Yonsei Med J.  2010 Sep;51(5):661-671. 10.3349/ymj.2010.51.5.661.

Acute Stress and Chronic Stress Change Brain-Derived Neurotrophic Factor (BDNF) and Tyrosine Kinase-Coupled Receptor (TrkB) Expression in Both Young and Aged Rat Hippocampus

• Shi SS
• Shao SH
• Yuan BP
• Pan F
• Li ZL

• KMID: 1071416
• DOI: http://doi.org/10.3349/ymj.2010.51.5.661

Abstract

PURPOSE
The purpose of this study is to explore the dynamic change of brain-derived neurotrophic factor (BDNF) mRNA, protein, and tyrosine kinase-coupled receptor (TrkB) mRNA of the rat hippocampus under different stress conditions and to explore the influence of senescence on the productions expression.
MATERIALS AND METHODS
By using forced-swimming in 4degrees C cold ice water and 25degrees C warm water, young and aged male rats were randomly divided into acute stress (AS) and chronic mild repeated stress (CMRS) subgroups, respectively. BDNF productions and TrkB mRNA in the hippocampus were detected by using Western-blotting and reverse transcription-polymerase chain reaction (RT-PCR), separately, at 15, 30, 60, 180, and 720 min after the last stress session.
RESULTS
The short AS induced a significant increase in BDNF mRNA and protein in both age groups, but the changes in the young group were substantially greater than those of the aged group (p < 0.005). The CMRS resulted in a decrease in BDNF mRNA and protein, but a significant increase in TrkB mRNA in both young and age groups. The expression of BDNF mRNA and protein in the AS groups were higher than in the CMRS groups at 15, 30, and 60 min after stress.
CONCLUSION
The results indicated that the up/down-regulation of BDNF and TrkB were affected by aging and the stimulus paradigm, which might reflect important mechanisms by which the hippocampus copes with stressful stimuli.

Keyword

BDNF; chronic stress; senescence; behavior; plasma corticosterone

MeSH Terms

Animals
Blotting, Western
Brain-Derived Neurotrophic Factor/genetics/*metabolism
Corticosterone/blood
*Gene Expression Regulation
Hippocampus/*metabolism
Male
Radioimmunoassay
Random Allocation
Rats
Rats, Wistar
Receptor, trkB/genetics/*metabolism
Reverse Transcriptase Polymerase Chain Reaction
Stress, Physiological/genetics/*physiology

Figure

• 

  Fig. 1 Diagrammatic representation of the protocols for AS and CMRS exposures. (A) The AS protocol. (B) The CMRS protocol. AS, acute stress; CMRS, chronic mild repeated stress.

• 

  Fig. 2 The results of the open field test for young CMRS group rats (Mean ± SD) (n = 6). *p < 0.05, **p < 0.001, different from control groups (Student's t-test). (A) Number of square crossings in different stress periods. (B) Vertical movements in different stress periods. (C) Number of grooming events in different stress periods. (D) Time spent in the center square during a stress course. CMRS, chronic mild repeated stress.

• 

  Fig. 3 The results of an open field test for aged CMRS group rats (Mean ± SD) (n = 6). *p < 0.05, **p < 0.001, †p < 0.05, ‡p < 0.001, different from control groups (Student's t-test). (A) Number of square crossings in different stress periods. (B) Vertical movements in different stress periods. (C) Number of grooming events in different stress periods. (D) Time spent in the center square during a stress course. CMRS, chronic mild repeated stress.

• 

  Fig. 4 Temporal profile of plasma corticosterone (ng/mL) levels of young and aged AS group rats. Values are shown for control animals (0 min, n = 6) and after 15, 30, 60, 180 and 720 min of stress. The results are expressed as mean ± SEM. *p < 0.001 vs. young control group. **p < 0.001 vs. aged control group. AS, acute stress.

• 

  Fig. 5 Temporal profile of plasma corticosterone (ng/mL) levels in young and aged CMRS rat groups. Values are shown for control animals (0 min) and after 15, 30, 60, 180 and 720 min of stress. The results are expressed as mean ± SEM. *p < 0.001 vs. young control group. **p < 0.01 and †p < 0.001 vs. aged control group. CMRS, chronic mild repeated stress.

• 

  Fig. 6 Time course of BDNF protein expression in the hippocampus measured by Western blotting after acute stress (AS). (A) Representative photographic film, illustrating the dynamic changes in BDNF protein from young and aged AS groups. (B) Results of statistical analysis of BDNF protein expression in control groups (unstressed, 0 min) and stressed animals after 15, 30, 60, 180, 720 min of stress (following acute or chronic stress imposition). The results were expressed as mean ± SEM. *p < 0.05 and **p < 0.001 vs. young control group. †p < 0.05 and ‡p < 0.001 vs. aged control group. n = 5-6 rats per each time point studied in two independent stress conditions. BDNF, brain-derived neurotrophic factor; AS, acute stress.

• 

  Fig. 7 Time course of BDNF protein expression in the hippocampus measured by Western blotting after chronic repeated stress. (A) Representative photographic film, illustrating the dynamic changes in BDNF protein from the young and aged CMRS groups. (B) Results of statistical analysis of BDNF protein expression in control groups (unstressed, 0 min) and stressed animals after 15, 30, 60, 180, 720 min of stress (in two stress conditions). The results were expressed as mean ± SEM. *p < 0.05 and **p < 0.001 vs. young control group. †p < 0.05 and ‡p < 0.001 vs. aged control group. n = 5-6 rats per each time point studied in two independent stress conditions. BDNF, brain-derived neurotrophic factor; CMRS, chronic mild repeated stress.

• 

  Fig. 8 BDNF mRNA expression detected by semiquantitative RT-PCR in control groups (unstressed, 0 min) and young and aged AS groups after a period of different stress performance. Total RNA was isolated from hippocampus and assayed for BDNF at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms illustrating the expression of BDNF mRNA in the young control and AS groups. (B) Expression of BDNF mRNA in the aged AS group. (C) Quantitative analysis of BDNF mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of the β-actin (internal standard) band and expressed as the mean ± SEM. *p < 0.05 and **p < 0.001 vs. young control group. †p < 0.05 and ‡p < 0.001 vs. aged control group. n = 5-6 rats per each time point studied in two independent stress conditions. BDNF, brain-derived neurotrophic factor; AS, acute stress; RT-PCR, reverse transcription-polymerase chain reaction.

• 

  Fig. 9 TrkB mRNA expression measured by semiquantitative RT-PCR in the control groups (unstressed, 0 min) and young and aged AS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for TrkB mRNA at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of TrkB mRNA in the young control (0 min) and AS groups. (B) TrkB mRNA expression in the aged control and AS groups. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. n = 5-6 rats per each time point studied. TrkB, tyrosine kinase-coupled receptor; RT-PCR, reverse transcription-polymerase chain reaction; AS, acute stress.

• 

  Fig. 10 BDNF mRNA expression detected by RT-PCR in the control groups (unstressed, 0 min) and young and aged CMRS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for BDNF at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of BDNF mRNA in the young control and CMRS groups. (B) BDNF mRNA expression in the aged control and CMRS groups. (C) Quantitative analysis of BDNF mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. *p < 0.05 and **p < 0.001 vs. young control group. †p < 0.05 and ‡p < 0.001 vs. aged control group. n = 5-6 rats per each time point studied in two independent stress conditions. BDNF, brain-derived neurotrophic factor; CMRS, chronic mild repeated stress; RT-PCR, reverse transcription-polymerase chain reaction.

• 

  Fig. 11 TrkB mRNA expression measured by semiquantitative RT-PCR in the control groups (unstressed, 0 min) and young and aged CMRS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for TrkB at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of TrkB mRNA in the young control and CMRS groups. (B) TrkB mRNA expression in the aged control and CMRS groups. (C) Line chart represented the results of quantitative analysis of TrkB mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. *p < 0.05 and **p < 0.001 vs. young control group. †p < 0.001 vs. aged control group. n = 5-6 rats per each time point studied in two independent stress conditions. TrkB, tyrosine kinase-coupled receptor; CMRS, chronic mild repeated stress; RT-PCR, reverse transcription-polymerase chain reaction.

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