting increased DNA damage
• no obvious RAD51 foci are detected in oocyte nuclei, indicating failure to initiate DNA damage repair
• gamma-H2AX-positive oocytes are RAD51 negative, unlike in heterozygous controls
• RAD51 protein abundance is severely reduced in P0 ovaries
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homeostasis/metabolism
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• at P0, 64.1% of oocytes stain positive for gamma-H2AX (a marker of DNA double stranded breaks) versus 24.8% in heterozygous controls, suggesting increased DNA damage
• no obvious RAD51 foci are detected in oocyte nuclei, indicating failure to initiate DNA damage repair
• gamma-H2AX-positive oocytes are RAD51 negative, unlike in heterozygous controls
• RAD51 protein abundance is severely reduced in P0 ovaries
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endocrine/exocrine glands
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• at P0 and P1, ovaries exhibit significantly more TUNEL-positive cells than heterozygous control ovaries
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• absence of primordial follicles
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small ovary
(
J:64677
)
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• 1/20 the size of wild-type
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