The formation of the mesoderm in urodelean amphibians

Summary Xenoplastic recombinations of animal and vegetative parts ofAmbystoma mexicanum and Triturus alpestris blastulae, and similar recombinations of parts of³H-thymidinelabelled and unlabelledAmbystoma mexicanum blastulae demonstrate convincingly that the vegetative part (zone IV, see Nieuwkoop, 1969a) of such a recombinate does not contribute to mesoderm formation, but exclusively forms endodermal derivatives. In contrast, the animal cap of the blastula (zones I.II)—which only gives rise to atypical ectoderm if isolated—not only furnishesall the ecto-neurodermal derivatives, butall the mesodermal structures of the developing recombinate as well, and finally to a varying extent forms additional endodermal structures in the recombinate.In the recombinates endodermization of the ectodermal cap occurred at the anterior end of the invaginated archenteron—corresponding to the presumptive pharyngeal endoderm —, and along the dorsal side of the endodermal tube, while an endoderm-like epithelium is formed at the boundary between the caudal endoderm and the ectoderm (proctodaeum formation). These results suggest that in normal development also endodermization occurs in the ectodermal half of the egg. This occurs particularly on the dorsal side, leading to the formation of the presumptive pharyngeal endoderm situated above the dorsal blastoporal groove.These experiments show that the vegetative half of the amphibian blastula is firmly determined as the future endoderm, whereas the animal half is still virtually undetermined and pluripotent.     

The formation of the mesoderm in urodelean amphibians

Summary Dorsal (D), lateral (L and R), and ventral (V) portions of the endoderm of blastulae ofAmbystoma mexicanum of different age (stages 8⁺ to 10^–) were combined with ectodermal caps of stage 8⁺ blastulae. All V and most L and R portions induced only ventrocaudal mesodermal structures — ventral type of mesoderm induction. Almost all D portions induced much more voluminous structures of predominantly axial character — dorsal type of mesoderm induction. The difference in mesoderm-inducing capacity of the dorsal as against the lateral and ventral endoderm is probably purely quantitative in character. The dorsal endoderm exhibits a pronounced dominance in mesoderm-inducing capacity. During the early symmetrization of the amphibian egg it is apparently especially the presumptive dorsal endoderm that becomes endowed with strong mesoderm-inducing properties.A comparison of the results obtained with endodermal portions of blastulae of different age showed that the mesoderm-inducing capacity first begins to decline in the dorsal endoderm (around stage 9), subsequently in the lateral, and finally in the ventral endoderm (at stage 10^–). At stage 10^– the dorsal endoderm no longer has mesoderm-inducing capacities.In the recombinates there is a striking correspondence between the regional differentiation of the mesoderm and that of the endoderm. The latter differs markedly from the presumptive significance of the various endodermal regions in the normal embryo.Primordial germ cells, which constitute a characteristic component of the ventral type of mesoderm induction, can be induced not only by ventral, but also by lateral and to some extent even by dorsal endoderm. The development of primordial germ cells from the ectodermal component of the various recombinates indicates that in the urodeles the origin of the primordial germ cells differs markedly from that in the anurans.The authors want to thank Miss A. de wit for expert technical assistance, Miss E. Bartová for making the drawings, and Dr. J. Faber for editorial help.     

The formation of the mesoderm in urodelean amphibians

Summary Experiments are described in which in early to late blastulae ofAmbystoma mexicanum (stages 7–8/9 Harrison) the animal, ectodermal half (zones I.II) was combined with the vegetative, endodermal yolk mass (zone IV) in various orientations, viz. in random orientation or with the dorso-ventral axes of the two components in identical, opposite or perpendicular orientation (0°, 180°, or 90° translocation respectively). The results demonstrate unequivocally that the dorso-ventral polarity of the induced mesoderm, and thus of the embryo, depends exclusively upon the inherent dorso-ventral polarity of the endoderm, whereas the grey crescent, a considerable part of which is located in the animal, ectodermal half, plays no causal role whatsoever.The results also show that the dorso-ventral polarity is inherent in the entire endodermal mass, but that the subsequent regional differentiation of the endoderm depends upon stimulating influences emanating from the surrounding mesoderm, the later nutritive yolk representing that part of the endoderm which normally does not come under the influence of the mesoderm, and therefore fails to receive the necessary stimulus for further differentiation.On the basis of these findings Schultze's Umkehrexperiment as studied byPenners andSchleip, Penners, andPasteels are reinterpreted, whileDalcq andPasteels' general developmental theory as well asCurtis' cortical grafting experiments are critically discussed.

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Zusammenfassung Es werden Experimente beschrieben, in denen in frühen bis späten Blastulae vonAmbystoma mexicanum (Stadien 7–8/9 Harrison) die animale, ektodermale Hälfte (Zonen I.II) mit der vegetativen, entodermalen Dottermasse (Zone IV) kombiniert wurde, und zwar in verschiedener Orientierung, d. h. in willkürlicher Orientierung oder mit den Dorsoventralachsen der beiden Komponenten identisch, entgegengesetzt oder senkrecht zueinander orientiert (0°, bzw. 180° oder 90° transloziert). Die Ergebnisse zeigen eindeutig, daß die Dorsoventralpolarität des induzierten Mesoderms, und damit die des Embryos, ausschließlich von der inhärenten Dorsoventralpolarität des Entoderms bestimmt wird, während der graue Halbmond, der zu einem beträchtlichen Teil in der animalen, ektodermalen Hälfte liegt, überhaupt keine kausale Rolle spielt.Außerdem zeigen die Ergebnisse, daß die Dorsoventralpolarität der ganzen Entodermmasse inhärent ist, daß aber die spätere regionale Differenzierung des Entoderms von stimulierenden Einflüssen seitens des umgebenden Mesoderms abhängig ist; der spätere Nährdotter ist derjenige Teil des Entoderms der normalerweise außerhalb des Wirkungsbereiches des Mesoderms liegt, und infolgedessen den für seine weitere Differenzierung benötigten Reiz nicht erhält.Angesichts dieser Befunde wird das Schultzesche Umkehrexperiment, welches vonPenners undSchleip, Penners, undPasteels näher untersucht worden ist, neu interpretiert, während die allgemeine Entwicklungstheorie vonDalcq u.Pasteels sowie die Cortextransplantationen vonCurtis kritisch diskutiert werden.

     

SEM analysis of amphibian mesodermal migration

Summary At the end of gastrulation, the lateral mesoderm of amphibian embryos migrates ventrally between the ectoderm and the endoderm. The present study is an examination of the morphology of the leading cells of the mesodermal sheet and of the substratum over which they move (the inner surface of the ectoderm). The cells of the leading edge of the mesoderm are generally round, with very short and narrow flattened projections in the forward direction. These projections do not have a ruffled morphology, regardless of whether fixation is carried out before or after the ectoderm and mesoderm are dissected away from the endoderm. The inner surface of the ectoderm is covered with fine (450–500A) filamentous extracellular material and the ectoderm cells sometimes extend cytoplasmic processes (approx. 0.1 wide) onto the leading surface of the mesoderm or onto adjacent ectoderm cells. These studies indicate that the morphology of cell migration in amphibians is closer to that seen inFundulus than to that characteristic of chick or mammalian cells.This paper is dedicated to the memory of Mac V. Edds, Jr., who warmly encouraged the developmental biologists of the Pioneer Valley     

The formation of the mesoderm in urodelean amphibians

Summary Strong Li treatment leads to auto-activation of blastula and gastrula ectoderm due to lethal and sublethal cytolysis, resulting in adirect meso- and endodermization of the ectoderm. In contrast, a mild Li treatment-0.04 M LiCl in Holtfreter solution for only two hours—which does not show cytolizing effects, has no or only a weak mesodermizing effect upon blastula ectoderm. However, when the explant contains endoderm as well the same treatment leads to a marked transformation of ectoderm into mesoderm and subsequently into endoderm (vegetalization). Thisindirect action of the Li ion under physiological conditions apparently represents an enhancement of the normally occurring mesoderm formation from the ectodermal component of the blastula under the inducing action of the endodermal component and its subsequent transformation into endodermal structures, probably due to a rise in competence. The question is raised in how far the direct meso-and endodermization of gastrula ectoderm by heterogeneous inductors is open to the same criticsm as in the case of auto-activation of the ectoderm by a strong Li treatment. Finally, the experiments of Ave, Kawakami and Sameshima (1968) are briefly discussed.

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Zusammenfassung Eine starke Li-Behandlung führt im Blastula- und Gastrula-Ektoderm durch letale und subletale Zytolyse zu einer Selbstaktivierung, die sich auswirkt in einerdirekten Meso-und Entodermisierung des Ektoderms. Im Gegensatz dazu zeigt eine gelinde Li-Behandlung — 0,04 M LiCl in Holtfreter-Lösung während nur 2 Std —, die keine zytolyzierende Wirkung hat, keinen oder nur einen schwachen Einflu\ auf isoliertes Blastula-Ektoderm. Wenn dagegen das Explantat auch Entoderm enthlt, führt die gleiche Behandlung zu einer betrchlichen Umbildung von Ektoderm in Mesoderm und weiter in Entoderm (Vegetalisierung). Dieseindirekte Wirkung des Li-Ions unter physiologischen Verhltnissen stellt also offenbar eine Verstrkung dar der im normalen Keim vor sich gehenden Mesodermbildung aus der ektodermalen Komponente unter dem Einflu\ der induzierenden Wirkung der entodermalen Komponente und der darauffolgenden Transformierung in entodermale Strukturen, wsch. durch Kompetenzsteigerung. Es wird die Frage aufgeworfen, inwiefern die unmittelbare Meso- und Entodermisierung von Gastrula-Ektoderm durch heterogene Induktoren der gleichen Beurteilung zugnglich ist wie der Fall der Selbstaktivierung des Ektoderms durch starke Li-Behandlung. Schlie\lich werden die Experimente von Ave, Kawakami u. Sameshima (1968) kurz diskutiert.

     

Head and trunk-tail organizing effects of the gastrula ectoderm of Cynops pyrrhogaster after treatment with activin A

Differentiation tendency and the inducing ability of the presumptive ectoderm of newt early gastrulae were examined after treatment with activin A at a high concentration (100 ng/ml). The activin-treated ectoderm differentiated preferentially into yolk-rich endodermal cells. Combination explants consisting of three pieces of activin-treated ectoderm formed neural tissues and axial mesoderm along with endodermal cells. However, the neural tissue was poorly organized and never showed any central nervous system characteristics. When the activin-treated ectoderm was sandwiched between two sheets of untreated ectoderm, the sandwich explants differentiated into trunk-tail or head structures depending on the duration of preculture of activin-treated ectoderm in Holtfreter's solution. Short-term (0–5 h) precultured ectoderm induced trunk-tail structures accompanied by axial organs, alimentary canal and beating heart. The arrangement of the explant tissues and organs was similar to that of normal embryos. However, archencephalic structures, such as forebrain and eye, were lacking or deficient. On the other hand, long-term (10–25 h) precultured ectoderm induced archencephalic structures in addition to axial organs. Lineage analysis of the sandwich explants using fluorescent dyes revealed that the activin-treated ectoderm mainly differentiated into endodermal cells and induced axial mesoderm and central nervous system in the untreated ectoderm. These results suggest that activin A is one of the substances involved in triggering endodermal differentiation and that the presumptive ectoderm induced to form endoderm displays trunk-tail organizer or head organizer effects, depending on the duration of preculture.     

Establishment and movement of egg regions revealed by the size class of yolk platelets in Xenopus laevis

Sizes of yolk platelets were measured in sections of oocytes and embryos in Xenopus. It was found that the average size of the largest group of platelets in cells differed between germ layers of neurulae. It was small (3 to 5 m) in the ectoderm, medium-sized (5 to 8 µm) in the mesoderm, and large (over 8 m) in the endoderm. Platelets of these size classes formed layers in egg, the yolk gradient, by the end of oocyte maturation. The yolk gradient contained products of the mitochondrial cloud and a part of the germinal vesicle material at certain positions. The layers of small, medium and large platelets in the egg changed their locations to distribute to the ectoderm, mesoderm and endoderm of neurulae, respectively. The yolk layers in the egg thus represented different prospective fates, and a figure describing the locations of these layers could be regarded as a fate map for the one-cell stage. Most of the marginal blastomeres of embryos at cleavage stages consisted of a few parts with different prospective fates. Results were discussed with reference to available fate maps for cleavage stage embryos.     

Polarities in Mammalian Evolution Seen Through Homologs of the Inner Cell Mass

Embryogenetic pathways differ markedly among monotremes, marsupials, and placentals, and their analysis provides information of fundamental importance to recognition of mammalian evolutionary directions. The cap of cuboidal cells of the marsupial late unilaminar blastocyst, generally known as the embryonic area, probably is induced to form (prior to origin of Hensen's node) by signals from earliest hypoblastic cells (anterior visceral endoderm). The thickened cap is a medullary plate of sauropsid terminology because it includes epiblastic cells presumptive to neurectoderm (including neural crest), Hensen's node, primitive streak, and gut endoderm. The remainder of the definitive embryo (i.e., parts of epidermal origin, including ectodermal placodes) derives from squamous ectoderm (surrounding the medullary plate) of the blastocyst's ill-named trophoblastic area. Amniotic ectoderm develops farther distally within the trophoblastic area. The autapomorphic inner cell mass (ICM) of placental mammals is homologous to medullary plate of the marsupial blastocyst plus morphologically undefined, proximal parts of surrounding ectoderm (of the trophoblastic area). Considerations of early cell lineages in marsupials are greatly affected by recognition that the boundary between future embryonic and extra-embryonic tissues does not match the margin of the medullary plate (i.e., embryonic area). Marsupials and monotremes largely conform to sauropsid early embryogenesis, but placentals express, at earliest developmental stages, innovations unique within Amniota that are linked to early establishment of the brain. Neonatal marsupials and hatchling monotremes are extremely altricial and closely comparable anatomically/physiologically; they share a temporal pattern in combining early morphogenesis of craniofacial features (related to suckling) with deferral of telencephalic completion into postnatal/posthatching life. Placentals contrast greatly in establishing the central nervous system prior to rudiments of the cranial skeleton and associated musculature, and they complete essentials of forebrain development before birth. Comparative evidence from transitory periderm suggests that primordial eutherians had extremely altricial hatchlings or newborns, whichever was the mode of early development. Details remain unknown about the origin of the unique specialization of ICM plus encapsulating trophoblast from the more generalized blastula of ancestral synapsids.     

Abdominal segment formation in Spirorbis moerchi (Polychaeta)

Summary The development of abdominal segments in Spirorbis moerchi (Polychaeta: Annelida) was studied by light and electron microscopy. Abdominal segments develop in strict succession from anterior to posterior. Segmentation is initiated in the mesoderm and is followed by segmentation of the ectoderm. The mesoderm of the abdominal segments arises entirely from pygidial residual mesoderm; inward migration of cells from the pygidial ectoderm to give rise to mesoderm does not occur. The primordial germ cells remain distinct from the residual mesoderm of the pygidial growth region. After several abdominal segments have developed, the primordial germ cells migrate posteriorly from the achaetous region, invade the abdominal segments, and give rise to the retroperitoneal gonads. Abdominal segment formation is discussed in terms of heteronomy, primordial germ cell origin, gonad formation, and development of the circulatory system.     

Ultrastructure of Bacillus pulvifaciens

The ultrastructure of vegetative cells and spores of Bacillus pulvifaciens was studied by CTEM and SEM methods. The vegetative cells are rods, 1.6–4.5 m long and 0.4–0.6 m wide, exhibiting typical ultrastructural features of Gram-positive bacteria. The spores are of ellipsoidal shape, 0.6×1.2 m in size, with six longitudinal ribs reaching up to 130 nm in height. There are satelite ribs on both sides of the longitudinal ribs, reaching up to 20 nm in height. Between the longitudinal ribs, additional transversal ribs were observed in SEM. A special tubular layer, separating the outer and inner coat of the spores, was revealed in ultrathin sections. This layer seems to be a typical ultrastructural feature of Bacillus pulvifaciens spores.     

Polarities in Mammalian Evolution Seen Through Homologs of the Inner Cell Mass

Embryogenetic pathways differ markedly among monotremes, marsupials, and placentals, and their analysis provides information of fundamental importance to recognition of mammalian evolutionary directions. The cap of cuboidal cells of the marsupial late unilaminar blastocyst, generally known as the embryonic area, probably is induced to form (prior to origin of Hensen's node) by signals from earliest hypoblastic cells (anterior visceral endoderm). The thickened cap is a medullary plate of sauropsid terminology because it includes epiblastic cells presumptive to neurectoderm (including neural crest), Hensen's node, primitive streak, and gut endoderm. The remainder of the definitive embryo (i.e., parts of epidermal origin, including ectodermal placodes) derives from squamous ectoderm (surrounding the medullary plate) of the blastocyst's ill-named trophoblastic area. Amniotic ectoderm develops farther distally within the trophoblastic area. The autapomorphic inner cell mass (ICM) of placental mammals is homologous to medullary plate of the marsupial blastocyst plus morphologically undefined, proximal parts of surrounding ectoderm (of the trophoblastic area). Considerations of early cell lineages in marsupials are greatly affected by recognition that the boundary between future embryonic and extra-embryonic tissues does not match the margin of the medullary plate (i.e., embryonic area). Marsupials and monotremes largely conform to sauropsid early embryogenesis, but placentals express, at earliest developmental stages, innovations unique within Amniota that are linked to early establishment of the brain. Neonatal marsupials and hatchling monotremes are extremely altricial and closely comparable anatomically/physiologically; they share a temporal pattern in combining early morphogenesis of craniofacial features (related to suckling) with deferral of telencephalic completion into postnatal/posthatching life. Placentals contrast greatly in establishing the central nervous system prior to rudiments of the cranial skeleton and associated musculature, and they complete essentials of forebrain development before birth. Comparative evidence from transitory periderm suggests that primordial eutherians had extremely altricial hatchlings or newborns, whichever was the mode of early development. Details remain unknown about the origin of the unique specialization of ICM plus encapsulating trophoblast from the more generalized blastula of ancestral synapsids.     

Schwermetallaffine Strukturen des peripheren Nerven

Zusammenfassung Konvertiertes Kupfer wurde an axonalen und Schwannschen Zellmembranen, vor allem im nodalen und paranodalen Bereich beobachtet. Kupfer wird offenbar in inner und outer leaflet der unit membrane ein- und an diese angelagert. Denn bei wenig Präzipitat oder geringer Elektronendichte des Niederschlags stellten sich die Membranen als dreischichtiger Komplex dar, der in den Dimensionen der unit membrane entsprach. Bei großen Präzipitatmengen oder bei hoher Elektronendichte des Niederschlags blieb nur die helle Mittelschicht von ca. 30 Å frei. Diese Metallaffinität wird im Zusammenhang mit den cytochemischen AChE-Nachweisverfahren diskutiert, weil bei diesen Methoden Schwermetallionen im Inkubationsmedium verwendet werden, und die Kupferbindungsstellen zum Teil mit den Lokalisationen der AChE identisch sind. Möglicherweise gibt beim cytochemischen AChE-Nachweis schon die Darstellung von unit membranes in unkontrastierten Präparaten Aufschluß auf erfolgte unspezifische Schwermetallbindung. Als weitere Kontrolle wird Inkubation in der verwendeten Schwermetallösung mit nachfolgender Konversion vorgeschlagen.

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Heavy metal affinity of peripheral nerve structuresI. Possibility of interference with cytochemical localisation of AChE

Summary The ultrastructural localisations of copper in ventral and dorsal roots of the rat were studied by varying fixation and converting medium. Precipitated copper was deposited in axonal membranes (axolemma, membranes of axonal organelles) and Schwann cell membranes (inner and outer plasmalemma, major and minor dense lines in the region of the splitting myelin sheath, terminal loops and Schwann cell interdigitations). Copper obviously was bound within and attached to inner and outer leaflet of these membranes. If there was little or less electron-dense precipitate membranes appeared three-layered, dimensions corresponding to those of the unit membrane. If there was electron-dense or plenty of precipitate only the middle electron-lucent layer of approximately 30 Å was revealed. The predominant deposition on nodal and paranodal membranes may be explained either by facilitated access to this region or by specific metal ion binding sites at those specialised membranes. Metal ion binding is discussed in relation to cytochemical demonstration of AChE activity, because these methods use incubation media containing heavy metal cations and copper binding sites partly correspond with the localisation of AChE activity. The appearance of unit membranes in unstained sections probably could be a hint at unspecific heavy metal ion binding ocouring in demonstration of specific AChE. Therefore control-incubation in the heavy-metal solution with subsequent conversion is suggested.

     

Über quantitative Veränderungen „gomoripositiver“ Substanzen in Infundibulum und Hypophysenhinterlappen der Ratte nach beidseitiger Adrenalektomie

Zusammenfassung Bei der normalen weißen Ratte verhält sich die Zona externa des Infundibulum im Gegensatz zur Zona interna infundibuligomorinegativ.Am 7. Tag nach beidseitiger Adrenalektomie treten in der Zona externa gomoripositive Substanzen auf, deren Menge über den 14. Tag nach der Operation hinaus zunimmt.Im supraoptico-hypophysären System ergibt die quantitative Messung eine Entleerung des neurosekretorischen Materials am 3. Tage nach beidseitiger Adrenalektomie. Im Verlauf von 14 Tagen nach dem operativen Eingriff füllt sich das neurosekretorische System wieder bis zu den beim Normaltier ermittelten Werten auf.Die gomoripositiven Granula in der Zona externa inf. sind möglicherweise das morphologische Äquivalent eines Corticotropin-releasing factors.

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On quantitative changes of Gomori-positive substances in the median eminence and neural lobe of the rat hypophysis after bilateral adrenalectomy

Summary In the normal albino rat, the external layer of the median eminence shows a Gomori-negative histochemical reaction as opposed to the internal layer.On the 7th day after bilateral adrenalectomy, Gomori-positive substances appear in the external layer, their amount increasing up to and beyond the 14th day p.o.In the supraoptico-hypophyseal system, quantitative measurements show a depletion of the neurosecretory substance on the 3rd day following bilateral adrenalectomy. This loss of neurosecretory material is restored during the course of 14 days p.o., the amount then corresponding to that found in the normal rat.It is conceivable that the Gomori-positive granules in the outer layer of the median eminence are the morphological equivalent of a CRF.

     

L'histogenèse régénératrice chez les phoronidiens

Summary The regeneration (organogenesis was studied by Emig, 1972 a, b) of Phoronida can be divided into three phases: the first one, cicatrisation, is characterized by a provisional mesodermal scar-tissue, later the old epidermis cover this scar-tissue. The regenerating blastema, second phase, takes place by cellular dedifferentiation processes; each germ layer (ectoderm, mesoderm, endoderm) regenerates itself from its own elements. One exception only seems to be oesophagel regeneration by metaplasia of the prestomacal cells during the asexual reproduction. The differentiation of the amputated structures (third phase) appears submitted to the inductive influence of the mesoderm and to the trophic action of the nervous system (especially the epithelial plexus). The polarity in regeneration sets a problem in Phoronida.

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Ce travail a été effectué dans le cadre du contrat L. A. n 41 au C. N. R. S.     

Experimental analyses of the rearrangement of ectodermal cells during gastrulation and neurulation in avian embryos

The rearrangement of ectodermal cells was studied in chimeras in which grafts were transplanted during late gastrula and early neurula stages to heterotopic locations in avian embryos. Three types of experiments were done. In all experiments, Hensen's node was extirpated completely and replaced with an epithelial plug derived from 1 of 3 regions of the prospective ectoderm. In type-1 experiments, Hensen's node was replaced with a plug consisting of precursor cells of the floor plate of the neural tube. In type-2 experiments, Hensen's node was replaced with a plug consisting of precursor cells of the lateral wall of the neural tube. In type-3 experiments, Hensen's node was replaced with a plug consisting of precursor cells of the epidermal ectoderm. In all experiments, the amount and direction of cell rearrangement that occurred in the transplanted ectodermal plug was essentially typical for prospective ectodermal cells normally residing within Hensen's node. That is, transplanted ectodermal cells underwent lateralto-medial cell-cell intercalation and contributed to the ventral midline of the neural tube along its entire rostrocaudal extent. In most embryos, a notochord was reconstituted from host cells, despite the fact that Hensen's node — the prime source of prospective notochordal cells in intact embryos — was extirpated completely; however, a few embryos had long notochordal gaps. In such essentially notochordless embryos, the ventral midline of the neural tube still derived from grafted cells, but it failed to form a floor plate, providing further confirmation of the results of several previous studies that the notochord is required to induce the floor plate. Collectively, our results provide evidence that the rearrangement of ectodermal cells does not require the presence of a trail of prospective floor plate cells (laid down by the regressing Hensen's node), or of a notochordal substrate, and that the continued presence of an organizer per se, ostensibly Hensen's node, is not required. In addition, our results demonstrate that the rearrangement of cells still occurs in the absence of boundaries between ectodermal cells of different phenotypes (e.g., between cells of the floor plate and lateral walls of the neural tube). Finally, our results reveal further that the amount and direction of cellular rearrangement is not regulated in a cell-autonomous fashion, but rather it is determined by the overall magnitude and vector of the displacement of the community of rearranging cells within a developmental field.     

Endoderm differentiation and inductive effect of activin-treated ectoderm in Xenopus

When presumptive ectoderm is treated with high concentrations of activin A, it mainly differentiates into axial mesoderm (notochord, muscle) in Xenopus and into yolk-rich endodermal cells in newt (Cynops pyrrhogaster). Xenopus ectoderm consists of multiple layers, different from the single layer of Cynops ectoderm. This multilayer structure of Xenopus ectoderm may prevent complete treatment of activin A and subsequent whole differentiation into endoderm. In the present study, therefore, Xenopus ectoderm was separated into an outer layer and an inner layer, which were individually treated with a high concentration of activin A (100 ng/mL). Then the differentiation and inductive activity of these ectodermal cells were examined in explantation and transplantation experiments. In isolation culture, ectoderm treated with activin A formed endoderm. Ectodermal and mesodermal tissues were seldom found in these explants. The activin-treated ectoderm induced axial mesoderm and neural tissues, and differentiated into endoderm when it was sandwiched between two sheets of ectoderm or was transplanted into the ventral marginal zone of other blastulae. These findings suggest that Xenopus ectoderm treated with a high concentration of activin A forms endoderm and mimics the properties of the organizer as in Cynops.     

Scanning electron microscopy of the rat retina

Summary Fixed and unfixed, freeze-dried pieces of isolated retina and the posterior part of the eye bulb from adult rats were examined in a scanning electron microscope.The inner limiting membrane shows distinct cell borders, protrusions, and scattered microvilli-like structures. Different types of nerve cells are observed in the ganglion cell layer and the inner nuclear layer. They all lack synaptic boutons on the surface of their perikarya. There is an intercellular space between the processes in the nerve fiber layer.The inner and outer segments are surrounded by a space with extracellular material. Their surface is smooth or slightly undulated. There is no evidence indicating the existence of basal infoldings continuous with the membraneous structures inside the rod outer segments. The connecting piece between the inner and outer segments resemble a symmetrically shaped hour-glass. The surface of the epithelial cells is covered by microvilli forming a honeycomb-like structure and each outer segment is surrounded by several microvilli.The results obtained are discussed in relation to those obtained by transmission electron microscopy. The probable existence of a significant extracellular space and the distribution of extracellular material between the segments and the microvilli are discussed.Supported by grants from H. Jeanssons Stiftelse Riksföreningen mot Cancer (265-B69-OIX) and the Swedish Medical Research Council (B70-12X-2534-02). I would like to thank the Swedish Silicate Research Institute, Göteborg, for using their scanning electron microscope, and Miss. M. Persson for skilful technical assistance.     

Synaptische Strukturen und Elementargranula in der Neurohypophyse des Meerschweinchens

Zusammenfassung Die Ultrastruktur des Nucleus infundibularis tuberis, der Zona interna, der Zona externa und des Hypophysenhinterlappens von Meerschweinchen wird beschrieben. Vergleichende Beobachtungen an den Nervenfasern der genannten Gebiete zeitigen folgende Ergebnisse:Die Axone der Zona interna und des Hypophysenhinterlappens enthalten gleichgeartete, große neurosekretorische Elementargranula (1,400 Å). Die Axonquerschnitte im Bereich des Nucleus infundibularis tuberis und der Zona externa weisen Granula gleichen Kalibers (800 Å) auf und lassen damit auf einen funktionell-morphologischen Zusammenhang des Nucleus infundiburis und der Zona externa schließen. Ob die kleinen Elementargranula das morphologische Substrat der sogenannten releasing factors darstellen oder ob es sich um adrenerge Axone und Endigungen mit typischen Katecholamingranula handelt, ist nicht zu klären.Es wird unterschieden zwischen echten Synapsen und synapsenähnlichen Kontakten. Nur erstere zeigen alle klassischen Synapsenmerkmale. Ihnen sind im Bereich der Neurohypophyse axosomatische und interaxonale Synapsen zuzuordnen. Zona externa und Hypophysenhinterlappen stellen neurohämale Kontaktzonen dar und zeichnen sich durch gleichartig konstruierte Synapsen aus. Neben interaxonalen Synapsen treten vor allem neurokapilläre Kontakte in den Vordergrund. In der Zona externa sind die Tanyzyten synaptisch mit Axonen verknüpft, so wie im Hypophysenhinterlappen die Pituizyten synaptische Verbindungen mit Nervenfasern eingehen. Die synapsenähnlichen Bildungen im Verlauf und an den Enden der Axone können somit neurokapillärer, neurotanyzytärer und neuropituizytärer Natur sein. Sie enthalten neben den synaptischen Vesikeln auch meist kleine oder große Elementargranula. Die ähnliche Morphologie der synaptischen Bildungen in der Zona externa und im Hinterlappen läßt einen gleichgearteten Inkretionsmechanismus beider neurohypophysärer Abschnitte vermuten. Tanyzyten und Pituizyten könnten dabei eine Überträgerfunktion ausüben. Eine rezeptorische Funktion der Tanyzyten wird gleichfalls diskutiert.Die Drüsenzellen der Pars intermedia der Hypophyse sind durch direkte neuroglanduläre Kontakte mit den Neuronen des Hypothalamus verknüpft. Die Axone stülpen sich hierbei tief in das Zytoplasma der Drüsenzellen ein. Es entstehen dadurch beim Meerschweinchen drei nervöse Endigungsformen, die Synapsen ausbilden: 1. Endigungen, die nur synaptische Vesikel enthalten; 2. Endigungen mit synaptischen Vesikeln und kleinen Granula; 3. Endigungen mit synaptischen Vesikeln und großen Granula.Zwischen den Drüsenzellen der Pars infundibularis der Adenohypophyse sind ebenfalls Axonquerschnitte zu beobachten. Es handelt sich dabei entweder um vegetative Faserbündel oder Nervenfasern mit kleinen Granula wie in der Zona externa (800 Å).

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Summary The ultrastructure of the nucleus infundibularis tuberis, the inner and outer layer of the median eminence and the posterior lobe of the guinea pig are described. Comparative studies on the nerve fibres of these regions lead to the following conclusions:The axons of the inner layer of the median eminence and of the posterior lobe contain similar types of large, neurosecretory elementary granules (Diameter 1400 Å).The cross-sections of the axons in the area of the nucleus infundibularis tuberis and of the outer layer of the median eminence exhibit the same sized granules (Diameter 800 Å), which points to a functional-morphological relationship between the two. Whether the small elementary granules represent the morphological substrate of the so-called releasing factors, or whether it is a question of adrenergic axons and terminations with typical catecholamine granules, is undecided.A distinction is made between true synapses and synapse-like contacts. Only the former display all the classical characteristics of synapses. To this sort must be classed the axosomatic and interaxonal synapses in the neurohypophysis. The external layer of the median eminence and the posterior lobe are distinguished by neurohaemal contact areas and show similarly constructed synapses. Of particular prominence, besides interaxonal synapses, are neurocapillary contiguities. Just as in the posterior lobe the pituicytes show synaptic connections with nerve fibres, so are the tanycytes in the outer layer of the median eminence in synaptical contact with axons. Thus the synapse-like formations along and at the endings of the axons can be of a neurocapillary, neurotanycytial and neuropituicytial nature. They contain, apart from the synaptic vesicles, mainly small but also large elementary granules. The similar morphology of the synaptic formations in the outer layer of the median eminence and the posterior lobe suggests the existence of a similar mechanism of internal secretion in both these neurohypophysial areas. Tanycytes and pituicytes could exercise a carrier function. A possible receptor-function of the tanycytes is considered.The glandular cells of the pars intermedia of the hypophysis are linked by means of direct neuroglandular contacts with the neurons of the hypothalamus, whereby the axons make deep incursions into the cytoplasm of the glandular cells. In the guinea pig three types of termination can be observed that form synapses: 1. terminations that contain only synaptic vesicles; 2. terminations with small granules; 3. terminations with large granules.Cross-sections of axons can also be observed between the glandular cells of the pars infundibularis of the adenohypophysis. Here they can be identified either as vegetative fibre-bundles or as nerve fibres with small granules, such as occur in the outer layer of the median eminence (Diameter 800 Å).

     

Comparative study of cholinergic cells in retinas of various mouse strains

We examined cholinergic cells in the retinas of BALB/C albino, C57BL/6J black, and 129/SvJ light chinchilla mice by using immunocytochemistry with specific antisera against choline acetyltransferase (ChAT). Two types of ChAT-immunoreactive amacrine cell bodies were found in the inner nuclear layer (INL) and ganglion cell layer in the retinas of all three mouse strains. They were distributed with mirror-image symmetry and their processes ramified in strata 2 and 4 of the inner plexiform layer. A distinct type of ChAT-immunoreactive cell was found only in C57BL/6J mouse retina. The somata of this third type of ChAT-immunoreactive cell were located in the outermost part of the INL, with their processes extending toward the outer plexiform layer. Double-labeling experiments demonstrated that these were not horizontal cells and that they were GABA-immunoreactive. The results suggested that these cells were probably misplaced cholinergic amacrine cells showing GABA immunoreactivity. This feature of the C57BL/6J mouse retina should be taken into account in studies of mutant mice having a mixed genetic background with a C57BL/6J contribution.Tae-Hoon Kang, Young-Han Ryu and In-Beom Kim contributed equally to this study.This work was supported by Neurobiology Support Grant (M1-0108-00-0059) of the Ministry of Science and Technology, Korea     

Campaniform sensilla of Calliphora vicina (Insecta,Diptera)

Summary A classification scheme of campaniform sensilla using morphological criteria was developed. All variations of the two most important outer structural elements, the cuticular cap and the cuticular collar, were taken into consideration: (a) the external shape of the cuticular cap; (b) the position of the cuticular cap in relation to the remaining cuticle; (c) the position of the cuticular collar in relation to the cuticular cap. This resulted in a classification of campaniform sensilla into 24 types. This typology was applied to the campaniform sensilla of Calliphora, which show considerable variations in their outer structures. According to SEM (scanning electron microscope) pictures and TEM (transmission electron microscope) sections we found only 9 out of 24 different types of campaniform sensilla in the fly.