Dopamine modulates S-potential amplitude and dye-coupling between external horizontal cells in carp retina

Horizontal cells in the fish retina are electrically coupled and possess gap junctions so that intracellularly injected dye normally diffuses freely to neighbouring cells. Applied dopamine (DA) alters the spatial properties of the horizontal cell responses to light, increasing the amplitude of photopic L-type S-potentials but decreasing their lateral spread. These effects have been attributed to the action of DA on horizontal cell membrane resistance, particularly at the gap junctions, and our present study on the carp retina agrees with this in showing that DA also restricts intracellular Lucifer yellow (LY) to single injected horizontal cells, an effect, like those of DA on the S-potentials, which is antagonized by the dopamine blocker haloperidol. In addition, we present evidence that dopaminergic interplexiform cells in fish normally function to regulate the spatial properties of responses in horizontal cells, possibly acting on their junctional resistance via a DA-receptor-mediated mechanism. Previous destruction of the interplexiform cells with 6-hydroxydopamine (6-OHDA) resulted in much reduced L-type S-potentials to centred lights but wider lateral spread of these responses, while the dye injected spread extensively to neighbouring cells. After 6-OHDA treatment, however, applied DA retained its normal activity, restoring large-amplitude, narrow receptive-field S-potentials and restricting LY to the injected cells, effects which were both closely mimicked by dibutyryl cyclic AMP.     

A sensitive and inexpensive high-performance liquid chromatographic assay for tyrosine hydroxylase

We describe a highly sensitive assay method for tyrosine hydroxylase (TH) using high-performance liquid chromatography with amperometric determination. This assay method could be applicable to any tissues with low enzyme activity, such as rat cerebellum. We also describe the kinetic properties of TH in rat cerebral cortex.     

Gramicidin S analogs containing N-methylleucine in place of leucine

Zusammenfassung Nach Methoden der konventionellen Peptidsynthese wurden zwei Gramicidin S Analoga synthetisiert, in denen Leucin durch N-Methylleucin ersetzt ist. Beim antibakteriellen Test sind diese beiden Analoga praktisch gleich wirksam wie Gramicidin S und auch die Konformation dieser zwei Analoga ist derjenigen von Gramidicin S ähnlich.     

Oncogenic mutations of ALK kinase in neuroblastoma

Neuroblastoma in advanced stages is one of the most intractable paediatric cancers, even with recent therapeutic advances. Neuroblastoma harbours a variety of genetic changes, including a high frequency of MYCN amplification, loss of heterozygosity at 1p36 and 11q, and gain of genetic material from 17q, all of which have been implicated in the pathogenesis of neuroblastoma. However, the scarcity of reliable molecular targets has hampered the development of effective therapeutic agents targeting neuroblastoma. Here we show that the anaplastic lymphoma kinase (ALK), originally identified as a fusion kinase in a subtype of non-Hodgkin's lymphoma (NPM-ALK) and more recently in adenocarcinoma of lung (EML4-ALK), is also a frequent target of genetic alteration in advanced neuroblastoma. According to our genome-wide scans of genetic lesions in 215 primary neuroblastoma samples using high-density single-nucleotide polymorphism genotyping microarrays, the ALK locus, centromeric to the MYCN locus, was identified as a recurrent target of copy number gain and gene amplification. Furthermore, DNA sequencing of ALK revealed eight novel missense mutations in 13 out of 215 (6.1%) fresh tumours and 8 out of 24 (33%) neuroblastoma-derived cell lines. All but one mutation in the primary samples (12 out of 13) were found in stages 3-4 of the disease and were harboured in the kinase domain. The mutated kinases were autophosphorylated and displayed increased kinase activity compared with the wild-type kinase. They were able to transform NIH3T3 fibroblasts as shown by their colony formation ability in soft agar and their capacity to form tumours in nude mice. Furthermore, we demonstrate that downregulation of ALK through RNA interference suppresses proliferation of neuroblastoma cells harbouring mutated ALK. We anticipate that our findings will provide new insights into the pathogenesis of advanced neuroblastoma and that ALK-specific kinase inhibitors might improve its clinical outcome.     

Genetic dissection of the circuit for hand dexterity in primates

It is generally accepted that the direct connection from the motor cortex to spinal motor neurons is responsible for dexterous hand movements in primates. However, the role of the 'phylogenetically older' indirect pathways from the motor cortex to motor neurons, mediated by spinal interneurons, remains elusive. Here we used a novel double-infection technique to interrupt the transmission through the propriospinal neurons (PNs), which act as a relay of the indirect pathway in macaque monkeys (Macaca fuscata and Macaca mulatta). The PNs were double infected by injection of a highly efficient retrograde gene-transfer vector into their target area and subsequent injection of adeno-associated viral vector at the location of cell somata. This method enabled reversible expression of green fluorescent protein (GFP)-tagged tetanus neurotoxin, thereby permitting the selective and temporal blockade of the motor cortex–PN–motor neuron pathway. This treatment impaired reach and grasp movements, revealing a critical role for the PN-mediated pathway in the control of hand dexterity. Anti-GFP immunohistochemistry visualized the cell bodies and axonal trajectories of the blocked PNs, which confirmed their anatomical connection to motor neurons. This pathway-selective and reversible technique for blocking neural transmission does not depend on cell-specific promoters or transgenic techniques, and is a new and powerful tool for functional dissection in system-level neuroscience studies.     

分阶式双渐开线齿轮轮齿刚度的研究

用有限元方法首次计算分析了分阶式双渐开线齿轮轮齿的刚度、啮合刚度和载荷分配系数,结果表明双渐开线齿轮的刚度、啮合刚度均提高了,而载荷分配系数基本不变;提出直线逼近齿廓曲线近似求解该齿轮刚度的能量积分法。     

Mutations affecting components of the SWI/SNF complex cause Coffin-Siris syndrome

By exome sequencing, we found de novo SMARCB1 mutations in two of five individuals with typical Coffin-Siris syndrome (CSS), a rare autosomal dominant anomaly syndrome. As SMARCB1 encodes a subunit of the SWItch/Sucrose NonFermenting (SWI/SNF) complex, we screened 15 other genes encoding subunits of this complex in 23 individuals with CSS. Twenty affected individuals (87%) each had a germline mutation in one of six SWI/SNF subunit genes, including SMARCB1, SMARCA4, SMARCA2, SMARCE1, ARID1A and ARID1B.