Preparation of isolated cells from rat heart

Summary Suspensions of isolated cells from rat heart were prepared and the data for viability and yield are given. Glucose uptake by the cells was mediated by a carrier system.The technical assistance of Mr.E. T. Potter, Mr.K. D. Patel and Mr.M. Griffin is gratefully acknowledged.     

Effect of the juvenoid methoprene on the hemolymph composition of the cabbage maggotDelia radicum (Diptera: Anthomyiidae)

Summary Treatment of post-feeding larvae of the cabbage maggotDelia radicum with methoprene did not affect the capacity of the insect to pupate, but suppressed eclosion to the adult stage. The concentration of hemolymph trehalose was significantly decreased by methoprene treatment, although hemolymph protein and amino nitrogen levels were unaffected.19 December 1986The authors are grateful to D. C. Read of the Research Centre, Agriculture Canada, Charlottetown, P.E.I., Canada for supplying eggs ofD. radicum for the stock colony. TheD. radicum colony was maintained at the Agriculture Canada Research Station, St. John's West, Newfoundland, Canada. The sample of methoprene was donated by Zoecon Corp., Palo Alto, California, U.S.A. This research was supported by a grant from the Natural Sciences and Engineering Research Council of Canada (Operating grant A6679).     

Transposon-dependent mutant phenotypes at the Notch locus of Drosophila

Many mutations at complex genetic loci in the fruitfly Drosophila melanogaster are associated with insertions of transposable elements. At the Notch locus, members of one class of insertion-associated mutations, termed glossy-like, produce a recessive viable, smooth-eye phenotype with mottled pigmentation. Members of a second class, facet, produce a recessive viable, rough-eye phenotype with homogeneous pigmentation. Both classes of mutations fail to complement Notch lethal mutations, so they behave as Notch alleles. Here we report that each glossy-like mutation is associated with an insertion of the same transposable element (flea). Each flea insertion occurs in the same orientation, but at different locations within intervening sequences of the Notch locus. In contrast, each facet mutation is associated with insertion of a unique, non-flea, transposable element. Insertions producing a facet phenotype and insertions causing a glossy-like phenotype can break Notch intervening sequences at precisely the same location. This suggests that the type of insertion element rather than its position within an affected gene is the primary determinant of the phenotype observed.     

Electrolyte transport in the seminiferous tubules of the rat studied by the stopped-flow microperfusion technique

Zusammenfassung Mit Hilfe der «stopped-flow»-Mikroperfusionstechnik wurden am Samenkanälchen Elektrolyttransportprozesse untersucht. Es wurde gefunden, dass die Kanälchen ein kaliumreiches Primärsekret bilden. Dieses unterscheidet sich in seiner Zusammensetzung von dem Sekret, das man gewöhnlich unter ungestörten Fliessbedingungen findet.

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This project was supported by the Rural Bank of Australia. One of us (R.D.H.) thanks the National Health and Medical Research Council of Australia for the award of a B.Sc. (med.) studentship for 1970.     

Intragenic amplification and divergence in the mouse alpha-fetoprotein gene

The DNA sequences of the 14 exon junctions in the murine alpha-fetoprotein gene were determined using cloned genomic DNA. When these exons were examined with respect to the polypeptide segments they encoded, a direct correspondence between a threefold repeat of four exons and three protein domains was observed. Nucleotide sequence comparisons among the four exons of each domain were used to deduce the likely structure of the primordial domain, and the order and mechanism of its triplication to form the tripartite ancestral gene from which both alpha-fetoprotein and serum albumin arose. Sequence homologies among the four exons that constitute a single domain also suggest that they were derived, at least in part, from a common sequence which underwent successive amplification and divergence.     

Structural mechanism for sterol sensing and transport by OSBP-related proteins

The oxysterol-binding-protein (OSBP)-related proteins (ORPs) are conserved from yeast to humans, and are implicated in the regulation of sterol homeostasis and in signal transduction pathways. Here we report the structure of the full-length yeast ORP Osh4 (also known as Kes1) at 1.5-1.9 A resolution in complexes with ergosterol, cholesterol, and 7-, 20- and 25-hydroxycholesterol. We find that a single sterol molecule binds within a hydrophobic tunnel in a manner consistent with a transport function for ORPs. The entrance is blocked by a flexible amino-terminal lid and surrounded by basic residues that are critical for Osh4 function. The structure of the open state of a lid-truncated form of Osh4 was determined at 2.5 A resolution. Structural analysis and limited proteolysis show that sterol binding closes the lid and stabilizes a conformation favouring transport across aqueous barriers and signal transmission. The structure of Osh4 in the absence of ligand exposes potential phospholipid-binding sites that are positioned for membrane docking and sterol exchange. On the basis of these observations, we propose a model in which sterol and membrane binding promote reciprocal conformational changes that facilitate a sterol transfer and signalling cycle.