【摘 要】
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Objective:Although the localization of orexin-containing cell bodies is restricted to the hypothalamus,orexincontaining fibers and their receptors are widely distributed in the brain.The present study
【机 构】
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Department of Physiology and Pathophysiology, Shanghai Medical College, Fudan University, Shanghai 2
【出 处】
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中国神经科学学会第四次会员代表大会暨第七届全国学术会议(The 7th Biennial Meeting and the
论文部分内容阅读
Objective:Although the localization of orexin-containing cell bodies is restricted to the hypothalamus,orexincontaining fibers and their receptors are widely distributed in the brain.The present study was to examine the expression of orexinA (OXA) and orexin type 1 receptor (OX1R) in medulla of neonatal and adult SD rats.The differential expression of both groups was observed with maturation.Methods (1) Tissue preparation:neonatal (1-5d,n=5) and adult (6-8w,n=5) SD rats were perfused transcardially via the left ventricle with 4% paraformaldehyde in PBS (pH 7.4) under deep anesthesia.After postfixation,30 μm thick sections were cut serially on a freezing microtome (Leica CM 1900).(2) Immunohistochemistry:free-floating sections were washed (5 min,3 times) before incubated with 0.3% Triton-100 (30 min),0.3% H2O2 (10 min),blocking (goat serum,20 min at 37℃),and incubated with anti-OXA or anti-OX 1R antibody (rabbit-anti-rat IgG,1∶500,1∶ 100,both Sigma) ovemight at 4℃,and were washed (5 min,3 times) before secondary antibody (bio-goat-anti-rabbit IgG,both Huamei,20 min,37℃),ABC complexation (20 min at 37℃).
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