The ultrastructure of the female accessory gland,the cement gland,in the insect Rhodnius prolixus

The cement gland of Rhodnius prolixus is an epidermally derived tubular gland consisting of a distal synthetic region and a proximal muscular duct region. The synthetic region consists of numerous secretory units joined to a central chitinous duct via cuticular ductules. Proteinaceous secretion, synthesized by the goblet-shaped secretory cell, passes through the delicate cuticular lattice of a ductule-end apparatus and out through fine ductules to the central duct. Secretory cells are rich in rough endoplasmic reticulum and mitochondria. Light microscopy, SEM and TEM reveal the delicate lattice-like end apparatus structure, its formation and relationship to the secretory cell. The secretory cell associates via septate junctions with a tubular ductule cell that encloses a cuticle-lined ductule by forming an elaborate septate junction with itself. The ductules are continuous with the cuticle lining of the large central duct that conveys secretion to the proximal area. The proximal muscular duct has a corrugated cuticular lining, a thin epithelium rich in microtubules and thick longitudinal, striated muscles which contract during oviposition, forcing the secretion out. Histochemistry and electrophoresis reveal the secretion as proteinaceous.     

Development and cellular differentiation of an insect telotrophic ovary (Rhodnius prolixus)

Differentiation events accompanying the larval-adult ovarian transformation in Rhodnius prolixus can be divided into three phases: proliferative phase (unfed to 3 days post-feed or DPF), early differentiation phase (9–15 DPF) and late differentiation phase (16 DPF to moult at 21 DPF). Ovarioles remain morphologically larval until feeding initiates development. The unfed ovariole contains germ cells surrounding a central trophic core region with the ‘germarial lumen’ occupying the basal region of the tropharium immediately above the pre-follicular tissue. Mitosis of germ cells during the proliferative phase results in a progressive increase in tropharial size with no differentiation of tissues. Regional specialization within the ovariole marks the beginning of the early differentiation phase. A zone of oocytes is established at the base of the tropharium with nuclei containing synaptonemal complexes and condensing chromosomes. Nurse cell differentiation is characterized by nucleolar elaboration and nucleo-cytoplasmic transport, the cytoplasm becoming rich in ribosomes. Autoradiographic results suggest that functional nurse cell-oocyte divergence occurs concurrently with morphological divergence. Pre-follicular tissue is divided into apical and basal zones with apical zone differentiation occurring during early and late differentiation phases.     

Development of nurse cell-oocyte interactions in the insect telotrophic ovary (Rhodnius prolixus)

The establishment and reorganization of intercellular bridges during larval-adult ovarian differentiation is the basis of the syncytial nature of the adult hemipteran telotrophic ovary. The formation, in the late differentiation phase, of groups of closely arranged nurse cell nuclei occupying a common cytoplasm results from membrane fusions. Oocyte-oocyte intercellular bridge systems later are modified to form the trophic cords. The trophic core, which undergoes a restructuring during the late differentiation phase, mediates nurse cell-oocyte interactions in this system. Material, transported to and accumulated by late differentiation phase pre-vitellogenic oocytes, originates from trophic core restructuring and zone III nurse cell production.     

Luteolytic potencin in a novel prostaglandin analogue

Estrus induction in cycling sows with exogenous prostaglandin is hindered by the extended refractory period and resistance of swine to the luteolytic action of PG's. The luteolytic potency of a novel PGF_2α analogue, Schering ZK 71677, was assessed at four different dosages in chronically cannulated cycling swine. Dosages totalling 1, 1.5, 3 and 10 mg of ZK 71677 were split into two injections given intramuscularly on day 13 of the estrous cycle. Onset of behavioural estrus and serum levels of progesterone and PGF metabolite were monitored. The three lowest doses did not advance estrus but did cause a transitory decline in serum progesterone concentration on day 13. The 10 mg level advanced estrus in three of four sows although the overall mean cycle length did not differ from pretreatment cycles (18.5 ± 1.3 vs 21.3 ± 0.6 days). Progesterone values droppd steadily in all four sows in the 10 mg treatment group. No level of ZK 71677 affected serum levels of PGF metabolite. The highest level tested of PGF_2α analogue ZK 71677 elicited an uninterrupted decline in serum progesterone concentrations but was inconsistent in its ability to promote early onset of behavioural estrus in swine.     

Structural changes in light- and dark-adapted compound eyes of the australian earwig Labidura riparia truncata (dermaptera)

The apposition acone eye of Labidura is relatively small—550–600 facets—with a thick corneal lens and shallow retina. The retinula cell columns are each formed of six peripheral cells plus two central cells, a partially fused rhabdom, and dense pigment in two or three cell types. Upon adaptation from light to dark, the most striking photomechanical response is a proximal broadening of the cone cells, which results in a 38-fold increase in cross-sectional area of the aperture. While longitudinal rhabdom movement is small, microvillar diameters swell in response to light and contract in the dark. Irregularities of facet pattern and shape, and in ommatidial alignment were found, particularly towards eye margins. Three types of interommatidial sense organs on the eye surface are described, one of which has not been previously reported. An argument is presented to explain how the field of view and sensitivity are both apparently decreased in the acone eye by exposure to light.     

The suppression of experimental allergic encephalomyelitis in Lewis rats by treatment with myelin basic protein-cell conjugates

Pretreatment of Lewis rats with guinea pig (GP) myelin basic protein (MBP) coupled to syngeneic spleen leukocytes (SL) suppressed the subsequent induction of experimental allegic encephalomyelitis (EAE) with GP-MBP in Freund's complete adjuvant. The degree of suppression correlated positively with the amount of antigen coupled to the SL. GP-MBP coupled to syngeneic red blood cells (RBC) also resulted in suppression of EAE and the extent of the suppression was related to the dose of cells. These regimens of pretreatment also resulted in a decrease in the in vitro lymphocyte proliferative response to GP-MBP and in the extent of perivascular cuffing in the spinal cord. No decrease in the anti-MBP antibody response was detected in rats pretreated with either GP-MBP-SL or GP-MBP-RBC conjugates. Transfer of lymph node cells from rats pretreated with GP-MBP-RBC resulted in a decrease in disease severity in recipients. It is concluded that prior administration of MBP-cell conjugates is an effective way of suppressing the symptoms of EAE.     

Human monocyte chemiluminescence triggered by IgG aggregates. Requirement of phospholipase activation and modulation by Fc receptor ligands

Human mononuclear cells stimulated with soluble IgG aggregates generated chemiluminescence, a response attributable to monocytes. Some requirements of this reaction were examined by preincubation of the cells with a variety of inhibitors. The protease antagonists TPCK and TLCK, the phospholipase inhibitors quinacrine and BPB, and the calcium channel blocker verapamil were all inhibitory at micromolar concentrations. The oxygen metabolite scavengers SOD and catalase were less inhibitory. These findings are consistent with a major role for arachidonic acid metabolites in the generation of light. Modulation of monocyte Fc-mediated chemiluminescence also occurred by preincubation of the cells with Fc ligands. While IgG aggregates and monomeric IgG blocked Fc-dependent chemiluminescence, IgG Fc fragments were stimulatory of this response.     

Changes of immunoreactive somatostatin and beta-endorphin content in rat brain after amygdaloid kindling

A possible contribution of brain beta-endorphin and somatostatin to the epileptogenicity established by amygdaloid kindling was investigated in rats. Fourteen male rats were chronically implanted with electrodes placed bilaterally into the amygdala. The rats received 1 sec of electrical stimulation to the left amygdala each day. Generalized seizures were observed on average 10 days after initiation of kindling and the electrical stimulation was continued up to twenty-one days. Two months after the completion of the kindling procedure, each kindled and control rat was killed by microwave irradiation and the brains were dissected on ice into thirteen subregions. Each region was homogenized and centrifuged twice in 0.1 N acetic acid. The supernatant extracts were decanted and stored at - 20 degrees C until assay. Immunoreactive beta-endorphin and somatostatin were measured by radioimmunoassays. There were no significant differences in brain beta-endorphin contents between the two groups. In kindled rats, immunoreactive somatostatin was increased significantly in amygdala, sensorimotor, piriform, and entorhinal cortex. The results suggest that changes in somatostatin may be associated with epileptic susceptibility induced by the electrical kindling procedure.     

Insights into the RNA quadruplex binding specificity of DDX21

Guanine quadruplexes can form in both DNA and RNA and influence many biological processes through various protein interactions. The DEAD-box RNA helicase protein DDX21 has been shown to bind and remodel RNA quadruplexes but little is known about its specificity for different quadruplex species. Previous reports have suggested DDX21 may interact with telomeric repeat containing RNA quadruplex (TERRA), an integral component of the telomere that contributes to telomeric heterochromatin formation and telomere length regulation. Here we report that the C-terminus of DDX21 directly interacts with TERRA. We use, for the first time, 2D saturation transfer difference NMR to map the protein binding site on a ribonucleic acid species and show that the quadruplex binding domain of DDX21 interacts primarily with the phosphoribose backbone of quadruplexes. Furthermore, by mutating the 2′OH of loop nucleotides we can drastically reduce DDX21's affinity for quadruplex, indicating that the recognition of quadruplex and specificity for TERRA is mediated by interactions with the 2′OH of loop nucleotides.     

A comparison between the synthesis of contractile proteins and the accumulation of their translatable mRNAs during calf myoblast differentiation

The morphogenesis of muscle spindles in the mouse extensor digitorum longus was studied in closely spaced, serial, ultrathin transverse sections, permitting evaluation of the developing spindles' three-dimensional cytoarchitecture. Afferent nerve terminals are identifiable as early as 15 days in utero, and the adult number of spindles is present at 17 days in utero. By establishing continuity between the clearly identifiable equatorial regions and the polar regions, which are morphologically indistinguishable from surrounding extrafusal fibers, it could be determined that fusimotor innervation was present by the 19th day of the in utero development. In all spindles examined, the efferent innervation occurred extracapsularly. At birth, the adult number (four or five) of intrafusal fibers are found in each spindle, and fusimotor innervation is frequently intracapsular. Evidence is presented supporting the hypothesis that successive generations of intrafusal myotubes are formed by the fusion of mononucleated cells. Newly formed myotubes are preferentially distributed in close relationship to the sensory-innervated region of the primary intrafusal myotube. Analyses of growth parameters indicate that the difference in length and diameter between intrafusal and extrafusal fibers is principally a postnatal phenomenon.     

Monoclonal antibody-directed immunopurification and identification of cytochromes P-450

A 28 amino acid peptide with diuretic and natriuretic activity has been purified from rat atrial muscle. The primary structure of this atrial peptide is H-Ser-Leu-Arg-Arg-Ser-Ser-Cys-Phe-Gly-Gly-Arg-Ile-Asp-Arg-Ile-Gly- (sequence in text) Ala-Gln-Ser-Gly-Leu-Gly-Cys-Asn-Ser-Phe-(Arg)-Tyr-OH. The biological activity of this peptide is identical to that of atrial natriuretic factor and cardionatrin I isolated from rat atria.     

The thermodynamics of lanthanide ion binding to adriamycin

We have examined the thermodynamics of lanthanide ion binding to adriamycin by monitoring the effects of variations in temperature on the dissociation constants of various lanthanide ion complexes of the drug. These constants were obtained by analyzing the extent of quenching of the fluorecence of adriamycin in the presence of lanthanide ions in terms of an equilibrium binding process. Our binding model included the following features, all of which are supported by evidence derived from previous published reports, vide infra. The lanthanides form 1:1 complexes with adriamycin. The binding is dependent on the pH of the solution, indicating that only the nonprotonated amine form of the drug participates in lanthanide ion binding. And finally the drug self-associates in solution to for a dimeric species. Our present results indicate that the binding process is almost completely independent of temperature, indicating that the enthalpy of complex formation is extremely small. The entropy terms are consistent with the formation of a complex in which the adriamycin acts as a bidentate ligand. Our results suggest that the lanthanide complexes are isostructural, at least as far as the adriamycin is concerned, throughout the lanthanide series.     

Metal ion-biomolecule interactions. XII. 1H and 13C NMR evidence for the preferred reaction of thymidine over guanosine in exchange and competition reactions with Mercury(II) and Methylmercury(II)

Mercury(II) bridge complexes of the type [Nuc-Hg-Nuc] (Nuc = thymidine or guanosine), and methylmercury(II) complexes of thymidine and guanosine of the type [CH₃Hg(Nuc)], have been prepared under appropriate conditions of pH and reactant's stochiometry in acqueous soluton. The various complexes have been characterized by ¹H and ¹³C NMR and used as probes, in competition and exchange studies, to establish the relative affinities of Hg(II) and CH₃Hg(II) towards the nucleosides guanosine and thymidine. These studies have confirmed that Hg(II) and CH₃Hg(II) bind to N₃ of thymidine in preference to N₁ of guanosine. The studies further show that reactions of mercury(II) with the nucleosides are thermodynamically controlled; the preperential binding reflects the relative stabilities of the respective complexes.     

Chromosome condensation activity in Rana pipiens eggs matured in vivo and in blastomeres arrested by cytostatic factor (CSF)

The ability of brain nuclei to give rise to condensed chromosomes was studied inRana pipiens eggs which had undergone meiotic maturation in vivo, in blastomeres of two-cell embryos which had been arrested at metaphase by the injection of cytostatic factor (CSF) from mature eggs, and in immature fully grown ovarian oocytes with and without prior CSF injection. Chromosomes from brain nuclei were found to condense within 4 h in mature eggs and this chromosome condensation activity was enhanced by the chelation of free Ca²⁺ in the nuclear isolation medium. Chromosomes also condensed in CSF-arrested blastomeres whether they were placed in the blastomere 30 min before the CSF injection or as long as 22 h after the CSF. Both the Ca²⁺-sensitive CSF, 1CSF, and the Ca²⁺-insensitive CSF, 2CSF, resulted in chromosome condensation within arrested blastomeres. The condensation was accompanied by the formation of multipolar spindles and asters. However, it was found that cytoplasm in CSF-arrested blastomeres does not arrest mitosis at metaphase when transferred into a cleaving blastomere. Other experiments demonstrated that chromosome condensation does not occur in ovarian oocytes even when supplied with CSF. The results are interpreted as indicating that CSF does not directly bring about chromosome condensation, but arrests the cell cycle at metaphase and stabilizes the cytoplasmic conditions of metaphase which, in turn, induce chromosome condensation in foreign nuclei as well as spindle and aster formation.     

Preparation of radioactive iodinated cholylhistamine for use in the radioimmunoassay of cholic acid

A major handicap in the development of simple and accurate radioimmunoassay procedures for bile acids has been the lack of a radioactive standard of high specific activity. To provide such a compound, we first synthesized cholylhistamine using the carbodiimide reaction. The hypothesized structure was confirmed by elemental analysis, thin-layer chromatography, infrared and mass spectral analysis. The cholylhistamine was then iodinated with 125I, using the choloramine-T method. The 125I-cholylhistamine was bound by antisera raised against a cholic acid-bovine serum albumin conjugate. This procedure should prove useful in preparing radioactive conjugates for all of the bile acids.     

The lambda head-tail joining reaction: purification, properties and structure of biologically active heads and tails

Procedures were developed to obtain biologically active lambda heads and tails at high purity with 20 to 40% recovery. Free heads, free tails and phage particles differ markedly in stability. Phage are stable in solutions containing Mg²⁺ but tails are not. The protein subunits which form the shaft of the tail dissociate in the presence of Mg²⁺ and form multisubunit spherical structures. EDTA protects free tails against inactivation but disrupts heads and phage particles. The four carbon diamine, putrescine, stabilizes heads against inactivation; the three and five carbon diamines are less effective. Electron micrographs reveal a new “knob” structure at the distal end of the tail fiber of phage and of free tails. Tails released from EDTA-disrupted phage possess a “head-tail connector”, a structure not present on the tail before its joining with a head.     

Development of the chick embryo mesoblast. Formation of the embryonic axis and establishment of the metameric pattern

The mesoblast of the primary organizer region of the developing chick embryo at the early head process stage was examined with the scanning electron microscope. It was found that the mesoblast layer is patterned from its inception at the primitive streak. Viewed dorsally, the mesoblast region most recently traversed by Hensen's node is metameric. Each segment consists of two 175-μm-diameter circular buttons of paraxial mesoblast (somitomeres) and an enclosed axial region. These tripartite segments are stacked tandemly and mark precisely, in the ectoderm above, the limit of neural plate formation. Viewed ventrally, the metameric pattern of the mesoblast is most closely mimicked by underlying endoblast, which shows corresponding radially arranged wedge-shaped cells in somitomere-sized circular patches. At this stage of development, each paraxial somitomere is a slightly hollowed, squat cylinder, composed of tapering mesenchymal cells whose long axes are directed toward the core center. Closely timed with neurulation, somitomeres undergo morphogenesis, being first converted to triangular wedges and, finally, condensed into cubes. Anteriorly, somitomeres participate in branchiomeric development, while posteriorly, they develop into somites. Examination of segmental plates shows that they consist of about 11 tandem somitomeres not visible by light microscopy. The most mature somitomeres, closest to the emerging somites, are delineated from one another by cellular orientations and the progressive buildup of fibrous extracellular matrix. The least mature somitomeres are not as well defined, but appear initially just posterior to Hensen's node and merge medially with the notochordal process. The observations suggest that the emergence of somitomeres from the paraxial mesoblast of the primitive streak is the result of its association with nodal cells. Further, this combined association of the mesoblast heralds primary induction and establishes the metameric pattern of the basic body plan.     

Activation of the contractile system in crustacean muscle: Ultrastructural evidence for the role of the T system

Freeze-fracture and thin sections of lobster abdominal fast flexor muscle were used to study the morphology of the sarcoplasmic reticulum (SR) and T system of crustacean muscle. Tannic acid mordanting, which can result in a dense black deposit in the T system lumen, was used to distinguish T system from SR membranes. Ferritin was also used as an extracellular tracer to confirm the tannic acid method. The T system consists of an extensive network of flattened sacs which fills most of the space between the mycfibrils and is in close contact with them. The SR also appears as flattened sacs, sometimes with fenestrations. There is extensive junctional contact between the SR and T system. Quantitative estimates of the volume and surface area of the membranes show that the T system has about 50 % more surface area than the SR. The intramembrane particle (IMP) density of the PF face of the T system is about 1100/ μm² membrane, while the IMP density of the PF face of the SR is about 4800/ μm² membrane. In morphology, extent, and IMP density, the T system of lobster abdominal fast flexor muscle appears (AFF) adapted to provide at least part of the Ca²⁺ for muscle activation and the transport system for relaxation.