Molecular Characterization of the D Surface Protein Gene Subfamily in Paramecium tetraurelia

ABSTRACT. When Paramecium tetraurelia expresses the D serotype, detectable by serum tests, high molecular mRNA could be isolated, which corresponds to the molecular mass of the D surface protein. Using this D specific mRNA as a probe for screenings in different genomic libraries a subfamily of five very similar genes was found, named α-51D, _γ1-51D, _γ2-51D, δ-51D and ε-51D. Each of them is about 8-kb long, they show regions of identity to each other, and there is no evidence that any are defective genes or pseudogenes. Up to now serotype D is the only known serotype showing this phenomenon. Another novel feature is that two of the D isogenes are closely linked. The sequence for the entire coding region of the α-51D gene has been determined, as well as the upstream and downstream noncoding regions. Its deduced amino acid sequence shows the same characteristic cysteine periodicity displayed by all other immobilization antigen (i-ag) genes from Paramecium. However, in contrast to most other such genes, tandem repeats are missing from the 7599-bp long coding region of the α-51D gene. When the sequences of the type 51D genes are compared to each other, the similarity is very high and extends to coding as well as to noncoding regions. Similarity within noncoding regions is usually only observed for allelic i-ag genes. We conclude that the type D genes constitute a family of isogenes that are nonallelic. They contain slightly different consensus sequences with possible functions as regulatory regions.     

Cladistic analysis of the Cirripedia Thoracica

We present a cladistic analysis of the Cirripedia Thoracica using morphological characters and the Acrothoracica and Ascothoracida as outgroups. The list of characters comprised 32 shell and soft body features. The operational taxonomic units (OTUs) comprised 26 well-studied fossil and extant taxa, principally genera, since uncertainty about monophyly exists for most higher ranking taxonomic units. Parsimony analyses using PAUP 3.1.1 and Hennig86 produced 189 trees of assured minimal length. We also examined character evolution in the consensus trees using MacClade and Clados. The monophyly of the Balanomorpha and the Verrucomorpha sensu stricto is confirmed, and all trees featured a sister group relationship between the ‘living fossil Neoverruca and me Brachylepadomorpha. In the consensus trees the sequential progression of ‘pedunculate‘sister groups up to a node containing Neolepas also conforms to current views, but certain well-established taxa based solely on plesiomorphies stand out as paraphyletic, such as Pedunculata (= Lepadomorpha); Eolepadinae, Scalpellomorpha and Chthamaloidea. The 189 trees differed principally in the position of shell-less pedunculates, Neoverruca, the scalpelloid Capitulum, and the interrelationships within the Balanomorpha, although the 50% majority rule consensus tree almost fully resolved the latter. A monophyletic Sessilia comprising both Verrucomorpha and Balanomorpha appeared among the shortest trees, but not in the consensus. A tree with a monophyletic Verrucomorpha including Neoverruca had a tree length two steps longer than the consensus trees. Deletion of all extinct OTUs produced a radically different tree, which highlights the importance of fossils in estimating cirripede phylogeny. Mapping of our character set onto a manually constructed cladogram reflecting die most recent scenario of cirripede evolution resulted in a tree length five steps longer than any of our shortest trees. Our analysis reveals that several key questions in cirripede phylogeny remain unsolved, notably the position of shell-less forms and the transition from ‘pedunculate‘to ‘sessile‘barnacles. The inclusion of more fossil species at this point in our understanding of cirripede phylogeny will only result in even greater levels of uncertainty. When constructing the character list we also identified numerous uncertainties in the homology of traits commonly used in discussing cirripede evolution. Our study highlights larval ultrastructure, detailed studies of early ontogeny, and molecular data as the most promising areas for future research.     

Palaeobotryllus from the Upper Cambrian of Nevada — a probable ascidian

Phosphatic microfossils etched from Upper Cambrian (Trempealeauan Stage) Limestones of the Whipple Cave Formation, Nevada, and other contemporaneous formations in western North America, are colonies, the individuals of which are arranged irregularly around one or more nuclei. No secondary growth has occurred. During their lifetime, however, the individuals partially filled their chambers with phosphatic matter. The highly differentiated morphology of the colonies is strikingly similar to the living ascidian tunicate Botryllus. Ecological conditions and size range of the fossils are likewise similar to those of the Ascidiacea. If recognized as an ascidian, Palaeobotryllus taylori n.g., n.sp. extends the known occurrence of Tunicata into the Upper Cambrian.     

Heterogeneity of Intermediate Filament Expression in Vascular Smooth Muscle: A Gradient in Desmin Positive Cells from the Rat Aortic Arch to the Level of the Arteria Iliaca Communis

The display of the two distinct intermediate filament proteins, desmin and vimentin, in rat vascular smooth muscle tissue was studied by immunofluorescence microscopy on frozen sections of aorta and other blood vessels. Vascular smooth muscle cells present in these vessels always appeared rich in vimentin. However, staining of sections covering six distinct but contiguous parts of the aorta showed that the number of desmin containing cells was low distal to the truncus brachiocephalicus, but increases until in distal parts of the aorta and in the arteria iliaca communis almost all cells appear positive for desmin. Thus blood vessels show heterogeneity of intermediate filament expression not only in cross-section but can also display heterogeneity along their length. Muscular arteries such as the renal artery and the arteria femoralis, as well as arterioles and veins including the vena jugularis and the vena cava also contain desmin. Thus it may be that low numbers of desmin-positive cells are typical of elastic arteries, while muscular arteries and other blood vessels are characterized by large numbers of desmin-positive cells. We discuss whether desmin-positive and desmin-negative vascular smooth muscle cells may perform different functions and raise the possibility that desmin expression may coincide with the turn on of a specially regulated contractility program.     

Ecdysteroid-Responsive Genes in a Drosophila Cell Line

SYNOPSIS. In the Drosophila Kc-H cell line ecdysteroids causerapid increases in the synthesis of two groups of polypeptideswhich we call ecdysteroid-inducible polypeptides. We have shownthat these changes occur at least in part at the level of mRNAtiters and we have isolated cDNA clones corresponding to themRNAs for these polypeptides. The increases in these mRNAs maywell be among the primary hormone effects.     

Regulation of Internodal Extension in Avena Shoots by the Inflorescence, Nodes, Leaves, and Intercalary Meristem

It has been found in recent studies that the inflorescence and nodes (node-pulvini) are the primary sources for native gibberellins in the Avena shoot, and that GA₃ is the predominant gibberellin in the inflorescence. In the present work, linear growth of next-to-last internode is drastically reduced by removal of the inflorescence and last leaf. This growth is completely abolished when the nodes are also excised. It is restored fully by the addition of GA₃ when the nodes are present, and restored only partially when the nodes are deleted. Internodal growth in Avena stem segments with basal node present is also restored by native GA₃-like substances extracted from Avena inflorescences and partially purified by silica gel partition column chromatography. Evidence from these studies, taken in toto, indicates that the inflorescence, nodes, and leaves supply gibberellins, leaves supply substrate, and nodes modulate the gibberellin growth response in next-to-last Avena internodes.     

Timing and Regulation of Nuclear and Cortical Events in the Cell Cycle of Tetrahymena pyriformis*

SYNOPSIS. Using continuous flow cultures based on the chemostat principle, we varied the cell generation times of the ciliate Tetrahymena pyriformis strain GL, from 4.9 to 22.2 hr and studied various parameters of the cell cycle at 28 C. These included: the duration of the periods required for oral morphogenesis, macronuclear division, cell division, G₁ S, and G₂. The size of individual cells was also measured. Independent of the growth rate, the period of oral morphogenesis occurred during the last 90 min of the cell cycle. In all cases macronuclear and cell divisions took place during the last part of these 90 min, and the final macronuclear separation occurred just before final cell separation. The S-period increased slightly, while the G₁ and G₂ both increased in roughly the same relative proportion to the increasing generation times. Slowly growing cells (generation time 20.5 hr) were shorter but broader and somewhat larger in volume than quickly growing cells (generation time 4.9 hr).     

Nuclease Digestion of Chromatin from the Eukaryotic Algae Olisthodiscus luteus,Peridinium balticum,and Crypthecodinium cohnii 1, 2

Chromatin from a uninucleate dinoflagellate, Crypthecodinium cohnii, a binucleate dinoflagellate, Peridinium balticum, and a chromophyte, Olisthodiscus luteus, was examined by nuclease digestion and the results were compared to those from vertebrates. Gel analysis of the products of staphylococcal (micrococcal) nuclease digestion revealed a DNA repeat unit of 220(±5) base pairs for O. luteus and 215(±5) for P. balticum. Limit digestion gave a core particle of 140 base pairs, revealing that these longer repeat sizes are due to longer linker regions. No repeating subunit structure was found upon electrophoresis of digests of C. cohnii nuclei. Examination of the DNA fragments produced by DNAse I digestion of nuclei isolated from P. balticum and O. luteus showed the same ladder of ten base multiples as seen in chromatin from other eukaryotes. Examination of the kinetics of digestion by DNAse II of Peridinium chromatin revealed less susceptibility when compared to DNAse I digestions while 70% of Olisthodiscus chromatin and 35% of C. cohnii chromatin was sensitive to DNAse II. These data, taken together with previous results from Euglena, indicate that while algal chromatin is similar to that of higher eukaryotes in regard to DNAse I and II action, it differs in that the linker DNA is longer. In addition, the Hl-like histone from O. luteus and P. balticum is located in the linker DNA as in higher eukaryotes.     

Seasonal and diurnal variations in the energetics and foraging activities of the brown honey eater,Lichmera indistincta

In general, the brown honeyeater, Lichmera indistincta, flies more and expends more energy when visiting flowers in the early morning than at other times. Floral nectar is most abundant early in the day, and nectar energy intake by L. indistincta is greatest at that time. It appears as if birds can only store excess energy during the early part of the day, incurring energy deficits at other times. Unit perching and flight costs are greatest for L. indistincta in winter and spring. As a result, peak total energy expenditures and requirements occur at these times, birds apparently being unable to reduce energy demands by employing nocturnal torpor. Estimates of probable energy uptake by L. indistincta from the plant species studied exceed predicted requirements in all seasons except winter, when it is apparent that birds would need to be especially selective in the plants that they used as major energy sources.