A cytoplasmic region determines single-channel conductance in 5-HT3 receptors

5-hydroxytryptamine type 3 (5-HT3) receptors are cation-selective transmitter-gated ion channels of the Cys-loop superfamily. The single-channel conductance of human recombinant 5-HT3 receptors assembled as homomers of 5-HT3A subunits, or heteromers of 5-HT3A and 5-HT3B subunits, are markedly different, being 0.4 pS (refs 6, 9) and 16 pS (ref. 7), respectively. Paradoxically, the channel-lining M2 domain of the 5-HT3A subunit would be predicted to promote cation conduction, whereas that of the 5-HT3B subunit would not. Here we describe a determinant of single-channel conductance that can explain these observations. By constructing chimaeric 5-HT3A and 5-HT3B subunits we identified a region (the 'HA-stretch') within the large cytoplasmic loop of the receptor that markedly influences channel conductance. Replacement of three arginine residues unique to the HA-stretch of the 5-HT3A subunit by their 5-HT3B subunit counterparts increased single-channel conductance 28-fold. Significantly, ultrastructural studies of the Torpedo nicotinic acetylcholine receptor indicate that the key residues might frame narrow openings that contribute to the permeation pathway. Our findings solve the conundrum of the anomalously low conductance of homomeric 5-HT3A receptors and indicate an important function for the HA-stretch in Cys-loop transmitter-gated ion channels.     

Post-translational disruption of dystroglycan-ligand interactions in congenital muscular dystrophies

Muscle eye brain disease (MEB) and Fukuyama congenital muscular dystrophy (FCMD) are congenital muscular dystrophies with associated, similar brain malformations. The FCMD gene, fukutin, shares some homology with fringe-like glycosyltransferases, and the MEB gene, POMGnT1, seems to be a new glycosyltransferase. Here we show, in both MEB and FCMD patients, that alpha-dystroglycan is expressed at the muscle membrane, but similar hypoglycosylation in the diseases directly abolishes binding activity of dystroglycan for the ligands laminin, neurexin and agrin. We show that this post-translational biochemical and functional disruption of alpha-dystroglycan is recapitulated in the muscle and central nervous system of mutant myodystrophy (myd) mice. We demonstrate that myd mice have abnormal neuronal migration in cerebral cortex, cerebellum and hippocampus, and show disruption of the basal lamina. In addition, myd mice reveal that dystroglycan targets proteins to functional sites in brain through its interactions with extracellular matrix proteins. These results suggest that at least three distinct mammalian genes function within a convergent post-translational processing pathway during the biosynthesis of dystroglycan, and that abnormal dystroglycan-ligand interactions underlie the pathogenic mechanism of muscular dystrophy with brain abnormalities.     

Automated DNA sequencing and analysis of 106 kilobases from human chromosome 19q13.3.

A total of 116,118 basepairs (bp) derived from three cosmids spanning the ERCC1 locus of human chromosome 19q13.3 have been sequenced with automated fluorescence-based sequencers and analysed by polymerase chain reaction amplification and computer methods. The assembled sequence forms two contigs totalling 105,831 bp, which contain a human fosB proto-oncogene, a gene encoding a protein phosphatase, two genes of unknown function and the previously-characterized ERCC1 DNA repair gene. This light band region has a high average density of 1.4 Alu repeats per kilobase. Human chromosome light bands could therefore contain up to 75,000 genes and 1.5 million Alu repeats.     

Dietary fat and tissue cholesterol in female rats

Zusammenfassung Junge, ausgewachsene weibliche Ratten nahmen nach 8 Wochen fettreicher Diät mehr Fett auf als solche mit einer fettarmen Diät, aber gleichviel Kalorien. Bei Weibchen mit körperlicher Bewegung war der Cholesteringehalt des Blutes und des Gewebes höher als bei eingesperrten, sitzenden Tieren.

---

---

Financial support for this study was provided by Nutrition Foundation Grant No. 383.     

The gene mutated in bare patches and striated mice encodes a novel 3beta-hydroxysteroid dehydrogenase.

X-linked dominant disorders that are exclusively lethal prenatally in hemizygous males have been described in human and mouse. None of the genes responsible has been isolated in either species. The bare patches (Bpa) and striated (Str) mouse mutations were originally identified in female offspring of X-irradiated males. Subsequently, additional independent alleles were described. We have previously mapped these X-linked dominant, male-lethal mutations to an overlapping region of 600 kb that is homologous to human Xq28 (ref. 4) and identified several candidate genes in this interval. Here we report mutations in one of these genes, Nsdhl, encoding an NAD(P)H steroid dehydrogenase-like protein, in two independent Bpa and three independent Str alleles. Quantitative analysis of sterols from tissues of affected Bpa mice support a role for Nsdhl in cholesterol biosynthesis. Our results demonstrate that Bpa and Str are allelic mutations and identify the first mammalian locus associated with an X-linked dominant, male-lethal phenotype. They also expand the spectrum of phenotypes associated with abnormalities of cholesterol metabolism.     

Sequence identification of 2,375 human brain genes.

We recently described a new approach for the rapid characterization of expressed genes by partial DNA sequencing to generate 'expressed sequence tags'. From a set of 600 human brain complementary DNA clones, 348 were informative nuclear-encoded messenger RNAs. We have now partially sequenced 2,672 new, independent cDNA clones isolated from four human brain cDNA libraries to generate 2,375 expressed sequence tags to nuclear-encoded genes. These sequences, together with 348 brain expressed sequence tags from our previous study, comprise more than 2,500 new human genes and 870,769 base pairs of DNA sequence. These data represent an approximate doubling of the number of human genes identified by DNA sequencing and may represent as many as 5% of the genes in the human genome.     

基于热压块的不锈钢粉尘还原实验

为了从不锈钢粉尘中回收利用Fe,Cr和Ni等,对不锈钢粉尘热压块制备及其自还原过程进行了研究.在热压温度为200℃,热压压力为35 MPa条件下,抗压强度达到900 N/个以上.高温条件下,煤热解产生的挥发分可参与不锈钢粉尘还原反应,当还原温度为1 400,1 450℃时,挥发分还原作用率达到0.4.据XRD分析和热力学计算,自还原过程中含铬物质的物相转变顺序为Fe Cr2O4,Cr2O3,Cr7C3,[Cr]Fe-Cr-Ni-C.当还原温度为1 450℃,烟煤中固定碳与粉尘中可去除氧的物质量的比(xc/xo)为0.72时,不锈钢粉尘热压块不能完全还原;当xc/xo大于0.8,还原20 min时,不锈钢粉尘热压块能完全还原.     

NaOH-Ca(OH)2分解独居石精矿的研究

为降低独居石分解工艺的加碱量,并提高独居石分解率,本研究在NaOH-Ca(OH)₂体系中对独居石精矿进行焙烧分解,采用XRD对焙烧产物进行物相分析,并结合焙烧矿中稀土元素在盐酸中的浸出率判断独居石分解效果.实验分别研究了NaOH添加量、Ca(OH)₂添加量、焙烧温度以及焙烧时间对独居石精矿分解的影响.结果表明,在NaOH-Ca(OH)₂体系中,独居石精矿分解的最佳工艺条件:NaOH添加量为25%,Ca(OH)₂添加量为20%,焙烧温度为800℃,焙烧时间为1.5h.该焙烧条件下独居石全部分解为稀土氧化物,浓盐酸对稀土浸出率可达到98%左右.与现有工业生产工艺相比,本研究工艺中碱添加量降低55%左右,独居石分解率提高2%左右.     

低迟滞PMN-PT叠层型驱动器件的制备

研究了PMN-PT电致应变陶瓷材料的铌铁矿法制备及其电致应变性能,以及该陶瓷膜叠层型驱动器件的制备条件及性能.结果表明,0.94PMN-0.06PT陶瓷的应变迟滞最小;利用压延法可以提高叠层陶瓷膜的密度等特性;叠层型驱动器件的成型压力、预烧和烧结阶段的压力分别为200和0.01 MPa时,驱动器件的应变迟滞小于百分之一,并且驱动特性优异.