6E10+ signal also increased significantly in the hippocampus from day 30 to day 90 (cysts, confirming that induces A immunoreactivity in infected mice but not in mock-infected controls (Fig.?2d). Open in a separate window Fig. behavioral and anatomical effects using immunohistochemistry, immunofluorescence, Western blotting, cell culture assays, as well as an array of mouse behavior assessments. Results We show that contamination induced two major hallmarks of AD in the brains of C57BL/6 male and female mice: beta-amyloid (A) immunoreactivity and hyperphosphorylated Tau. Infected mice showed significant neuronal death, loss of contamination also caused anxiety-like behavior, altered acknowledgement of interpersonal novelty, altered spatial memory, and reduced olfactory sensitivity. This last obtaining was unique to male mice, as infected females showed intact olfactory sensitivity. Conclusions These results demonstrate that can induce advanced indicators of AD in wild-type mice and that it may induce AD in some individuals with underlying health problems. Electronic supplementary material The online version of this article (10.1186/s12974-018-1086-8) contains supplementary material, which is available to authorized users. (is an obligate intracellular protozoan pathogen that enters the central nervous system (CNS) following its initial invasion and replication in the gut [5, 6]. infects an estimated one third of the adult populace worldwide, although seroprevalence varies significantly by country [7, 8] and?is usually highly dependent on place of birth, educational level, living conditions, occupation, race, and ethnicity [9, 10]. Emerging studies implicate as a contributor to Parkinsons disease, schizophrenia, obsessive compulsive disorder, and Tourette syndrome [11]. Little is known about the long-term impact of contamination on neuronal cells, and their receptors such as the contamination induces indicators of AD in C57BL/6 (wild-type) mice including A production, which is known to alter NMDAR signaling [19]. Here, we show that causes indicators of neurodegeneration in wild-type mice. We demonstrate that as contamination progresses, induces loss of NMDAR transmission with concomitant induction of AD pathology including production of hyperphosphorylated Tau and A immunoreactivity in the brain. We also observed neuronal death in the olfactory bulb accompanied by alterations in olfactory sensitivity which is known to occur in patients with AD [20]. The effects around the latter varied depending on sex: While olfactory sensitivity was reduced in male Mouse monoclonal to Human Albumin mice, females showed intact olfactory ability. Infected mice also showed alterations in interpersonal behavior, anxiety-like symptoms, and alterations in spatial learning. Thus, can directly confer neurodegenerative and behavioral indicators of AD in infected wild-type mice. Methods Mice Eight- to 12-week-old C57BL/6 2”-O-Galloylhyperin (wild-type) male and female mice were purchased from Jackson Laboratories. Mice were bred and housed under specific 2”-O-Galloylhyperin pathogen-free conditions in the animal facility at Cornell University or college. contamination in mice Mice were randomly assigned to mock-infected and was orally infected following the contamination protocol from Mahamed et al. [6]. Briefly, wild-type mice were infected with 10 ME49 cysts which were managed in Swiss Webster mice and isolated before contamination. Survival rates and excess weight loss were monitored daily. For our experiments, groups of mice were euthanized at 15, 30, 60, and 90?days post contamination and tissues (brain, spleen, spinal cord) were collected for further processing. Antibodies and reagents Anti-APP antibody (6E10) was purchased from Covance (Cat# SIG-39320-1000). Anti-beta amyloid was purchased from Abcam (Cat#2539). Anti-antibody was purchased from US biologicals (Cat# T8075-01). BAG1 was a gift from Dr. Dubey. Normal mouse IgG (SC-2025) and normal rabbit IgG (SC-2027) were used as isotype controls to test the specificity of the 6E10 and BAG1 antibodies, respectively. Anti-phospho-Tau antibody (AT8) was purchased from Thermofisher (Cat# MN1020). Anti-NeuN was purchased from Cell Signaling (Cat# 12943S) or Abcam (ab104224). Anti-VGLUT2 (Cat#71555S), anti-GAPDH (Cat# 2118), and anti-Tau (Cat#4019) antibodies were purchased from Cell Signaling. Anti-NMDAR (Cat# ab17345), anti-VGLUT1 (Cat# ab134283), and anti-GAD67 (Cat# ab26116) antibodies were purchased from Abcam. 2”-O-Galloylhyperin Thioflavin S was purchased from (Sigma Aldrich) and neuro-tracer from Life Technologies. Western blot Immunoblots were performed on whole brain lysates from mock-infected and main antibodies, anti-RH strain 2”-O-Galloylhyperin or anti-BAG1 (1:1000), anti-NMDAR (1:200), anti-VGLUT1 (1:100), anti-VGLUT2 (1:50) or anti-GAD67 (1:100), anti-6E10 (1:200), anti-beta amyloid (1:200), anti-pTau (AT8) (1:1000), and anti-NeuN (1:200) overnight at 4?C. Samples were washed twice with PBS and incubated with fluorochrome-conjugated secondary antibodies or horseradish peroxidase for 1?h for immunofluorescence assay (IFA) or 20?min for immunohistochemistry (IHC). For neurotracer staining, sections were incubated with neurotracer (1:500) for 30?min at room heat. For IFA, samples were washed with PBS twice and coverslipped using vectashield with 4,6-diamidino-2-phenylindole (DAPI) mounting media. For IHC, samples were washed with PBS twice then developed with AEC developing kit (Invitrogen) before counterstaining with hematoxylin. Slides were then.