Lab Anim Res.  2018 Dec;34(4):317-328. 10.5625/lar.2018.34.4.317.

Comparison of scopolamine-induced cognitive impairment responses in three different ICR stocks

• Yoon WB
• Choi HJ
• Kim JE
• Park JW
• Kang MJ
• Bae SJ
• Lee YJ
• Choi YS
• Kim KS
• Jung YS
• Cho JY
• Hwang DY
• Song HK

• KMID: 2459311
• DOI: http://doi.org/10.5625/lar.2018.34.4.317

Abstract

Cognitive impairment responses are important research topics in the study of degenerative brain diseases as well as in understanding of human mental activities. To compare response to scopolamine (SPL)-induced cognitive impairment, we measured altered parameters for learning and memory ability, inflammatory response, oxidative stress, cholinergic dysfunction and neuronal cell damages, in Korl:ICR stock and two commercial breeder stocks (A:ICR and B:ICR) after relevant SPL exposure. In the water maze test, Korl:ICR showed no significant difference in SPL-induced learning and memory impairment compared to the two different ICRs, although escape latency was increased after SPL exposure. Although behavioral assessment using the manual avoidance test revealed reduced latency in all ICR mice after SPL treatment as compared to Vehicle, no differences were observed between the three ICR stocks. To determine cholinergic dysfunction induction by SPL exposure, activity of acetylcholinesterase (AChE) assessed in the three ICR stocks revealed no difference of acetylcholinesterase activity. Furthermore, low levels of superoxide dismutase (SOD) activity and high levels of inflammatory cytokines in SPL-treated group were maintained in all three ICR stocks, although some variations were observed between the SPLtreated groups. Neuronal cell damages induced by SPL showed similar response in all three ICR stocks, as assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, Nissl staining analysis and expression analyses of apoptosis-related proteins. Thus, the results of this study provide strong evidence that Korl:ICR is similar to the other two ICR. Stocks in response to learning and memory capacity.

Keyword

ICR; Korl:ICR; scopolamine; learning and memory

MeSH Terms

Acetylcholinesterase
Animals
Brain Diseases
Cognition Disorders*
Cytokines
DNA Nucleotidylexotransferase
Humans
Learning
Memory
Mice
Mice, Inbred ICR
Neurons
Oxidative Stress
Scopolamine Hydrobromide
Superoxide Dismutase
United Nations
Acetylcholinesterase
Cytokines
DNA Nucleotidylexotransferase
Scopolamine Hydrobromide
Superoxide Dismutase

Figure

• 

  Figure 1 Comparison of the three ICR stocks on SPL-induced impairment of learning and memory in the Morris water maze. The first reaching time was evaluated in the target quadrant of the pool among Korl:ICR (A), A:ICR (B), and B:ICR (C) mice after exposure to SPL or Vehicle. Each first reaching time was evaluated as described in Materials and Methods. Data represents the mean±SEM of n=8/group (*P<0.05 versus Vehicle control group).

• 

  Figure 2 Comparison of behavioral responses of the three ICR stocks towards SPL-induced learning and memory impairment in the passive avoidance test. Panel represents the latency period into the dark chamber for Korl:ICR (A), A:ICR (B), and B:ICR (C) mice after treatment with SPL or Vehicle. Each latency period was evaluated as described in Materials and Methods. Data represents the mean±SEM of n=8/group (*P<0.05 versus Vehicle control group).

• 

  Figure 3 Comparison of AChE activity in SPL treated brain samples among the Korl:ICR, A:ICR, and B:ICR mice. Relative AChE activity levels were measured using the Acetylchol-inesterase Assay Kit, according to the experimental procedure described in the Materials and Methods. A, B, and C plots present the relative AChE activity of brain tissue from Korl:ICR, A:ICR, and B:ICR stocks, respectively. Data represents the mean±SEM of n=8/group (*P<0.05 versus Vehicle control group).

• 

  Figure 4 SOD activity was measured in the homogenate of the brain tissue collected from SPL treated ICR stocks. Each panel represents the SOD activity of Korl:ICR (A), A:ICR (B), and B:ICR (C) mice after treatment with SPL or Vehicle. Data represents the mean±SEM of n=8/group (*P<0.05 versus Vehicle control group).

• 

  Figure 5 Differing inflammatory responses of SPL-treated brain tissue among the three ICR stocks. The mRNA levels of inflammation related proteins (IL-1β and COX-2) were measured by RT-PCR using specific primers. Amplified DNA products were electrophoresed on the agarose gel and the intensity of the specific bands was recorded by a gel documentation system. Each panel represents the mRNA expression level of inflammation related proteins among Korl:ICR (A), A:ICR (B), and B:ICR (C) mice after treatment with SPL or Vehicle (*P<0.05 versus Vehicle control group).

• 

  Figure 6 Determination of SPL-induced apoptosis among the three ICR stocks using TUNEL assay. Representative images show staining of TUNEL in brain tissues treated with Vehicle or SPL, as indicated. (A), (B), and (C) panel display fluorescence of FITC-BrdU on brain tissue from Korl:ICR, A:ICR, and B:ICR stocks, respectively.

• 

  Figure 7 Comparison of SPL-induced apoptosis among Korl:ICR, A:ICR, and B:ICR mice using Nissl staining. This staining represent apoptotic cells in the hippocampus of the brain tissue from each ICR stock. Low intensity was observed in the hippocampus (4× magnification). Detailed histological features of several regions of the hippocampus are shown at 400× magnification. (A), (B), and (C) panels display the Nissl staining outcome on brain tissues from Korl:ICR, A:ICR, and B:ICR stocks, respectively.

• 

  Figure 8 Determination of SPL-induced apoptosis among the three ICR stocks by comparing the Bax/Bcl-2 protein ratio. Total tissue lysates were prepared from the brain tissues of SPL-treated Korl:ICR (A), B:ICR (B), and B:ICR (C) mice of each group, as described in Materials and Methods. A total of 50 µg of protein per sample was immunoblotted with antibodies for each protein. Three samples were assayed in triplicate by western blotting. Data represents the means±SEM of three replicates (*P<0.05 versus Vehicle control group).

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