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.     

Elektronenmikroskopische Untersuchungen zur Dynamik neurosekretorischer Zellen von Enchytraeus (Oligochaeta)

Zusammenfassung Von lichtmikroskopischen Befunden der Neurosekretion bei dem Oligochaeten Enchytraeus ausgehend, haben wir an zwei elektronenmikroskopisch genau bezeichneten und festgelegten Zellen bzw. Zelltypen, die bereits lichtmikroskopisch charakterisiert worden waren, Untersuchungen über die submikroskopisch faßbare Zelldynamik durchgeführt. Die beiden Arten neurosekretorischer Zellen (Q-Zelle und P-Zellen) sind elektronen-mikroskopisch schon durch ihre Lage zu erfassen. Sie können nicht nur durch die Zellgröße, sondern auch durch ihre Elementargranula, den Aufbau des endoplasmatischen Retikulums und den Golgi-Apparat eindeutig unterschieden werden. Wie schon in den lichtmikroskopischen Untersuchungen wurde die Sekretionsaktivität mit der Amputation ausgelöst. Sowohl in der Q -als auch in der P-Zelle bewirkt die Amputation eine unmittelbare Sekretentleerung. Die darauf einsetzende Phase der Sekretproduktion ist submikroskopisch durch eine erhöhte Zahl von Golgistrukturen in diesen Zellen, durch das deutlich in Erscheinung tretende granuläre endoplasmatische Retikulum und durch eine fortschreitende Vergrößerung und Verdichtung von Lysosomen in beiden Zelltypen gekennzeichnet. Für die Q-Zelle sind weiterhin die Verstärkung des diesen Zelltyp besonders noch charakterisierenden Bereiches von Membranzisternen und die dortige Ribosomenbildung typisch. Auf Grund der Feststellungen wird die Frage der Beziehung einzelner Strukturen in diesen beiden Zelltypen zur Produktion des Neuro-sekrets diskutiert. Die elektronenmikroskopische Untersuchung führte zur Entdeckung eines weiteren Zelltyps, der im Lichtmikroskop bisher nicht erkannt worden war und der sich durch besonderen Reichtum an Mitochondrien und großen Lipoid (?)-Komplexen auszeichnet (M-Zelle). Über seine Bedeutung ist jedoch noch keine Aussage möglich.

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Summary The cytophysiology of two types of neurosecretory cells (Q and P cell) in the brain of the oligochete Enchytraeus was studied at the ultrastructural level. These cell types can be identified by their location, and particularly by the size difference of their elementary granules. Amputation of the last ten segments caused a release of secretory product followed by a phase of renewed production. This was characterized by changes in the endoplasmic reticulum, the Golgi apparatus, and the lysosomes. The role of these structures in the production of neurosecretory material was discussed. Furthermore, a cell type with extraordinarily numerous mitochondria, hitherto unknown in Enchytraeus, was described. Its function has not yet been determined.

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Mit Unterstützung durch die Sächsische Akademie der Wissenschaften zu Leipzig.

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Herrn Prof. Dr. F. Seidel, Marburg, zum 70. Geburtstag gewidmet.     

Identification of molecular markers in soybean comparing RFLP,RAPD and AFLP DNA mapping techniques

Three different DNA mapping techniques—RFLP, RAPD and AFLP—were used on identical soybean germplasm to compare their ability to identify markers in the development of a genetic linkage map. Polymorphisms present in fourteen different soybean cultivars were demonstrated using all three techniques. AFLP, a novel PCR-based technique, was able to identify multiple polymorphic bands in a denaturing gel using 60 of 64 primer pairs tested. AFLP relies on primers designed in part on sequences for endonuclease restriction sites and on three selective nucleotides. The 60 diagnostic primer pairs tested for AFLP analysis each distinguished on average six polymorphic bands. Using specific primers designed for soybean fromEco RI andMse I restriction site sequences and three selective nucleotides, as many as 12 polymorphic bands per primer could be obtained with AFLP techniques. Only 35% of the RAPD reactions identified a polymorphic band using the same soybean cultivars, and in those positive reactions, typically only one or two polymorphic bands per gel were found. Identification of polymorphic bands using RFLP techniques was the most cumbersome, because Southern blotting and probe hybridization were required. Over 50% of the soybean RFLP probes examined failed to distinguish even a single polymorphic band, and the RFLP probes that did distinguish polymorphic bands seldom identified more than one polymorphic band. We conclude that, among the three techniques tested, AFLP is the most useful.     

Ultrastructure of the prothoracic gland cells of the last instar of Galleria mellonella in relation to the state of development

The prothoracic glands of the last instar of Galleria mellonella undergo characteristic alterations of their cellular fine structure closely related to cellular activity. During progressive secretory activity of the gland cells there are extensive plasmalemmal infoldings and formation of a pronounced lacunar system. Mitochondria of the active cell phase are characterized by a specific increase in size and paler colour of the matrix. In contrast to the alterations, nuclei, ER and Golgi cisterns do not undergo any submicroscopic changes during the different phases of cellular activity. The relationship between the substructural phenomena and the specific phases of cellular activity are discussed.     

Genetic Diversity in Musa acuminata Colla and Musa balbisiana Colla and some of their natural hybrids using AFLP Markers

Genetic diversity and relationships were assessed in 28 accessions of Musa acuminata (AA) Colla and Musa balbisiana (BB) Colla, and some of their natural hybrids, using the amplified fragment length polymorphisms (AFLP) technique. Fifteen AFLP +3 primer pairs produced 527 polymorphic bands among the accessions. Neighbor-joining and principal co-ordinate (PCO) analyses using Jaccard's similarity coefficient produced four major clusters that closely corresponded with the genome composition of the accessions (AA, BB, AAB and ABB). The AFLP data distinguished between the wild diploid accessions and suggested new subspecies relationships in the M. acuminata complex that are different from those based on morphological data. The data suggested that there are three subspecies within the M. acuminata complex (ssp. burmannica Simmonds, malaccensis Simmonds, and microcarpa Simmonds). 'Tjau Lagada' (ssp. microcarpa), 'Truncata' [ssp truncata (Ridl.) Shepherd] and 'SF247' [ssp. banksii (F.Muell) Simmonds] clustered very closely with 'Gros Michel' and 'Km 5', indicating that more than one M. acuminata subspecies may be involved in the origin of triploid AAA bananas. 'Calcutta 4' (ssp. burmannicoides De Langhe &; Devreux) and 'Long Tavoy' (ssp. burmannica) were closely related and could be together in the same subspecies. This study also showed that there is much more genetic diversity within M. balbisiana that was split into two groups: (1) 'I-63' and 'HND' and (2) 'Los Banos', 'MPL' (Montpellier), '10852', 'Singapuri', 'Etikehel', and 'Butohan 1' as the other.     

Genetic diversity in an African plantain core collection using AFLP and RAPD markers

Fifteen AFLP primer pairs (EcoRI+3 and MseI+3) and 60 10-mer RAPD primers were used to detect polymorphisms and assess genetic relationships in a sample of 25 plantains from diverse parts of Western and Central Africa. The discriminatory power of the AFLP technique was greater than that of the RAPD technique, since the former produced markers with greater polymorphic information content (PIC) than the latter. Hence, AFLP analysis appeared to be a more-powerful approach for identifying genetic differences among plantain accessions. In this regard, significant genetic diversity within the plantains was shown by the unweighted pair-group method of arithmetic averages (UPGMA) and the multidimensional principal coordinate (PCO) analyses. The AFLP-derived clusters indicated closer relationships between similar inflorescence types than the RAPD-derived clusters. A small group of cultivars from Cameroon were separated from the bulk of other plantains, suggesting that Cameroon may harbour accessions with useful or rare genes for widening the genetic base of breeding populations derived from the plantains. A greater effort should be directed at collecting and characterizing plantain cultivars from Cameroon.     

Osmoregulation, immunolocalization of Na+/K+-ATPase, and ultrastructure of branchial epithelia in the developing brown shrimp, Crangon crangon (Decapoda, Caridea)

Aspects of osmoregulation including salinity tolerance, osmoregulatory capacity, location of transporting epithelia, and the expression of the enzyme Na+/K+-ATPase were investigated in the developing brown shrimp, Crangon crangon (L.), from the North Sea. Early developmental stages and large juveniles were exposed to a wide range of salinities for measurement of hemolymph osmolality and survival rates. In media ranging from 17.0 per thousand to 32.2 per thousand, salinity tolerance was generally high (survival rates: 70%-100%) in all developmental stages, but it decreased in media <10.2 per thousand. Zoeal stages and decapodids slightly hyperregulated at 17.0 per thousand and osmoconformed in media > or =25.5 per thousand. At 10.2 per thousand, these stages showed high mortality, and only juveniles survived at 5.3 per thousand. Juveniles hyperregulated at 10.2 per thousand and 17.0 per thousand, osmoconformed at 25.5 per thousand, and hyporegulated in media > or =32.2 per thousand. Large juveniles hyperregulated also at 5.3 per thousand. Expression of the Na+/K+-ATPase and ion-transporting cells was located through immunofluorescence microscopy and transmission electron microscopy. In zoeae I and VI, a strong immunoreactivity was observed in cells of the inner epithelia of the branchiostegites and in epithelial cells lining the pleurae. Their ultrastructure showed typical features of ion-transporting cells. In decapodids and juveniles, ionocytes and expression of Na+/K+-ATPase remained located in the branchiostegite epithelium, but they disappeared from the pleurae and appeared in the epipodites. In large juveniles, the cells of the gill shaft showed positive immunolabeling and ultrastructural features of ionocytes. In summary, the adult pattern of osmoregulation in C. crangon is accomplished after metamorphosis from a moderately hyperosmoconforming decapodid to an effectively hyper-/hyporegulating juvenile stage. Salinity tolerance and osmoregulatory capacity are closely correlated with the development of ion-transporting cells and the expression of Na+/K+-ATPase.     

Genetic Diversity and Population Assessment of Musa L. (Musaceae) Employing CDDP Markers

Plant Molecular Biology Reporter - Sixty-six accessions of Musa genus with different genomic groups that consisted of wild relatives and cultivated lines were obtained from the International...