Anat Cell Biol.  2010 Mar;43(1):54-63. 10.5115/acb.2010.43.1.54.

Effects of repeated restraint stress on platelet endothelial cell adhesion molecule-1 immunoreactivity and protein levels in the gerbil hippocampus after transient cerebral ischemia

• Park OK
• Lee CH
• Hwang IK
• Yoo KY
• Choi JH
• Won MH

• KMID: 2168897
• DOI: http://doi.org/10.5115/acb.2010.43.1.54

Abstract

Stress has long been known to be a causative factor of various disease states. In this study, we investigated the effects of repeated restraint stress on platelet endothelial cell adhesion molecule-1 (PECAM-1), a very important mediator in inflammation, immunoreactivity and protein levels as well as neuronal damage, in the gerbil hippocampus after 5 minutes of transient cerebral ischemia. Transient ischemia-induced neuronal death was shown in CA1 pyramidal cells 4 days after ischemia/reperfusion. However, repeated restraint stress protected neuronal death induced by ischemic damage. In the ischemia-group, PECAM-1 immunoreactivity and its protein levels were significantly increased in all the hippocampal subregions 4 days after ischemia/reperfusion. However, PECAM-1 immunoreactivity and its protein levels did not change significantly in the hippocampus of the stress-ischemia-group compared to the sham-groups. These results indicate that repeated restraint stress protects neuronal damage induced by transient cerebral ischemia, and this may be associated with maintenance of PECAM-1levels.

Keyword

Repeated restraint stress; cerebral ischemia; platelet endothelial cell adhesion molecule-1; gerbil; hippocampus

MeSH Terms

Antigens, CD31
Blood Platelets
Brain Ischemia
Gerbillinae
Hippocampus
Inflammation
Ischemic Attack, Transient
Neurons
Pyramidal Cells
Antigens, CD31

Figure

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  Fig. 1 CV staining in the sham- (A and B), ischemia- (C and D), stress-sham- (E and F) and stress-ischemia- (G and H) groups. Only a few CV+ cells are detected in the stratum pyramidale (SP) 4 days after ischemia/reperfusion (I/R) in the ischemia-group. However, in the stress-ischemia-group, many CV+ cells are found in the SP 4 days after I/R. CA, conus ammonis; DG, dentate gyrus; SO, stratum oriens; SR, stratum radiatum. Bar = 800 µm (A, C, E and G), 50 µm (B, D, F and H).

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  Fig. 2 Relative analysis of the number of CV+ cells 4 days after I/R (*P<0.05, significantly different from the sham-groups, †P<0.05, significantly different from the ischemia-group). Bars indicate means±SEM.

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  Fig. 3 Immunohistochemical staining for PECAM-1 in the CA1 region (A~D), CA2/3 region (E and F) and dentate gyrus (G and H) in the sham- (A and B) and ischemia-operated (C~H) gerbils of the control- (A, C, E and G) and stress- (B, D, F and H) groups 4 days after I/R. PECAM-1 immunoreaction (arrows) is detected in blood vessels located in strata oriens (SO) and radiatum (SR) in both the sham-groups (A and B). PECAM-1 immunoreactivity in the ischemia-group is markedly increased 4 days after I/R (C). However, in the stress-ischemia-group, PECAM-1 immunoreactivity is much lower than that in the ischemia-group (D). Immunohistochemical staining pattern for PECAM-1 in the CA2/3 region and dentate gyrus is similar to that in the CA1 region. SP, stratum pyramidale; ML, molecular layer; GCL, granule cell layer; PL, polynmorphic layer. Bar=50 µm.

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  Fig. 4 Relative optical density as % of PECAM-1 immunoreactive structures in the sham-, ischemia-, stress-sham-, and stress-ischemia-groups (*P<0.05, significantly different from the sham-group, †P<0.05, significantly different from the ischemia-group). Bars indicate means±SEM.

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  Fig. 5 (A) Western blot analysis of PECAM-1 (approximately 130 kDa) 4 days after I/R. (B) Relative optical density as % of immunoblot band is represented (*P<0.05, significantly different from the sham-group, †P<0.05, significantly different from the ischemia-group). Bars indicate means±SEM.

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