Neuronal Damage Using Fluoro-Jade B Histofluorescence and Gliosis in the Striatum After Various Durations of Transient Cerebral Ischemia in Gerbils

Abstract

Ischemic damage occurs well in vulnerable regions of the brain, including the hippocampus and striatum. In the present study, we examined neuronal damage/death and glial changes in the striatum 4 days after 5, 10, 15 and 20 min of transient cerebral ischemia using the gerbil. Spontaneous motor activity was increased with the duration time of ischemia-reperfusion (I-R). To examine neuronal damage, we used Fluoro-Jade B (F-J B, a marker for neuronal degeneration) histofluorescence staining. F-J B positive cells were detected only in the 20 min ischemia-group, not in the other groups. In addition, we examined gliosis of astrocytes and microglia using anti-glial fibrillary acidic protein (GFAP) and anti- ionized calcium-binding adapter molecule 1 (Iba-1), respectively. In the 5 min ischemia-group, GFAP-immunoreactive astrocytes were distinctively increased in number, and the immunoreactivity was stronger than that in the sham-group. In the 10, 15 and 20 min ischemia-groups, GFAP-immunoreactivity was more increased with the duration of I-R. On the other hand, the immunoreactivity and the number of Iba-1-immunoreactive microglia were distinctively increased in the 5 and 10 min ischemia-groups. In the 15 min ischemia-group, cell bodies of microglia were largest, and the immunoreactivity was highest; however, in the 20 min ischemia-group, the immunoreactivity was low compared to the 15 min ischemia-group. The results of western blotting for GFAP and Iba-1 were similar to the immunohistochemical data. In brief, these findings showed that neuronal death could be detected only in the 20 min ischemia-group 4 days after I-R, and the change pattern of astrocytes and microglia were apparently different according to the duration time of I-R.

Hyperthermic preconditioning severely accelerates neuronal damage in the gerbil ischemic hippocampal dentate gyrus via decreasing SODs expressions

“…This finding is supported by previous studies that showed that intensely increased microglia activation appeared in the hippocampal CA1 region and striatum with severe neuronal damage following prolonged ischemic duration [11,31]. Therefore, we suggest that the excess of microglia activation in the (H + ischemia)-groups might be related with the accelerated neuronal death in the polymorphic layer of the dentate gyrus.…”

Aggrecan, link protein and tenascin-R are essential components of the perineuronal net to protect neurons against iron-induced oxidative stress

“…Fluoro-Jade B as anionic dye specifically stains degenerating neurons by binding to a currently unknown, most likely a poly-amine, basic substance in the neuron. 59, 60 For counting procedures, Fluoro-Jade B was chosen because of its high technical feasibility and reproducibility. To evaluate a potential contribution of PV to the neuroprotective properties of the PNs, sections were also stained with an antibody against PV (PV25, 1 : 500, Swant, Marly, Switzerland) in combination with WFA and Fluoro-Jade B as well as HAG and Fluoro-Jade B.…”

Alpha7 nicotinic acetylcholine receptor is required for blood-brain barrier injury-related CNS disorders caused by Cryptococcus neoformans and HIV-1 associated comorbidity factors

“…Mice brains were harvested for fluoro-Jade B (FJB) staining [ 28 , 29 ] and immunostaining with an anti-NeuN antibody [ 30 ] after infection, fixed in 10 % buffered formalin for 24 h, embedded in paraffin, and sections with 5 μm thickness were prepared. Dried mounted brain sections were fixed with 4 % PFA vapors in a closed container placed in a water bath heated at 37 °C for 2.5 h. Slides were dehydrated and rehydrated through graded concentrations of alcohol (50, 70, 100, 70, and 50 % alcohol, 1 min each), and rinsed with distilled water for 1 min.…”