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为进一步探讨视网膜节细胞的轴突终末与视交叉上核（SCN）内VIP和AVP能神经元间的联系，采用假狂犬病毒（PRV）跨神经元顺行追踪及免疫荧光双标法。对成年大鼠单侧眼球内缓慢注入PRV（Bartha株、5X108PFU／ml）3PI，留针10分钟。动物分别存活56、72和96小时。1％戊巴比妥钠腹腔注射麻醉，经左心左灌注生理盐水（每100ml含肝素100单位）持续5分钟，流速30—40ml／分钟，冷固定液（含4％多聚甲醛、15％苦味酸，用0IM／L磷酸缓冲液配制、PH7‘4），开始流速为朋～40ml分钟，持续5～7分钟，然后降慢到10ml／分钟，维持半小时。开颅取材、修块、入上述固定液后固定一小时，再转入ZO％蔗糖磷酸缓冲液过夜、恒冷箱切片，片厚30Pm。免疫荧光双标法：含03％Triton、2％驴血清（NDS）的001PBS（pH7．4）稀释混合两种一抗，羊抗PRV1：2000，兔抗VIP或AVP1：2000，室温24～48h。二抗分别为驴抗羊IgG（IRITC标记）1：100，驴抗兔IgG（FITC标记）1：50，用2％NDS的0．01MPBS稀释，室温1．sh。以上各步间均用0．01MPBS漂洗10’X3.裱片，PBS甘油封片。免疫荧光双标操作在暗室里进行。结果；被PRV标记的细胞呈现出，绿色荧光，含VIP或AVP能神经元呈现出红色荧光，既含PRV又含VIP或AVP的神经元即双标细胞呈现出桔黄色荧光?
To further explore the relationship between the terminal axon of retinal ganglion cells and the VIP and AVP neurons in the suprachiasmatic nucleus (SCN), a double-stranded immunofluorescence double-stranded immunoassay with pseudorabies virus (PRV) was performed. PRV (Bartha strain, 5 × 10 8 PFU / ml) 3 PI was slowly infused into unilateral eyeballs of adult rats, and the needles were left for 10 minutes. Animals survived 56, 72 and 96 hours respectively. 1% pentobarbital sodium intraperitoneal anesthesia, left heart left perfusion saline (100 units per 100ml containing heparin 100) for 5 minutes, the flow rate of 30-40ml / min, cold fixative (containing 4% paraformaldehyde, 15% Picric acid, prepared with 0 IM / L phosphate buffer, PH7’4), the initial flow rate of friends ~ 40ml minutes for 5 to 7 minutes, then slowed down to 10ml / min, maintained for half an hour. Craniotomy, repair block, into the fixed solution for one hour, then transferred to ZO% sucrose phosphate buffer overnight, constant cold box section, the thickness of 30Pm. Immunofluorescence double standard method: 001PBS (pH 7.4) containing 03% Triton and 2% Donkey Serum (NDS) diluted and mixed with two primary antibodies, goat anti-PRV1: 2000, rabbit anti-VIP or AVP1: 2000, . Secondary antibodies were donkey anti-goat IgG (IRITC marker) 1: 100, donkey anti-rabbit IgG (FITC marker) 1:50, diluted with 0.01% MBS in 2% NDS, sh. The above steps were rinsed with 0.01MPBS 10’X3. Mounting piece, PBS glycerol packing. Double-labeled immunofluorescence operation in the dark room. Results; PRV-labeled cells showed green fluorescence, VIP or AVP-containing neurons showed red fluorescence, both PRV and VIP or AVP-containing neurons that double-labeled cells showed orange fluorescence?