FUNCTIONAL MORPHOLOGY AND FEEDING OF DONAX SERRA ROODING AND DONAX SORDIDUS HANLEY (BIVALVIA: DONACIDAE)

The functional morphology of Donax serra and D. sordidus fromSouth African beaches was examined, and comparative measurementsmade of the rate of water filtration. The two species differin the structure and ciliation of the ctenidia, the number andspacing of the ridges on the labial palps, the direction ofciliary currents on the surface of the mantle and visceral mass,and the length and coiling of the mid-gut. The maximum ratesof water filtration recorded showed similar relationships tosize for the two species, but in D. sordidus the rate declinedrapidly with time during the observations. These differencesindicate differences in feeding strategy related to differencesin distribution and behaviour of the two species on beaches.     

MICROEVOLUTION AND PHENOTYPIC PLASTICITY IN DONAX SERRA RODING (BIVALVIA: DONACIDAE) ON HIGH ENERGY SANDY BEACHES

Morphological differences between populations of the wedge clamDonax serra inhabiting two different coasts and biogeographicregions of South Africa were investigated. Both adults and juvenilesoccupied different positions along the beach gradient dependingon the coast: on the southeast coast adults occurred in themid intertidal and juveniles and recruits were low intertidalto subtidal; on the west coast the zonation pattern was reversed.Not only adults but also juvenile clams had shapes differingsignificantly between the two coasts; west coast clams werethinner, rounder and had a higher body density than the southeastones-recruits were less dense in the former coast. Differencesin shell shape between coasts are probably the result of directionalselection on the adults with the microevolutionary changes beingmaintained by geographical isolation. Shell density, on theother hand, seems to be environmentally determined through physiologicalcontrol of shell calcification, i.e. more mobile intertidalclams having lower shell density than less mobile subtidal clams.Ontogenetic changes in shape and density are presumably adaptiveand appear to be related to mobility, i.e. the larger, heavierand denser adults being more stable in the substrate, and thesmaller, thinner and less dense juveniles being more mobileand dispersive. Phenotypic plasticity in present D. serra populationsis an important factor that enabled this species to occupy differenthabitats and biogeographic regions and to survive 5 millionyears of environmental changes. ¹ Present address: Laboratory for Ecology and Aquaculture, ZoologicalInstitute, Katholic Universiteit, Leuven, Naamsestraat 59 B-3000,Leuven, Belgium. E-mail: agsoares{at}eudoramail.com (Received 15 October 1997; accepted 5 January 1998)     

丙烯酰胺非整倍体诱发效应的荧光原位杂交和CREST染色的研究

本文以小鼠着丝粒次要卫星DNA探针FISH和抗着丝粒CREST染色,研究了可疑的非整倍体毒剂丙烯酰胺(AA)诱导的小鼠NIH3T3细胞微核(MN)的着丝粒组成情况和小鼠骨髓染色体畸变(CA)情况。结果发现AA在100—400μg/ml诱导的MN约52.7％—71.6％为FISH阳性,60.5％—68.2％的MN为CR-EST阳性,两种结果均显示AA具有较强的非整倍体诱发效应。小鼠骨髓CA的FISH表明,AA既能诱导染色体结构畸变,又能诱导非整倍体形成,而以非整倍体诱发效应更为明显。     

RAPID DETECTION OF GERMINATING BACILLUS CEREUS CELLS USING FLUORESCENT IN SITU HYBRIDIZATION

Methods for the specific detection of Bacillus spores are needed in many situations such as the recognition of food poisoning. This study presents an experimental design in order to find the best combination of germination conditions leading to a rapid and detectable fluorescent in situ hybridization (FISH) signal from Bacillus cereus spores present in pure cultures and milk samples.
B. cereus ATCC 14579 and HER 1414 were incubated in 20 different growth media by using a combination of various germinants such as sugars, amino acids and dipicolinic acid. Also, three different germination factors were tested: incubation temperature, inoculum concentration and a heat shock treatment. Permeabilization procedure and hybridization time were optimized on the best germination condition found. B. cereus -specific FISH probes were validated under the optimized condition and in detection of spiked B. cereus spores in 1% ultra heat-treated milk samples. FISH-labeled cells were detected by using flow cytometry, and the results were confirmed by fluorescence microscopy. The optimal condition allows the detection of B. cereus spores in less than 2 h. Overall, a ninefold reduction in total time for detection was achieved when comparing with previous works. Therefore, the permeabilization and hybridization optimizations mentioned in this study are major improvements for the detection time of B. cereus spores.

PRACTICAL APPLICATIONS

By using the optimized conditions of germination/outgrowth, permeabilization and hybridization, the detection of 10³ cfu/mL of Bacillus cereus spores using fluorescent in situ hybridization is possible within 2 h in milk sample.     

利用原位杂交技术检测免疫球蛋白重链mRNA在T和B淋巴瘤内的表达

应用原位杂交技术和免疫组织化学法对８例Ｂ淋巴瘤和４例Ｔ淋巴瘤进行了ＩｇＨｍＲＮＡ和ＩｇＭ,κ,λ的检测。结果显示,８例Ｂ淋巴瘤ＩｇＨｍＲＮＡ均为阳性,位于胞核周围,有的ｍＲＮＡ阳性反应成点状位于胞质的一侧。７例胞质中ＩｇＭ,κ或λ为阳性。１例胞质内无ＩｇＭ,κ或λ,但胞核内ＩｇＨｍＲＮＡ阳性反应成点状分布,可能是初级ＲＮＡ转录。在Ｔ淋巴瘤中,只有一例胞核内出现初级ＲＮＡ转录,因为早期Ｔ淋巴瘤,也有Ｄ、Ｊ基因片段的重排     

转基因水稻中外源基因的荧光原位杂交(FISH)分析

利用荧光原位杂交(FISH),在供试8个转基因水稻株系的染色体上检出了转入的外源基因barnase-ps1片段.其中两株系各检出了两个不同的信号位点,而其他株系均只检出一个信号位点.所有染色体着丝粒区都没有杂交信号.结合Southern杂交及表达分析发现,大多数株系中,整合在染色体端部区的barnase基因表现为高水平表达,而靠近着丝粒区的则表现为低水平表达,表明表达水平与转基因整合位置可能存在一定程度的相关性.表达水平与拷贝数无明显相关.本文还利用多色荧光原位杂交(Multi-color FISH)技术,进行了barnaseps1片段与报道基因pHctinG的共杂交,多数情况下,barnase-ps1片段与报道基因整合在染色体的同一位置.     

原位杂交和原位PCR技术在鱼类基因定位中的应用

染色体原位杂交（ ｉｎ ｓｉｔｕ ｈｙｂｒｉｄｉｚａｔｉｏｎ, ＩＳＨ）是基因物理定位的主要方法之一,它根据核酸分子碱基互补配对的原理,将标记的外源核酸探针与染色体上变性处理后的单链ＤＮＡ互补配对,再经检测而将靶顺序在染色体上的位置显示出来。 原位ＰＣＲ技术是将原位杂交与ＰＣＲ技术有机的结合,在原位扩增目的ＤＮＡ片段,并在原位检测其扩增产物,因而兼有ＰＣＲ及原位杂交技术二者的优越性：既可以同时获得组织及细胞的形态结构信息与分子信息,进行定性、定位分析,又具有比原位杂交更好的敏感性与专一性。 染色体原位…     

COMPARATIVE SPERMATOLOGY OF THREE SPECIES OF DONAX (BIVALVIA) FROM SOUTH AFRICA

The fine structure of the sperm and spermatogenesis in threespecies of Donax (D. madagascariensis, D. sordidus and D. serra)are described. Although the morphology of the sperm of all speciesis very similar, each has unique features. Donax madagascariensisand D. sordidus reportedly hybridize in regions of sympatryand their spermatozoa are morphologically closer to one anotherthan to D. serra. All sperm are of the primitive type with ahead(about 2 µmu; long), mid-piece of four mitochondria andtail. The head comprises a barrel-shaped nucleus which is cappedby a small, complex acrosome. The structure of the acrosomeis typical of heterodont bivalves. During spermatogenesis thepattern of nuclear chromatin condensation is granular. Glycogenfirst appears in the cytoplasm of spermatids, and in the maturesperm is sited in the mid-piece and base of the acrosome. (Received 15 May 1989; accepted 25 June 1989)     

FISH技术在微生物生态学中的研究及进展

分子生物学技术在微生物生态学研究中具有灵敏、精确和快速的优势,但不能提供微生物的形态学、数量性状、空间分布等信息。荧光原位杂交技术结合了分子生物学的精确性和显微镜的可视性信息,可以在自然生境中监测和鉴定不同的微生物个体,尤其是对难培养和未被培养的微生物进行检测。荧光原位杂交技术被广泛用于微生物群落结构诊断和评价,现已成为微生物分子生态学研究中的热点技术。对荧光原位杂交技术的发展和在微生物分子生态学中的应用进行了综述,探讨了该技术应用中存在的问题和发展前景。     

神经坏死病毒在赤点石斑鱼组织中的分布

研究采用地高辛原位杂交和免疫荧光检测技术分别检测病毒RNA2和衣壳蛋白在患病赤点石斑鱼Epinephelus akaara稚鱼中分布。地高辛检测结果表明病毒RNA2主要分布在脑、脊髓、视网膜和鳃上; 免疫荧光检测结果和地高辛检测结果一致, 表明病毒靶器官主要也是脑、脊髓、视网膜和鳃。肠道中几乎检测不到阳性信号, 可能不是病毒的靶器官。因此可以推测神经坏死病毒感染赤点石斑鱼的主要途径是鳃, 而不是肠道。       

食管鳞癌人乳头状瘤病毒感染的原位杂交检测和观察

目的 :检测食管鳞癌中人乳头状瘤病毒 (HPV)的感染情况 ,探讨 HPV与食管癌发病的关系。方法 :用原位杂交技术及免疫组化方法对 33例食管鳞癌手术切除标本进行 HPV检测。结果 :用原位杂交方法检测食管鳞癌中 HPV DNA检出率为 4 5 .5 % (15 / 33)。HPV DNA阳性与食管鳞癌肿瘤细胞的分化程度无关 (P>0 .0 5 )。HPV衣壳蛋白的免疫组化检测均为阴性。结论 :HPV感染可能是引起食管上皮癌变的原因之一 ,与食管鳞癌的发生有关。即使在存在 HPV DNA的病例中 ,HPV衣壳蛋白亦可以不表达     

肾上腺髓质素在正常大鼠脑中的分布一免疫组化及原位杂交分析

目的验证大鼠脑内是否存在ADM及其mRNA。方法取10只健康雄性SD大鼠,体重200-250g,应用免疫组织化学法和原位杂交组织化学法检测正常大鼠脑内ADM及其mRNA的表达情况。结果在大鼠脑内ADM及其mRNA阳性细胞主要表达在大脑皮质、海马结构、齿状回、丘脑、室旁组织、脉络丛、室管膜细胞、基底节、血管内皮细胞,其中脉络丛、室旁组织、丘脑为高表达区,其次为大脑皮质、海马。在大鼠大脑内ADMmRNA的表达与ADM阳性细胞的表达相一致。结论ADM及其mRNA在大脑内广泛表达,提示ADM在中枢神经系统内具有重要的作用。     

应用原位杂交技术检测鼻咽癌组织中bcl-2基因断裂点

为探讨ｂｃｌ┐２基因断裂与鼻咽癌发生的关系,采用分子原位杂交技术检测４１例鼻咽癌组织ｂｃｌ┐２基因的两个断裂热点。结果显示４１例鼻咽癌发生ｂｃｌ┐２基因断裂有４例,阳性率为９．８％,各组织学分级间无明显差异。鼻咽良性病变为阴性。结果说明ｂｃｌ┐２基因断裂非淋巴瘤的特异性改变,ｂｃｌ┐２基因断裂与鼻咽癌细胞的分化程度和恶性生物学行为间无明显关系,在鼻咽癌发生过程中的作用有限     

用碘标记核酸检查鼻咽癌上皮细胞中的EB病毒基因

用同位素碘化钠直接标记核酸。以细胞内原位杂交实验检查鼻咽癌上皮细胞中的EB病毒基因。共检查病理确诊的10例鼻咽癌活检标本的细胞涂片,其中9例直接证实有EB病毒核酸。本文建立并改进了细胞内原位核酸杂交技术,首先使用重组的单链DNA作探针,并将碘标记的核酸用于原位核酸杂交实验。     

植物染色体原位杂交技术的发展与应用

植物染色体原位杂交技术的发展与应用奇文清,李懋学（北京大学生命科学学院北京１００８７１）ＴＨＥＤＥＶＥＬＯＰＭＥＮＴＡＮＤＡＰＰＬＩＣＡＴＩＯＮＳＯＦＩＮＳＩＴＵＨＹＢＲＩＤＩＺＡＴＩＯＮＴＥＣＨＮＩＱＵＥＩＮＰＬＡＮＴＣＨＲＯＭＯＳＯＭＥＳ￥ＱｉＷ...     

用非放射性原位杂交和免疫金染色在豌豆间期染色质内显示的rDNA的分布

以异羟基洋地黄毒甙标记的ｒＲＮＡ为探针,通过与植物根尖分生组织区振动切片中的ｒＤＮＡ原位杂交,结合免疫金染色及银加强金处理研究了豌豆间期染色质中ｒＤＮＡ的分布。研究结果清楚地显示供试植物间期细胞内有１－５个显著的ｒＲＮＡ探针结合位点,其中显示４个结合位点的细胞占６０．６％,少于和多于４个结合位点的细胞分别上３７．９％和１．１４％。在显示４个结合位点的细胞核内,它们分别按照２：１、３：１和２：１：１     

大鼠颌下腺促性腺激素释放激素及其mRNA的免疫组织化学与原位杂交研究

用免疫组织化学及原位杂交法,研究了促性腺激素释放激素及其ｍＲＮＡ在大鼠颌下腺的分布。结果显示,大鼠颌下腺的浆液性腺泡的上皮细胞,各级导管的上皮细胞及副交感神经节细胞均呈促性腺激素释放激素免疫反应阳性,阳性反应物质分布在胞质,胞核呈阴性反应。颌下腺的浆液性腺泡上皮细胞,各级导管上皮细胞同样被检测到很强的促性腺激素释放激素ｍＲＮＡ杂交信号。以上结果提示,大鼠颌下腺能自身合成促性腺激素释放激素,促性腺激素释放激素对消化功能可能有重要调节作用。     

大鼠甲状腺促性腺激素释放激素受体的免疫组织化学及原位杂交研究

目的 探讨大鼠甲状腺中是否存在促性腺激素释放激素受体（GnRH-R)及其细胞定位,方法 收集15例雄性SD大鼠甲状腺,分别制成石蜡切片和冰冻切片,采用免疫组织化学ABC法和原位杂交技术。确定GnRH-R在其中的表达与定位。结果 大鼠甲状腺中,GnRH-R呈较强的免疫反应阳性,阳性物质分布在胞持,胞核呈阴性,原位杂交也检测到较强的GnRH-RmRNA阳性杂交信号,亦分布在胞质,胞核未见表达,结论 大鼠甲状腺可能自身合成GnRH-R。由此推断GnRH可能参与大鼠甲总而言之 腺功能的调节。     

原位杂交检测石蜡包埋组织中丙型肝炎病毒RNA

为了提高ＨＣＶＲＮＡ的原位杂交检出率,我们应用地高辛标记ＨＣＶ５＇非编码区ｃＤＮＡ作探针,采用原位分子杂交法对９６Ｎ肝硬变,１０２例肝细胞肝癌进行了ＨＣＶＲＮＡ检测,并结合不同年份标本中ＨＣＶＲＮＡ的检出率,探讨ＨＣＶＲＮＡ在肝脏的分布及其丢失问题．结果显示ＨＣＶＲＮＡ定位于肝细胞和肝癌细胞胞浆内,肝脏其它细胞未见明确阳性杂交信号。ＨＣＶＲＮＡ杂交阳性细胞呈灶状和弥漫分布,正常对照、替代、空白对照为阴性,在不同年份肝硬变及肝细胞肝癌中两两比较表明新近包埋蜡块组织较陈旧蜡块组织ＨＣＶ　ＲＮＡ检出率明显高。本文结果证实ＨＣＶＲＮＡ存在于肝细胞和癌细胞的胞浆内,未见肝内其它细胞阳性,还发现ＨＣＶＲＮＡ在肝硬变、肝细胞癌中的检出率随保存时间的延长,其阳性检出率明显降低,表明ＨＣＶＲＮＡ在蜡块中存在明显的降解,应尽可能选用近期标本、为临床分析ＨＣＶ感染,ＨＣＶ与肝硬变、肝细胞肝癌的关系提供更客观的数据。     

用非放射性原位杂交在同一张切片上同时在光学和电子显微镜下显示的豌豆rDNA的分布(简报)

原位杂交目前已成为研究各类组织或细胞内DNA或RNA定位最有效的手段,特别是在发育生物学领城,对于研究基周的表达和修饰更具有重要意义。旱期的原位杂交主要是在