Myosin heavy chain composition of single cells from avian slow skeletal muscle is strongly correlated with velocity of shortening during development

We have determined the myosin heavy chain (MHC) composition (using a sensitive sodium dodecyl sulfate-polyacrylamide gel electrophoresis system) and the maximal velocity of shortening (Vmax) of single cells from neonatal and adult chicken anterior latissimus dorsi (ALD) muscles. In addition, the MHC, myosin light chain, and regulatory protein (i.e., troponin and tropomyosin subunits) compositions of bundles of ALD fibers were determined at late embryonic, neonatal, and adult ages. At young ages, there are two MHCs in ALD muscle, SM1 and SM2, with SM1 decreasing in relative amount with increasing age, as shown previously by others. The mean Vmax of single fibers also decreases from neonatal to adult ages. A strong quantitative correlation is demonstrated between the specific MHC composition and Vmax among individual cells of the ALD muscle at several ages. Since virtually no changes occur in the regulatory protein and myosin light chain compositions of the ALD muscle between late embryonic and adult ages, it appears that the MHC composition of an individual cell in this muscle is the primary determinant of the maximal shortening velocity. These results are the first to illustrate the functional significance of the developmental transition in myosin heavy chain composition of an avian slow skeletal muscle, consistent with our previous findings on mammalian muscle.     

T cell-activating protein on murine lymphocytes

A functional T cell surface molecule, T cell-activating protein (TAP), has been identified on murine lymphocytes. TAP is a protein with an apparent molecular mass of 10-12 kilodaltons (kDa) nonreduced, 16-17 kDa reduced. Cross-linking of TAP can result in profound activation of T lymphocytes to produce lymphokines and to enter the cell cycle. Furthermore, antibody binding to TAP can modulate antigen-driven T cell stimulation. Current data suggest that TAP is physically distinct from the T cell receptor complex. On unstimulated lymphocytes, TAP is expressed on T cells and defines heterogeneity within the major T cell subsets. Its profile of expression is rapidly altered on cell activation. Ontologically, it is first detected in the thymus, where it is found on both the most immature and the most mature cell subsets, and it is functional on both. Together, these TAP+ cells constitute a small fraction of thymocytes. TAP expression, however, defines the immunocompetent compartment of the thymus, and thus could be involved in functional maturation. Finally, the gene controlling TAP expression has been mapped, and is tightly linked to a locus of hematopoietic antigens (Ly-6). TAP is molecularly distinct from these antigens. Furthermore, all of these proteins show a markedly distinct developmental regulation in their cell surface expression.     

The expression, function, and ontogeny of a novel T cell-activating protein, TAP, in the thymus

The TAP molecule is an allelic 12,000 m.w. membrane protein that participates in T cell activation. This report analyzes the expression, function, and ontogeny of this molecule in the thymus. TAP is expressed on a small subset (10 to 20%) of thymocytes which is distinct from its expression on a majority (70%) of peripheral T lymphocytes. In the adult thymus, the majority of the TAP-bearing thymocytes are cortisone-resistant, Thy-1+, TL-, J11D-, and PNA-, which localizes TAP expression to medullary thymocytes. Cortical thymocytes do not bear this determinant. Parallel functional studies demonstrated that TAP+ thymocytes are required for Con A and MLR responsiveness. Anti-TAP MAb plus PMA specifically induces proliferation of mature thymocytes comparable in magnitude to the Con A response. These results demonstrate that TAP expression defines the immunocompetent thymocyte compartment and, further, that this molecule is functional on these cells. The ontogeny of TAP expression was also analyzed. TAP is expressed early in fetal thymic development at a time when most T cell markers (except Thy-1 and the iL2-R) are absent. The small sub-population of adult L3T4- and Lyt-2- thymocytes, which resemble early fetal thymocytes, also express TAP. These early thymocytes are capable of being activated through the TAP molecule. The implications of these findings for T cell development and, in particular, the relationship of TAP to T cell receptor expression and acquisition of immunocompetence are discussed.     

Isolation of cDNA clones for fourteen nuclear-encoded thylakoid membrane proteins

Summary Spinach cDNA libraries, made from polyadenylated seedling RNA, have been constructed in pBR322 and the expression vector gt11. Recombinant plasmids or phage for 14 intrinsic and peripheral thylakoid membrane proteins and one stromal protein have been identified. They encode components containing antigenic determinants against the lysine-rich 34 kd, the 23 kd and 16 kd proteins all associated with the water-splitting apparatus of the photosystem II reaction center, the ATP synthase subunits gamma, delta and CF_o-II, the Rieske Fe/S protein of the cytochrome b/f complex, subunits 2, 3, 5 and 6 of the photosystem I reaction center, plastocyanin, ferredoxin oxidoreductase, chlorophyll a/b-binding apoproteins of the lightharvesting complex associated with photosystem II, and the small subunit of the stromal enzyme ribulose bisphosphate corboxylase/oxygenase. The cDNA inserts lack complementarity to plastid DNA but hybridize to restricted nuclear DNA as well as to discrete poly A⁺-mRNA species. The precursor products obtained after translation of hybrid selected RNA fractions in a wheat germ assay are imported and processed by isolated unbroken spinach chloroplasts. The imported components comigrate with the respective authentic proteins.     

Immunoelectron microscopic studies on the localization of peptidoglycan peptide subunit pentapeptides in bacterial cell walls

The peptide subunit pentapeptide H-L-Ala-D-Glu(L-Lys-D-Ala-D-Ala-OH)-NH₂ of peptidoglycan was localized in the cell walls of several Gram-positive bacteria employing the indirect immunoferritin technique. Specific antibodies to the D-alanyl-D-alanine moiety of non-crosslinked peptide subunit pentapeptide were raised in rabbits by immunization with synthetic immunogen albumin-(CH₂CO-Gly-L-Ala-L-Ala-D-Ala-D-Ala-OH)₃₉. Specificity of these antibodies for the peptide subunit pentapeptide and not for the peptide subunit tetrapeptide was corroborated in a Farr-type radio-active hapten binding assay. Specificity of labelling with ferritin was established by immunoelectron microscopic controls, and by the excellent correlation between specific labelling of cells with ferritin and the particular peptidoglycan primary structure of bacterial strains investigated. Cells of Lactobacillus gasseri, Streptococcus pyogenes and Staphylococcus aureus revealing non-crosslinked peptide subunit pentapeptides in their peptidoglycans could specifically be labelled. Lactobacillus acidophilus and Bacillus subtilis, on the contrary, missing such pentapeptides, failed in labelling.The implication of this method to possibly localize the points of attack of penicillin or cycloserine is discussed.Abbreviations used meso-A₂pm meso-diaminopimelic acid - DSM Deutsche Sammlung für Mikroorganismen, Göttingen, FRG This paper is dedicated to Professor Gerhart Drews on the occasion of his 60th birthday     

Ultrastruktur und Bildungsweise penialer Hartstrukturen bei freilebenden Plathelminthen

Zusammenfassung Ultrastruktur und Differenzierung von Penisstiletten bzw. Stilettnadeln wurden an Vertretern verschiedener Taxa freilebender Plathelminthen untersucht. Die Ausdifferenzierung der Hartstrukturen erfolgt auf unterschiedliche Weise, jedoch stets intrazellulär. Die Stilettnadeln von Philocelis cellata (Acoela) bestehen aus Mikrofibrillen und werden sukzessiv gebildet, mit der Spitze beginnend und basalwärts fortschreitend. Eine ebenfalls sukzessive Bildungsweise zeigen die Stilettapparaturen des Taxons Paromalostomum (Macrostomida); doch bestehen die Hartstrukturen hier aus Mikrotubuli mit angelagertem elektronendichtem Material und nicht aus Mikrofibrillen. Die sukzessive Ausdifferenzierung des Stiletts von Ciliopharyngiella intermedia erfolgt ähnlich wie bei den Hartstrukturen bestimmter Proseriaten mit der Anlage von Mikrofibrillen, die mit elektronendichtem Material verkleidet werden, jedoch weist C. intermedia zusätzlich eine innere und äußere Glättungsschicht auf. Dagegen erfolgt die Stilettbildung bei Adenorhynchus balticus (Typhloplanoida), Marirhynchus longasaeta (Kalyptorhynchia) und Provortex psammophilus und P. tubiferus (Dalyellioida) simultan und ohne Anlage einer Rohform mit einem fibrillären oder tubulären Gerüst. Strukturell sehr ähnlich wie bei A. balticus und M. longasaeta, jedoch sukzessiv erfolgt die Bildung der langen Stilette bei verschiedenen Species des Taxons Promesostoma (Typhloplanoida).Die bisher bekannten feinstrukturellen Organisationsmerkmale und die verschiedenen Bildungsmodi von penialen Hartstrukturen werden unter phylogenetischen Gesichtspunkten diskutiert.

---

Ultrastructure and differentiation of penial hard structures in free-living plathelminths

Summary Ultrastructure and differentiation of penis stylets and stylet needles have been investigated in several representatives of different groups of the free-living plathelminths. The formation of the hard structures occurs in different ways, but always intracellularly. The stylet needles of Philocelis cellata (Acoela) consist of microfibrils and are formed successively, beginning with the distal tip. The species of the taxon Paromalostomum (Macrostomida) have stylet apparatuses which also show a successive formation mode; however, the hard elements are built of microtubules and enveloped by electron-dense material. The successive differentiation of the stylet of Ciliopharyngiella intermedia occurs similarly to the hard structure formation of certain proseriates by building a framework of microfibrils which becomes enveloped by electron-dense material; in addition to this rough-form, in C. intermedia an intracellular smooth layer is formed on the inner and outer side of the stylet. In contrast, the stylet formation of Adenorhynchus balticus (Typhloplanoida), Marirhynchus longasaeta (Kalypthorhynchia), Provortex psammophilus and P. tubiferus (Dalyellioida) occurs synchronously and without a roughform, containing a fibrillar or tubular framework. The stylets in several species of the taxon Promesostoma (Typhloplanoida) are built in a way very similar to that in A. balticus and M. longasaeta, but successively.The fine structural properties known at present and the various formation modes of penial hard structures are discussed from the phylogenetic aspect.

---

Abkürzungen ae Atriumepithelzelle - ag Atrium genitale - am Atriummuskulatur - ba Bakterium - bl Basallamina oder vergleichbare Interzellularsubstanz - bm Bulbusmuskulatur - bz Stilettbildungszelle - cw Cilienwurzel - de Ductus ejaculatorius - dr Drüsenrohr - dz Ductuszelle - fz Füllzelle - hd Hemidesmosom - hz Hüllzelle - k Kern einer Stilett- bzw. Nadelbildungszelle - m entstehende Muskulatur - ma Macula adhaerens - mc Muskelzylinder - mf Mikrofibrillen - mv Mikrovilli - n Nerv - nz Nadelbildungszelle - pm äußere Penismuskulatur - rm Ringmuskulatur - s Stilett - sd Septatdesmosom - sg Sekretgangzelle - sgz Sekretgangzelle - skg Spermakornsekretgang - skr Spermakornsekretrohr - sm stilettumhüllende Muskulatur - sn Stilettnadel - sna Stilettnaht - sp Spermium - sz Sekretzelle - vg Vesicula granulorum - vgm Muskulatur der Vesicula granulorum - za Zonula adhaerens     

Tactile localisation: the function of active antennal movements in the crayfishCherax destructor

Video recordings and single frame analysis were used to study the function of the second antennae of crayfish (Cherax destructor) as a sensory system in freely behaving animals. Walking crayfish move their antennae back and forth through horizontal angles of 100 degrees and more, relative to the body long axis. At rest, animals tend to hold their antennae at angular positions between 20 and 50 degrees. Movements of the two antennae are largely independent of each other. Before and during a turn of the body the ipsilateral antenna is moved into the direction of the turn. Solid objects are explored by repeatedly moving the antennae towards and across them. Both seeing and blinded crayfish can locate stationary objects following antennal contact. On antennal contact with a small novel object, a moving animal withdraws its antenna and attacks the object. When the antenna of a blinded crayfish is lightly touched with a brush the animal turns and attacks the point of stimulation. The direction taken and the distance covered during an attack can be correlated with: the angle at which the antenna is held at the moment of contact and the distance along the antennal flagellum at which the stimulus is applied. From behavioural evidence we conclude that crayfish use information about the angular position of their antennae and about the position of stimulated mechanoreceptors along the antennal flagellum to locate objects in their environment. We suggest ways in which an active tactile system-like the crayfish's antennae--could supply animals with information about the three-dimensional layout of their environment.     

Sexual dimorphism in the visual system of flies: The divided brain of male Bibionidae (Diptera)

Summary The mapping of the compound eyes onto the visual neuropils and the cell types in the lamina and the lobula complex of Bibionidae (Diptera) were studied by means of extracellular cobalt injections and Golgi impregnations. Dorsal and ventral eyes in males map into separate dorsal-and ventral neuropils up to the level of the lobula complex. The dorsal-eye lamina is unilayered, while the ventral-eye lamina in males and the lamina in females are multilayered: layers A and C are invaded by en-passant terminals of long visual fibres, layer B by the terminals of short visual fibres. Long visual fibres have a short and a long terminal in the ventral medulla with terminal specialisations in three distinct layers. Only one type of receptor ending exists in the dorsal medulla, the terminal branches of which are restricted to one layer only. Arrays of contralateral neurones are found in the medial part of the dorsal lobula, which receives input from the zone of binocular vision of the ipsilateral dorsal eye, and in the posterior dorsal lobula and lobula plate. The dorsal lobula plate contains large tangential neurones, the dendritic arborisations of which are revealed by cobalt injection into the thoracic ganglia. The divided brain of male bibionids offers the opportunity to investigate separately the nervous systems involved in sex-specific visually guided flight behaviour and in general visually guided flight control.     

Mechanisms of cell differentiation: Affinity of a morphogenetic factor to DNA

Summary A morphogenetic factor which induces inTriturus gastrula ectoderm tissues which are derived from mesoderm and endoderm has been extracted from chicken and amphibian embryos. The factor which is protein in nature has been obtained from chicken embryos in a highly purified state.The biological activity of the chicken factor is partially inhibited when the factor is combined with chicken DNA or sonicated chicken DNA.When the 3H-labelled factor is combined with sonicated DNA and then centrifuged on a sucrose gradient the factor migrates in part with the DNA. This indicates that the factor is bound to DNA.The inferences from these results are discussed with regard to the possible mechanism of action of the factor and the molecular mechanism of differentiation.