5-氟尿嘧啶多克隆抗体的制备和鉴定

目的制备5-氟尿嘧啶(5-FU)免疫原并制备其多克隆抗体。方法采用化学合成法对5-FU进行修饰制备5-氟尿嘧啶-1-基乙酸半抗原(5-FUAA),并经碳酰二亚胺盐法将其分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)连接以制备免疫原。采用5-FU与BSA结合的免疫原(5-FUAA-BSA)免疫BALB/c小鼠制备多克隆抗体,5-FU与OVA结合物(5-FUAA-OVA)包被酶标板经间接ELISA检测抗血清效价,并采用ELISA与Western blot法检测抗血清的特异性。结果成功合成半抗原5-FUAA,并分别制备5-FUAA-BSA和5-FUAA-OVA完全抗原。将5-FUAA-BSA免疫BALB/c小鼠制备的免疫血清效价达1∶1 280 000,且该多克隆抗体能特异地结合5-FU半抗原。结论成功制备了5-FU的多克隆抗体。     

5-氟尿嘧啶缓释剂制备及体外释药性能比较*

背景：5-氟尿嘧啶-聚乳酸-乙醇酸共聚物缓释微球在青光眼滤过术后抑制滤过泡的瘢痕化具有潜在应用价值,但微球制备程序复杂,微球载药量一般较低,且药物突释现象明显。 目的：比较乳化溶剂挥发法制备的5-氟尿嘧啶-聚乳酸-乙醇酸共聚物微球和喷雾成膜法制备的5-氟尿嘧啶-聚乳酸-乙醇酸共聚物缓释膜两种缓释剂的形态、载药量、体外释放规律,以探讨获得缓释效果较佳的5-氟尿嘧啶缓释剂制备方法。 方法：以聚乳酸-乙醇酸共聚物为载体,采用乳化溶剂挥发法制备5-氟尿嘧啶-聚乳酸-乙醇酸共聚物微球;用喷雾成膜法制备5-氟尿嘧啶-聚乳酸-乙醇酸共聚物缓释膜。 结果与结论：用乳化溶剂挥发法制备的微球外观圆整,粒径为(4 447.4±359.8) nm,载药量(8.67±0.37)%,包封率为(86.68± 1.92)%;用喷雾成膜法制备的缓释膜表面光滑平整,质量为(13.76±0.26) mg ,直径为6 mm ,厚度为(0.24±0.005) mm,载药量(23.76±0.37)%,包封率为(95.04±1.36)%。缓释剂制备过程未影响5-氟尿嘧啶的药物性能。微球体外释放突释明显,缓释膜的体外释放平稳持久,释放曲线符合Higuchi方程。结果表明缓释膜制备方法更简单易行,且能明显提高缓释剂的载药量,降低突释现象,同时延长药物的缓释时间。 关键词：聚乳酸-聚乙醇酸;5-氟尿嘧啶;微球;缓释膜;体外释放 doi:10.3969/j.issn.1673-8225.2012.08.022     

靶向抑制晶状体上皮细胞增殖的载药毫微球的研究

采用碳二亚胺法使抗人晶状体上皮细胞单克隆抗体与聚乳酸载 5 -氟尿嘧啶毫微球偶联 ,制备出抗人晶状体免疫毫微球。以牛血清白蛋白作乳化剂采用复乳法制备聚乳酸载 5 -氟尿嘧啶毫微球 ,使毫微球表面吸附牛血清白蛋白 ,再将抗人晶状体上皮细胞单克隆抗体与聚乳酸载 5 -氟尿嘧啶毫微球偶联。采用扫描电镜和透射电镜观察微球形貌和结构 ,用动态光散射粒径分析仪测载药毫微球粒径。ELISA法检测偶联后的抗体活性 ,MTT法检测免疫毫微球对晶状体上皮细胞的抑制作用 ,间接免疫荧光法检测该免疫毫微球与晶状体上皮细胞的特异性结合能力和细胞内化过程。结果显示载药毫微球表面光滑 ,平均粒径为 191.0± 0 .2 0 2nm ,其载药率为 8.2 % ,药物利用率为2 4 .6 %。免疫载药毫微球中的单克隆抗体HILE6保留达到原免疫活性 84 % ,2 4h对兔晶状体上皮细胞抑制率可达6 8.4 2 %。该免疫载药毫微球能与晶状体上皮细胞特异性结合 ,30min后可吸附到晶状体上皮细胞上 ,在 4h内进入细胞质最后进入细胞核。该实验为特异性抑制晶状体上皮细胞增殖 ,预防白内障术后并发症 -后囊混浊提供了重要的科学依据。     

乳酸-羟基乙酸共聚物载药纳米粒的制备及体外释药性

背景：医用纳米粒作为药物传递的新型载体,目前已经成为医药领域研究的重点。 目的：构建以生物可降解材料乳酸-羟基乙酸共聚物为载体,负载抗肿瘤药物5-氟尿嘧啶的载药纳米粒。 方法：利用复乳-溶剂挥发法制备乳酸-羟基乙酸共聚物载药纳米粒。场发射扫描电子显微镜观察纳米粒表面形态;激光粒度分析仪测定粒径分布并计算成球率;紫外分光光度计测定5-氟尿嘧啶载药量、包封率,并对体外释药进行评估。 结果与结论：纳米粒呈球性,平均粒径为(186±14) nm,成球率、载药量和包封率分别为70.8%、6.6%、28.1%,体外释药有突释现象,24 h内5-氟尿嘧啶累积释药量达36.2%,10 d达83.6%。提示成功制备乳酸-羟基乙酸共聚物载药纳米粒,其具有缓释效应。     

磁性碳纳米管载附5-氟尿嘧啶对结肠癌淋巴结转移的靶向抑制作用☆

背景：将碳纳米管制备成磁靶向药物载体后,在磁场作用下能更好地把药物运送至体内靶器官或靶组织。 目的：观察磁性碳纳米管新化疗载体对结肠癌淋巴结转移的抑制效果。 方法：MTT法检测5-氟尿嘧啶、磁性多壁碳纳米管-5-氟尿嘧啶、磁性多壁碳纳米管(每种药物分别含0.000 3,0.003,0.03,0.3,3 g/L 5-氟尿嘧啶)对结肠癌SW480细胞的抑制作用。将含相同浓度5-氟尿嘧啶的5-氟尿嘧啶、磁性多壁碳纳米管-5-氟尿嘧啶、磁性多壁碳纳米管分散液分别注入SD大鼠足垫皮下及结肠癌淋巴结转移裸鼠体内。 结果与结论：体外各质量浓度的5-氟尿嘧啶和磁性多壁碳纳米管-5-氟尿嘧啶对癌细胞的毒性存在剂量依赖性,相同5-氟尿嘧啶质量浓度时两者对体外SW480细胞的抑制作用无明显差异,说明磁性多壁碳纳米管-5-氟尿嘧啶的主要药效成分为5-氟尿嘧啶。体内研究高效液相检测显示磁性多壁碳纳米管-5-氟尿嘧啶能有效聚集在淋巴结,长时间持久释放,淋巴结浓集效果明显优于5-氟尿嘧啶(P < 0.05),且不良反应小,肉眼容易辨识,细胞穿透性好;TUNNEL检测见磁性多壁碳纳米管-5-氟尿嘧啶化疗后结肠癌淋巴结转移灶细胞有明显凋亡现象,在磁场作用下效果更显著。说明磁性多壁碳纳米管-5-氟尿嘧啶对结肠癌SW480细胞淋巴结转移有明显抑制作用。关键词：磁性碳纳米管;5-氟尿嘧啶;淋巴化疗;结肠癌;淋巴结转移;生物材料与纳米技术 缩略语注释：5-FU：5-fluorouracil,5-氟尿嘧啶;mMWNTs：multi walled carbon nanotubes,磁性多壁碳纳米管 doi:10.3969/j.issn.1673-8225.2012.16.008      

二氢青蒿素通过抑制SIRT1的表达而增强5-氟尿嘧啶对胃癌的抗肿瘤活性

目的:探讨二氢青蒿素对5-氟尿嘧啶治疗胃癌的辅助作用并研究其机制。方法:实验分为对照组、二氢青蒿素组、5-氟尿嘧啶组、5-氟尿嘧啶联合二氢青蒿素组和5-氟尿嘧啶+二氢青蒿素+SIRT1质粒组。MTT法检测胃癌细胞系BGC-823在5-氟尿嘧啶联合二氢青蒿素处理下的细胞活力。Western blot实验检测5-氟尿嘧啶联合二氢青蒿素对BGC-823细胞SIRT1和NADPH氧化酶表达水平,caspase-9和caspase-3活化水平及凋亡信号调节激酶1(ASK1)和c-Jun氨基末端激酶(JNK)蛋白磷酸化水平的影响。流式细胞术检测BGC-823细胞在5-氟尿嘧啶和二氢青蒿素联合处理下的活性氧簇(ROS)生成水平和细胞凋亡率。结果:二氢青蒿素处理能显著抑制BGC-823细胞SIRT1的表达并增加NADPH氧化酶的蛋白水平,明显提高BGC-823细胞对5-氟尿嘧啶的敏感性,降低5-氟尿嘧啶的半数抑制浓度;转染SIRT1表达质粒后,二氢青蒿素联合5-氟尿嘧啶对BGC-823细胞的杀伤活性受到显著抑制(P0.05)。二氢青蒿素能明显促进5-氟尿嘧啶对BGC-823细胞生成ROS的诱导效应和ASK1及JNK的磷酸化(P0.05)。用ROS清除剂N-乙酰半胱氨酸(NAC)或JNK特异性抑制剂SP600125处理后,二氢青蒿素联合5-氟尿嘧啶对BGC-823细胞的杀伤活性和caspase-9及caspase-3的活化均受到明显抑制(P0.05)。另外,NAC能显著抑制二氢青蒿素联合5-氟尿嘧啶对JNK磷酸化的促进作用,而SP600125却不能影响BGC-823细胞ROS的产生,表明JNK是ROS的下游分子。结论:二氢青蒿素联合5-氟尿嘧啶通过SIRT1/NADPH氧化酶/ROS/JNK通路诱导胃癌细胞发生caspase依赖的凋亡。     

胃癌单克隆抗体-氨甲喋呤交联物的制备及其细胞毒作用的研究

本文用本室已研制出12株鼠抗人胃癌单抗中一株MGb_2制备抗体-BSA-氨甲喋呤结合物,探讨所选单抗在胃癌导向治疗的可行性。 BSA与5倍过量的SPDP反应,每克分子BSA中引入3克分子二硫吡啶基团(PDP)。PDP-BSA通过碳二亚胺与氨甲喋呤连接。PDP-BSA-MTX以二硫代苏糖醇还原得到HS-BSA-MTX。     

miR-449b介导5-氟尿嘧啶/顺铂抑制肝癌细胞的迁移

目的研究化疗药5-氟尿嘧啶和顺铂对miR-449b在肝癌细胞中表达的影响及其抑制肝癌细胞迁移能力的分子机制。方法收集肝癌患者的组织样本,提取总RNA,用real-time PCR法检测肝癌组织样本中miR-449b的表达;5-氟尿嘧啶和顺铂处理肝癌细胞,检测miR-449b的表达;在肝癌细胞中过表达miR-449b或用5-氟尿嘧啶和顺铂分别处理肝癌细胞,利用细胞划痕实验检测肝癌细胞的迁移能力变化;设计拯救实验,探究化疗药5-氟尿嘧啶和顺铂通过诱导miR-449b表达,参与肝癌细胞迁移能力调控;软件预测结合Western blot实验,鉴定miR-449b的功能靶基因。结果 miR-449b在肝癌患者组织中表达下调(P0.001);5-氟尿嘧啶和顺铂诱导肝癌细胞内源性miR-449b的表达上调(P0.001);过表达miR-449b可以抑制肝癌细胞的迁移(P0.001);敲低miR-449b的表达可以抑制5-氟尿嘧啶和顺铂对肝癌细胞迁移能力的抑制作用(P0.05);catenin-δ是miR-449b的直接靶基因。结论5-氟尿嘧啶和顺铂通过诱导miR-449b的表达抑制肝癌细胞的迁移。     

14型肺炎球菌荚膜多糖-破伤风类毒素结合疫苗研究

目的：探讨制备肺炎球菌荚膜多糖（PNCPS）－蛋白结合疫苗适宜条件。方法：采用碳二亚胺法将14型PNCPS与破伤风类毒素（TT）结合，将结合物免疫NIH小鼠，用ELISA法检测小鼠血清中抗14例PNCPS的IgG抗体滴度。结果：结合反应产率和结合物中多糖，蛋白含量的测定显示试验成功合成了14型PNCPS－TT结合物，该结合物具有14型PNCPS的血清学特异性，将之免疫小鼠诱生的抗14型PNCPS特异性IgG抗水平显著高于单纯14型PNCPS。结论：试验成功地合成了14型PNCPS－TT结合物，本试验采用的结合方法可行。     

白藜芦醇通过促进Apaf-1/caspase-9复合物的形成增强5-氟尿嘧啶对骨肉瘤CD133~+细胞亚群凋亡的诱导

目的:探讨白藜芦醇联合化疗药物5-氟尿嘧啶对骨肉瘤CD133~+细胞亚群的协同杀伤效应及机制。方法:将人骨肉瘤细胞系MG-63 CD133~+细胞亚群及相应CD133~-细胞亚群用5-氟尿嘧啶及白藜芦醇进行体外处理。MG-63细胞的相对细胞活力用MTT法进行检测;细胞凋亡率用流式细胞术进行检测;用Western blot实验检测caspase-9和caspase-3的活化、Apaf-1的表达水平及细胞色素C的释放;用免疫共沉淀法检测Apaf-1与caspase-9前体的相互作用。结果:5-氟尿嘧啶对MG-63 CD133~+细胞亚群的杀伤活性和凋亡诱导活性均显著低于MG-63 CD133~-细胞亚群。但联用白藜芦醇能显著提高5-氟尿嘧啶对MG-63 CD133~+细胞亚群的细胞活力抑制率。白藜芦醇处理能显著上调MG-63 CD133~+细胞亚群中Apaf-1的表达水平,在MG-63 CD133~+细胞亚群中转染Apaf-1siRNA后,5-氟尿嘧啶联用白藜芦醇的协同效应受到显著抑制。另外,免疫共沉淀实验结果表明白藜芦醇联合5-氟尿嘧啶能显著诱导MG-63 CD133~+细胞亚群中Apaf-1/caspase-9复合物的形成,从而诱导caspase-9发生活化。结论:白藜芦醇通过促进Apaf-1/caspase-9复合物的形成,增强5-氟尿嘧啶对骨肉瘤CD133~+细胞亚群凋亡的诱导。     

The exposed carbohydrates of schistosomula of Schistosoma mansoni and their modification during maturation in vivo

Lectins labeled with 125I or conjugated with fluorescein were employed to study the carbohydrates on the surface of different stages of schistosomula of Schistosoma mansoni. Newly transformed schistosomula were shown to bind concanavalin A; the 60 000 and 120 000 dalton agglutinins from Ricinus communis; the fucose-binding protein from Lotus tetragonolobus; wheat germ agglutinin and peanut agglutinin. Soybean agglutinin, Ulex europaeus agglutinin and Dolichos biflorus agglutinin, on the other hand, failed to bind to the schistosomulum surface. The binding of peanut and soybean agglutinin was unaffected by pretreatment of the parasites with neuraminidase. Binding of concanavalin A, the 120 000 dalton agglutinin from Ricinus communis, wheat germ agglutinin and peanut agglutinin to the surface of 5-day schistosomula, recovered from the lungs of mice, was also demonstrated. In each case, however, the level of binding was approximately 70% less than that observed with newly transformed schistosomula and the binding of the fucose-binding protein from L. tetragonolobus practically disappeared. In contrast with newly transformed schistosomula, lung stage schistosomula, pretreated with neuraminidase, displayed a significant increase in the binding of peanut and soybean agglutinin. The results indicate that a significant alteration in the surface carbohydrates of S. mansoni occurs during in vivo maturation of the parasite. This change may contribute to the organism's ability to survive in the vertebrate host.     

The Binding of Different Lectins on Peripheral Blood Mononuclear Cells from Patients with Chronic Inflammatory and Malignant Diseases

Peripheral blood mononuclear cells from patients with multiple myeloma, gastrointestinal tumors, and inflammatory bowel disease were analyzed for binding of various lectins. The results demonstrated that in most of the patients with multiple myeloma a significantly increased percentage of cells positive for Lotus tetragonolobus agglutinin (LTA), peanut agglutinin (PNA), soybean agglutinin (SBA), and wheat germ agglutinin (WGA), and a decreased number of Agaricus bisporus agglutinin (ABA) positive cells were present as compared to a normal control group. This could not be shown in malignant or inflammatory disorders of the gastrointestinal tract where only some patients exhibited an increased PNA and LTA binding, respectively. Patients with the systemic malignant disease differed from patients with solid localized tumors by a significantly altered number of ABA, LTA and SBA-positive peripheral blood mononuclear cells. Double fluorescence studies using monoclonal antibodies and lectins revealed that most of the cells expressing receptors for ABA had also receptors for OKT3, whereas most of the cells with receptors for LTA, PNA SBA, and WGA were found to be positive for OKM.     

Lectin binding to glycoproteins in the surface membrane of Schistosoma mansoni

A number of lectins were assessed for their ability to bind to glycoproteins in the surface membrane of Schistosoma mansoni. The membrane polypeptides were separated by SDS-PAGE and the glycoproteins visualised by incubating the gel with radio-iodinated lectin followed by autoradiography. Most of the individual lectins bound to a variety of glycoproteins but peanut agglutinin and Dolichos biflorus agglutinin bound preferentially to a single glycoprotein of apparent molecular weight 170 000. This glycoprotein was subsequently shown to be exposed at the surface of the parasite and localised at the tubercles.     

Lectin-HPMA copolymer conjugates: potential oral drug carriers for targeting diseased tissues

N-(2-Hydroxypropyl)methacrylamide (HPMA) copolymer-lectin conjugates were investigated for potential use as targeted oral drug carriers for treatment of inflammatory conditions such as colitis. Wheat germ agglutinin (WGA)-HPMA copolymer and peanut agglutinin (PNA)-HPMA copolymer and fluorescein isothiocyanate (FITC)-labeled WGA- and PNA-HPMA copolymer conjugates were synthesized. Conjugate dissociation constants (K_d) for lectin-carbohydrate binding determined by frontal affinity chromatography indicated that no activity reduction of the lectins occurred during the synthesis of these conjugates. K_d values measured were in good agreement with literature findings for similar lectin-carbohydrate interactions, on the order of 10⁻⁵ M⁻¹. Biorecognition of these conjugates by healthy rat intestinal tissue resulted in differential HPMA copolymer-lectin conjugate binding patterns in the same tissue. HPMA copolymer-WGA conjugate showed strong binding in the healthy rat intestinal tissues, while the HPMA copolymer-PNA conjugate showed minimal, but specific binding. This differential binding suggests that site-specific drug delivery via specific lectin recognition may be feasible for treatment of colon inflammation or cancer.     

Concanavalin A binding sites of rough endoplasmic reticulum containing intracisternal microtubules in canine neurones

Lectin histochemistry was used to identify sugar residues of IM-containing RER in elderly canine sympathetic ganglionic neurones. IM-inclusions stained with ConA-peroxidase conjugate, but not with soybean agglutinin (SBA), wheat germ agglutinin (WGA), peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA), Ricinus communis agglutinin (RCA I) and Ulex europaeus agglutinin (UEA I). ConA-binding sites were visualized within cisternae of RER containing IM; reaction product was localized in IM-containing RER cisternae and IM. Inhibition with specific sugars (0.1 M alpha-methyl-D-mannoside or 0.5 M D-glucose) blocked the binding of ConA to IM-inclusions and normal Nissl substance. When a low sugar concentration (5 x 10(-3) M alpha-methyl-D-mannoside or 0.2 M D-glucose) was employed, IM-inclusions were still strongly ConA-positive, but normal Nissl substance was not. These results demonstrate that IM-containing RER have an excessive amount of carbohydrates (mannose or glucose-rich sugars) which are essentially detected in flattened RER under normal conditions and further indicate that glycoproteins in IM differ from those in cytoplasmic microtubules.     

5-Fluorouracil–lipid conjugate: Potential candidate for drug delivery through encapsulation in hydrophobic polyester-based nanoparticles

The encapsulation of 5-fluorouracil (5-FU) in hydrophobic polymeric materials is made feasible by a lipid-based prodrug approach. A lipid–5-FU conjugate of 5-FU with palmitic acid was synthesized in two-step process. A synthesized dipalmitoyl derivative (5-FUDIPAL) was characterized using Fourier transform infrared spectroscopy and ¹H-nuclear magnetic resonance. The 5-FUDIPAL was encapsulated in polyester-based polymers by the double emulsion–solvent evaporation method. The nanoparticles were characterized by scanning electron microscopy, transmission electron microscopy and dynamic light scattering. The thermal stability was assessed by differential scanning calorimetry data. In vitro release kinetics measurements of the drug from nanoparticles showed the controlled release pattern over a period of time. Cytotoxicity measurements by MTT assay confirmed that dipalmitoyl derivative in nano formulation successfully inhibited the cell growth. Thus the combined physical and biological evaluation of the different polyester-based nanoparticle containing the modified drug showed a facile approach to delivering 5-FU to the tumour site with enhanced efficacy.     

Adenosine deaminase activity in lymphocyte subpopulations of B-16 melanoma and normal C57BL bearing mice

The activity of adenosine deaminase (ADA) was measured in thymus and spleen subpopulations separated by peanut agglutinin (PNA) of melanoma B-16 C57BL bearing mice and normal age-matched C57BL mice. Groups of 10 mice were used each time and the experiments were repeated 6 times. The adenosine deaminase activity in the PNA+ thymocytes of B-16 bearing mice was about 2.5 times lower than that of the normal C57BL mice while the ADA activity in the PNA+ fraction of spleen of the B-16 melanoma bearing mice was 2.5 times higher. These results demonstrate that the tumor burden probably induces a different redistribution and traffic of lymphocytes from one lymphopoietic organ to another. This traffic can also explain the thymus involution and spleen enlargement found in the B-16 mice.     

Cholinesterases and peanut agglutinin binding related to cell proliferation and axonal growth in embryonic chick limbs

Embryonic cholinesterases are assigned important functions during morphogenesis. Here we describe the expression of butyrylcholinesterase and acetylcholinesterase, and the binding of peanut agglutinin, and relate the results to mitotic activity in chick wing and leg buds from embryonic day 4 to embryonic day 9. During early stages, butyrylcholinesterase is elevated in cells under the apical ectodermal ridge and around invading motoraxons, while acetylcholinesterase is found in the chondrogenic core, on motoraxons and along the ectoderm. Peanut agglutinin binds to the apical ectodermal ridge and most prominently to the chondrogenic core. Measurements of thymidine incorporation and enzyme activities were consistent with our histological findings. Butyrylcholinesterase is concentrated near proliferative zones and periods, while acetylcholinesterase is associated with low proliferative activity. At late stages of limb development, acetylcholinesterase is concentrated in muscles and nonexistent within bones, while butyrylcholinesterase shows an inverse pattern. Thus, as in other systems, in limb formation butyrylcholinesterase is a transmitotic marker preceding differentiation, acetylcholinesterase is found on navigating axons, while peanut agglutinin appears in non-invaded regions. These data suggest roles for cholinesterases as positive regulators and peanut-agglutinin-binding proteins as negative regulators of neural differentiation.     

人红细胞老化过程中花生凝集素和Con A受体电镜细胞化学研究

目的　观察人血液中 ,老化红细胞膜上花生凝集素 (PNA)和刀豆凝集素 (Con A)受体细胞化学反应的变化。　方法　应用金标花生凝集素 (PNAg)和刀豆凝集素 (Con Ag) ,对早期和老化红细胞膜上 PNA和 Con A受体进行电镜细胞化学显示 ,并用计算机图像分析系统对电镜照片进行定量处理。　结果　与早期红细胞相比 ,老化红细胞膜上 PNAg颗粒和 Con Ag颗粒均有明显增多。　结论　老化红细胞膜上唾液酸的丢失导致半乳糖基和甘露糖基暴露的相应增加 ,这种变化可能与巨噬细胞对老化红细胞的识别和吞噬机制有关     

Lectin staining in the basal nucleus (Meynert) and the hypothalamic tuberomamillary nucleus of the developing human prosencephalon

Previous studies have demonstrated that extracellular matrix glycoconjugates, shown by lectin-histochemistry with Vicia villosa agglutinin (VVA) and peanut agglutinin (PNA) as so-called perineuronal nets, play an important role in brain maturation. Concanavalin A (ConA) binding to neuronal surface glycoconjugates may be a marker of synaptic junctions. The present study was done to demonstrate the binding sites of these lectins in two functionally related nuclei of the prosencephalon, the basal nucleus (Meynert) and the hypothalamic tuberomamillary nucleus. Fetal brains of 16–36 weeks of gestation were examined by using VVA, PNA, and ConA to determine appearance and distribution patterns of specific lectin-binding sites on glycoconjugates during fetal brain development. The basal nucleus and the tuberomamillary nucleus showed a characteristic “cellular staining” that may have been due to cytoplasmatic labeling, surface labeling, or both. Lectin-staining occurred much earlier in the basal nucleus than in the tuberomamillary nucleus. Although all three lectins were bound to neurons of the basal nucleus, only ConA-positive neurons were observed in the tuberomamillary nucleus. In conclusion, lectin-labeled cells most probably represent projection neurons that are GABAergic (tuberomamillary nucleus) or cholinergic (basal nucleus). Labeling with the three lectins demonstrated nuclear-specific staining patterns that occur early in fetal development and gradually increase. Binding sites for lectins characterizing perineuronal nets (VVA, PNA) occurred only in the basal nucleus, whereas binding sites for ConA on neuronal-surface glycoconjugates, which seem to play a role in early synaptogenesis, were present in the basal and the tuberomamillary nucleus. The basal nucleus, however, expressed ConA binding sites distinctly earlier, probably indicating early arriving afferents. Anat. Rec. 252:149–159, 1998. © 1998 Wiley-Liss, Inc.