Immunocytochemical evidence for intragranular processing of pro-opiomelanocortin in the melanotropic cells of the rabbit

Summary The immunogold technique, employing antisera with clear-cut specificities, was used to localise different processing stages of pro-opiomelanocortin (POMC) in rabbit melanotropic cells. While the antiserum against ₃-MSH labelled all the secretory granules including intrasaccular condensations in the Golgi apparatus, antisera against -MSH only labelled extra-Golgi secretory vesicles (SV). All extra-Golgi SV were likewise labelled with the three antisera against -MSH used, despite their different specificities for the desacetylated, N-acetylated or C-amidated forms of the peptide. The antibody against -endorphin also labelled the extra-Golgi SV, while only some SV were labelled with the antibody against -endorphin. These results correlate with biochemical data in favour of mainly — if not exclusively — intragranular processing of POMC. Except for ₃-MSH, the cleavage of which could coincide with Golgi packaging of secretory material, other post-translational modifications of the precursor seem to occur when SV are discharged outside the Golgi area. The cleavage of -endorphin appears to be a later step in POMC processing, occurring in some mature SV.     

Occurrence of unidentified bodies that resemble “plastid initials” in poplar cells after breaking of dormancy in midwinter

Summary Electron-microscopic studies of plastids in cortical cells of poplar (Populus euramericana cv. gelrica) were carried out to examine whether any structural changes were initiated after breaking of dormancy in midwinter under non-growing conditions. After the breaking of dormancy, ultrastructural changes became evident and the profiles of plastids became heterogeneous. Organelles resembling the plastid initials proposed by Mühlenthaler and Frey-Wyssling in 1965 were frequently observed concomitant with changes in the plastid envelope. The formation of plastid initials appeared to be initiated by the formation of septa in pre-existing plastids. After this stage, narrow connections appeared between the initials and the parent plastids. Approximately 50 days after the breaking of dormancy in late March, further heterogeneity in the profiles of plastids was observed. At this stage, young plastids (plastids without starch granules) were frequently observed and the formation of plastid initials was hardly ever observed. These observations suggest that the plastid initials may be present for only a limited period in the cortical cells of the poplar and may be the precursors of the proplastids. Similar ultrastructural profiles were found in cortical cells of mulberry and in leaf buds of apple trees, suggesting that such changes in the ultrastructure of plastids are a general feature of perennials.     

Dynamics of localization and protein composition of plastid nucleoids in light-grown pea seedlings

Summary Localization and protein composition of plastid nucleoids was analyzed in light-grown pea seedlings at various stages of leaf development. In young plastids of unopened leaf buds, nucleoids were abundant and localized in the periphery of plastids, whereas, in mature leaves, chloroplasts contained nucleoids within narrow spaces restricted by thylakoids or grana. The migration of nucleoids into the interior of plastids preceded the formation of grana, and hence, the maturation of the photosynthetic apparatus. The protein composition of nucleoids was considerably different in young plastids and mature chloroplasts. Polypeptides with a molecular mass of 70–100 kDa predominated in the nucleoids of young plastids, whereas polypeptides with molecular mass of 20–30 kDa were abundant in the nucleoids of mature chloroplasts. Immuno-blot analysis with antibodies against the nucleoids of young plastids identified various polypeptides that were significantly more abundant in the nucleoids of young plastids than in the nucleoids of mature chloroplasts. These results demonstrate that plastid nucleoids are subject to dynamic changes in both localization and composition during the normal development of chloroplasts in the light.Abbreviations DAPI 4,6-diamidino-2-phenylindol - DiOC₆ 3,3-dihexyloxacarbocyanine iodide     

Characterization of glycoconjugates of guinea pig seminal vesicle by lectin histochemistry

In the present study, the expression of glycoconjugates in the guinea pig seminal vesicle was localized and partially characterized by lectin histochemistry using a battery of 30 different lectins specific for different carbohydrate residues. The results indicate that the glandular epithelium of the guinea pig seminal vesicle exhibits complex glycoconjugates rich in Man, -GlcNAc, -Gal, /-GalNAc, Fuc and complex NeuAc(2,6)Gal/GalNAc residues, as shown by its positive reactions to most lectins used. The Golgi region of the luminal secretory epithelial cells expresses a complex glycoconjugate pattern, as shown by its strong reactions to Man-(PSA, GNA), -GlcNAc-(S-WGA, PWA, DSA, UDA), -Gal- (RCA-I and -II), /-GalNAc-(SBA, Jac, VVA, BPA) and complex NeuAc-(SNA) specific lectins, indicating that the secretory epithelial cells are active in glycosylation and secretion process. It was also shown in the present study that the basal and luminal epithelial cells are different in their glycoconjugates. The basal epithelial cells are rich in NeuAc(2,3)Gal residues as they are stained specifically by MAA. The fibroblasts in the epithelial-smooth muscle interface and the smooth muscle cells close to the glandular epithelium are shown to express more glycoconjugates as they are stained intensely by GS-I-B4, GS-II and SBA. However, their role in the epithelial-stromal interaction in the seminal vesicle remains to be elucidated. In summary, the present study reports for the first time on the lectin binding patterns of the guinea pig seminal vesicle, and the results show that the seminal vesicle epithelium elaborates and secretes glycoconjugates in a complex pattern. Some of the lectins might be useful as histochemical markers for the secretory activity and specific structural components in the guinea pig seminal vesicle. © 1998 Chapman & Hall     

Transmission of paternal chloroplasts in Nicotiana

Summary Transmission of paternal chloroplasts was observed in Nicotiana, considered to inherit organelles in a strictly maternal way. Plants carrying streptomycin resistant plastids were used as pollen donors. Cell lines with paternal plastids in the offspring were selected as green (resistant) sectors on calli induced from the seedlings on streptomycin-containing media. The presence of paternal plastids in the regenerated plants was confirmed by restriction analysis. In the Nicotiana plumbaginifolia xN. plumbaginifolia Np(SR1)3 and the N. plumbaginifolia Np(gos)29 xN. tabacum SR1 crosses 2.5% and 0.07% of the offspring were found to contain paternal (tabacum) plastids, respectively. These plants, however, carried maternal mitochondria exclusively. This sexual cybridization method offers a simple way to transfer chloroplasts solely, a goal not accessible by protoplast fusion.     

The Dream as Text, The Dream as Narrative

The concepts of text and narrative are reviewed in terms of their appropriate application to the study of dreams. It is proposed that, once experienced, all dreams are texts, but that not all of these texts are narratives. Blending Jung's proposal for the form of the average dream with basic terms from narratology, dream data are employed in order to examine the narrativity of dreams, and by way of inquiring into cognitive aspects of narrative structure. Questions about the relation between language and narrative structure on the one hand, and between dreaming and language on the other are also briefly taken up.     

Mode of uptake of sterol by Arthrobacter simplex

The mechanism of uptake of water-insoluble -sitosterol by a newly isolated strain of Arthrobacter simplex SS-7 was studied. The production of an extracellular sterol-pseudosolubilizing protein during growth of A. simplex on -sitosterol was demonstrated by isolating the factor from the cell-free supernatant and its subsequent purification by Sephadex G-150 column chromatography. The M _r of the purified sterol-pseudosolubilizing protein determined by SDS–PAGE was 19kDa. The rate of sterol pseudosolubilization (5.2×10^–3g l^–1h^–1) could not adequately account for the rate of sterol uptake (72×10^–3g l^–1h^–1) and the specific growth rate (56×10^–3 h^–1). However in the unfavourable growth condition, when the cells were treated with sodium azide at the level of 30–60% of MIC, the sterol pseudosolubilization accounted for nearly 74% of the total growth containing 96% free cells. Cellular adherence to substrate particles was found to play an active role in the normal growth of the strain on -sitosterol. Unlike sodium acetate-grown cells, whose surface activity was negligible (60mNm^–1), the sterol-grown cells had strong surface activity (40mNm^–1). The high lipid content and long chain fatty acids in the cell-wall of -sitosterol-grown cells probably contribute to the high sterol adherence activity of the cells.     

Discrimination of LH,FSH, TSH and ACTH in dissociated porcine anterior pituitary cells by light and electron microscope immunocytochemistry

Summary Using the PAP unlabelled antibody method, LH, FSH, TSH and ACTH were localized at light microscope level in cultured cells dissociated from the porcine adenohypophysis. Antisera were shown to be specific for the subunits of the porcine glycoprotein hormones by radioimmunoassay and absorption studies. Using these antisera, it was found that LH and FSH were contained within the same cell, with TSH in a separate cell. When absorbed with LH, anti-porcine ACTH stained a separate distinct population of ACTH cells.Adjacent ultra-thin sections stained with anti-pLH and anti-pFSH, and examined at electron microscope level, showed that the ovoid, 150–400 nm secretory granules of the LH/FSH gonadotrophs contained both LH and FSH.The authors gratefully acknowledge the technical assistance of Carole Smith and Adrian Walsh     

The Sertoli cell of the water buffalo (Bubalus bubalis) during the spermatogenic cycle

Summary Ultrastructural features and morphometric evaluations of buffalo Sertoli cells are reported for the six phases of the spermatogenic cycle. The phases of the tubular seminiferous epithelium are identified according to characteristic cellular associations with completed spermiation as demarcation between two cycles. Average tubular diameter (245 m) and epithelial height (61 m) do not vary significantly during the cycle. The relative Sertoli cell volume in the seminiferous epithelium varies between 30% (phase 4) and 39% (phase 8). The calculated volume of a single Sertoli cell increases from a nadir of 7118 m³ in phase 3 abruptly to a maximum of 8968 m³ in phase 4 and is then gradually reduced during the following phases. The Sertoli cell surface area shows a similar trend: it amounts to 11105 m² in phase 3 and to 14260 m² in phase 4. The contact area of the Sertoli cell with adjacent cells and structures is subject to characteristic changes; from the expansion of basal Sertoli-Sertoli contacts it is concluded that the blood-testis barrier in the buffalo is particularly tight during phases 8, 1 and 2. The irregularly contoured nucleus contains a vesicular nucleolus, has a calculated volume from 465 m³ to 543 m³ and occupies 5 to 7% of the cell. Volume percentages of mitochondria (4%), Golgi apparatus and lysosomal bodies are rather constant during the cycle. Whorls and orderly arranged aggregates of the smooth endoplasmic reticulum occur in basal location as well as in close association with elongating spermatids. Smooth ER is the organelle that exhibits the most prominent changes during the Sertoli cell cycle: it occupies 5.79% in phase 3 and 20.9% in phase 4 of the total cellular volume. Phagocytosis of residual bodies is insignificant in this species and a lipid cycle is absent in buffalo Sertoli cells.     

Plastid development and floridean starch grain formation during carposporogenesis in the coralline red algaLithothrix aspergillum Gray

Summary The development of the chloroplasts and the formation of floridean starch grains during carposporegenesis in the coralline algaLithothrix aspergillum Gray, was studied by electron microscopy. Proplastids are first recognized next to the plasmalemma in the youngest cell of the carposporangial filament.The peripheral thylakoid is the first internal membrane system observed in the proplastid. As the pro-plastid increases in size an irregular branched tubular membrane system, made up of membranous tubules 30–35 nm in diameter, forms in the DNA region of the plastid. This membrane system appears to function as a pro-lamellar body as the internal thylakoids are connected to it and appear ot form from it. As thylakoid development continues, the plastids enlarge and may divide, leaving one of the daughter plastids with the pro-lamellar body. Thylakoids develop either from the pro-lamellar body, or by extension and possibly duplication of existing thylakoids. At about the time that starch grain formation commences the thylakoids have arranged themselves into parallel groups and the single DNA containing region of the plastid has separated into a number of small areas of DNA fibrils destributed throughout the plastid. The earliest signs of phycobilisomes on the outside of the thylakoids are also seen at this stage.Starch grains develop in the cytoplasm, but in intimate association with the endoplasmic reticulum (ER) which passes on both sides of the developing starch grain, and which appears to be involved in starch grain formation. This close ER-starch grain association, can persist into the mature carpospore. At no stage has any close association between the ER and the chloroplasts been observed.     

Effect of culture media on lymphokine-activated killer effector phenotype and lytic capacity

Summary We compared the ability of murine lymphokine-activated killer (LAK) cells grown in either a serum-supplemented standard medium (MEM plus fetal calf serum) or a serum-free medium (AIM-V) to lyse a range of tumour targets. LAK cells grown in either of the media killed a cultured murine tumour line (YAC-1 lymphoma) well and spared syngeneic self cells (concanavalin-A-stimulated splenocytes). However, a striking difference was noted in the ability of LAK cells grown in MEM plus fetal calf serum (as opposed to AIM-V) to kill modified self cells (trinitrophenol-modified concanavalin A blasts); LAK cells grown in the former always killed modified self cells better than those grown in the latter. This pattern held under a broad range of experimental manipulations and was found to be related to a relative increase in CD3-bearing LAK cells grown in the standard medium. These data suggest that the two media cannot be used interchangeably. This conclusion may have clinical implications for the use of LAK cells, as animal studies have been done using LAK cells generated in serum-containing medium and clinical studies have used LAK cells generated in serum-free medium.     

Purification of an X-prolyl-dipeptidyl aminopeptidase from the cell wall proteolytic system of Lactococcus lactis subsp. cremoris

Summary The -casein specific cell wall proteolytic system of Lactococcus lactis subsp. cremoris P8-2-47 contains a metal-independent X-prolyl-dipeptidyl-aminopeptidase. Suitable substrates for its assay are Gly-Pro-nitroanilide and Ala-Pronitroanilide. It is suggested that the function of the enzyme is to cleave the proline-rich sequences of -casein, as shown by the degradation of -casomorphin. It is a serine proteinase with a monomer molecular mass of about 90 000 daltons, a temperature optimum of 45°–50°C, and a pH optimum of about 7.     

Transmission of plastids by pollen in Antirrhinum majus

Summary Up to now Antirrhinum was classified as a typical example for a uniparentalmaternal inheritance of the plastids. However, the findings reported here prove that also the male gametophyte of Antirrhinum may occasionally transmit plastids into the egg. This conclusion is based on genetic experiments involving a form of the plastom mutant prasinizans which is described as gelbgrüne prasinizans. In contrast to all other plastid mutations known in Antirrhinum majus this mutant originated in Sippe 50 is completely viable. In plants containing plastids of this mutant type only, the mutant character is manifested during early growth stages. Cotyledons and first foliage leaves which are initially white or white yellow, slowly turn green and become indistinguishable from normal Sippe 50. Reciprocal crosses of green Sippe 50 with gelbgrüne prasinizans gave few variegated descendants; the others were exclusively plants identical with the maternal parent as far as leaf colour is concerned (Table). The variegated individuals cannot be gene mutants since selfing and crossing experiments showed non-mendelian inheritance. Furthermore it could be ruled out that in the cross Sippe 50 x gelbgrüne prasinizans the three variegated descendants represent spontaneous new plastom mutants because the pale tissue in these plants turned green in the same way as the paternal parent. Because of the typical greening of this mutant and since plastid mutations could be ruled out we have to conclude that plastids were transmitted by the pollen parent into the egg. There these plastids multiplied together with the maternal plastids giving rise to the chimeras after sorting-out of the two plastid types. This interpretation is supported by the observation of mixed cells in tissues where the leaf variegation is finely mosaiced. The results were possible only because the plastids of the pollen parent can be unequivocally recognised.     

Transfer of defined numbers of chloroplasts into albino protoplasts by subprotoplast/protoplast microfusion: chloroplasts can be “cloned”, by using suitable plastome combinations or selective pressure

Summary Defined numbers (1–5) of (donor) chloroplasts were transferred into (acceptor) protoplasts of plastid albino mutants by subprotoplast/protoplast microfusion. Single transferred plastids gave rise to new organelle populations in the progeny of the fusion products when suitable combinations of plastomes were used or when selective pressure for the plastome transferred was applied. This process is termed chloroplast cloning and is the first reported case of cloning a cell organelle. The plastome combination and the presence or absence of selective pressure were found to influence the frequencies with which cell lines, containing both plastomes or acceptor or donor only, were obtained, and the number of cell generations needed for complete segregation — as measured by the duration of culture before the green donor plastome could be detected. The high frequency of cell lines and regenerated shoots recovered with donor plastome only, even when only a single chloroplast was transferred, leads to the conclusion that all organelles present in the fusion product contribute to the organelle population of the progeny, i.e. organelle death or loss are not regularly occurring events during plant regeneration from protoplasts in Nicotiana tabacum.Some of the results reported here were presented at the 8th International Protoplast Symposium, Uppsala 1991     

Über die Hemmung der lambda-Phagen-Produktion in Nährbouillon

Zusammenfassung Escherichia coli K 12-Zellen produzieren in Nährbouillon in Abhängigkeit vom Pepton-Gehalt einen NB-Faktor, der durch Methotrexat induzierte lysogene Zellen an der Phagenproduktion hindert. Der NB-Faktor interferiert offenbar mit einem direkt oder indirekt bei der Phagen-Morphogenese beteiligten Schritt.     

Differences in N-acetyllactosamine synthesis between β-1,4-galactosyltransferases I and V

Unlike classical -1,4-galactosyltransferase (-1,4-GalT I), -1,4-GalT V (formerly IV*) has little activity towards 1 mM N-acetylglucosamine [Sato et al. (1998) Proc Natl Acad Sci USA 95: 472-477]. The human -1,4-GalTs I and V were expressed individually in Sf-9 cells by transfection of the full coding sequences, and their N-acetyllactosamine synthetase activities were determined towards different N-acetylglucosamine concentrations. Kinetic studies using the cell homogenates as an enzyme source revealed that -1,4-GalTs I and V possess Km values of 0.6 mM and 33 mM towards N-acetylglucosamine, and of 48 µM and 41 µM towards UDPGal, respectively. No significant inhibition of N-acetyllactosamine synthesis with -lactalbumin was observed for -1,4-GalT V but the significant inhibition with -lactalbumin was observed for -1,4-GalT I.     

Improved visualization of plastid fine structure: Plastid microtubules

Summary Fixation of leaf segments with caffeine added to the glutaraldehyde and wash buffers dramatically improves the resolution of plastid fine structure. Control preparations, fixed without the addition of caffeine, have plastids with extremely electron dense stroma material, indicative of the indiscriminate movement of phenols from the vacuole into the plastid. No structural detail may be observed in the stroma and the thylakoids stand out in negative relief. However, after caffeine fixation, electron micrographs reveal an extensive array of plastid microtubules. These plastid microtubules appear to connect the thylakoid apparatus to the plastid envelope and may even allow for changes in plastid morphology.     

Three dimensional reconstruction of the X-Y pair during pachytene in the rat (Rattus norvegicus)

The ultrastructure of the X-Y pair from rat spermatocytes has been reconstructed sterically by the study of serial sections. The X-Y pair of the rat at early pachytene contains two dense cores, a long and a short one, which form a synaptonemal complex 1.7 long at their common end. The long core (10.6 ) and the short core (4.5 ) correspond to X and Y, respectively. There is no RNA histochemically detectable in the X-Y pair. Nucleoli are independent of the X-Y pair. A low number of cells show nucleoli very near the X-Y pair but no continuity exists between these structures.     

Probable male heterogamety in the deep-sea fish Bathylagus wesethi (Teleostei: Bathylagidae)

A presumed XY chromosome pair is described fromt estis squashes from the mesopelagic deep-sea fish Bathylagus wesethi, whose 2N chromosome number was determined as 34-XY. Although the metacentric X-chromosome is the largest in the entire compliment, the Y is the smallest and only acrocentric element. The positive heteropycnosis of the sex elements was not easily distinguishable in the nuclei of first meiotic prophase. Tetraploid nuclei were observed in peripheral supporting cells of the testis. Males of at least two other congeners have similar karyotypes.     

Zur Feinstruktur der Elektrozeptorepidermis der Mormyridae (Teleostei,Pisces)

Zusammenfassung Die schwachelektrischen Mormyridae haben eine dreischichtige Epidermis, deren innere Schicht aus nur etwa 0,22 m dicken sechseckigen Zellen von ca. 60 m Durchmesser besteht. Die etwa 2 m dicken, linsenförmigen Kerne von 7,6 m Durchmesser liegen am Zellrand. Die Zellen sind zu Säulen aufgeschichtet. Ihr Rand ist ausgezackt und dort, wo er die Säulengrenze erreicht, auf etwa 0,34 m verdickt. In der Nähe der Säulengrenzen sind die Zellen über Desmosomen mit den Nachbarn in der eigenen und in der angrenzenden Säule verbunden. Diese Epidermisschicht ist auf die Körperpartien beschränkt, in denen auch Elektrorezeptoren ausgebildet sind.Die beiden anderen Epidermisschichten haben den üblichen Aufbau einer Fischepidermis, abgesehen vom Fehlen der Becherzellen.

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Ultrastructure of the electroceptor epidermis of the Mormyridae (Teleostei, Pisces)

Summary The weakly electric fish of the family Mormyridae have a three layered epidermis, with a medium layer consisting of hexagonal cells of only 0.22 m in thickness and about 60 m in diameter. The lens-shaped nuclei are about 2 m thick and 7.6 m in diameter and are situated near the border of the cells. The cells are piled up to hexagonal columns. Their margin is serrate and where it reaches the boundary of the column, it has a thickness of about 0.34 m. Close to the boundaries of the columns, the cells are linked to their neighbours within the column and of the adjoining column by desmosomes. This layer of the epidermis is confined to those regions of the body surface which also contain electroreceptors.The other layers of the epidermis have a structure as usual in fish, except for the lack of goblet cells.