Km670 671nl

The University of Kansas Institute of Animal Use and Care Committee has authorized all animal procedures and complies with the NIH standards for the use and care of vertebrate animals. Transgenic hAPP mice overexpressing a mutant human form of amyloid precursor protein (APP) that encodes hAPP695, hAPP751, and hAPP770 bearing mutations linked to familial AD [APPV717F (Indiana), KM670/671NL(Swedish)], J-20 line were obtained from Jackson Laboratory). J20 mice and wild-type (WT) females from the same background were bred to generate heterozygotes for the hAPP transgene. J20 males and C57BL/6J-Tg (Thy1-GCaMP6s) GP4.3Dkim/J (Strain: 024275, JAX; (Chen et al., 2013 (link))) females were bred to generate J20-GCaMP6s mice. Five mice at most were kept in a cage, but following cranial window surgery, they were individually housed on a 12h light/12h dark cycle or 24 hours darkness for visual deprivation experiments. Both genders were used in the study.

Male 3xTg-AD mice harboring APP (KM670/671NL), PS1 (M146V), and tau (P301L) protein transgenes [B6;129-Psen1^tm1MpmTg(APPSwe,tauP301L)1Lfa/J]; non-transgenic-control mice [B6129SF2/J]; Thy1.1 mice [B6.PL-Thy1/CyJ]; and C57BL/6 mice were obtained from The Jackson Laboratory (Bar Harbor, ME, USA). Animals were housed under a 12 h light/dark cycle and had free access to food and water. All animal experiments were conducted according to the guidelines for animal care and the guiding principles for experiments using animals and were approved by the University of Kyung Hee Animal Care and Use Committee, [KHUASP(SE)-21-102] and [KHUASP(SE)-20-240]. All animal studies were reviewed and performed in compliance with the ARRIVE guidelines.

5xFAD transgenic mice overexpressing mutant human APP (770) with the Swedish (KM670/671NL), Florida (I716V), and London (V717I) familial Alzheimer’s disease (FAD) mutations, and human PS1 with FAD mutations (M146L and L285V) were purchased from The Jackson Laboratory (34840-JAX, Bar Harbor, ME, USA). Both the APP and PS1 transgenes were under the control of the mouse Thy1 promoter^{70 (link)}. The backgrounds of the mice were C57BL/SJL, which was produced by crossbreeding C57BL/6J female and SJL/J male mice. APP-KI mice possess a single human APP gene with the Swedish (KM670/671NL), Arctic (E693G), and Beyreuther/Iberian (I716F) mutations^{45 (link)}.

5xFAD transgenic mice overexpressing mutant human APP (770) with the Swedish (KM670/671NL), Florida (I716V), and London (V717I) familial AD (FAD) mutations and human PS1 with two FAD mutations (M146L and L285V) were purchased from The Jackson Laboratory (CA, USA). Both the APP and PS1 transgenes were under the control of the mouse Thy1 promoter^{93 (link)}. The background of the mice was C57BL/SJL, which was generated by crossbreeding C57BL/6J female and SJL/J male mice. A comprehensive proteomics analysis was performed with brain tissue samples from male 5xFAD mice at 1, 3, 6, and 12 months of age, as previously described^{40 (link)}. APP-KI (App^{NL-G-F/NL-G-F}) mice^{94 (link)} carry Swedish (KM670/671NL), Beyreuther/Iberian (I716F), and Arctic (E693G) mutations in humanized APP mouse gene. PKC inhibitor (Go6976, 6.6 μM) was administered to the subarachnoid space of mouse cerebral cortex at 0.15 μl/h from 1.5 to 6 months of age by using osmotic pump (ALZET Osmotic Pumps #2006, Cupertino, CA, USA).

All mice were housed at the Wake Forest School of Medicine barrier facility under the supervision of the Animal Research Program, in compliance with the NIH Guide for the Care and Use of Laboratory Animals. Mice adhered to a 12-h light/12-h dark cycle, with regular feeding, cage cleaning, and 24-h food and water access. Both male and female mice were used at the age of 12–16 months for experimentation. A total of 51 mice were used for experiments (WT, n = 17; APP, n = 9; eEF2K^–/–, n = 16; APP/eEF2K^–/–, n = 9). Breeders of the APP/PS1 mice were purchased from the Jackson Laboratory and expressed human transgenes for APP (KM670/671NL) and presenilin-1 (PSEN1 L166P) (Radde et al., 2006 (link)). Homozygous eEF2K^–/– mice were a generous gift from Dr. Alexey Ryazanov (Rutgers University) and used to generate double mutant APP/PS1/eEF2K^–/– animals. APP/PS1 mice were crossbred with the eEF2K^± mice to generate littermate groups: wild type (WT), APP/PS1 (APP), eEF2K^±, and APP/PS1/eEF2K^–± double-mutant mice. Further, eEF2K^± mice were crossbred with APP/eEF2K^± mice to generate the four experimental genotypes: WT, APP, eEF2K^–/–, and APP/eEF2K^–/– double mutant mice. All genotyping was done by polymerase chain reaction (PCR). All protocols involving animals were approved by the Institutional Animal Care and Use Committee of Wake Forest University School of Medicine.

AD model (3xTg-AD) mice harboring transgenes encoding APP (KM670/671NL), PS1 (M146V), and tau (P301L) proteins [B6;129-Psen1^tm1Mpm Tg(APPSwe,tauP301L)1Lfa/J] were obtained from Jackson Laboratory (Bar Harbor, ME, USA). Age-matched male C57BL/6 mice were purchased from Charles River Korea (OrientBio, Sungnam, Korea). All animals were maintained under specific pathogen-free conditions and a 12-hour light/dark cycle. All mice had free access to food and water during the experiments. All animal experiments were conducted in accordance with the Rules for Animal Care and the Guiding Principles for Experiments Using Animals and were approved by the University of Kyung Hee Animal Care and Use Committee [KHUASP(SE)-16-085].

The 3xTg-AD mice harboring a mutant APP (KM670/671NL), which is a human mutant PS1 (M146V) knock-in, and tau (P301L) transgenes (B6;129-Psen1^tm1Mpm Tg(APPSwe, tauP301L)1Lfa/J) were obtained from Jackson Laboratory [18 (link)]. Male C57BL/6 mice were purchased from Charles River Korea (OrientBio, Sungnam, Korea). All mice were maintained under pathogen-free conditions with air conditioning and a 12-h light/dark cycle. All of the mice had free access to food and water during the experiments. The study was conducted in accordance with the Rules for Animal Care and the Guiding Principles for Animal Experiment Using Animals and was approved by the University of Kyung Hee Animal Care and Use Committee (KHUASP(SE)-13-015).