Scanning electron microscopy of the lens of the adult rat

Summary The lenses of albino rats were studied after freeze-drying with scanning electron microscopic technique.The multilamellated lens capsule is covered by adhering fibers in the equatorial region but looks otherwise almost smooth. The cuboidal, polyhedronal cells on the anterior surface of the lens are characterized by their complex interdigitations and their paucity of extracellular space. The cortical polyhedronal, mostly hexagonal lens fibers have several processes interconnecting neighbouring cells. The plasma membrane appears slightly undulated. It is covered by extracellular material. There is a well-delimited extracellular space between the lens fibers in the cortical layer. The results are discussed in relation to previous studies based upon light and electron microscopy.Supported by grants from the Swedish Medical Research Council (B70-12X-2543-02) and Magn. Bergwalls Stiftelse.     

Monoamine storage sites in the rat superior cervical ganglion following synthesis inhibition

Summary Monoamine storage sites in paraganglionic (PG-)cells of the rat superior cervical ganglion were investigated by electron and fluorescence microscopy following treatment with p-chlorophenylalanine (pCPA), disulfiram or guanethidine respectively.Dense core vesicles in PG-cells are significantly decreased (p< 0.001) in number following pCPA, and in the majority of these cells following disulfiram and guanethidine. However in a minor portion of PG-cells the latter agents cause an increase in number and in size of dense core vesicles, in parallel with structural alterations. In agreement with these electron microscopic findings fluorescence microscopic and cytophotometric evaluations reveal a general decrease in catecholamine content with few cells showing an increase.The findings provide a morphological basis for the assumption, that monoamine storage sites in PG-cells can be decreased by inhibition of monoamine synthesis, following administration of pCPA, disulfiram and guanethidine. However the two types of responses of PG-cells which occur after disulfiram and guanethidine demonstrate a functional heterogeneity of this cell system in the rat superior cervical ganglion which is discussed.Supported by Deutsche Forschungsgemeinschaft — Grant He 919/1.I like to thank Prof. Arnold, Tübingen, for the kind disposal of cytophotometric equipment.     

The fine structure of the branchial heart appendage of the cephalopod Octopus dofleini martini

Summary The branchial heart appendage of Octopus dofleini martini has been investigated electron microscopically. This organ is dominated by peripherally lobed blood sinuses. It contains free hemocyanin (often aligned in rows), amoebocytes, endothelial cells, and muscle cells which occur mainly in connection with neurons. The neurons are often exposed to the blood. The blood sinuses are enclosed by a basement membrane which contains collagen equivalents and fine fibrillar elements. The sinuses are covered by two different epithelia: 1) the epithelium in the caoity of the appendage consisting of irregularly shaped cells with processes, the so called ( 30 high) podocytes, and 2) the epithelium ( 40 in height) on the surface of the organ, which is composed of two parts: a) a lacuna-forming portion directly adjacent to the basement membrane, which is topped by b) a continuous tissue portion with occasional lacuna-canals. The intercellular spaces of the inner and outer epithelium are connected. The structures of these epithelial cells are discussed in relation to the formation of the pericardial fluid.Our thanks are given to Professor Dr. Georg Kümmel, Freie Universität Berlin, for suggesting the theme and his scientific guidance; to Dr. Kenneth M. Towe, Smithsonian Institution, Washington D.C., for generously allowing the use of his instruments and for his technical assistance; to Mr. Frank Denys, Medical Dental School of Georgetown University, Washington D.C., for sharing with me his technical skills and for making possible the occasional use of a dark room; and finally to Dr. Fred E. Witmer, Office of Saline Water, U.S. Dept. of Interior, Washington D.C., for his help with the translation, and for taking all the side effects of this study as patiently as he did.Supported in part by Grant number GB 17539 of the National Science Foundation. Used as part of a thesis submitted to the Freie Universität Berlin.     

Ultrahistochemical study on the ruthenium red surface staining

Summary The cell coat picture effect which is usually obtained with the conventional RR method, that is with the RR/OsO₄ coupled reaction, is investigated. In this first paper, each of conceivable events which might take place between RR, OsO₄ and cell surface membrane is discussed or studied. Various tests are carried out on ascites Ehrlich carcinoma cells and Zajdela ascites hepatoma cells treated with numerous chemical reagents, as also on a few pure proteins. The set of data supports the concept that the staining pattern is due to the combination in surface membranes of RR with a colloidal-like form of OsO₂. The latter might occur during the formation of stable cyclic osmic acid diesters between OsO₄ and membrane unsaturated lipids. A possibility by which the resulting marker is though also to be in a colloidal-like state is put forward. A next report will deal with this problem.     

Ultrastructure of the surface of the epithelial cells in the rat retina

Summary The epithelial cells in the retina of rats were examined by scanning and transmission electron microscopy.The apical surface of the epithelial cells is covered by a large number of microvilli, which are embedded in an extracellular material. Distinct holes remaining after detached photoreceptor cell segments are numerous. The cells are polyhedronal, usually hexagonal and holes are scattered in their plasma membrane. Many of the epithelial cells are binucleated. The borders between adjacent cells are always close to each other. Numerous cytoplasmic folds or foot processes occur at the base of the cells as well as an extracellular space.The characteristic surface structures are discussed in relation to the function of the retinal epithelial cells.Supported by grants from the Swedish Medical Research Council (B70-12X-2543-02) and Riksföreningen mot Cancer (69171, 265-B69-01X).     

Die Dolipore der Basidiomyceten

Zusammenfassung Bei den Doliporen einiger Basidiomyceten sind weitere Differenzierungen bekannt geworden. 1. Der Porenkanal kann durch 1–3 Membranen oder durch Pfropfen verschlossen werden. 2. Das Parenthesom ist von einem ribosomenreichen Material bedeckt, das durch endoplasmatisches Reticulum vom übrigen Cytoplasma abgegrenzt wird. Diese Abßenkappe kontrolliert vielleicht den Stofftransport.

---

The dolipore of the basidiomycetes

Summary Using electron microscopy, more details have been found in the fine structure of dolipores in some basidiomycetes: 1. The pore channel may be closed by 1–3 membranes or plugs. 2. The parenthesome is covered by large plugs of material rich in ribosomes which is separated from the cytoplasm by endoplasmic reticulum. It is suggested that this outer cap controls the intercellular transport.

     

Fine structure of sensory tubes on the antennule of Conchoecia spinirostris (Ostracoda,Crustacea)

Summary In addition to setae, the first antennae of Conchoecia spinirostris also bear soft sensory tubes (4 tubes + 1 seta; 2 tubes + 3 setae). These tubes were examined electron microscopically. Each tube is divided into 4 regions: the stem, the bulbous region, the main region, and the tip. A tube contains 40–60 multiciliated dendrites, some hypodermal cells, and nonneuronal cells, and it has a specialized cuticle. Each dendrite develops within the tube, on the terminal 5–8 m of its inner dendritic segment, approx. 25 cilia in a 9 × 2 + 0 pattern, whose rootlets are absent or only poorly developed. Each cilium splits up into 9 ramifications which extend into the tip. These ramifications partly take a spirallike course and form a ring in the distal main part beneath the cuticle. Their membranes often dilate into spindleshaped swellings. In the center of the middle and distal parts of the main region approx. 7 dendrites without cilia are located, one of them reaches into the tip. The poreless cuticle is extremely delicate and electron lucid. In contrast to the cuticle of the setae it is elastic and soft. Special substructures are described. The tubes are completely covered by a filamentous surface coat. Because of the structure and the thin walled nature of the cuticle, permeability for dissolved substances is assumed. The ciliary ramifications are likely to represent the receptive apparatus. The sensory tubes are interpreted as chemoreceptors. They can best be compared with the chemoreceptors of certain crustaceans, but differ strongly from the types of sensilla found in insects.Supported by project 3540 of the Fonds zur Förderung der wissenschaftlichen Forschung in Österreich. The author is deeply indebted to Prof. Gamulin (Dubrovnik) for his support and to Prof. R. Riedl and Dr. W. Klepal for helpful discussions     

Physical mechanisms by which low-frequency magnetic fields can affect the distribution of counterions on cylindrical biological cell surfaces

Effects of dc and low-frequency ac magnetic fields on the motion and distribution of counterions on surfaces of cylindrical biological cells are examined. Magnetic fields along the cell axis as well as perpendicular to it are considered. When a dc magnetic field of any physically realizable magnitude is parallel to the cell axis it has no effect on ion motion, since the resulting Lorentz force is much smaller than the counterion-to-ion attractive force. However a dc magnetic field perpendicular to the cell surface will distort any preexisting ion motion and the resulting current (i) perpendicular to the original motion will be much larger than any current induced by a low-frequency ac magnetic field of the same magnitude as the dc field and parallel to it. Nevertheless i will still be much smaller than the current i_o constituting the original ion motion since (i/i_o)=/, where is the ion cyclotron frequency and the effective counterion collision frequency. With no preexisting coherent ion motion (i_o=0) the circulating current induced by a sinusoidally time-varying magnetic field parallel to the cell axis will be well below thermal fluctuation noise as long as only a single cell is considered; however when even an infinitesimal exchange of ions between adjacent cells is assumed the magnetic field will cause a redistribution of counterions on the cell surface. The resulting steady-state distribution becomes independent of the frequency of the applied magnetic field () when , where is 1/2 of the relaxation frequency for counterion diffusion. On the basis of these results it is suggested that whenever modification of cell behavior in response to application of a low-frequency magnetic field is established, measurements of dielectric permittivity versus frequency of the cell preparation be performed. Redistribution of counterions on the cell surface would be a likely cause if the noted effect becomes independent of the frequency of the applied magnetic field above the counterion dielectric relaxation frequency. It is also suggested that in magnetic field exposure of cell preparations the size of the sample (e.g. diameter of Petri dish) and direction of the magnetic field relative to average cell orientation can critically affect experimental results.     

Characterization of the fibrillar layer at the epithelial-mesenchymal junction in tooth germs

A characteristic layer containing numerous fibrils is associated with the basement membrane of the inner enamel epithelium during the early stages of odontogenesis. However, its nature is not well understood. In this study, the layer was examined with high-resolution electron microscopy and immuno-histochemical staining. Tooth germs of monkeys (Macaca fuscata) were studied and each fibril in the layer was found to be a tubular structure, 8–9 nm in width, resembling a basotubule, the tubular structure previously observed in various basement membranes. The space between the fibrils was filled with a network formed by irregular anastomosing strands with an average thickness of 4 nm; these strands resembled the cords forming the network in the lamina densa of basement membranes. After immunoperoxidase staining, fine threads immunoreactive for laminin staining were seen winding along the strands of the network, and 1.5-nm wide filaments, immunoreactive for type IV collagen, took the form of a network arrangement. The 5-nm-wide ribbon-like structures associated with the strands were identified as heparan sulfate proteoglycan by immunostaining. These results are similar to those obtained for the cord network of the lamina densa. The fibrillar layer therefore represents a highly specialized lamina fibroreticularis of the basement membrane of the inner enamel epithelium, and rich in basotubules.     

Scanning electron microscopic studies on the synaptic bodies in the rat retina

Summary The ultrastructure of the synaptic bodies in the outer and inner plexiform layers of the rat retina was studied with scanning and transmission electron microscopy.The synaptic bodies in the outer plexiform layer are pear-shaped and their vitreal pole invaginated by processes from nerve cells. Their surfaces are covered with extracellular material, which is partly dissolved or redistributed during the fixation and rinsing procedure. The internal structure of the synaptic bodies is described.The synaptic bodies in the inner retinal plexiform layer are more difficult to identify with the scanning electron microscope. They are polyhedronal and also covered with extracellular material.The observations are discussed. The value of the application of two different preparation and analyzing methods, i. e. the scanning and the transmission electron microscopy, is stressed.Supported by grants from the Swedish Medical Research Council (B70-12X-2543-02), Expressens prenatalforskningsfond and Riksföreningen mot Cancer (265-B69-01X).     

Integral and peripheral protein composition of the apical and basolateral membrane domains in MDCK cells

Summary Selective biotinylation of the apical or basolateral domains of confluent MDCK monolayers grown on polycarbonate filters with a water soluble biotin analog, sulfo-NHS-biotin, was employed to reveal strikingly distinct patterns of endogenous peripheral and integral membrane proteins. Peripheral proteins were found to be approximately fivefold more abundant with this procedure than integral membrane proteins, both on the apical and on the basolateral surface. The distinct apical and basal patterns were shown to depend upon the integrity of the monolayer; when the tight junctions were disrupted by preincubation in calcium-depleted medium, the patterns appeared practically indistinguishable. Two-dimensional gel electrophoresis demonstrated that only a very small percentage of the biotinylated proteins were found in similar amounts on both apical and basolateral domains. These results indicate that the sorting mechanisms that segregate apical and basolateral epithelial proteins are very strict. The simple procedure described here has clear advantages over other methods available to label apical and basal epithelial surface domains, namely, higher accessibility of the biotin probe to the basolateral membrane, possibility of purifying biotinylated proteins via immobilized streptavidin and minimal exposure of the researcher to isotopes. It should be very useful in characterizing the apical and basolateral protein compositions of other epithelial cells and in studies on the development of epithelial cell polarity.     

Ultrastruktur und Bildung von Poren im Endothel von porösen und geschlossenen Kapillaren

Zusammenfassung An einer Reihe von normalen und pathologisch veränderten Organen wird die Porenstruktur, Porenbildung und Porenrückbildung elektronenmikroskopisch untersucht. Die Endothelpore ist eine Diskontinuität in der Endothelwand mit einem sehr konstanten Durchmesser von 500 Å. Das Diaphragma ist nur mit der äußeren Membranlamelle am Porenrand verbunden. Diaphragmalose Poren sind etwas größer (Ø650 Å), zeigen einen glatten Porenrand und kommen besonders in verdichteten Endothelteilen vor. Frustrane Poren münden blind in Vakuolen oder liegen in porösen Endothelfalten im Gefäßlumen. Die eigentliche Bildung der Poren geschieht immer in abgeflachten ( 800 Å) Endothelteilen. Die vorbereitende Abflachung geht jedoch in den verschiedenen Endothelzonen (Periksryon, dicker Endothelwand, Cytoplasmainseln) unterschiedliche Wege. Alle diese Vorgänge stellen Vesikulation in besonderer Lage und mit besonderer Fusionsrichtung der Vesikel dar. Wegen dieser Unterschiede wird die Endothelwand in 4 Zonen eingeteilt: Perikaryon; dicke, porenlose Wandteile mit cytoplasmatischen Vesikeln; dünne, porenhaltige Teile ohne Zellorganellen; dicke Cytoplasmainseln, die die porösen Wandteile voneinander trennen. Der Vorgang der Porenrückbildung bleibt unklar. Vielleicht besteht er in der Faltung des Endothels, die zu porösen Vakuolen führt. Die Porenbildung verändert die Endotheloberfläche nicht, kann aber das Cytoplasmavolumen vermindern. Der Aufbau des Diaphragmas sowie der Mechanismus und die auslösenden Faktoren der Porenbildung werden diskutiert.

---

Summary Ultrastructure, formation and disappearance of endothelial pores was studied electron microscopically in several normal or experimentally changed organs. Pores being discontinuies in the endothelial wall have a very constant diameter of 500 Å. The diaphragm at the margin of the pore is in contact with only the outer lamella of the unit membrane. Pores without a diaphragm being somewhat larger (Ø 650 Å) show a smoother margin and are to be found in endothelia with dense cytoplasm. Frustrated pores form blind openings in vacuoles or are situated in porous endothelial folds within the vessel lumen. Pores alway are formed in flattened parts of endothelium ( 800 Å). In thick capillary walls the endothelium previously is flattened by vesiculation, which differs in different zones of the wall (Perikaryon, thick continuous endothelium, and cytoplasmic islands in porous capillaries) in location and direction of vesicle fusion. Because of these differences the endothelial wall is divided in 4 zones: Perikaryon; thick parts without pores containing cytoplasmic vesicles etc.; flattened, porous parts containing no cytoplasmic organelles; thick cytoplasmic islands which separate porous parts. The process removing pores is not clear. Perhaps they are removed by folding of the endothelial surface and formation porous vacuoles. Formation of pores dosn't enlarge or reduce the surface, but may reduce the cytoplasmic volume of endothelium. The nature of diaphragm as well as the mechanism and releasing factors of the formation of pores are discussed.

---

---

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft     

Potentiation of growth suppression and modulation of the antigenic phenotype in human melanoma cells by the combination of recombinant human fibroblast and immune interferons

Summary Administration of interferon as a single therapeutic regimen in cancer patients with various neoplasias has had only limited efficacy in ameliorating the negative clinical course of their disease. In the present study, we have evaluated the effect of recombinant human fibroblast (IFN) and immune (IFN) interferon, alone and in combination, on growth, differentiation and the expression of class I and II histocompatibility locus antigens (HLA) and melanoma-associated antigens on the human melanoma cell line H0-1. The effect of combinations of interferons on the antigenic profile of human melanoma cells displaying different organ colonization and spontaneous metastatic potential in athymic nude mice was also determined. H0-1 cells were more sensitive to the antiproliferative activity of IFN than to IFN and the combination of interferons resulted in a potentiation of growth suppression. The antiproliferative effect of both interferons was greater in later-passage than in earlier-passage H0-1 cells, possibly reflecting alterations in the evolving tumor cell population as a result of long-term in vitro propagation and/or the selective outgrowth of cells with an increased growth rate. The enhanced growth suppression observed in H0-1 cells treated with the combination of IFN plus IFN was not associated with a significant increase in the level of melanin, a marker of melanoma differentiation, above that observed with either interferon used alone. IFN and IFN differentially modulated the expression of class I and II HLA and melanoma-associated antigens in H0-1 cells and a series of melanoma cells with different organ colonization and metastatic potential, including MeWo, MeM 50-10, MeM 50-17, 3S5 and 70W. No consistent potentiation or antagonism in the expression of any specific antigen was observed in any of the melanoma cell lines exposed to the combination of interferons. The present study demonstrates that the combination of IFN plus IFN can potentiate growth suppression in H0-1 human melanoma cells and that this effect is not associated with an increase in differentiation or a potentiation in antigenic modulation. In addition, no direct correlation between the expression of any specific antigen or its modulation by IFN or IFN, alone or in combination, and organ colonization and metastatic potential in nude mice was observed in the different melanoma cell lines.     

Hydranth structure and digestion rate as a function of temperature and salinity inClava multicornis (Cnidaria,Hydrozoa)

Summary 1. Responses to 12 different combinations of constant levels of temperature and salinity were tested in the colonial athecate hydroidClava multicornis. Criteria measured were: (a) length and width of hydranth bodies, (b) number and length of tentacles and (c) rate of digestion.2. Test colonies were obtained by allowing single hydranths — cut off from an individual primary colony — to regenerate via asexual reproduction into new secondary colonies.3. In the resulting — genetically identical — material, all criteria tested vary as a function of the different environments offered.4. In 16, 24 and 32 S hydranth length reaches maximum values at 12°C, followed by 17° and then 22°C. With increasing salinity, hydranth length declines markedly at 12°C, while there is little or no decline at 17°C and a definite increase from 24 to 32 S at 22°C.5. Hydranth width varies less extensively; in general, it follows similar patterns as does hydranth length.6. Tentacle number per hydranth tends to be positively related to the size of the hydranth body; it decreases with increasing temperature. Salinity levels producing maximum tentacle numbers vary with temperature; maxima are found at 12°C in 16, at 17°C in 24 and at 22°C at 32 S.7. Tentacle length — although a more variable criterion — is affected similarly to tentacle number; it attains, however, relatively higher values at 17°C.8. In regard to the hydranth dimensions measured, combinations of low temperatures/low salinities and of high temperatures/high salinities tend to produce maximum values.9. Rate of digestion is taken here to be identical to the time elapsing between completed food intake and defecation. This time span is reduced with increasing temperature. In all temperature levels, digestion time is shortest in 32 S.10. The results presented above compare well with those obtained earlier under similar conditions onCordylophora caspia.11. It is assumed that the structural modifications of hydranths affect rate and efficiency of exchanges between organism and environment and may thus represent a means of metabolic adjustment.

---

Hydranthenstruktur und Verdauungsrate als Funktion von Temperatur und Salzgehalt beiClava multicornis (Cnidaria, Hydrozoa)

Kurzfassung Erbgleiche Polypen vonClava multicornis Forskål wurden in 12 verschiedenen Temperatur-Salzgehalts-Kombinationen durch asexuelle Vermehrung zu neuen Kolonien herangezogen und an den adulten Hydranthen dieser erbgleichen Sekundärkolonien Länge und Breite des Hydranthenkörpers, Anzahl und Länge der Tentakel sowie die Verdauungsgeschwindigkeit ermittelt. Alle gemessenen Parameter verändern sich — zum Teil sogar erheblich — als Funktion von Temperatur und Salzgehalt. Die Veränderungen der Hydranthendimensionen führen zu Verschiebungen der Oberfläche-Volumen-Quotienten. Da der Stoffaustausch zwischen Kolonie und Umwelt primär im Bereich der Hydranthen erfolgt, kommt diesen Verschiebungen vermutlich eine Bedeutung zu im Rahmen der Stresskompensation (nicht-genetische Adaptation). Die erzielten Ergebnisse bestätigen frühere Untersuchungen vonKinne an dem naheverwandten HydroidpolypenCordylophora caspia.

     

The ultrastructure of the eye of the mosquitofishGambusia affinis

Summary The ultrastructure of the tissue components of the eye ofGambusia affinis, excluding the sensory cells, is described. The cornea consists of two different sections of collagenous layers of different density. The choroid includes an argentea composed of- and-melanophores, lipopterinophores and a choriocapillaris associated with the rete mirabile of the choroid body. Bruch's membrane, underlying the retinal pigment layer, can develop complex associations with fibroblasts delimiting the choriocapillaris. The outer section (stroma) of the iris includes several cell types that are not found in the inner or vitread section. In adultGambusia the lens capsule is well developed, but in twoweek-oldSarotherodon larvae the lens epithelium is covered only by a glycocalyx.     

Electron microscopic investigation of the surface membrane structures of the fat-cell and of their changes during depletion of the cell

Summary The main purpose of this paper was to investigate with the electron microscope the structural relationship between the fat cell and the surrounding connective tissue under various functional conditions and thereby to solve questions not decided in the past which concern the membrane of the fat-cell. Outside the well defined plasma membrane two layers were observed, one of lesser electron density next to the plasma membrane and another denser line which separates the former from the connective tissue ground substance. Fundamentally this three-layered surface membrane complex (Robertson) is the same as described by numerous authors in other cells bordering on connective tissue. However, the changes occurring in the surface membrane complex during depletion of fat cells are of special interest. The numerous long processes formed by the cell during the loss of fat in starvation are retracted in extreme depletion. At this time a pericellular space opens between the outer lamella and the plasma membrane. While the less dense material apparently becomes liquified the outer lamella of the surface membrane complex remains in contact with the connective tissue ground substance. These observations made it possible to interpret the surface membrane structures of the fat-cell as consisting, beside the plasma membrane, of a material derived from the ground substance, which is analogous to Robertson's gap substance at the surface of the Schwann cell, and of a limiting membrane toward the ground substance. The nature and possible derivation of the extracellular layers are discussed and the general functional significance of the surface membrane complex is emphasized. These considerations support the repeatedly raised objection against the use of the histological term basement membrane for the submicroscopic structures. During the depletion of the fat cell intensive micropinocytosis occurs regularly in the plasma membrane. It is suggested that the pinching off of numerous pockets may effect the elimination of membrane material in conjunction with the decrease in the surface area which has been found to take place in extreme depletion of the fat cell.This work was performed under the auspices of the U.S. Atomic Energy Commission.     

Distribution of pectic polysaccharides throughout walls of suspension-cultured carrot cells

Summary A monoclonal antibody (2 F 4) recognizing a conformational epitope of polygalacturonic acid was used for immunogold localization of pectins in walls of suspension-cultured carrot (D. carota L.) cells at the electron microscopic level. In microcolonies of young or mature cells, polygalacturonic acid was essentially located on the middle lamella material expanded at three-way junctions between cells or lining intercellular spaces but was not found in primary walls. Middle lamellae far from junction zones and intercellular spaces were not recognized. Largely esterified pectic polymers, only detected by the 2 F 4 antibodies after on-grid de-esterification treatment by pectin methyl esterases, were present within all primary cell walls. Golgi bodies and associated vesicles were also labeled by the 2 F 4 antibodies only after de-esterification treatment, which indicates that pectic polymers are synthesized and secreted in a highly esterified form. A decrease of pectin esterification, which results probably from an in situ enzymatic de-esterification of the pectic polymers of the primary walls, was observed in senescent cells. These results are discussed in relation to biochemical analyses showing changes of the methyl ester content of pectins during the cell-wall growth.     

Studies on the acetylcholinesterase (ache)-positive and -negative autonomic axons supplying smooth muscle in the normal and 6-hydroxydopamine (6-OHDA)treated rat iris

Summary The adrenergic and cholinergic innervation to the rat iris has been studied at a light and electron microscopic level. Catecholamine fluorescence histochemistry showed adrenergic nerves to be present in both the dilatator and the constrictor pupillae regions. At a fine structural level the terminal innervation of the iris was studied and criteria for the differentiation between presumptive adrenergic and presumptive cholinergic axon terminals were examined. To aid this examination presumptive adrenergic axons were either labelled with the false adrenergic transmitter, 5-hydroxydopamine, or chemical sympathectomy performed using 6-hydroxydopamine. The value of using acetylcholinesterase staining as a marker for cholinergic nerve terminals was also studied.Results showed a mixed adrenergic/cholinergic innervation to the dilatator pupillae. In the constrictor pupillae an exclusively cholinergic innervation was found although adrenergic and cholinergic nerves were found supplying the blood vessels and at the dilatator-constrictor interface. These findings are discussed with regard to innervation-function relationships in the iris.     

βs Haplotypes in various world populations

Summary We have determined the ^s haplotypes in 709 patients with sickle cell anemia, 30 with SC disease, 91 with S--thalassemia, and in 322 Hb S heterozygotes from different countries. The methodology concerned the detection of mutations in the promoter sequences of the ^G- and ^A-globin genes through dot blot analysis of amplified DNA with ³²P-labeled probes, and an analysis of isolated Hb F by reversed phase high performance liquid chromatography to detect the presence of the ^AT chain [^A75 (E19) IleThr] that is characteristic for haplotype 17 (Cameroon). The results support previously published data obtained with conventional methodology that indicates that the ^s gene arose separately in different locations. The present methodology has the advantage of being relatively inexpensive and fast, allowing the collection of a vast body of data in a short period of time. It also offers the opportunity of identifying unusual ^s haplotypes that may be associated with a milder expression of the disease. The numerous blood samples obtained from many SS patients living in different countries made it possible to compare their hematological data. Such information is included (as average values) for 395 SS patients with haplotype 19/19, for 2 with haplotype 17/17, for 50 with haplotype 20/20, for 2 with haplotype 3/3, and for 37 with haplotype 31/31. Some information on haplotype characteristics of normal ^A chromosomes is also presented.     

The relationship between nerves and smooth muscle cells in the rat iris

Summary The dilatator muscle cells form short projections into the stroma of the iris. Close to these projections run several nerve bundles. The unmyelinated axons often show enlargements (varicosities) containing mitochondria and vesicles. Several of the varicosities are partly denuded of the Schwann cell and are covered only by a basement membrane. The varicosities are then separated from the muscle cells only by basement membranes and a 0.1–1 stromal space. The ultrastructure of the iris dilatator muscle thus also fits the view that the autonomic ground plexus with its varicosities forms the real innervation apparatus.The smallest space between axon and muscle has a width of 700–900 Å and is cemented with basement membrane material. It is suggested that the main function of these contact sites is not to transmit a nerve impulse but to anchor the nerves to their effector organ.This study has been supported by grants from the Swedish State Research Council (U 267) and the United States Public Health Service (N B 2854-04).