Analysis Tools
mortality/aging
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• although present at normal Mendelian ratios through E18.5, no homozygotes are recovered at birth (P0)
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growth/size/body
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• homozygotes exhibit a growth delay starting at ~E13.5
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embryo
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• at E13.5, homozygotes display generalized tissue fragility
• however, no major defects in gross organ or tissue histology are noted at E15.5
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craniofacial
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• by E18.5, homozygotes display flattened craniofacial features
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limbs/digits/tail
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• by E18.5, homozygotes show a downward facing orientation of the forelimbs
• however, no skeletal patterning defects are observed at E17.5
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cardiovascular system
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• at E18.5, a reduction in the width of both the ascending and descending aortic walls is observed
• however, the organization of elastin fibers and collagen appears normal
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skeleton
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• mass spectrometry analysis revealed that type I collagen isolated from E18.5 mutant calvarial bone shows a reduction of telopeptide hydroxylysine-aldehyde crosslinking; both the amino- and carboxyl telopeptide lysines are underhydroxylated
• mutant type I collagen is more soluble, consistent with fewer stable crosslinks; the prominence of beta dimers is more characteristic of skin than bone collagen
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cellular
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• MEFs isolated from E13.5 mutant embryos exhibit dilated ER with embedded matrix as well as intracytoplasmic collagen fibers
• upon stimulation of collagen synthesis with ascorbic acid, mutant MEFs show retention of procollagen in the cell layer, with no apparent changes in procollagen secretion as determined by pulse chase analysis
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• MEFs isolated from E13.5 mutant embryos exhibit dilated, irregular rough ER engorged with abnormal, poorly formed collagen matrix
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• MEFs isolated from E13.5 mutant embryos exhibit a dilated ER with an improperly formed matrix in both the ER and cytoplasm
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