Behavioral and electrophysiological analysis of Ca-activated K-channel transgenes in Drosophila.

The slowpoke gene of Drosophila melanogaster encodes a Ca-activated K channel. This gene is expressed in neurons, muscles, tracheal cells, and the copper and iron cells of the midgut. The gene produces a large number of alternative products using tissue-specific transcriptional promoters and alternative mRNA splicing.

Calcium-activated potassium channel gene expression in the midgut of Drosophila.

The slowpoke gene of Drosophila encodes a pore-forming subunit of a BK-type Ca(2+)-activated K+ channel. The gene is expressed in neurons, muscles, tracheal cells and in the midgut. The P1 transgene gene contains the entire slowpoke transcriptional control region and drives the expression of a reporter protein comprised of slowpoke amino terminal sequences fused to beta-galactosidase.

STAR, a gene family involved in signal transduction and activation of RNA.

A new subfamily of KH-domain-containing RNA-binding proteins is encoded by genes that are conserved from yeast to humans. Mutations with interesting developmental phenotypes have been identified in Caenorhabditis elegans, Drosophila and mouse. It is hypothesized that these bifunctional proteins provide a rich source of interesting molecular information about development and define a new cellular pathway that links signal transduction directly to RNA metabolism.

Isolation and characterization of Melanoplus sanguinipes adipokinetic hormone: a new member of the AKH/RPCH family.

A neuropeptide hormone isolated from corpora cardiaca of Melanoplus sanguinipes was purified by HPLC. The HPLC fractions were examined for adipokinetic activity with an in vivo bioassay. A single large UV absorbent peak was active in the mobilization of lipid while the other HPLC fractions showed no detectable activity.

Cyclosporin A retards the wallerian degeneration of peripheral mammalian axons.

The distal (anucleate) segments of mammalian peripheral axons typically undergo complete Wallerian degeneration within 1-3 days after severance from their cell bodies, unlike invertebrates and lower vertebrates, where anucleate axons do not degenerate for weeks to months. This rapid Wallerian degeneration in mammals could be due to a more efficient immune system and/or to differences in calcium-dependent pathways relative to invertebrates and lower vertebrates. To suppress the immune system and to inhibit calcium-dependent pathways in axons, we gave daily subcutaneous injections of cyclosporin A (CsA: 10 mg/kg) to Sprague-Dawley rats for 7 days before, and 5 days after, severing their right ventral tail nerves.

Novel embryonic regulation of Ca(2+)-activated K+ channel expression in Drosophila.

The slowpoke gene of Drosophila melanogaster encodes a Ca(2+)-activated K+ channel that is expressed in neurons, muscles, tracheal cells and the middle midgut. The entire transcriptional control region of slowpoke is contained in 11 kb of genomic DNA. Previous work has identified four different tissue-specific promoters (Promoters C1, C1b, C1c and C2) and sequences that regulate their activity.

Medium weight neurofilament mRNA in goldfish Mauthner axoplasm.

Although axons are generally considered to lack the ability to synthesize proteins, the Mauthner axon (M-axon) of the goldfish has been reported to contain some of the basic components of the translational machinery, such as transfer RNA (tRNA), ribosomal RNA (rRNA), and ribosomes. To determine if the M-axon also contains mRNA, we isolated samples of M-axoplasm free of glial contamination as demonstrated by the absence of glial-specific mRNA and protein. Reverse transcription-polymerase chain reaction (RT-PCR) of M-axoplasmic cDNA in the presence of primers for the goldfish medium-weight neurofilament (NF-M) gene produced a single product of the expected length for RT-PCR amplification of goldfish NF-M mRNA.

Developmental- and eye-specific transcriptional control elements in an intronic region of a Ca(2+)-activated K+ channel gene.

The range of electrical properties that a neuron or muscle cell can manifest is determined by which ion channel genes it expresses and in what amounts. The Drosophila slowpoke Ca(2+)-activated K+ channel gene has four distinct promoters. Here we assess the role that a downstream intronic region, called the C2/C3 region, plays in modulating Promoter C1 and Promoter C2 activity.

Tissue-specific expression of a Ca(2+)-activated K+ channel is controlled by multiple upstream regulatory elements.

The electrical properties of a cell are produced by the complement of ion channels that it expresses. To understand how ion-channel gene expression is regulated, we are studying the tissue-specific regulation of the slowpoke (slo) Ca(2+)-activated K+ channel gene. This gene is expressed in the central and peripheral nervous system, in midgut and tracheal cells, and in the musculature of Drosophila melanogaster.

Mechanisms for the maintenance and eventual degradation of neurofilament proteins in the distal segments of severed goldfish mauthner axons.

Cellular mechanisms that might affect the degradation of neurofilament proteins (NFPs) were examined in the distal segments of severed goldfish Mauthner axons (M-axons), which do not degenerate for more than 2 months after severance. Calpain levels, as determined by reactivity to a polyclonal antibody, remained constant for 80 d postseverance in distal segments of M-axons and then declined from 80 to 85 d postseverance. Calpain activity in rat brain, as determined by a spectrophotometric assay, was much higher than calpain activity in control and severed goldfish brain, spinal cord, muscle, or M-axons.

Multiple interspecies transmissions of human and simian T-cell leukemia/lymphoma virus type I sequences.

Using two sets of nucleotide sequences of the human and simian T-cell leukemia/lymphoma virus type I (HTLV-I/STLV-I), one consisting of 522 bp of the env gene from 70 viral strains and the other a 140-bp segment from the pol gene of 52 viral strains, I estimated cladograms based on a statistical parsimony procedure that was developed specifically to estimate within-species gene trees. An extension of a nesting procedure is offered for sequence data that forms nested clades used in hypothesis testing. The nested clades were used to test three hypotheses relating to transmission of HTLV/STLV sequences: (1) Have cross-species transmissions occurred and, if so, how many? (2) In what direction have they occurred? (3) What are the geographic relationships of these transmission events? The analyses support a range of 11-16 cross-species transmissions throughout the history of these sequences.

New fluorescent calcium indicators designed for cytosolic retention or measuring calcium near membranes.

A new family of fluorescent calcium indicators has been developed based on a new analog of BAPTA called FF6. This new BAPTA analog serves as a versatile synthetic intermediate for developing Ca2+ indicators targeted to specific intracellular environments. Two of these new Ca2+ indicators, fura-PE3 and fura-FFP18, are described in this report.

Is there a Brachyury the Second? Analysis of a transgenic mutation involved in notochord maintenance in mice.

A new phenotype mapping to the t-complex, which is designated Brachyury the Second (T2), is characterized by a slightly shortened tail in heterozygotes and homozygous failure to form an organized notochord with subsequent abnormal development of posterior somites and neural tube. The phenotype of T2 superficially resembles that of Brachyury; however, there are several important differences. Brachyury homozygotes fail to make posterior somites, notochord, floor plate, and a placental connection, resulting in death by 10.

Tissue-specific expression of a Drosophila calcium-activated potassium channel.

The Drosophila slowpoke (slo) gene encodes a subunit of a CAK channel homologous to the vertebrate BK channel. We have examined slo expression throughout development. It is expressed in muscle cells, neurons of the CNS and PNS, mushroom bodies, a limited number of cells in embryonic and larval midgut and in epithelial-derived tracheal cells.

Tctex2: a sperm tail surface protein mapping to the t-complex.

Transmission ratio distortion (TRD) in mouse t-haplotypes remains the most significant example of meiotic drive in vertebrates. While the underlying mechanism that fuels it is still mysterious, TRD is clearly a complex multigene phenomenon. The characterization of Tctex2 (t-complex testis expressed 2) shows it to be one of several candidates for involvement in TRD.

Electrolytic lesions to the ventromedial hypothalamus abolish receptivity in female whiptail lizards, Cnemidophorus uniparens.

Lesions to the ventromedial hypothalamus (VMH) inhibit receptive sexual behavior in the whiptail lizard, Cnemidophorus uniparens, an all-female species. All lesions to the VMH that effectively abolished receptivity specifically damaged the dorsal lateral VMH, an area containing high concentrations of estrogen receptor in this species. These data further emphasize conservation of the VMH as a brain area critical to the expression of female sexual behavior in vertebrates.

Cooling of peripheral myelinated axons retards Wallerian degeneration.

The histological and ultrastructural status of intact and severed axons was examined in the ventral tail nerve of rats whose tails were maintained at 32, 23, and 13 degrees C. Compared to contralateral intact nerves, distal (anucleate) portions of severed myelinated axons morphologically and ultrastructurally degenerated within 3 days at 32 degrees C and within 6 days at 23 degrees C. In contrast, anucleate myelinated axons in ventral tail nerves maintained at 13 degrees C did not degenerate for at least 10 days.

Intraspecific phylogenetics: support for dental transmission of human immunodeficiency virus.

A new method to estimate within-species gene genealogies was used to establish linkages among individuals associated with the Florida dental human immunodeficiency virus transmission case. Phylogenetic relationships were estimated from 103 nucleotide sequences from the V3 region of the env gene representing the Florida dentist, eight of his seropositive patients, and many local controls. The cladogram estimation procedure supports linkages among individuals within the previously described dental clade, whereas local controls and other patients form independent networks or are outliers in the main network, indicating more distant evolutionary relationships.

Calcium-activated proteolysis of neurofilament proteins in goldfish Mauthner axons.

We have examined the proteolytic breakdown of neurofilament proteins (NFPs) in isolated Mauthner axoplasm (M-axoplasm). Documentation of proteolytic breakdown of NFPs in M-axoplasm is important because NFPs are not degraded in distal segments of severed Mauthner axons (M-axons) maintained in vivo for up to 62 days at 20 degrees C. By incubating M-axoplasm with 2 mM calcium in vitro, we have demonstrated that M-axoplasm contains an endogenous calcium-activated neutral protease that degrades NFPs.

Toward a characterization of landscapes of combinatorial optimization problems, with special attention to the phylogeny problem.

This article introduces a coherent language base for describing and working with characteristics of combinatorial optimization problems, which is at once general enough to be used in all such problems and precise enough to allow subtle concepts in this field to be discussed unambiguously. An example is provided of how this nomenclature is applied to an instance of the phylogeny problem. Also noted is the beneficial effect, on the landscape of the solution space, of transforming the observed data to account for multiple changes of character state.

Long-term survival followed by degradation of neurofilament proteins in severed mauthner axons of goldfish.

The morphology and protein composition of intact and severed Mauthner axons (M-axons) from goldfish were examined on electron micrographs, sodium dodecyl sulfate gels, and immunoblots. Neurofilaments were the most common cytoskeletal element on electron micrographs, and neurofilament proteins (NFPs) were the most intensely silver-stained bands in M-axoplasm microdissected from control M-axons. NFPs at about 235, 145, 123, 105, 80, and 60 kD in M-axoplasm were identified with four monoclonal and three polyclonal antibodies.

Genomic organization and expressed sequences of the mouse extended H-2K region.

The mouse major histocompatibility complex (MHC) has long been of great interest to many biologists because of not only its critical role in the immune system, but also its association with at least three embryonic lethal genes. Here, we present an analysis of the mouse extended H-2K region using YAC technology. Six new expressed sequences were identified, demonstrating that the high gene density previously described continues.

Molecular analysis of mouse Rab11b: a new type of mammalian YPT/Rab protein.

Since the realization that the vesicular transport machinery is conserved from yeast to vertebrate neurons, much interest in the proteins involved has been generated. Here we describe a new type of mammalian YPT/Rab protein, named Rab11b, that is most abundantly expressed in brain, heart, and testis. It has all the hallmarks of a YPT3/Rab11 protein, but is more closely related to a ypt3-related gene isolated from fish (97% homology) than to the previously described four mammalian Rab11 genes (89% homology).

Normal neurulation in amphibians.

How does cell behaviour accomplish neurulation in amphibian embryos? During neurulation, the neural plate (while preserving the same volume) doubles its length, triples its thickness, narrows 10-fold, greatly decreases its surface and rolls into a tube. Cells that compose the neural plate produce these changes in three ways. They change shape, change neighbours and attempt to crawl beneath the contiguous epidermis.