TY - JOUR
T1 - Photocoagulation-induced retinal gliosis is inhibited by systemically expressed soluble TGF-β receptor type II via adenovirus mediated gene transfer
AU - Hisatomi, Toshio
AU - Sakamoto, Taiji
AU - Yamanaka, Ichiro
AU - Sassa, Yukio
AU - Kubota, Toshiaki
AU - Ueno, Hikaru
AU - Ohnishi, Yoshitaka
AU - Ishibashi, Tatsuro
N1 - Funding Information:
This work was supported in part by grant-in-Aid 09671804 and 09307040 for Scientific Research from the Ministry of Education, Science, Sports, and Culture of the Japanese Government, the Japan National Society for the Prevention of Blindness (Tokyo), the Fukuoka Anti-Cancer Association (Fukuoka), the Kaibara Morikazu Medical Science Promotion Foundation (Fukuoka), and the Casio Science Promotion Foundation (Tokyo). Address reprint requests to: Dr. Toshio Hisatomi, Department of Ophthalmology, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. E-mail: hisatomi@med.kyushu-u.ac.jp
PY - 2002
Y1 - 2002
N2 - Retinal gliosis is one of the major causes of visual dysfunction due to the loss of the retinal regular structure and function in various diseases, including diabetic retinopathy, retinal detachment, and glaucoma. Transforming growth factor-β (TGF-β) is assumed to play an important role in this disease process. In the present study, we determined whether the systemically expressed extracellular domain of the TGF-β type II receptor by adenovirus-mediated gene delivery could inhibit experimental retinal gliosis both in vitro and in vivo. Cultured bovine retinal glial cells, Müller cells, were stimulated by recombinant TGF-β and the expression of the glial marker, glial fibrillary acidic protein (GFAP), was evaluated by immunohistochemistry, semiquantitative RT-PCR, and Western blotting. In cultured Müller cells, TGF-β stimulated the GFAP expression in a dose-dependent fashion, and the conditioned medium from 293 cells transfected with adenovirus encoding for a soluble form TGF-β type II receptor (AdTβ-ExR) inhibited the expression of GFAP stimulated by exogenous TGF-β (p < 0.05). In this process, Smad4 protein, which plays a key role in intracellular signaling after cell surface receptors, actually translocated from cytosol to nucleus with TGF-β stimulation. The conditioned medium from AdTβ-ExR also inhibited the cytosol-nuclear translocation of Smad4. For in vivo studies, AdTβ-ExR was injected into the femoral muscles of Brown Norway rats and retinal photocoagulation was subsequently carried out. Immunohistochemical studies revealed that GFAP was strongly expressed around the photocoagulation spots after 12 days and these phenomena were inhibited by AdTβ-ExR. Western blotting of total retinal extract demonstrated the same results as those observed after immunohistochemistry. Our results suggest that TGF-β plays a pivotal role in the pathologic processes in retinal gliosis, and that the systemically expressed soluble TGF receptor by gene delivery may thus have a potential therapeutic value by inhibiting excessive retinal gliosis in various ocular diseases.
AB - Retinal gliosis is one of the major causes of visual dysfunction due to the loss of the retinal regular structure and function in various diseases, including diabetic retinopathy, retinal detachment, and glaucoma. Transforming growth factor-β (TGF-β) is assumed to play an important role in this disease process. In the present study, we determined whether the systemically expressed extracellular domain of the TGF-β type II receptor by adenovirus-mediated gene delivery could inhibit experimental retinal gliosis both in vitro and in vivo. Cultured bovine retinal glial cells, Müller cells, were stimulated by recombinant TGF-β and the expression of the glial marker, glial fibrillary acidic protein (GFAP), was evaluated by immunohistochemistry, semiquantitative RT-PCR, and Western blotting. In cultured Müller cells, TGF-β stimulated the GFAP expression in a dose-dependent fashion, and the conditioned medium from 293 cells transfected with adenovirus encoding for a soluble form TGF-β type II receptor (AdTβ-ExR) inhibited the expression of GFAP stimulated by exogenous TGF-β (p < 0.05). In this process, Smad4 protein, which plays a key role in intracellular signaling after cell surface receptors, actually translocated from cytosol to nucleus with TGF-β stimulation. The conditioned medium from AdTβ-ExR also inhibited the cytosol-nuclear translocation of Smad4. For in vivo studies, AdTβ-ExR was injected into the femoral muscles of Brown Norway rats and retinal photocoagulation was subsequently carried out. Immunohistochemical studies revealed that GFAP was strongly expressed around the photocoagulation spots after 12 days and these phenomena were inhibited by AdTβ-ExR. Western blotting of total retinal extract demonstrated the same results as those observed after immunohistochemistry. Our results suggest that TGF-β plays a pivotal role in the pathologic processes in retinal gliosis, and that the systemically expressed soluble TGF receptor by gene delivery may thus have a potential therapeutic value by inhibiting excessive retinal gliosis in various ocular diseases.
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U2 - 10.1097/01.LAB.0000018829.49754.DD
DO - 10.1097/01.LAB.0000018829.49754.DD
M3 - Article
C2 - 12118088
AN - SCOPUS:0036318031
SN - 0023-6837
VL - 82
SP - 863
EP - 870
JO - Laboratory Investigation
JF - Laboratory Investigation
IS - 7
ER -
