Limits...
Murine Aβ over-production produces diffuse and compact Alzheimer-type amyloid deposits.

Xu G, Ran Y, Fromholt SE, Fu C, Yachnis AT, Golde TE, Borchelt DR - Acta Neuropathol Commun (2015)

Bottom Line: Transgenic overexpression of amyloid precursor protein (APP) genes that are either entirely human in sequence or have humanized Aβ sequences can produce Alzheimer-type amyloidosis in mice, provided the transgenes also encode mutations linked to familial Alzheimer's Disease (FAD).Both lines of mice that produce mouse Aβ develop amyloid deposits, with the moAPPswe/PS1dE9 micedeveloping extracellular compact, cored, neuritic deposits that primarily localize to white matter tracts andmeningial layers, whereas the tet.moAPPsi mice developed extracellular diffuse cortical/hippocampal deposits distributed throughout the parenchyma.These findings demonstrate that murine Aβ peptides have the capacity to produce amyloid deposits that are morphologically similar to deposits found in human AD provided the murine APP gene harbors mutations linked to human FAD.

View Article: PubMed Central - PubMed

Affiliation: Center for Translational Research in Neurodegenerative Disease, University of Florida, Gainesville, FL, 32610, USA.

ABSTRACT

Introduction: Transgenic overexpression of amyloid precursor protein (APP) genes that are either entirely human in sequence or have humanized Aβ sequences can produce Alzheimer-type amyloidosis in mice, provided the transgenes also encode mutations linked to familial Alzheimer's Disease (FAD). Although transgenic mice have been produced that overexpress wild-type mouse APP, no mice have been generated that express mouse APP with FAD mutations. Here we describe two different versions of such mice that produce amyloid deposits consisting of entirely of mouse Aβ peptides. One line of mice co-expresses mouse APP-Swedish (moAPPswe) with a human presenilin exon-9 deleted variant (PS1dE9) and another line expresses mouse APP-Swedish/Indiana (APPsi) using tetracycline-regulated vectors (tet.moAPPsi).

Results: Both lines of mice that produce mouse Aβ develop amyloid deposits, with the moAPPswe/PS1dE9 micedeveloping extracellular compact, cored, neuritic deposits that primarily localize to white matter tracts andmeningial layers, whereas the tet.moAPPsi mice developed extracellular diffuse cortical/hippocampal deposits distributed throughout the parenchyma.

Conclusions: These findings demonstrate that murine Aβ peptides have the capacity to produce amyloid deposits that are morphologically similar to deposits found in human AD provided the murine APP gene harbors mutations linked to human FAD.

No MeSH data available.


Related in: MedlinePlus

Comparison of APP and PS1 expression levels in PrP.MoAβ/PS1 and PrP.HuAβ/PS1 mice. The brains of 2 non-transgenic (age 1.6–2.4 months) and 3 each PrP.MoAβ/PS1 (Line D-943; aged 2.5–3.3 months) and PrP.HuAβ/PS1 (Line 85, aged 1.6–2.4 months) mice were homogenized in RIPA buffer, the supernatants were analyzed by immunoblotting 4–20% SDS-PAGE gels with antibodies to PS1 (1:5000, top row), human APP (mAb 6E10, 1:1500, second row), total APP (mAb 22C11, 1:1000, third row), or mouse SOD1 (1:2500, fourth row). The mouse SOD1 immunoblot served as a loading control. The levels of PS1dE9 and total APP were similar in the two lines of mice. Each lane contains 50 μg of total protein
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4644287&req=5

Fig3: Comparison of APP and PS1 expression levels in PrP.MoAβ/PS1 and PrP.HuAβ/PS1 mice. The brains of 2 non-transgenic (age 1.6–2.4 months) and 3 each PrP.MoAβ/PS1 (Line D-943; aged 2.5–3.3 months) and PrP.HuAβ/PS1 (Line 85, aged 1.6–2.4 months) mice were homogenized in RIPA buffer, the supernatants were analyzed by immunoblotting 4–20% SDS-PAGE gels with antibodies to PS1 (1:5000, top row), human APP (mAb 6E10, 1:1500, second row), total APP (mAb 22C11, 1:1000, third row), or mouse SOD1 (1:2500, fourth row). The mouse SOD1 immunoblot served as a loading control. The levels of PS1dE9 and total APP were similar in the two lines of mice. Each lane contains 50 μg of total protein

Mentions: A line of moAPPswe/PS1dE9 mice designated as line D-943 (which we identify from here on as PrP.MoAβ/PS1) (Additional file 1: Table S1) was found to express total APP (transgene plus endogenous) at levels comparable to the previously described MoHuAPPswe/PS1dE9-Line 85 mice [24] (from here on identified as PrP.HuAβ/PS1) (Fig. 3). Brain homogenates of 3 different PrP.MoAβ/PS1 mice were compared to brain homogenates of 3 different PrP.HuAβ/PS1 mice by immunoblot with antibodies to human presenilin, the monoclonal antibody 6E10 (prefers human APP/Aβ), the monoclonal antibody 22C11 (recognizes an N-terminal epitope that is shared by all APP constructs used), and SOD1 (a loading control). The immunoblots probed with 22C11 demonstrated similar levels of total APP in the PrP.MoAβ/PS1 and PrP.HuAβ/PS1 mice (Fig. 3). Similarly, the levels of human PS1dE9 protein were similar in the two lines of mice (Fig. 3). The APP expressed in PrP.HuAβ/PS1 mice was much more reactive to the monoclonal antibody 6E10 as expected. Immunoblots probed with the SOD1 antibody demonstrated equal loading (Fig. 3). These data demonstrate that our newly developed PrP.MoAβ/PS1 mice express the transgene at levels that are similar to our previously characterized PrP.HuAβ/PS1 mice [24].Fig. 3


Murine Aβ over-production produces diffuse and compact Alzheimer-type amyloid deposits.

Xu G, Ran Y, Fromholt SE, Fu C, Yachnis AT, Golde TE, Borchelt DR - Acta Neuropathol Commun (2015)

Comparison of APP and PS1 expression levels in PrP.MoAβ/PS1 and PrP.HuAβ/PS1 mice. The brains of 2 non-transgenic (age 1.6–2.4 months) and 3 each PrP.MoAβ/PS1 (Line D-943; aged 2.5–3.3 months) and PrP.HuAβ/PS1 (Line 85, aged 1.6–2.4 months) mice were homogenized in RIPA buffer, the supernatants were analyzed by immunoblotting 4–20% SDS-PAGE gels with antibodies to PS1 (1:5000, top row), human APP (mAb 6E10, 1:1500, second row), total APP (mAb 22C11, 1:1000, third row), or mouse SOD1 (1:2500, fourth row). The mouse SOD1 immunoblot served as a loading control. The levels of PS1dE9 and total APP were similar in the two lines of mice. Each lane contains 50 μg of total protein
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4644287&req=5

Fig3: Comparison of APP and PS1 expression levels in PrP.MoAβ/PS1 and PrP.HuAβ/PS1 mice. The brains of 2 non-transgenic (age 1.6–2.4 months) and 3 each PrP.MoAβ/PS1 (Line D-943; aged 2.5–3.3 months) and PrP.HuAβ/PS1 (Line 85, aged 1.6–2.4 months) mice were homogenized in RIPA buffer, the supernatants were analyzed by immunoblotting 4–20% SDS-PAGE gels with antibodies to PS1 (1:5000, top row), human APP (mAb 6E10, 1:1500, second row), total APP (mAb 22C11, 1:1000, third row), or mouse SOD1 (1:2500, fourth row). The mouse SOD1 immunoblot served as a loading control. The levels of PS1dE9 and total APP were similar in the two lines of mice. Each lane contains 50 μg of total protein
Mentions: A line of moAPPswe/PS1dE9 mice designated as line D-943 (which we identify from here on as PrP.MoAβ/PS1) (Additional file 1: Table S1) was found to express total APP (transgene plus endogenous) at levels comparable to the previously described MoHuAPPswe/PS1dE9-Line 85 mice [24] (from here on identified as PrP.HuAβ/PS1) (Fig. 3). Brain homogenates of 3 different PrP.MoAβ/PS1 mice were compared to brain homogenates of 3 different PrP.HuAβ/PS1 mice by immunoblot with antibodies to human presenilin, the monoclonal antibody 6E10 (prefers human APP/Aβ), the monoclonal antibody 22C11 (recognizes an N-terminal epitope that is shared by all APP constructs used), and SOD1 (a loading control). The immunoblots probed with 22C11 demonstrated similar levels of total APP in the PrP.MoAβ/PS1 and PrP.HuAβ/PS1 mice (Fig. 3). Similarly, the levels of human PS1dE9 protein were similar in the two lines of mice (Fig. 3). The APP expressed in PrP.HuAβ/PS1 mice was much more reactive to the monoclonal antibody 6E10 as expected. Immunoblots probed with the SOD1 antibody demonstrated equal loading (Fig. 3). These data demonstrate that our newly developed PrP.MoAβ/PS1 mice express the transgene at levels that are similar to our previously characterized PrP.HuAβ/PS1 mice [24].Fig. 3

Bottom Line: Transgenic overexpression of amyloid precursor protein (APP) genes that are either entirely human in sequence or have humanized Aβ sequences can produce Alzheimer-type amyloidosis in mice, provided the transgenes also encode mutations linked to familial Alzheimer's Disease (FAD).Both lines of mice that produce mouse Aβ develop amyloid deposits, with the moAPPswe/PS1dE9 micedeveloping extracellular compact, cored, neuritic deposits that primarily localize to white matter tracts andmeningial layers, whereas the tet.moAPPsi mice developed extracellular diffuse cortical/hippocampal deposits distributed throughout the parenchyma.These findings demonstrate that murine Aβ peptides have the capacity to produce amyloid deposits that are morphologically similar to deposits found in human AD provided the murine APP gene harbors mutations linked to human FAD.

View Article: PubMed Central - PubMed

Affiliation: Center for Translational Research in Neurodegenerative Disease, University of Florida, Gainesville, FL, 32610, USA.

ABSTRACT

Introduction: Transgenic overexpression of amyloid precursor protein (APP) genes that are either entirely human in sequence or have humanized Aβ sequences can produce Alzheimer-type amyloidosis in mice, provided the transgenes also encode mutations linked to familial Alzheimer's Disease (FAD). Although transgenic mice have been produced that overexpress wild-type mouse APP, no mice have been generated that express mouse APP with FAD mutations. Here we describe two different versions of such mice that produce amyloid deposits consisting of entirely of mouse Aβ peptides. One line of mice co-expresses mouse APP-Swedish (moAPPswe) with a human presenilin exon-9 deleted variant (PS1dE9) and another line expresses mouse APP-Swedish/Indiana (APPsi) using tetracycline-regulated vectors (tet.moAPPsi).

Results: Both lines of mice that produce mouse Aβ develop amyloid deposits, with the moAPPswe/PS1dE9 micedeveloping extracellular compact, cored, neuritic deposits that primarily localize to white matter tracts andmeningial layers, whereas the tet.moAPPsi mice developed extracellular diffuse cortical/hippocampal deposits distributed throughout the parenchyma.

Conclusions: These findings demonstrate that murine Aβ peptides have the capacity to produce amyloid deposits that are morphologically similar to deposits found in human AD provided the murine APP gene harbors mutations linked to human FAD.

No MeSH data available.


Related in: MedlinePlus