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Improved Expression of {gamma}-Aminobutyric Acid Receptor in Mice with Cerebral Infarct and Transplanted Bone Marrow Stromal Cells: An Autoradiographic and Histologic Analysis

Hideo Shichinohe, MD, PhD1, Satoshi Kuroda, MD, PhD1, Shunsuke Yano, MD, PhD1, Takako Ohnishi, MSc2, Hiroshi Tamagami, MSc2, Kazutoshi Hida, MD, PhD1 and Yoshinobu Iwasaki, MD, PhD1

1 Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan; and 2 Research and Development Division, Research Center, Nihon Medi-Physics Co. Ltd., Sodegaura, Japan


Figure 1
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  		FIGURE 1.  Representative findings for 125I-iomazenil in vitro autoradiography of control mouse. Two coronal slices through striatum (A) and hippocampus (B) are shown. In this study, ROIs are placed to semiquantitatively evaluate binding for central-type benzodiazepine receptor in neocortex in anterior cerebral artery territory, dorsal neocortex adjacent to cerebral infarct, cerebral infarct, ventral neocortex adjacent to cerebral infarct, striatum, and hippocampus. Same ROIs are also placed in contralateral hemisphere as a control.

 

Figure 2
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  		FIGURE 2.  Representative findings for 125I-iomazenil in vitro autoradiography of mice subjected to permanent middle cerebral artery occlusion. As shown with arrows, binding for central-type benzodiazepine receptor in dorsal neocortex adjacent to cerebral infarct remained subnormal in BMSC-transplanted mice (C and D), compared with results in vehicle-transplanted mice (A and B).

 

Figure 3
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  		FIGURE 3.  Bar graph showing results of semiquantitative analysis of 125I-iomazenil in vitro autoradiography. Values are ratios of radioactivity in each ROI to radioactivity in corresponding ROI of contralateral hemisphere. As shown in Figure 2, binding for central-type benzodiazepine receptor in dorsal neocortex adjacent to cerebral infarct was significantly higher in BMSC-transplanted mice than in vehicle-transplanted mice (P = 0.0053).

 

Figure 4
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  		FIGURE 4.  (A–C) Photomicrographs (original magnification, x400) of fluorescence immunostaining using antibody against GABAA receptor. Images are of dorsal neocortex adjacent to cerebral infarct of BMSC-transplanted mice (A) or vehicle-transplanted mice (B) or from contralateral neocortex of vehicle-transplanted mice (C). (D) Average numbers of cells positive for GABAA receptor in each area under x1,000 magnification. Dorsal neocortex adjacent to cerebral infarct had significantly more positive cells in BMSC-transplanted mice than in vehicle-transplanted mice (P = 0.0049). Bars represent SD of each value.

 

Figure 5
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  		FIGURE 5.  (A and B) Photomicrographs (original magnification, x400) of double fluorescence immunostaining using primary antibodies against GFP and GABAA receptor in dorsal neocortex adjacent to cerebral infarct (A) and striatum (B) of BMSC-transplanted mice. Majority of GFP-positive cells express GABAA receptor in dorsal neocortex adjacent to cerebral infarct (arrows) but not in striatum (arrows). (C) Photomicrographs (original magnification, x400) of double fluorescence immunostaining using primary antibodies against GFP and MAP2 reveal that majority of GFP-positive cells also express neuron-specific protein, MAP2, in dorsal neocortex adjacent to cerebral infarct (arrows) but not in striatum (data not shown).

 




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