Hippocampal Amyloid Precursor Protein Research Articles

Molecular cloning, expression, and regulation of hippocampal amyloid precursor protein of senescence accelerated mouse (SAMP8)

Alzheimer's disease (AD) is associated with increased expression of amyloid precursor protein (APP) with a consequent deposition of amyloid beta peptide (Aβ) which forms characteristic senile plaques. We have noticed that the senescence accelerated mouse (SAMP8), a strain of mouse that exhibits age-dependent defects such as loss of memory and retention at an early age of 8-12 months, also produces increased amounts of APP and Aβ similar to those observed in Alzheimer's disease (AD). In order to investigate if this is due to mutations in APP similar to those observed in AD, and to develop molecular probes that regulate its expression, APP cDNA was cloned from the hippocampus of 8-month-old SAMP8 mouse. The nucleotide sequence is 99.7% homologous with that of mouse and rat, 88.7% with monkey, and 89.2% with human homologues. At the amino acid level, the homology was 99.2% and 97.6% with rodent and primate sequences, respectively. A single amino acid substitution of Alanine instead of Valine at position 300 was unique to SAMP8 mouse APP. However, no mutations similar to those reported in human familial AD were observed. When the cDNA was expressed in HeLa cells, glycosylated mature APP could be detected by immunoblotting technique. The expression could be regulated in a time- and concentration-dependent manner by using an antisense oligonucleotide specific to APP mRNA. Such regulation of APP expression may have a therapeutic application in vivo.Key words: cloning, amyloid precursor protein, transfection, expression, and antisense oligo.

Molecular cloning, expression, and regulation of hippocampal amyloid precursor protein of senescence accelerated mouse (SAMP8)

Alzheimer's disease (AD) is associated with increased expression of amyloid precursor protein (APP) with a consequent deposition of amyloid beta peptide (Abeta) which forms characteristic senile plaques. We have noticed that the senescence accelerated mouse (SAMP8), a strain of mouse that exhibits age-dependent defects such as loss of memory and retention at an early age of 8-12 months, also produces increased amounts of APP and Abeta similar to those observed in Alzheimer's disease (AD). In order to investigate if this is due to mutations in APP similar to those observed in AD, and to develop molecular probes that regulate its expression, APP cDNA was cloned from the hippocampus of 8-month-old SAMP8 mouse. The nucleotide sequence is 99.7% homologous with that of mouse and rat, 88.7% with monkey, and 89.2% with human homologues. At the amino acid level, the homology was 99.2% and 97.6% with rodent and primate sequences, respectively. A single amino acid substitution of Alanine instead of Valine at position 300 was unique to SAMP8 mouse APP. However, no mutations similar to those reported in human familial AD were observed. When the cDNA was expressed in HeLa cells, glycosylated mature APP could be detected by immunoblotting technique. The expression could be regulated in a time- and concentration-dependent manner by using an antisense oligonucleotide specific to APP mRNA. Such regulation of APP expression may have a therapeutic application in vivo.

726 Age related changes of Alzheimer's amyloid precursor protein in hippocampus of senescence accelerated mouse (SAM)

726 Age related changes of Alzheimer's amyloid precursor protein in hippocampus of senescence accelerated mouse (SAM)

Lack of long-term effects after β-amyloid protein injections in rat brain

Rat β(1–42) peptide (β/A4) or phosphate buffered saline (PBS) was bilaterally injected into the hippocampus (HIP) or the lateral ventricle (ICV) of 3-month-old Fischer-344 rats. Fifteen months later, the animal's ability to learn a spatial memory task was tested using the Morris water maze. Acquisition of the task was impaired by the bilateral injection of either peptide or PBS into the hippocampus. Hippocampal-injected animals showed an increased average latency to find the platform by approximately 6 s ( p < 0.05). However, injection of rat β-peptide into the hippocampus or lateral ventricles failed to induce behavioral impairment when compared to vehicle injected controls. Retention of this task was not significantly impaired in any group. The spatial acuity test, a trial without the platform, revealed that both groups of animals that received hippocampal injections were impaired, spending 23% less time in the target quadrant compared to ICV-injected animals ( p < 0.005). Hippocampal ChAT activity was decreased in β/A4-injected animals but not significantly ( p < 0.06). β/A4-immunoreactivity was detected at the bottom of the needle track and the adjacent parenchyma of β/A4 hippocampal injected animals after 16 months. However, long-term in vivo deposition of β/A4 in both regions did not result in an upregulation of hippocampal amyloid precursor protein (APP) expression and there was no qualitative neuronal loss in the hippocampus. These data suggest that even small CA1 lesions in aging rats caused by two injections using a blunt 26-gauge needle produce a significant spatial navigation deficit in the Morris water maze, but in vivo rat β/A4 (1–42) deposition into the hippocampus or the lateral ventricles failed to induce specific long-term behavioral, biochemical, or histological effects.

Processing of β/A4 Amyloid Precursor Protein Is Altered in the Hippocampus of Reserpinized Rat-Brain

Immunoreactivities of β/A4 amyloid precursor protein (APP) and soluble derivative of APP (APPs) were determined in the hippocampus and the striatum of reserpinized rat brains. Significant decrease of APPs immunoreactivity was observed in the cytosolic fraction of hippocampus, but not of striatum, whereas APP immunoreactivity was found to be increased in the membrane fraction of hippocampus. These findings suggest that processing of APP in hippocampus may be regulated through the monoaminergic signal transduction system.