Auerbach's plexus of mammals and man: Electron microscopic identification of three different types of neuronal processes in myenteric ganglia of the large intestine from rhesus monkeys,guinea-pigs and man

Summary Ganglia from Auerbach's plexus of the large intestine (caecum, appendix vermiformis, colon transversum and rectum) in man, rhesus monkey and guinea-pig are composed of nerve cells and their processes, typical Schwann cells and a vast neuropil. The neuropil consists of dendrites and axons of intrinsic nerve cell perikarya and axons of extrinsic neurons. Axonal profiles in large nerve fibre bundles are of uniform size and appearance, embedded in infoldings of Schwann cell cytoplasm and contain occasional large granular vesicles, mitochondria and neurotubules. Preterminal axons widen into vesicle filled varicosities, some of which establish synaptic contact with intrinsic nerve cell bodies.At least three different types of neuronal processes can be distinguished in the myenteric neuropil according to the size, appearance and commutual proportion of vesicles present in axonal varicosities, and their ability to accumulate exogenous 5- and 6-hydroxydopamine and 5-hydroxydopa: 1. Axonal enlargements containing a major population of small electron lucent synaptic vesicles (350–600 Å in diameter) together with a small number of membrane-bound, opaque granules (800–1,100 Å). These profiles have been identified as cholinergic axons. The boutons establish synaptic contacts with dendritic processes of intrinsic nerve cell bodies; membrane specializations are found at the preand postsynaptic sites. 2. Axonal beads of sometimes very large diameter, containing an approximately equal amount of large granular vesicles (850–1,600 Å) and small, electron lucent or faintly opaque vesicles (400–600 Å). The granular core of the large vesicles is of medium electron density and may either fill the entire vesicle or is separated from the limiting membrane by a more or less clear interspace. The fibres probably belong to intrinsic neurons, and because of the similarity of the large, membrane-bound vesicles with neurosecretory elementary granules, they have been designated p-type fibres (polypeptide fibres). The granular core of the vesicles in these fibres becomes more electron dense after treatment with 5-OH-dopa. The accumulation of an amine precursor analogue in combination with a possible storage of a polypeptide substance (or an ATP-like substance) resembles the situation in several diffusely distributed endocrine cell systems. 3. Varicosities of axons equipped with small (400–600 Å) empty or sometimes granular vesicles, medium sized (500–900 Å) vesicles with highly electron dense cores and occasional large (900–1,300 Å) granular vesicles. Pretreatment with 5-OH-dopamine increases the electron density in almost all medium-sized granular vesicles and some of the large granular vesicles; an osmiophilic core develops in some small vesicles. 6-hydroxydopamine results in degenerative changes in the varicosities of this type of neurons. Concomitantly, both catecholamine analogues markedly reduce neuronal noradrenaline in the large intestine, as demonstrated by fluorescence histochemistry and in fluorimetric determinations. The ultrastructural features of these varicosities and their reaction to 5- and 6-OH-dopamine indicate that they belong to adrenergic, sympathetic nerves. No membrane specializations could be detected at sites of close contact of the adrenergic boutons with dendrites and cell bodies of intrinsic nerve cells.Supported by grants from the Deutsche Forschungsgemeinschaft.Supported by a grant from Albert Pahlsson's Foundation, Sweden. The work was carried out within a research organization sponsored by the Swedish Medical Research Council (projects No. B70-14X-1007-05B, B70-14X-712-05, and B70-14X-56-06).     

Licht- und elektronenmikroskopische Untersuchungen über die Beeinflussung der Dynamik neurosekretorischer Zellen von Enchytraeus (Oligochaeta) durch Aktinomycin D

Zusammenfassung Die Wirkung von Aktinomycin auf die neurosekretorischen Q-und P-Zellen im Cerebralganglion von Enchytraeus wurde untersucht. Die Cytophotometrie lichtmikroskopischer Präparate von Q-Zellen ergab, daß in den ersten Stunden nach Aktinomycin-Behandlung eine deutliche Verminderung PAF-positiven Materials auftritt. Die ersten Veränderungen wurden elektronenmikroskopisch zwischen 1 und 4 Std nach Aktinomycin-Injektion beobachtet. Sie waren in beiden Zelltypen am eindeutigsten am Nucleolus. Es kommt zu einer Sonderung und räumlichen Trennung von granulärem und fibrillärem Material. Letzteres wird sehr stark vermehrt.In bezug auf Veränderungen der Strukturen des Cytoplasmas unterscheiden sich die Q-und P-Zellen besonders im Verhalten des Golgi-Apparates und der Ribosomen. Der Golgi-Apparat wird in den Q-Zellen kurze Zeit nach Applikation von Aktinomycin reduziert. In den P-Zellen persistiert er dagegen über alle beobachteten Zeitstufen hinweg. Die Ribosomen lösen sich von den Membranen in den Q-Zellen 4–8 Std nach Injektion, was in den P-Zellen nicht festzustellen ist. Diese Tatsachen führen zu der Annahme, daß das System der Proteinsynthese der P-Zellen relativ stabiler als das der Q-Zellen ist.Die in den späteren Zeitstufen beobachtete Normalisierung der Zellstrukturen läßt darauf schließen, daß die Wirkung des einmalig injizierten Aktinomycins 24 Std danach nachzulassen beginnt.

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Light and electron microscopic studies on the influence of actinomycin D on the dynamics of neurosecretory cells of Enchytraeus (Oligochaeta)

Summary The influence of actinomycin on the neurosecretory Q and P cells of the brain of Enchytraeus was studied. Cytophotometrical measurements of Q cells in light mirocscopic preparations showed a significant decrease of PAF-positive material in the first hours after actinomycin application. At the ultrastructural level primary changes were established one to four hours after injection of actinomycin: In the nucleolus granular and fibrillar material became separated; there was a substantial increase of the fibrillar component.Concerning structural changes of the cytoplasm, Q and P cells differed especially with respect to the Golgi apparatus and the ribosomes. In the Q cells the Golgi apparatus had become greatly reduced shortly after actinomycin treatment. However, it persisted in P cells during all stages examined. Ribosomes became detached from membranes only in Q cells between 4 and 8 hours after injection.These data indicate that protein synthesis in P cells shows greater stability than in Q cells. The restitution of normal ultrastruoture during subsequent stages indicates that effects begin to subside 24 hours after a single injection.

Für technische Unterstützung danken wir Frl. B. Reymann, Frl. A. Zinßer, Frau B. Cosack und Frau E. Wolschner.     

Melanogenesis in amphibians

Summary The pigmented epithelium of Rana pipiens tadpole eyes normally develops at least two types of melanosomes: (1) an elongated melanin granule of relatively homogeneous electron density, and (2) a complex melanosome which has an outer electrondense area and one or more less dense cores. Evidence indicates that complex melanosomes are formed by new melanin enclosing preexisting melanosomes. An organized fibrillar premelanosome is demonstrated with the aid of the antimelanogenic compound phenylthiourea (PTU). These premelanosomes are the developing forms of the elongated melanosomes. There is evidence that the premelanosomes originate in the smooth endoplasmic reticulum. Phenylthiourea blocks melanin synthesis in the premelanosomes; however, removal of the PTU allows pigment deposition. This finding of an organized, fibrillar premelanosome in an amphibian marks the lowest phylogenetic group in which these organelles have been described.An Oak Ridge Graduate Fellow from Catholic University of America, Washington, D.C., under appointment from Oak Ridge Associated Universities.The MAN Program is supported by the National Cancer Institute, the National Institute of General Medical Sciences, the National Institute of Allergy and Infectious Diseases, and the U.S. Atomic Energy Commission.Oak Ridge National Laboratory is operated by Union Carbide Corporation Nuclear Division for the U.S. Atomic Energy Commission.     

竹红菌甲素敏化嘌呤和嘧啶碱的光氧化作用及其影响因素

本工作利用光吸收和高效液相色谱(HPLC)技术研究了甲素对DNA分子中四种碱基A、G、C和T光氧化的敏化作用,发现在反应体系的pH为9.0、甲素浓度为3×10~(-5)mol/L、光照40分钟时,G和T紫外吸收明显降低;HPLC分析发现甲素敏化的G光氧化体系比对照体系多出现一组分峰(滞留时间0.927分钟),该峰用475nm波长检测比260nm波长检测灵敏。根据反应机制推测是G环破裂产物。在反应条件固定时,甲素敏化G的光氧化作用受pH、光照时间及甲素浓度影响极大。单线态氧淬灭剂——叠氮钠浓度在40—110mmol/L可部分抑制甲素敏化G的光氧化作用,>110mmol/L时反应完全被阻断,提示甲素对G光氧化的敏化作用主要通过单线态氧(~1O_2)即Ⅱ型机制起作用。本文还讨论了G光氧化的可能途径。     

Suspending disbelief—of Wynne-Edwards and his reception

Critique of Wynne-Edwards' views on population regulation and sociality suppose a population of discrete, mutually exclusive groups essential to his thought. Yet both his past and present work focus on continually distributed, philopatric populations; his critics have argued the untenability of a position never his own. Wynne-Edwardsian ‘group selection’ focuses on local population productivity under philopatry. A ‘group’ is a local confluence of genotypes which need not be reified, and group selection consists of the differential replication (hence heritability) of the local social environment in which a genotype is embedded. Differential productivity contingent on social environment can eliminate some relational structures on genotypes in favor of others, creating an expanding wave of population productivity as in Wright's shifting balance metaphor. Such a process is inherent in the evolution of reciprocity, where cooperators must cluster to successfully invade a population of defectors. Regulation of resource exploitation in continuously distributed populations may be modeled as overlapping n-person Prisoner's Dilemmas, where each individual participates in several distinct commons and defection represents local over-exploitation of resources.     

The androgen-dependent rat prostate protein,probasin, is a heparin-binding protein that co-purifies with heparin-binding growth factor-1

Summary Rat prostate extracts contain an abundant 20–22 kilodalton heparin-binding protein with near identical chromatographic properties, but only 0.2–1% of the mitogenic activity, of bovine brain heparin-binding growth factor-1 (acidic fibroblast growth factor). Amino terminal amino acid sequence (met-met-thr-asp-lys-asn-leu-lys-lys-lys-ile-glu-gly-asn-trp-arg-thr-val-tyr-leu-ala-ala-ser-?-val-glu-lys-ile-asn-glu-gly-ser-pro) and immunochemical analysis revealed that the protein is identical to the androgen-dependent protein “probasin”. This work was supported in part by NCI grant CA37589 (W. L. M., J. W. C.) and the Medical Research Council of Canada (R. J. M.).     

Regulation of insulin synthesis in an insulin-producing cell line (RINm5F): Long-term experiments

Summary The insulin-producing cell line RINm5F, has been used in short-term experiments to evaluate insulin secretion. We sought to maintain the responsiveness of these cells to stimuli for up to 2 days. We examined the course of new insulin synthesis over this period by measuring at intervals immunoreactive insulin (IRI) in two parts: IRI in the medium (M) and IRI extracted from the cells (C). Control cells were incubated in RPMI 1640/2.8 mM glucose/10% fetal bovine serum/200 μg/ml bacitracin (to prevent insulin degradation). The addition of dibutyryl cAMP 10 mM to the experimental dishes significantly increased total (M+C) IRI at 48 hr to 37% above the insulin content of the control dishes (p<0.01). Theophylline 10 mM increased total (M+C) IRI by 24% over control (p<0.05) after 24 hrs. Glucose, glyceraldehyde, leucine, arginine, glucagon and tolbutamide, other stimulants of insulin production, had no effect. Under the experimental conditions reported here, including the use of bacitracin, IRI synthesis can be studied for up to 48 hr. Portions of this study have been published in abstract form for the 47th Annual Meeting of the American Diabetes Association, Indianapolis, Indiana, 1987. Supported in part by the American Diabetic Association, Maryland Affiliate.     

Supercomputer analysis of purine and pyrimidine metabolism leading to DNA synthesis

A model-system is established to analyze purine and pyrimidine metabolism leading to DNA synthesis. The principal aim is to explore the flow and regulation of terminal deoxynucleoside triophosphates (dNTPs) in various input and parametric conditions. A series of flow equations are established, which are subsequently converted to differential equations. These are programmed (Fortran) and analyzed on a Cray X-MP/48 supercomputer. The pool concentrations are presented as a function of time in conditions in which various pertinent parameters of the system are modified. The system is formulated by 100 differential equations.     

Energetics of cell-cell and cell-biopolymer interactions

The energy vs distance balance of cell suspensions (in the presence and in the absence of extracellular biopolymer solutions) is studied, not only in the light of the classical Derjaguin-Landau-Verwey-Over-beek (DLVO) theory (which considered just the electrostatic (EL) and Lifshitz-van der Waals (LW) interactions), but also by taking electron-acceptor/electron-donor, or Lewis acid-base (AB) and osmotic (OS) interactions into account. Since cell surfaces, as well as many biopolymers tend to have strong monopolar electron-donor properties, they are able to engage in a strong mutual AB repulsion when immersed in a polar liquid such as water. The effects of that repulsion have been observed earlier in the guise of hydration pressure. The AB repulsion is, at close range, typically one or two orders of magnitude stronger than the EL repulsion, but its rate of decay is much steeper. In most cases, AB interactions are quantitatively the dominant factor in cell stability (when repulsive) and in “hydrophobic interactions” (when attractive). OS interactions exerted by extracellularly dissolved biopolymers are weak, but their rate of decay is very gradual, so OS repulsions engendered by biopolymer solutions may be of importance in certain long-range interactions. OS interactions exerted by biopolymers attached to cells or particles (e.g., by glycocalix glycoproteins), are very short-ranged and usually are negligibly small in comparison with the other interaction forces, in aqueous media.     

High-frequency1H NMR studies of the effects of growth factors and phorbol esters on normal syrian hamster diploid fibroblast cells

The nuclear magnetic resonance (NMR) parameters, spin-lattice (T₁), and spin-spin (T₂) relaxation time, are usually longer for neoplastic cells than for normal cells of the same cell type. This has generally been true at low NMR frequencies (≤100 MHz) when comparisons have been made between normal and neoplastic cells that have both spent a short time in culture. We have previously demonstrated that although the T₁ values of paired normal and neoplastic Syrian hamster (SH) fibroblastic cells in culture are not significantly different when measured at 300 MHz, the 300 MHz T₂ values for the neoplastic cells are smaller than those of the normal cells. (Xin et al. (1986),Cell Biophysics 8, 213.) Since treatment of normal diploid cells with polypeptide growth factors or tumor promoters frequently results in reversible expression of neoplasia-associated phenotypes, T₁ and T₂ were obtained at 300 MHz for treated and untreated SH cells to see if these compounds could also produce smaller 300 MHz T₂ values. Secondary culture SH fetal fibroblast cells were treated with epidermal growth factor (EGF), fibroblast growth factor (FGF), phorbol-12,13-didecanoate (PDD) and 4-α-phorbol-12,13-didecanoate (4αPDD). Treatment with either growth factor resulted in smaller T₂ values, but a statistically significant decrease was not observed for PDD or 4αPDD. The observed reductions in T₂ values were correlated with the morphological and growth-stimulatory effects of these compounds on the cells.