Date of Award
Doctor of Philosophy
Molecular Biology, Microbiology and Biochemistry
Decreased mitochondrial function is associated with aging and is an early step in Alzheimer's disease (AD). Autophagy also declines with age and is required for degradation of dysfunctional mitochondria but it is not known whether autophagosomal formation is overactive and/or degradation of autophagosomes is inhibited in AD. Although two-thirds of the 5 million Americans diagnosed with AD are women, without a clear mechanism. Since the role of gender and autophagy in AD is unclear, we wanted to know whether: A) decreased mitochondrial biogenesis precedes brain plaque formation, neuronal death and memory deficits B) autophagosomal formation is overactive and/or degradation of autophagosomes is inhibited in AD and C) deficits in mitochondrial biogenesis and autophagy occur earlier in life in the hippocampus/cortex of female AD-Tg, than male AD-Tg mice. To answer these questions, mitochondrial DNA (mtDNA), TFAM (transcription factor A mitochondrial) and two stages in autophagy, autophagosomal formation and degradation were measured in the hippocampus/cortex of non-transgenic (non-Tg) and the APP(Swe, DI)/NOS2-/- AD mouse model (AD-Tg) from 2 mo. through 12 mo. of age, when memory deficits develop. Male and female mice were evaluated for gender differences. By measuring mitochondrial gene copy number relative to nuclear gene copy number by qPCR, we found female specific decreases in mtDNA and TFAM protein levels in AD-Tg hippocampus/cortex, before 12 mo., when memory deficits develop. The male AD-Tg mice did not show any decline in the levels of mtDNA or TFAM protein through 12 mo. of age, indicating a decline in mitochondrial biogenesis earlier in life in female AD-Tg mice, than males. To determine whether autophagosomal formation is overactive and/or autophagosomal degradation is inhibited in AD, non-Tg and AD-Tg mouse neurons from 2-12 mo. of age were cultured and transfected with an adenovirus expressing the dual fusion protein, mCherry-GFP-LC3B. This system enables the distinction of early (autophagosomal formation) from late (autophagosomal degradation) stages of autophagy. Autophagosomal formation increased in female AD-Tg mice, while male AD-Tg neurons have decreased formation of autophagosomes, compared to non-Tg. Neurons from AD-Tg mice show deficits in degradation of autophagosomes by 2 mo. Our results show increased autophagosomal formation in female neurons with age and in our AD-like model without corresponding increases in degradation of autophagosomes. All together, our data indicates that the presence of familial APP mutations and knockout of NOS2 increases deficits in both mitochondrial biogenesis and autophagy in female mice, compared to males. These deficits occur before amyloid plaque formation and memory deficits, suggesting that mitochondrial and autophagic deficits may play a role in the higher incidence of AD in females.
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