Analysis Tools
pigmentation
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• cultured primary autophagy-deficient melanocytes stop proliferating after the third passage, unlike autophagy-competent melanocytes which can be maintained up to passage 5
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• mice show decreased pigmentation of dorsal hair; melanin content of dorsal hair is reduced by 10-15% relative to that in control mice
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• mice exhibit slight hypopigmentation of the tail epidermis
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• some melanocytes of tail skin contain misshapen, swollen, and possibly disintegrating mitochondria, not observed in control melanocytes
• however, when cultured in vitro, primary autophagy-deficient melanocytes isolated from body skin of newborn mice show no significant change of melanin content relative to autophagy-competent melanocytes
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• pigmentation of the feet is decreased but to a lesser extent than in tail epidermis
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• at 9 months of age, pigmentation of tail skin is consistently lower than that of control mice
• melanocyte counts are consistently lower but not significantly decreased in the tail epidermis
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• when cultured in vitro, primary autophagy-deficient melanocytes isolated from body skin of newborn mice show increased melanosome-associated vacuolation, unlike autophagy-competent melanocytes
• however, in vivo, both melanocytes and keratinocytes of tail skin contain mature melanosomes, suggesting normal melanosome formation, maturation, and transfer in the absence of an intact autophagy system
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integument
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• mice show decreased pigmentation of dorsal hair; melanin content of dorsal hair is reduced by 10-15% relative to that in control mice
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• mice exhibit slight hypopigmentation of the tail epidermis
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• some melanocytes of tail skin contain misshapen, swollen, and possibly disintegrating mitochondria, not observed in control melanocytes
• however, when cultured in vitro, primary autophagy-deficient melanocytes isolated from body skin of newborn mice show no significant change of melanin content relative to autophagy-competent melanocytes
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• pigmentation of the feet is decreased but to a lesser extent than in tail epidermis
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• at 9 months of age, pigmentation of tail skin is consistently lower than that of control mice
• melanocyte counts are consistently lower but not significantly decreased in the tail epidermis
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cellular
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• swollen mitochondria are observed in some melanocytes of tail skin, unlike in control mice
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• lipidation of microtubule-associated protein 1 light chain 3 beta (LC3), i.e. conversion of LC3-I to LC3-II, is completely blocked, indicating efficient disruption of autophagy in isolated melanocytes; in contrast, both LC3-I and LC3-II are detected in melanocytes of control mice
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• cultured primary autophagy-deficient melanocytes stop proliferating after the third passage, unlike autophagy-competent melanocytes which can be maintained up to passage 5
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• in culture, primary autophagy-deficient melanocytes show a strong reduction in proliferative capacity and start acquiring senescent morphotypes with distended cytoplasms early in the second passage
• the premature senescent phenotype becomes even more prominent at the end of passage 2 and beginning of passage 3
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• autophagy-deficient melanocytes accumulate reactive oxygen species damage, ubiquitinated proteins, and the multi-functional adapter protein SQSTM1/p62
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homeostasis/metabolism
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• lipidation of microtubule-associated protein 1 light chain 3 beta (LC3), i.e. conversion of LC3-I to LC3-II, is completely blocked, indicating efficient disruption of autophagy in isolated melanocytes; in contrast, both LC3-I and LC3-II are detected in melanocytes of control mice
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limbs/digits/tail
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• at 9 months of age, pigmentation of tail skin is consistently lower than that of control mice
• melanocyte counts are consistently lower but not significantly decreased in the tail epidermis
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