Journal: International Journal of Molecular Sciences

Article Title: Primary Cilia Structure Is Prolonged in Enteric Neurons of 5xFAD Alzheimer’s Disease Model Mice

doi: 10.3390/ijms222413564

Figure Lengend Snippet: Morphological and functional changes were observed in the gut of 5xFAD model mice. ( a ) Quantitative parameters were assessed within duodenal slices of mice. Gut slices were stained with HE (representative images for wild type and 5xFAD mice are shown) and respective structures were measured ( b ) The four anatomical structures villus length, crypt depth, submucosa and muscularis thickness were analyzed with four measurements of each duodenal structure in three slides per mouse ( n = 10 animals per group, aged 11 weeks, single values are shown) by using a clock-wise orientation in AD model mice and their respective wild type littermates. ( c ) Measurement of gut motility. Distal colonic sections ( n = 6 per group, mean age 25 weeks) were dissected and inserted into an organ bath with carbogen-gas perfused Krebs solution. A fecal pellet was inserted in the oral end and time measured for transport to the anal end. Statistical analysis was performed by using Student’s t -test ( n = 6 per group; * p < 0.05; ** p < 0.01).

Article Snippet: Male 5xFAD mice (APP K670N, M671L, I716V;PS1 M146L, L286V; Jackson Laboratory, Bar Harbor, ME, USA) were crossbred for maintenance with female C57B/6J mice (animal facility of the University Medical Center of Mainz).

Techniques: Functional Assay, Staining

Journal: International Journal of Molecular Sciences

Article Title: Primary Cilia Structure Is Prolonged in Enteric Neurons of 5xFAD Alzheimer’s Disease Model Mice

doi: 10.3390/ijms222413564

Figure Lengend Snippet: Primary enteric neuronal cultures of 5xFAD mice display increased neurite mass. ( a ) Preparations of myenteric plexus on longitudinal muscle (LMMP) were stripped off from colonic stretches and stained with βIII tubulin antibody. Procedure of quantitation is shown and one mesh area is labelled in red as an example. Representative pictures for wild type and 5xFAD are shown (scale bars: 100 µm). ( b ) Mesh area was calculated by ImageJ using two microscopic pictures per mouse. Meshes of the enteric network were subdivided in groups of 1500–4500 (small), 4500–10,000 (intermed. 1), 10,000–15,000 (intermed. 2), and 15,000–40,000 (large) µm 2 . Statistics were performed with One Way Anova (Fisher’s LSD test, n = 4 animals per group; aged 10–11 months, 10–36 meshes per animal; * p < 0.05). ( c ) Enteric neuron cultivation on transwell plates. Neurons were prepared from myenteric plexus and seeded on 3 µm transwell inserts. A representative microscopic picture of crystal violet stained neurons is shown. Neurites are seen as thin protrusions between cells or cell clusters. ( d ) On DIV4, cells were stained with crystal violet, cell bodies removed, neurite-attached stain extracted, and stain intensity measured. Cultures derived from 4 independent donor mice were analyzed with technical triplicates ( n = 12 per group). Values were normalized to values obtained for wild type and are presented in percentage as mean + SEM. Statistical analysis was performed by using Student’s t -test with Welch correction (* p < 0.05).

Article Snippet: Male 5xFAD mice (APP K670N, M671L, I716V;PS1 M146L, L286V; Jackson Laboratory, Bar Harbor, ME, USA) were crossbred for maintenance with female C57B/6J mice (animal facility of the University Medical Center of Mainz).

Techniques: Staining, Quantitation Assay, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Primary Cilia Structure Is Prolonged in Enteric Neurons of 5xFAD Alzheimer’s Disease Model Mice

doi: 10.3390/ijms222413564

Figure Lengend Snippet: Higher prevalence of elongated primary cilia in 5xFAD-derived enteric neurons. ( a ) Staining of cilia. Neurons were prepared as described before and seeded on cover slips. A representative microscopic image of neurons after staining for the ciliary membrane protein Arl13b (red) and the basal body marker Pericentrin (green) and counterstain with DAPI (blue) of both genotypes is shown (scale bars: 10 µm). ( b ) Quantification of ciliated cells. Number of cells carrying cilia was assessed and is presented in% of value obtained for wild type controls. The percentage of ciliated cells was calculated by determination of DAPI stained cells that showed Arl13b- and Pericentrin-double labelled primary cilia. Cultures derived from 4 independent donor mice in each group were analyzed with technical triplicates from 20 random visual fields with an average of 8 cells per image. ( c , d ) Cilia length was measured with ImageJ and mean length ( c ) or distribution in regard to length groups ( d ) was shown. Cultures derived from 3 (WT) and 4 (5xFAD) independent donors were analyzed with technical duplicates ( n = 159 cilia measured for WT and 260 for 5xFAD). Data are presented as mean + SEM. Statistical analysis was performed by using Student’s t -test or One Way Anova (post-test Fisher LSD; * p < 0.05).

Article Snippet: Male 5xFAD mice (APP K670N, M671L, I716V;PS1 M146L, L286V; Jackson Laboratory, Bar Harbor, ME, USA) were crossbred for maintenance with female C57B/6J mice (animal facility of the University Medical Center of Mainz).

Techniques: Derivative Assay, Staining, Membrane, Marker

Journal: International Journal of Molecular Sciences

Article Title: Primary Cilia Structure Is Prolonged in Enteric Neurons of 5xFAD Alzheimer’s Disease Model Mice

doi: 10.3390/ijms222413564

Figure Lengend Snippet: Effect of acute A-beta administration or of chronic exposure within transgenic enteric neurons on ciliary signaling pathways. ( a ) Enteric wild type neurons (DIV3) were treated for 24 h with A-beta peptides or scrambled control (sA-beta). mRNA was used for analysis of depicted gene products with Gapdh mRNA levels for normalization ( n = 6–8 per group). ( b ) Colonic mRNA of 8-month-old mice (WT or 5xFAD) was subjected to qPCR analysis. mRNA levels obtained were normalized to Gapdh mRNA levels ( n = 5–6 per group). Statistical analysis was performed by pairwise t -tests (* p < 0.05). All data are presented as mean + SEM.

Article Snippet: Male 5xFAD mice (APP K670N, M671L, I716V;PS1 M146L, L286V; Jackson Laboratory, Bar Harbor, ME, USA) were crossbred for maintenance with female C57B/6J mice (animal facility of the University Medical Center of Mainz).

Techniques: Transgenic Assay, Protein-Protein interactions, Control

Journal: Frontiers in Aging Neuroscience

Article Title: Comprehensive Evaluation of the 5XFAD Mouse Model for Preclinical Testing Applications: A MODEL-AD Study

doi: 10.3389/fnagi.2021.713726

Figure Lengend Snippet: Decreased total cholesterol and lipoproteins in 5XFAD males compared to wild-type males and decreased glucose levels in 5XFAD females compared to wild-type females. Non-fasted plasma was collected at the termination of the study, and total cholesterol (A) , low-density lipoproteins (B) , high-density lipoproteins (C) , triglycerides (D) , non-essential fatty acids (E) , and glucose (F) levels were measured.

Article Snippet: Male hemizygous 5XFAD mice (MMRRC stock #: 34848) were crossed with female C57BL6/J mice (JAX stock #: 000664) to maintain the 5XFAD and non-transgenic WT colonies at Indiana University and The Jackson Laboratory.

Techniques:

Journal: Frontiers in Aging Neuroscience

Article Title: Comprehensive Evaluation of the 5XFAD Mouse Model for Preclinical Testing Applications: A MODEL-AD Study

doi: 10.3389/fnagi.2021.713726

Figure Lengend Snippet: Functional assessment reveals hyperactivity associated with changes in frailty score in 5XFAD mice. Hyperactivity was observed in 5XFAD mice compared with controls was determined using open field and running wheels (A–D) . There were increases in distance traveled, vertical and horizonal activity and reductions in resting time. In the rotarod test (E) , both male and female 5XFAD mice maintained their balance for a greater period of time than the age-and sex-matched WT controls. Male and female 5XFAD mice and controls demonstrated aging-dependent increases in frailty scores from 6 months to 12 months of age (F) . 5XFAD male or female mice did not demonstrate deficits in percent alternation relative to age-and sex-matched controls at either 6 or 12 months of age (G) .

Article Snippet: Male hemizygous 5XFAD mice (MMRRC stock #: 34848) were crossed with female C57BL6/J mice (JAX stock #: 000664) to maintain the 5XFAD and non-transgenic WT colonies at Indiana University and The Jackson Laboratory.

Techniques: Functional Assay, Activity Assay

Journal: Frontiers in Aging Neuroscience

Article Title: Comprehensive Evaluation of the 5XFAD Mouse Model for Preclinical Testing Applications: A MODEL-AD Study

doi: 10.3389/fnagi.2021.713726

Figure Lengend Snippet: No emergent phenotypes of sleep or spike-wave activity in 5XFAD mice. No emergent phenotypes of sleep or spike wave activity in 5XFAD mice. Sleep and seizure activity was assessed by EEG in female and male 5XFAD mice ( n = 4 per group). Example traces during epochs of wake (A) , NREM sleep (B) , and REM sleep (C) are displayed across the top. No statistical differences in frequency are measured during wake (D) , NREM (E) , or REM (F) . Analysis of sleep phase distribution during each activity period throughout the 72 h recording does not reveal significant differences in time spent in each respective sleep phase. Increased wake (G) , and decreased NREM sleep (H) is witnessed in female mice during both activity phases compares to male controls. Differences in REM sleep (I) are not seen. Mean duration of wake epochs are significantly increased in females compared to male controls (J) . An example of a putative spike wave identified using pike sorting criteria (K) . No significant differences in spike wave activity (L) , spike wave duration (M) , or spikes per train (N) , are identified for sex or genotype.

Article Snippet: Male hemizygous 5XFAD mice (MMRRC stock #: 34848) were crossed with female C57BL6/J mice (JAX stock #: 000664) to maintain the 5XFAD and non-transgenic WT colonies at Indiana University and The Jackson Laboratory.

Techniques: Activity Assay

Journal: Frontiers in Aging Neuroscience

Article Title: Comprehensive Evaluation of the 5XFAD Mouse Model for Preclinical Testing Applications: A MODEL-AD Study

doi: 10.3389/fnagi.2021.713726

Figure Lengend Snippet: Transcriptomics identified genotype and sex being a major source of variation in between 5XFAD and WT mice and a positive correlation with most of the functionally distinct AMP-AD modules. (A) The first principal component accounted for 23% of the total variance and separated 5XFAD samples from WT animals. Female and male samples clustered distinctly at all ages in the second principal component (11% of total variance), suggesting the presence of sex-biased molecular changes in animals. Female and male samples are shown in red and green colors, respectively. 5XFAD and WT controls are represented as circles and triangles, respectively. Increasing point sizes represent the increasing age of the mice (4, 6, and 12 months, respectively). (B) KEGG enrichment analysis for genes significantly associated with sex (female), genotype (5XFAD) and sex by genotype interaction (5XFAD female) relative to age-matched male B6 mice. (C) Pearson correlation coefficients for gene expression changes in mice (log fold change of the 5XFAD mice minus age-matched B6 mice) and human disease (log fold change for cases minus controls). AMP-AD modules are grouped into five previously identified consensus clusters describing the major functional groups of AD-related alterations. Positive correlations are shown in blue and negative correlations in red. Color intensity and size of the circles are proportional to the correlation coefficient. Correlations with adjusted p -value > 0.05 are considered non-significant and are left blank.

Article Snippet: Male hemizygous 5XFAD mice (MMRRC stock #: 34848) were crossed with female C57BL6/J mice (JAX stock #: 000664) to maintain the 5XFAD and non-transgenic WT colonies at Indiana University and The Jackson Laboratory.

Techniques: Expressing, Functional Assay

Journal: Frontiers in Aging Neuroscience

Article Title: Comprehensive Evaluation of the 5XFAD Mouse Model for Preclinical Testing Applications: A MODEL-AD Study

doi: 10.3389/fnagi.2021.713726

Figure Lengend Snippet: Mapping the progression of methoxy-X04 labeled plaque deposition across brain regions. (A) Example brain section from 4-month-old female 5XFAD with methoxy-X04 labeled (gray) and segmentation masked plaques (red overlay). Zoom inset depicts (left to right) methoxy-X04 signal, mask (red), methoxy-X04 signal with mask overlay. (B) Plaque densities (fractional area) were quantified for all brain regions depicted as average density heat maps (red) mapped across the entire Allen Mouse Brain Reference Atlas for 6-month-old female 5XFAD, visualized as evenly spaced selected coronal sections (top) and 3D renderings (bottom). (C) Box plots of the plaque densities for 13 major brain regions from 2-, 3-, 4-, and 6-month-old female (top) to male (bottom) cohorts demonstrate regional plaque density progression, plots depict median and interquartile range (IQR-25:75) whiskers depict 1.5× IQR, and diamonds for values outside the range.

Article Snippet: Male hemizygous 5XFAD mice (MMRRC stock #: 34848) were crossed with female C57BL6/J mice (JAX stock #: 000664) to maintain the 5XFAD and non-transgenic WT colonies at Indiana University and The Jackson Laboratory.

Techniques: Labeling

Journal: Frontiers in Aging Neuroscience

Article Title: Comprehensive Evaluation of the 5XFAD Mouse Model for Preclinical Testing Applications: A MODEL-AD Study

doi: 10.3389/fnagi.2021.713726

Figure Lengend Snippet: NanoString GeoMX system indicates protein changes in cortex and hippocampus in 5XFAD mice compared to controls at 4 and 12 months. In the cortex (A) , several protein markers were upregulated in the 5XFAD mouse compared to controls at both time points. The greatest increases in the cortex were in Aββ1–42 and APP, while reductions were observed in NeuN by 12 months of age. In the hippocampus (B) , markers related to AD pathology (Aββ1–42, APP) as well as inflammation were upregulated as early as 4 months of age.

Article Snippet: Male hemizygous 5XFAD mice (MMRRC stock #: 34848) were crossed with female C57BL6/J mice (JAX stock #: 000664) to maintain the 5XFAD and non-transgenic WT colonies at Indiana University and The Jackson Laboratory.

Techniques:

Journal: Nutrients

Article Title: Intermittent Fasting on Neurologic Diseases: Potential Role of Gut Microbiota

doi: 10.3390/nu15234915

Figure Lengend Snippet: Fasting on intestinal flora in animal studies.

Article Snippet: Ruiyuan Pan, 2022 , 3-month-old 5xFAD mice , ADF until the mice grow to 5.5–6 months old , Feces , 16s rRNA gene , Firmicutes phylum ↑ Bacteroidetes ↓ Lactobacillus family ↑ [ ] .

Techniques: Western Blot, Expressing, Sequencing, Amplification, Control, Bacteria, Clinical Proteomics

Journal: Nutrients

Article Title: Intermittent Fasting on Neurologic Diseases: Potential Role of Gut Microbiota

doi: 10.3390/nu15234915

Figure Lengend Snippet: Impact of IF on various neurodegenerative diseases.

Article Snippet: Ruiyuan Pan, 2022 , 3-month-old 5xFAD mice , ADF until the mice grow to 5.5–6 months old , Feces , 16s rRNA gene , Firmicutes phylum ↑ Bacteroidetes ↓ Lactobacillus family ↑ [ ] .

Techniques: In Vitro, Activity Assay, Activation Assay, Expressing, Functional Assay