利用Pseudorabies病毒做透過神經運輸以探討中樞神經至精囊的神經支配

Abstract

HRP 分別注射到大白鼠精囊的壁上8-12 處，大白鼠則在72 小時後殺掉。我們 使用PBS 做心臟灌流然後使用1.25% glutaldehyde + 1% paraformaldehyde 做固 定。取出脊髓節T10-L2, L5-S3 及 major pelvic ganglia 固定在之前的固定液中並 且浸泡在phosphate buffer (30% PBS, 0.1 M, pH 7.4, 4 oC)3 天。冰凍的組織在做 HRP 組織化解反應之前用OCT 包埋及TMB 染色。經過酵素反應後的組織切片 用1 % neutral red 染色以做顯微鏡下觀察。 我們使用10 隻大白鼠做實驗；其中最後6 隻大白鼠中有2 隻大白鼠發現有模 糊的棕紅色顆粒物在major pelvic ganglia；但是在脊髓節T10-L2, L5-S3 責未發 現。

After exposing the seminal vesicle of the mature male Wistar rats, horseradish peroxidase (HRP) was injected into the wall of the seminal vesicle in 8-12 separated areas. The rats were sacrificed 72 hours later. We used phosphate buffered saline (PBS) for transcardial perfusion and followed by fixatives (1.25% glutaldehyde + 1% paraformaldehyde). The tissues of spinal segments T10-L2, L5-S3 and the major pelvic ganglia were removed and fixed in previous fixatives (1.25% glutaldehyde + 1% paraformaldehyde) and immersion in phosphate buffer (30% PBS, 0.1 M, pH 7.4, 4 oC) for another 3 days. The frozen specimens were cut into slices with OCT frozen medium and followed with tetramethylbenzidine (TMB) staining before HRP histochemical reaction. After treated with enzymatic reaction solution, the specimens were dried and stained with 1 % neutral red for 3 minutes and followed with light microscopic examination. 10 mature male Wistar rats were used in this study. Only 2 of the final six rats showed faint brownish red staining particles on the major pelvic ganglia. In the segments of spinal cord from T10 toL2 and L5 to S3, all of them did not show any evidence of horseradish peroxidase tracing.