Swelling and Ca2+-activated Anion Conductances in C127 Epithelial Cells Expressing WT and ΔF508-CFTR

CFTR is a chloride channel that is required for fluid secretion and salt absorption in many exocrine epithelia. Mutations in CFTR cause cystic fibrosis. CFTR expression influences some ion channels, but the range of channels influenced, the mechanism of the interaction and the significance for cystic fibrosis are not known. Possible interactions between CFTR and other ion channels were studied in C127 mouse mammary epithelial cell lines stably transfected with CFTR, ΔF508-CFTR, or vector. Cell lines were compared quantitatively using an ¹²⁵I efflux assay and qualitatively using whole-cell patch-clamp recording. As expected, ¹²⁵I efflux was significantly increased by forskolin only in the CFTR line, and forskolin-stimulated whole-cell currents were time- and voltage independent. All three lines responded to hypotonic challenge with large ¹²⁵I efflux responses of equivalent magnitude, and whole-cell currents were outwardly rectified and inactivated at positive voltages. Unexpectedly, basal ¹²⁵I efflux was significantly smaller in the ΔF508-CFTR cell line than in either the CFTR or control cell lines (P < 0.0001), and the magnitude of the efflux response to ionomycin was largest in the vector cell line and smallest in the cell line expressing ΔF508-CFTR (P < 0.01). Whole-cell responses to ionomycin had a linear instantaneous I-V relation and activated at depolarizing voltages. Forskolin responses showed simple summation with responses to ionomycin or hypotonic challenge. Thus, we found no evidence for interactions between CFTR and the channels responsible for swelling-mediated responses. Differences were found in basal and ionomycin-stimulated efflux, but these may arise from variations in the clonally selected cell lines that are unrelated to CFTR expression. Received: 15 November 1995/Revised: 16 February 1996     

Systematics of white-toothed shrews (Crocidura) (Mammalia: Insectivora: Soricidae) of Taiwan: Karyological and morphological Studies

We define the species boundaries of white-toothed shrews (genus Crocidura ) in Taiwan using karyological and morphological characteristics. Ninety-nine animals were obtained from all over Taiwan at capture rates usually less than 10%. Three species are recognized by distinct cytotypes: Crocidura attenuata tanakae 2n = 40, FN = 56; Crocidura suaveolens hosletti 2n = 40, FN = 50; Crocidura kurodai 2n = 40, FN = 54. A suite of six morphological characters diagnose the three species: shape of skull, position of incisive foramina, shape of fourth upper premolar, shape of pinna, tail vibrissae, and foot pads. A species key and notes on the life history of each species are provided. Finally, we discuss chromosomal evolution and biolgeography of Crocidura in East and South East Asia.     

广西爵床科植物新资料

本文报道广西爵床科３３个新分类群,４个新组合,８个中国新记录,１３个广西新记录,１种花的补充描述和１种花萼描述的修订     

黄鳝食性的初步研究

通过对不同产地的２７１尾野生黄鳝的食性研究表明,全长小于１００ｍｍ的稚鱼,其食性随着全长的生长而变化,稚鱼前期以摄食轮虫,枝角类为主,后期则以水生寡毛类,摇蚊幼虫为主,而全长１０１ｍｍ－２００ｍｍ的幼鳝及全长大于２００ｍｍ的成鳝的食性,随全长变化相对稳定,且细鳝和成鳝的食谱基本相同,其主要食物组成均包括摇蚊幼虫,水生寡毛类,蚯蚓,昆虫幼虫,枝角类和桡足类等。其肠内含物中的饵料生物种类含量均在季节变     

Diminishing adenovirus gene expression and viral replication by promoter replacement.

The adenovirus E4 promoter was replaced by a synthetic promoter composed of a minimal TATA box and five consensus 17-mer yeast GAL4-binding-site elements. The viral vectors, which also contained human factor IX (hFIX) cDNA driven by Rous sarcoma virus long terminal repeat in the E1 region, were then constructed and expanded in 293 cells permanently expressing GAL4/VP16 fusion protein. Viral replication and expression of adenovirus E4 genes and late genes (hexon and fiber) were evaluated in vitro in the human lung carcinoma cell line H1299. Viral replication and viral gene expression were dramatically reduced in the cells transduced by vectors with a replaced E4 promoter compared to the levels in the cells transduced by vectors with the wild-type E4 promoter. The levels of transgene (hFIX) expression remained similar between vectors with or without E4 promoter replacement. These results indicate that diminution of viral gene expression and viral replication is achievable by promoter replacement.     

Fingerprinting trifoliate orange germ plasm accessions with isozymes, RFLPs, and inter-simple sequence repeat markers

 Trifoliate orange [Poncirus trifoliata (L.) Raf.] is frequently used as a parent in citrus rootstock breeding, but the origin and amount of genetic diversity in germ plasm collections are poorly understood. Most accessions are self-compatible, but produce a mixture of sexual and apomictic seedlings. Variation among 48 vegetatively propagated trifoliate orange accessions was assessed at seven isozyme loci, together with the restriction fragment length polymorphisms (RFLPs) detected by 38 probe-enzyme combinations and the inter-simple sequence repeat (ISSR) markers generated by 11 primers. Isozymes and RFLPs detected few polymorphisms among accessions, although genetic analysis has shown that the common phenotype is heterozygous for four isozyme and at least four RFLP loci. ISSR amplification generated multiple banding profiles with an average of 58 fragments/primer/accession. These fragments were repeatable across DNA samples extracted from different trees of the same accession or extracted at different times, and across separate PCR runs. Seventeen unique marker phenotypes were identified. The 48 trifoliate orange accessions were classified into four major groups based on polymorphic ISSR markers. All large-flowered accessions are in group 4, while small-flowered accessions are in group 3. Many ISSR markers segregated in progeny derived by open-pollination (probably mostly selfing) of a common accession, indicating that these ISSR markers are also heterozygous. Accessions having identical genotypes for a large number of heterozygous markers are unlikely to have diverged by recombination. Thus the limited divergence we detected among most accessions most likely originated by mutation. ‘Monoembryonic’ and ‘Simmons’ differed from other accessions only in the loss of specific markers, indicating that they originated as zygotic seedlings of individuals similar to the common genotype. Three accessions recently introduced from China have relatively different fingerprints with 3–14 unique ISSR markers, and probably represent a much more divergent germ plasm that may be a valuable breeding resource. Received: 8 August 1996 / Accepted: 21 March 1997     

长江华溪蟹精子超微结构的研究

透射电镜下观察长江华溪蟹成熟精子的形态和超微结构。精子呈椭球形,无鞭毛,具数条由核延伸而成的辐射臂精子。由球形的顶体,膜复合体,核杯及辐射臂组成,顶体由头帽,亚帽带,丁体囊及中央管组成。在中央管的基部发现有中心粒。核膜与质膜紧贴在一起形成了精子的界膜。     

Altered Intermediate Filament Expression in Human Neuroblastoma Cells Transformed by a Growth-Promoting Agent Derived from Schizophrenic CSF

1. Transformed (TR) cell lines showed faster doubling times and higher cell densities at confluence, as well as altered morphology, changing from flat epitheloid to smaller round or bipolar shapes. Since such morphological changes are suggestive of alterations in intermediate filaments, we have analyzed the expression of both vimentin and neurofilament.2. Immunohistochemical analysis of vimentin showed a redistribution from a cytoplasmic network to a perinuclear accumulation in TR cell lines.3. Western blot analysis demonstrated that the vimentin content was decreased 60–90%. The content of the 70-kD neurofilament protein was also decreased in TR cells, but its intracellular distribution was indistinguishable from that in the control cell lines.