Ultrastructural localization of thyrotropin (TSH) in the porcine anterior pituitary

Summary Two different immunocytochemical techniques based on specific antibodies against -subunits of porcine, rat and bovine TSH were applied at the ultrastructural level to identify the TSH cells in the porcine anterior pituitary and to compare the subcellular localization of the hormone.The post-embedding method on serial ultrathin sections revealed the localization of TSH in the granules of a specific cell type, negative for the other hormones. TSH was found in polyhedral cells characterized (i) by their content of granules that were the smallest of all the cell types examined, and (ii) by their flattened or slightly dilated RER cisternae. The pre-embedding method applied to isolated cells permitted a good penetration of antisera and the maintenance of antigenicity in sites inaccessible to the post-embedding method. Thus, immunoreactivity of TSH was detected in the secretory granules, the cytoplasmic matrix and in portions of the rough endoplasmic reticulum, in association with some membranes and inside some saccular structures.     

Genetic heterogeneity of propionic acidemia: Analysis of 15 Japanese patients

Summary Propionic acidemia is an autosomal recessive metabolic disease resulting from a deficiency of propionyl CoA carboxylase (PCC) activity. We have analyzed the molecular heterogeneity of Japanese propionic acidemia patients using anti-human PCC antiserum and cDNA clones coding for the two protein subunits ( and ) of the enzyme. The steady state levels of both and subunits of PCC from 15 Japanese patients were determined by Western blot. Three patients had neither nor subunits, and the amounts of both and subunits were low in 3 other patients. According to our previous data, we classified these 6 patients as having subunit deficiency. In the remaining 8 patients, subunits were normal, but the subunits were aberrant. Two patients had low levels of normal-sized subunits and 6 had subunits smaller than normal in size and greatly reduced in quantity. These 8 patients were assigned to the subunit deficiency category. One patient had apparently normal and subunits. We could not determine this patient's primary defect. These data reveal the genetic heterogeneity of molecular defects causing propionic acidemia in the Japanese. Southern blot analysis did not reveal any gross alteration in gene structure when DNA was digested withHindIII,EcoRI andTaqI. However, DNA from 3 -subunit-deficient patients, when digested withMspI and probed with PCC cDNA, revealed a unique 2.7-kb band not observed in blots of DNA from any other patient or 15 normal controls. We conclude that this alteredMspI restriction map is the result of a mutation in the subunit gene of these patients.     

Influence of Light Preferences on the Avoidance Responses of Lake Whitefish, Coregonus clupeaformis, to Cadmium

The avoidance responses of lake whitefish to cadmium were studied in a counter-current trough where Cd was added to a shaded area to which whitefish were strongly attracted. In the first experiment, responses of individual fish to sequentially increasing Cd concentrations were monitored over a 1.6h period. Whitefish did not start to move out of the shaded, Cd-treated side until a concentration of 125µgl^-1 was reached, and did not abandon this side except at the highest test concentration (250µgl^-1). This was 50 times the concentration at which whitefish avoided Cd (5µgl^-1) under uniform light conditions. In a second experiment, responses to a single high Cd concentration (150µgl^-1) were monitored over a 18.5h period (4.5h light:12h dark:2h light) in the same shaded test arena and Cd exposure conditions as above. During the initial light period Cd was avoided to the same degree as that observed at the same concentration in the first experiment, i.e. while some Cd avoidance was evident, fish remained attracted to the shaded, but Cd-treated side; this avoidance remained consistent until darkness (4.5h). Once the light gradient was replaced with uniform darkness, avoidance of Cd became very strong, surpassing that observed under uniform lighted conditions. However, this response was transitory; after 10h, Cd avoidance under total darkness returned to levels near those observed under daylight conditions with no light gradient present. Once the lights came back on (light gradient reestablished), fish were again attracted to the shaded side, and avoidance of Cd diminished, returning to the same level as that observed at the start of exposure. The above results demonstrate that when whitefish encounter competing gradients of Cd and light, attraction to favourable light conditions can strongly suppress avoidance of toxic concentrations of Cd, even over an extended period.     

Determination of a complete set of coupling constants in 13C-labeled oligonucleotides

Summary Three experiments are introduced to determine a complete set of coupling constants in RNA oligomers. In the HCCH-E.COSY experiment, the vicinal proton-proton coupling constants can be measured with high accuracy. In the P-FIDS-CT-HSQC experiment, vicinal proton-phosphorus and carbon-phosphorus couplings are measured that depend on the phosphodiester backbone torsion angles and . In the refocussed HMBC experiment, vicinal carbon-proton couplings are measured that depend on the glycosidic torsion angle .     

Multiple rough endoplasmic cisternae in “C” cells

Summary Multiple rough endoplasmic cisternae were found in C cells of the adult rat, at interphase. They are considered to be normal constituents of C cells. Their morphological relation to rough endoplasmic reticulum and their close proximity to mitochondria, Golgi dictyosomes and secretory granules suggest that they may have a role in the secretory activity of this endocrine cell.     

Stress-induced peptide release from rat intermediate pituitary

Summary We tested the hypothesis that acute restraint stress results in ultrastructural evidence for enhanced release of alpha-melanocyte-stimulating hormone (-MSH) and -endorphin from the intermediate lobe (IL) of the rat pituitary. Measurements of plasma -MSH-and -endorphin-immunoreactivity (ir) were used to confirm ultrastructural findings. Plasma -MSH-ir was elevated after 20 and 30 min of restraint while plasma -endorphin-ir peaked 10 min after the onset of restraint. Ultrastructural analysis revealed a decrease in the content of secretory granules within IL cells of stressed rats. Analysis of Golgi-related immature secretory granules in IL cells indicated that new peptide synthesis was not enhanced after 30 min of restraint. These results confirm previous studies showing and elevation of plasma -endorphin and -MSH-ir during acute restraint. Furthermore, these results indicate that quantitative analysis at the ultrastructural level can be used to assess peptide release from IL secretory cells during stress.     

Histoenzymatic characterization of sub-types of Type I fibres in fast muscles of rats

Summary The histochemical activities of succinic dehydrogenase (SDH), myofibrillar Adenosine triphosphatase (ATPase) and alpha glycerophosphate dehydrogenase were studied in serial sections of rat vastus lateralis (red) (RVL), gastrocnemius and diaphragm muscles. Three main fibre-types were distinguished. The Type I fibres of RVL and gastrocnemius muscles fell into two distinct groups: one category-Type IA showed very low ATPase activity. The second category of Type IB fibres displayed moderate ATPase reaction. The Type IA fibres were divisible into two sub-groups when tested for SDH reaction. Type IA₁ fibres possessed a homogenous distribution of diformazan·granules throughout the fibre: Type IA₂ fibres displayed characteristic moth-eaten pattern of diformazan localization. The diaphragm muscle did not show either Type IB or Type IA₂ varieties. The great majority of Type I fibres were sub-type IA₁ in the three fast muscles studied. It is also demonstrated here that an inherent heterogeneity exists between Type I fibres of diaphragm and leg muscles in regard to -GPD localization. This histochemical data emphasizes the fact that subdivision of Type I striated muscle fibres of mammalian animals into two sub-types is only approximate and that a further subcategorization is possible.     

The pancreatic islets of the teleost Scorpaena scropha

Summary The principal pancreatic islets of the teleost Scorpaena scropha are found ultrastructurally to contain four different kinds of parenchymal cells, viz. ₁-(= D), ₂-, -and agranular cells. The -cells show considerable variations in the shape of the secretory granules. A peculiar feature is that many of these granules are composed of fibrillar subunits, often in parallel arrangement. All -granules are surrounded by membranes and between the membrane and the granule core there is a moderately wide electron lucent space. The electron density of the cytoplasm in the -cells varies somewhat. The ₂-cells possess typical secretory granules with an electron dense core and a closely applied membrane. The secretory granules in the ₁-cells show also a closely applied membrane but a less dense core. Also in the -cells the electron opacity of the cytoplasm varies. The agranular cells are mainly characterized by low cytoplasmic electron density, narrow cisterns of endoplasmic reticulum and sometimes a laminated Golgi complex. Small immature secretory granules are occasionally seen in the cytoplasm of these cells. The significance of the fibrillar -granules remains obscure.This work was supported by grants from the Nordic Insulin Fund, the Town of Umeå, the Swedish Medical Research Council (Project No. B69-12X-718-04A), and by a postdoctoral fellowship from the United States Public Health Service.     

Idioms of distress: Kinship and sickness among the people of the Kingdom of Tonga

Idioms of distress refers to the popular expression of emotional tension that arises in the relationship between sickness and kinship. By reference to case studies and discussions among the Polynesian people of Tonga, the author shows where such tension arises and how it influences the sickness process. Sickness is necessarily a collective phenomenon which can best be understood not simply as a clinical event, but as an experience that is part of the experience of family. Various ways of expressing distress as a reflexive encounter between personal and cultural meaning systems are reviewed, as are several new concepts such as doing sickness as kinship, and turning in the process of decision making in the kinship management of sickness. The explanatory models of sickness in Tonga are shown to encompass culturally sanctioned expressions of distress as part of the adaptive coping mechanisms in that society. Distress frequently emerges in somatic form, as a number of studies have shown. However, the author emphasizes the kinship meaning of sickness, kinship management and sickness therapy, the adaptive process of idiomatic expressions of distress, which are expanded here and offered as potential avenues for elaboration in other cultural milieu. Two aspects of the notion idioms of distress are noted, and the phenomenon is understood as a process which acts as a prime mover in social change.     

Possible sites of release of neurosecretory granules in the sinus gland of the crayfish,Orconectes nais

Summary The sinus gland is a neurohemal organ located in the crayfish eyestalk and represents a storage site for neurohormones prior to their release into the circulation. The sinus gland contains three classes of electron dense, membrane-limited granules. Class 3 granules are the largest and most electron dense of the granules found in the sinus gland. Granules of class 1 are the smallest, while those of class 2 are the most abundant. Generally, all granules undergo similar changes during their release.Release of neurosecretory material may be initiated by a preliminary fragmentation of the parent granules into smaller granules. Following the formation of numerous smaller granules, these move to the plasma membrane and their limiting membrane apparently fuses with it thus releasing its contents into the external lamina which is applied to the sinusoidal surface of the axon terminals. Granule release does not appear to occur along the entire plasma membrane adjacent to the blood sinus but, instead, probably occurs only at specific active sites on the membrane. The active sites are characterized in part by an accumulation of small granules and clear vesicles against the cytoplasmic side of the plasma membrane. At the site of release of the neurohormone, there is often an accumulation of dense homogeneous material beneath the axolemma.Occasionally, axon endings filled with large, electron lucent vesicles are seen. These clear granules vary from 1150–1750 Å in diameter and often exhibit broken limiting membranes. Few small vesicles are seen near the plasma membrane of these endings; however, instances of invaginations of the plasma membrane occur. The significance of endings filled with clear granules is discussed.Supported by a grant from the National Research Council of Canada (No. A-4675).     

An increase in the “inhibitor-β” content of detached wheat leaves following a period of wilting

Summary Wheat seedlings were grown under a 14-hour photoperiod and the first leaves excised at the end of the eighth dark period. The effect of treatments causing wilting on the inhibitor- content of such leaves was studied.When leaves were rapidly wilted (i.e. to a 6% fresh weight loss) and extracted immediately, the amount of inhibitor- per leaf was found to be the same as in fresh turgid leaves. However, when the leaves were maintained in a wilted condition in darkness for a period of 110 minutes, there was a marked increase in inhibitor- content.The greater the degree of wilting (i.e. up to about a 9% loss in fresh weight) the greater the eventual inhibitor- content. Moreover, the increment in inhibitor- was shown to be temperature dependent.The time lapse requirement and the temperature dependency of the inhibitor- formation suggest an enzymic conversion from a precursor.If a similar phenomenon occurs during the wilting of intact plants then the increase in this growth inhibitor might play a role in some of the physiological changes which accompany water stress.     

LH-producing cells in the ovine pituitary

Summary An immunocytochemical technique using the peroxidase-antiperoxidase complex (PAP) was applied to identify and characterize the LH-secreting cells in the ovine pituitary at the ultrastructural level. These cells, round or oval in shape, possessing flattened cisternae of the rough endoplasmic reticulum, contain one class of secretory granules (mean diameter 250 nm) and large dense bodies (600 to 800 nm in diameter). LH molecules and the two subunits LH and LH were localized on the secretory granules and on the small granules near the Golgi complex. The large dense bodies, the cisternae of the endoplasmic reticulum and the saccules of the Golgi complex showed no reaction product.Abbreviations used in this Article O-LH ovine luteinizing hormone - b-LH bovine luteinizing hormone - p-LH porcine luteinizing hormone - p-LH porcine LH subunit - p-LH porcine LH subunit - O-FSH ovine follicle stimulating hormone - b-TSH bovine thyrotropic hormone - A-b LH antiserum to bovine LH - A-pLH antiserum to porcine LH subunit - A-pLH antiserum to porcine LH subunit     

Über das Zusammenwirken des erregenden und des hemmenden Neurons des M. abductor der Krebsschere beim Ablauf von Reflexen des myotatischen Typus

Zusammenfassung Eine speziell für die Verwendung in strömenden Körperflüssigkeiten konstruierte Caissonelektrode ermöglichte an dem zum Abduktor des Scherenfingers ziehenden Paar motorischer Axone die oszillographische Analyse der bei Reflexvorgängen eintretenden peripheren Interferenz von erregenden und hemmenden Impulsen. Werden mit dem Fingerglied der festgeklemmten Schere entsprechende kurze Bewegungen vorgenommen, so treten myotatische Reflexe auf, und die Aktionen der erwähnten 2 motorischen Ganglienzellen entsprechen dabei formell sehr weitgehend jenem Schema Sherringtons, durch welches für das spinale Säugerpräparat die — dortselbst intrazentral erfolgende — Umschaltung solcher Reflexvorgänge auf die gemeinsame Endstrecke aufgehellt wurde: Wir registrierten einerseits autogene, auf der Seite des gedehnten Muskels selbst eintretende Effekte (nämlich eine Streuentladung erregender Impulse, welche von einer kürzer dauernden, inversen Hemmeraktivierung begleitet ist) und anderseits die durch Dehnung des Antagonisten auszulösende reziproke Reaktion, welche aus einer Streuentladung von Hemmerimpulsen samt schwächerer inverser Aktivierung des Erregerneurons besteht. Für die am häufigsten auftretende Form der reflektorischen Antwort — die Streuentladung — darf als charakteristisch gelten, daß die Impulsfrequenz der im betreffenden Versuchstypus dominierenden, also eigentlich myotatischen Reaktion beinahe übergangslos auf durchschnittlieh 70 Hz emporstieg, um nach einem oder zwei entsprechend kurzen Intervallen (von z. B. 17 msec) zunächst auf mäßige Werte wie etwa 10–15 Hz abzusinken und dann früher oder später zu erlöschen.     

Cytochemical localization of β-galactosidase in resident and inflammatory peritoneal macrophages from C57BL mice

Summary A cytochemical method for the detection of -galactosidase (-Gase) in mouse peritoneal macrophages was used to study the ultrastructural localization of this enzyme in these cells. It was found that the reaction product for -Gase was localized in the perinuclear cisternae, the endoplasmic reticulum, the Golgi complex, lysosomes, vesicles and on the cell surface of peritoneal macrophages from untreated C57BL mice. When examined by X-ray microanalysis the crystalline reaction product was found to contain bromine, an element present in the indolyl substrate which was used to identify -Gase. Injection of Proprionibacterium acnes (P. acnes) intraperitoneally or BCG intravenously caused a visible loss in -Gase from all the organelles and from the cell surface of the macrophages.Abbreviations used -Gase -galactosidase - RP reaction product - PNC perinuclear cisternae - RER rough endoplasmic reticulum     

Über den Eidimorphismus und die Oogenese von Dinophilus gyrociliatus (Archiannelida)

Zusammenfassung Dinophilus gyrociliatus bildet zwei Oocytentypen, einen größeren, aus dem hervorgehen (-Oocyte) und einen kleineren, der sich zu entwickelt (-Oocyte). Diese beiden Oocytentypen sind von frühen Stadien der Vitellogenese an durch ihr unterschiedliches Größenwachstum zu unterscheiden. Da bei Oogonien und prävitellogenen Oocyten keine zwei unterschiedlichen Zelltypen festzustellen sind, muß man annehmen, daß die Differenzierung in - und -Oocyten in einem zwischen der Prävitellogenese- und der Vitellogenesephase gelegenen Übergangsstadium beginnt. Während der Prävitellogenesephase finden Zellverschmelzungen statt, aber es konnten keine Beziehungen zwischen der Fusion von Oocyten und der späteren Differenzierung nachgewiesen werden.Die -Oocyte beginnt schon auf einem frühen Stadium der Vitellogenese mit der Produktion von Mucopolysaccharid-Granula, die -Oocyte erst später. Diese Granula bilden nach der Ablage der Eier die Ei- oder die Kokonhülle.Die -Oocyte bildet größere Proteindottergranula als die kleinere -Oocyte. Eine Trennung zweier Zellsorten nach Granulagrößen läßt sich schon auf dem Übergangsstadium durchführen. Der absolute RNS-Gehalt der reifen -Oocyte liegt wesentlich über dem der -Oocyte; dagegen ist die Konzentration der RNS in der -Oocyte höher. Die RNS-Synthese verläuft in beiden Oocytentypen parallel zur Volumenzunahme und dauert bis zum Ende der Vitellogenesephase.

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Egg dimorphism and oogenesis in the archiannelid Dinophilus gyrociliatus

Summary Dinophilus gyrociliatus produces two types of oocytes, a big, female producing -oocyte, and a smaller, male-producing -oocyte. They may be distinguished by their different volume from the beginning of the vitellogenic phase. Neither oogonia nor previtellogenic oocytes show two types of cells, and the beginning of differentiation in -oocytes and -oocytes has to be located in a connecting stage after the previtellogenic and before the vitellogenic phase. On previtellogenic stages the cells fuse and form bigger ones, but there is no connection to be found with the differentiation of the egg cells.The -oocyte starts the production of mucopolysaccharid granules at an early vitellogenic stage; the -oocyte does so only at later stages. These granules form the egg capsule after the eggs have been laid.The -oocyte contains bigger protein yolk granules than the smaller -oocyte. Already on the connecting stage it is possible to distinguish two groups of cells by the size of their granules.The ribonucleic acid content in the -oocyte exceeds greatly that of the -oocyte. The RNA-concentration, however, is higher in the latter one. During the vitellogenic stages the rate of RNA-synthesis in either type of oocytes parallels the increase in cell volume, the synthesis lasting up to the end of the vitellogenic phase.

Für die Aufenthalte in Lille standen mir ein Forschungsstipendium der französischen Regierung sowie eine Beihilfe der Gesellschaft der Freunde und Förderer der Universität des Saarlandes zur Verfügung.     

Taxol affects meiotic spindle function in locust spermatocytes

Summary Addition of 0.1 to 10 M taxol to meiotic spindles in locust spermatocytes leads to a concentration dependent promotion of MT assembly at the centrosomes and depletion of MTs at the kinetochores, leading to the formation of prominent asters. In anaphase spindles, the equatorial region of the interzone becomes partly depleted of MTs, too. Microcinematographically, cytostatic effects are highly concentration/time dependent, being most rapid and nearly complete at 10 M taxol, but even in 0.1 M and 1 M taxol anaphase A movement is clearly affected. The drug strongly reduces the rate of chromosome-to-pole movement (anaphase A), leading to an insufficient separation of the chromosomes which indirectly hampers cytokinesis. Obviously, the chromosomal movement seems to be ratelimited by the compactness of the centrosomal asters reaching the equatorial plane in meta- and anaphase. Although the interzonal MT-number has become strongly reduced, anaphase B is not seriously affected but appears even slightly accelerated. Together with an occasional broadening of the cell equator (transverse elongation) instead of normal elongation, these results could be taken as an indication of the previously suggested active role of the cell's cortex in spindle pole separation during anaphase B (Daub andHauser 1986).Prof. Dr. K.-E.Wohlfarth-Bottermann on the occasion of his 65th birthday.     

Computational constraints that may have favoured the lamination of sensory cortex

At the transition from early reptilian ancestors to primordial mammals, the areas of sensory cortex that process topographic modalities acquire the laminar structure of isocortex. A prominent step in lamination is granulation, whereby the formerly unique principal layer of pyramidal cells is split by the insertion of a new layer of excitatory, but intrinsic, granule cells, layer IV. I consider the hypothesis that granulation, and the differentiation between supra- and infra-granular pyramidal layers, may be advantageous to support fine topography in their sensory maps. Fine topography implies a generic distinction between where information, explicitly mapped on the cortical sheet, and what information, represented in a distributed fashion as a distinct firing pattern across neurons. These patterns can be stored on recurrent collaterals in the cortex, and such memory can help substantially in the analysis of current sensory input. The simulation of a simplified network model demonstrates that a non-laminated patch of cortex must compromise between transmitting where information or retrieving what information. The simulation of a modified model including differentiation of a granular layer shows a modest but significant quantitative advantage, expressed as a less severe trade-off between what and where. The further connectivity differentiation between infra-granular and supra-granular pyramidal layers is shown to match the mix of what and where information optimal for their respective target structures.     

Age-related effects in enzyme catalysis

Summary Rat muscle glyceraldehyde-3-phosphate dehydrogenase is one of several enzymes which have been found to undergo age-related modifications. While the amount of this enzyme in muscle tissue does not change with age, both its specific activity and affinity towards its co-enzyme are significantly reduced in the old tissue.Age-related structural changes were found to exist in the nicotinamide binding site of the enzyme and the reactions leading to the activity loss in old glyceraldehyde-3-phosphate dehydrogenase were shown to involve a reversible modification of the essential cysteine-149 residue at the active site of the enzyme. The aging effects were simulated by a controlled oxidation of cys-149 in samples of young glyceraldehyde-3-phosphate dehydrogenase and subsequent reduction of this residue by 2-mercaptoethanol. The enzyme modified in this way closely resembles native old glyceraldehyde-3-phosphate dehydrogenase, indicating that the structural modifications in the latter enzyme are indeed introduced by a post-translational process. The mechanism for aging of glyceraldehyde-3-phosphate dehydrogenase which is proposed, based on these observations, thus assumes an oxidation of cys-149 as its first step followed by irreversible conformational changes in the enzyme molecule. The aging of glyceraldehyde-3-phosphate dehydrogenase may thus be triggered by the reduced ability of old muscle tissue to protect its constituents against oxidation.Abbreviations CPL circular polarization of luminescence - DTNB 5,5-dithiobis (2-nitrobenzoic acid) - GPDH D-glyceraldehyde-3-phosphate dehydrogenase - ENAD⁺ nicotinamide 1,N⁶-ethenoadenine dinucleotide     

Conversion of free β-amylase to bound β-amylase on starch granules in the barley endosperm during desiccation phase of seed development

Summary Association of -amylase with starch granules in the starchy endosperm of barley (Hordeum vulgare L. cv. Menuet) grains was characterized biochemically. In whole homogenates of dry seeds, two forms of -amylase were detected: one is free -amylase extractable with saline solution and the other is bound -amylase extractable with saline solution containing a reducing agent. The two forms of -amylase were shown to be identical in terms of mobility on disc gels, antigenicity, and molecular specific activity, indicating that the -amylase molecules of the two forms are identical. The starch granules were isolated from either dry seeds or mature seeds harvested before the desiccation phase. Both starch granule preparations were morphologically identical by microscopic inspection. The bound -amylase was predominantly associated with starch granules isolated from dry seeds, whereas it was not associated with starch granules from mature seeds harvested before desiccation. Overall results show that the periphery of starch granules is the major site of deposition for bound -amylase in dry seeds. The association of -amylase with starch granules occurs during the desiccation phase of seed development, resulting in the conversion of free -amylase into a bound form.Recipient of an award from the Union Générale de la Brasserie Française (I. H.-N.) and from the Centre National de la Recherche Scientifique and the Japan Society for the Promotion of Science under the France-Japan Cooperative Science Programme, 1985 (M.N.).