Intercellular junctions in nerve-free hydra

Summary Epithelial cells of nerve-free hydra contain septate and gap junctions. In thin sections the gap junctions are characterized by a gap of 3–4 nm. Freeze-fracture demonstrates the presence of septate junctions and two further types of structures: (i) the E-type or inverted gap junctions with particles in an en plaque conformation appearing as a raised plateau on the E-face or as a depression on the P-face; (ii) structures morphologically similar to gap junctions in rat liver, containing particles on the P-face and corresponding pits on the E-face, both having hexagonal packing with a lattice constant of 8 nm. We propose that these structures are also gap junctions.     

Interneuronal and glial-neuronal gap junctions in the lamina ganglionaris of the compound eye of the housefly,Musca domestics

Summary The cell-body layer of the lamina ganglionaris of the housefly, Musca domestica, contains the perikarya of five types of monopolar interneuron (L1–L5) along with their enveloping neuroglia (Strausfeld 1971). We confirm previous reports (Trujillo-Cenóz 1965; Boschek 1971) that monopolar cell bodies in the lamina form three structural classes: Class I, Class II, and midget monopolar cells. Class-I cells (L1 and L2) have large (8–15 m) often crescentshaped cell bodies, much perinuclear cytoplasm and deep glial invaginations. Class-II cells (L3 and L4) have smaller perikarya (4–8 m) with little perinuclear cytoplasm and no glial invaginations. The midget monopolar cell (L5) resides at the base of the cell-body layer and has a cubshaped cell body. Though embedded within a reticulum of satellite glia, the L1–L4 monopolar perikarya and their immediately proximal neurites frequently appose each other directly. Typical arthropod (-type) gap junctions are routinely observed at these interfaces. These junctions can span up to 0.8 m with an intercellular space of 2–4 nm. The surrounding nonspecialized interspace is 12–20 nm. Freezefracture replicas of monopolar appositions confirm the presence of -type gap junctions, i.e., circular plaques (0.15–0.7 m diam.) of large (10–15 nm) E-face particles. Gap junctions are present between Class I somata and their proximal neurites, between Class I and Class II somata and proximal neurites, and between Class II somata. Intercartridge coupling may exist between such monopolar somata. The cell body and proximal neurite of L5 were not examined. We also find that Class I and Class II somata are extensively linked to their satellite glia via gap junctions. The gap width and nonjunctional interspace between neuron and glia are the same as those found between neurons. The particular arrangement and morphology of lamina monopolar neurons suggest that coupling or low resistance pathways between functionally distinct neurons and between neuron and glia are probably related to the metabolic requirements of the nuclear layer and may play a role in wide field signal averaging and light adaptation.     

Gap junctions and impulse propagation in embryonic epithelium of Amphibia

Summary Epithelium of amphibian embryos (Cynops orientalis, Xenopus laevis) was found in preceding experiments to generate and conduct impulses during a limited stage (26–37) of development. In order to elucidate the structural basis of impulse propagation, epithelial cells of four stages were examined by the freeze-etching method: (I) before and (II) during acquisition of conductivity; (III) when propagation was fully established, and (IV) when it was no longer present. Only few gap junctions (GJ) of small size were found in groups I and IV. GJ in epithelia of group III were increased in number and size, and appeared morphologically coupled, i.e., with more loosely arranged connexons. The size of gap-junctional particles did not differ significantly between coupled and uncoupled stages. Zonulae occludentes seemed leaky in stage I, and tight in stages II–IV. Thus, the morphological characteristics of specialized junctions between non excitable cells correlated with the opening and closing of low resistance intercellular current pathways during embryonic development.Gap junctions in particular seem to form an essential link in the non-neural stimulus-response system, which may facilitate the mobility of the embryo during early phases of aquatic life before the reflex pathways have been established. Coupling and uncoupling of gap junctions may also play an important role in the regulation of cell differentiation and morphogenetic movement. The experimental model used in this study provides a useful tool for further investigations of structural correlates of gap junctional permeability under physiological conditions.     

Detergent sensitivity and splitting of isolated liver gap junctions

Summary Isolated rat liver gap junctions were split by two methods. In the first method, isolated gap junctions were stabilized by cross-linking their cytoplasmic surfaces with glutaraldehyde under conditions that prevented the entry of glutaraldehyde into the gap region. The stabilized junctions were then split in the junctional gap with SDS. In the second procedure, unfixed gap junctions were split by incubation in ureacontaining solutions. Junctional splitting was monitored by electron microscopy of thin sectioned and freeze fractured membrane pellets. Sidedness of the split junctional membranes was defined by labeling their cytoplasmic surfaces with glutaraldehyde-activated ferritin before splitting with urea. Gap junctional splitting did not result in any loss of protein components as determined by SDS-gel electrophoresis. The glutaraldehyde cross-linking procedure was also used to determine the effects of various detergents on the protein-protein interactions in the gap region. Of the detergents tested, only SDS caused junctional splitting.     

The origin of skeleton forming cells in the sea urchin embryo

Summary In embryos of the modern sea urchin species, subclass Euechinoidea, primary mesenchyme cells are derived from the progeny of micromeres formed at the sixteen cell stage of embryogenesis. The micromeres reside within the vegetal plate epithelium and later ingress into the blastocoel as primary mesenchyme cells which form the larval skeleton. Embryos of Eucidaris tribuloides, a member of the primitive subclass Perischoechinoidea, exhibit several noteworthy differences from euechinoid primary mesenchyme cell lineage including variable numbers and sizes of micromeres, the absence of mesenchyme ingression, and the lack of any detectable primary mesenchyme although a larval skeleton forms. In the present study, the cell lineage of the spiculogenic mesenchyme has been studied in Eucidaris tribuloides and in the euechinoid Lytechinus pictus by microinjecting the fluorescent tracer, Lucifer Yellow, into individual blastomeres of the embryo. In addition, wheat germ agglutinin, a lectin which binds only to primary mesenchyme cells of the early euechinoid embryo, was injected into the blastocoel of embryos of both species in order to examine the distribution of cells which possess primary mesenchyme-specific cell surface markers. The results of these experiments demonstrate that the spiculogenic mesenchyme of both Lytechinus and Eucidaris arise from descendants of micromeres formed at the sixteen cell stage, although the temporal and spatial distribution of these mesenchyme cells varies considerably between species. Furthermore, the evidence obtained suggests that the information necessary for spicule formation is already segregated to the vegetal pole by the eight cell stage. The results also suggest that there are no gap junctions present between the blastomeres of the early sea urchin embryo.     

Cellular organization of the embryonic lobster heart

Summary The cellular organization of the embryonic heart of the lobster Homarus americanus was examined in 6-week and 6-month-old animals. The heart wall consists of an outer adventitial layer of fibroblast cells and an inner layer of transversely striated myocardial cells. Present in close association with the myocardium are cardiac neurons, hemocytes and so-called storage cells.Adjacent fibroblasts form fasciae adhaerentes and gap junctions. Adherent junctions also occur between fibroblasts and myocardial cells. Intercalated discs and differentiated membrane regions of close apposition (4 nm) occur between adjacent myocardial cells.The cardiac neurons form a ganglion that contains four small and five large somata. Regions of neuropil are present. Motor axons arising from the cardiac ganglion form neuromuscular synapses with the myocardial cells.The storage cells contain large inclusions and form gap junctions with the myocardial cells. They may supply nutritive material to the developing myocardium.The heart at 6 weeks is about 200 m long and 160 m wide. At 6 months, it is about 300 m long and 250 m wide. The myocardium at 6 weeks is one cell layer thick, and the cells are from 2–6 m in maximum width. At 6 months the myocardium is 2–4 cells thick, and the cells are from 6–12 m in width. Therefore, the myocardium grows by an increase in the number and size of the myocardial cells.     

Osmotic Forces and Gap Junctions in Spreading Depression: A Computational Model

In a computational model of spreading depression (SD), ionic movement through a neuronal syncytium of cells connected by gap junctions is described electrodiffusively. Simulations predict that SD will not occur unless cells are allowed to expand in response to osmotic pressure gradients and K⁺ is allowed to move through gap junctions. SD waves of [K⁺]_out 25 to 60 mM moving at 2 to 18 mm/min are predicted over the range of parametric values reported in gray matter, with extracellular space decreasing up to 50%. Predicted waveform shape is qualitatively similar to laboratory reports. The delayed-rectifier, NMDA, BK, and Na⁺ currents are predicted to facilitate SD, while SK and A-type K⁺ currents and glial activity impede SD. These predictions are consonant with recent findings that gap junction poisons block SD and support the theories that cytosolic diffusion via gap junctions and osmotic forces are important mechanisms underlying SD.     

Refinement of Harrison's normal table for the morula and blastula of the axolotl

Summary Films of cleaving embryos of the axolotl,Ambystoma mexicanum, taken by the double camera technique, were used in order to arrive at a more detailed staging based on quantitative criteria. Drawings were made of the animal hemispheres just prior to the start of each cleavage cycle from the 6th to the 15th cleavage. The number of cells intersected either by the (apparent) egg diameter (up to the 10th cleavage) or by half the diameter (from the 10th cleavage onwards) was determined. The cell numbers for each cleavage cycle did not overlap with those of the previous or succeeding cycles. On the basis of these cell numbers, in place of the four Harrison stages 6–9, 10 successive stages were established each of which corresponds to one cleavage cycle.     

Localization of Transforming Growth Factor β in Association with Neuromuscular Junctions in Adult Human Muscle

Transforming growth factors- 1, 2, and 3 are known for their regulatory function in embryogenesis, fibrogenesis, and tissue repair of different cell types. A trophic function of TGF- subclasses for motoneurons has been shown in vitro. TGF- 1 is a potent survival factor for cultured embryonic rat motoneurons. In addition, TGF- 1 stimulates proliferation of rat Schwann cells. Recently, TGF- 2 has been reported to be associated with the subsynaptic nuclei of mature rat neuromuscular junctions. In this study, we investigated the expression of TGF- 1, 2, and 3 at neuromuscular junctions in skeletal muscle of 11 adults without neuromuscular disease. On muscle biopsies, neuromuscular junctions were depicted by acetylcholine esterase reaction and acetylcholine receptor antibodies. TGF- 1, 2, and 3 were stained immunohistochemically with monoclonal antibodies. Some muscle fibers showed low levels of inhomogeneous immunoreactivity for both TGF- 1 and TGF- 3. Intense immunoreactivity of TGF- 1 and 3 was shown at the postsynaptic area of neuromuscular junctions. TGF- 2 was expressed in the same subcellular distribution, but less strongly. In conclusion, the colocalization of TGF- with neuromuscular junctions may suggest a significant function in neuromuscular communication.     

Effect of cell-surface binding on development of Ascidian egg

Summary The early development ofPhallusia mammillata eggs, dechorionated with trypsin and treated with Concanavalin A, was studied. Vital staining with a very dilute solution of acridine orange (0.01 g/ml) helped to visualize the mitochondrial crescent by fluorescence. At high concentrations of Concanavalin A (20–200 g/ml) fertilized eggs did not cleave, but went through early ooplasmic segregation movements (formation of the crescent) and multinuclear syncytia were formed. At lower concentrations of Concanavalin A (less than 10 g/ml), cleavage occurred, but the blastomeres remained rounded, leading to a grapelike embryo. Eggs attached to Concanavalin A treated nylon surfaces either did not cleave or produced grapelike embryos. Attachment of the eggs did not affect ooplasmic segregation. Considering modern theories of membrane structure it was concluded that Concanavalin A prevented cleavage either by immobilizing surface structures connected with microfilaments or by indirectly modifying other membrane structures. These structures could not have been involved in ooplasmic segregation, but their mobility was necessary for further morphogenesis.This work was performed at the Station Zoologique, Villefranche-sur-Mer and at the Station de Biologie Marine et Lagunaire. Sète     

The cleavage pattern of the axolotl egg studied by cinematography and cell counting

Summary The temporal pattern of cleavage in the egg of the axolotl,Ambystoma mexicanum, was studied 1. by time-lapse microcinematography, and 2. by counting the total number of blastomeres dissociated at successive stages.Eggs were filmed from the one-cell stage till the early gastrula either (A) simultaneously from above and below with a double-camera assembly, or (B) from the side with a single camera.The animal blastomeres divide synchronously from the 2nd up to and including the 10th cleavage. The cycle length is roughly constant from the 3rd till the 10th cleavage. The cycle from the 2nd to the 3rd cleavage is slightly longer, while that from the 1st to the 2nd cleavage is about 20% longer. After the 10th cleavage the synchrony of divisions is lost owing to variable lengthening of cell cycles in individual blastomeres. Gastrulation starts around the onset of the 15th cleavage in the animal blastomeres.The analysis of films taken in side view reveals seven recurring cleavage waves, from the 5th till the 11th cleavage. Cells in the animal, equatorial and vegetative regions in sequence repeatedly pass through the three successive phases of the cleavage cycle—rounding-up, division, and relaxation—but with a shift in phase. The start of the 10th cleavage division of the slowest vegetative cells more or less coincides with that of the 11th division of the animal cells; from then on the cleavage waves become increasingly obscured.Morulae and blastulae were dissociated by placing them in 1/15 M phosphate buffer (pH 7.8) for the duration of 2–3 cleavage cycles and then removing the vitelline membrane. In this solution cell divisions continued without disturbance of the temporal cleavage pattern. The dissociated cells were fixed either just prior to the onset of the next cleavage (up to the 10th cleavage) or at those times when cleavageswould have been expected, had there been no lenthening of cleavage cycles (beyond the 10th cleavage). The total cell number was counted, dividing cells being scored as two.Prior to the 11th cleavage the total cell number increased exponentially. Beyond the 10th cleavage the rate of increase was considerably lower. At the time when gastrulation would have started if the egg had not been dissociated, the total cell counts were 13,000–15,000, whereas the number anticipated without lengthening of cleavage cycles would be of the order of 130,000 (2¹⁷).The application of Balfour's rule to amphibian eggs is criticized.     

Die Dynamik der Frühentwicklung vonSagitta setosa

Zusammenfassung 1. Untersuchungsobjekt.Sagitta setosa Joh. Müll. Vergleichsobjekt:Sagitta elegans arctica Aurv. Methoden: Laufbild- und Teilbildanalyse von Mikrozeitrafferfilmen; kinematische Diagramme.2. Unter Zeittransformation (Zeitraffung, Z.R.) wird eine Ausdehnung des Keimes während der Teilungsphase, gefolgt von einer Kontraktion in der Interphase, im Z.R.-Laufbild erkennbar. Dieser Rhythmus ist etwa vom 32-Blastomerenstadium bis zur neunten Teilung (256/512 Zellen) zu beobachten. Er kommt zustande durch die Summation der nach jeder Teilung einsetzenden aktiven Aneinanderpressung der jeweils entstandenen beiden neuen Blastomere. Dieser Vorgang ist bei den beiden untersuchten Sagitten-Arten besonders intensiv und führt in der Kontraktionsphase zur fast völligen Abkugelung des Keimes.3. Die erste Andeutung des sehr kleinen Blastocoel wird bei der Aneinanderpressung der beiden ersten Blastomere in Gestalt eines in zwei Spitzen ausgezogenen Flüssigkeitstropfens wahrnehmbar. Die innerhalb der Berührungsfläche ausgepreßten kleineren Tropfen weisen zentripetale Ortsverlagerung auf und verschmelzen mit dem größeren Blastocoel-Tropfen. Der Keimbahnkörper ist im Leben bis zum 16-Zellenstadium feststellbar.4. Es folgt eine Wiederholung der Tropfenabsonderung nach jeder Teilung in der Pressungsphase bis zur neunten Teilung. Das Blastocoel vergrößert sich infolge der Flüssigkeitsaufnahme.5. Kinematische Diagramme der ersten Teilungen, durch Teilbild-Analyse aus Z.R.-Aufnahmen gewonnen, beweisen, daß die stärkste Tropfenabsonderung jeweils mit der Phase intensiver Abkugelung zusammenfällt.6. BeiSagitta elegans arctica Aurv. ist die Tropfenabsonderung in der Interphase (Abrundungs-Pressungsphase) erheblich größer, die Abrundung geringer.7. Die unbedeutende Vergrößerung des Blastocoel durch Flüssigkeitsaufnahme läßt keine direkte Entodermbildung durch Invagination zu. Der endgültigen Einstülpung gehen drei Versuche voraus.8. Die vom 32-Blastomerenstadium ab am vegetativen Pol teilweise herausragenden Urgeschlechtszellen weisen gegenüber den Somazellen Teilungsverzögerung auf. Bei der sechsten Teilung (32/64) zeigen die Urgeschlechtszellen unter Z.R. eine aktive zentripetale Bewegung; sie verschwinden am vegetativen Pol und drücken das Blastocoel mit ihren proximalen Enden ein: erster Invaginationsversuch. Zu Beginn der nächsten Teilung erfolgen rückläufige Bewegung und Wiedererscheinen am vegetativen Pol. Der zweite Versuch findet bei der siebten Teilung statt (128 Blastomere), der dritte bei der achten Teilung (256 Zellen). Die Aktivität der nunmehr vier Urgeschlechtszellen ist unverkennbar.9. Die sich zunächst nicht weiter teilenden vier Urgeschlechtszellen behalten etwa die Größe eines Blastomer des 64-Zellenstadium bei, während die Somazellen unterdessen wesentlich kleiner geworden sind. Gastrulation durch Invagination ist erst möglich, wenn das Größenverhältnis zwischen den beiden Zellgruppen eine Ortsverlagerung ektodermaler Zellen in das kleine Blastocoel dynamisch zuläßt.10. Auffallend ist unter Z.R. die Fähigkeit der Blastomere zu aktiv-passiven Bewegungen, vor allem in der Region des Prostoma, in der Umbiegungszone Ektoderm-Entoderm.11. Nach Invagination des Entoderm, die vier Urgeschlechtszellen an der Spitze, liegt zunächst ein schmales Urdarmlumen vor. Das Entoderm zeigt während der Interphase wieder eine rückläufige Bewegung, die als Restverhalten des vorausgegangenen Rhythmus aufzufassen ist.12. Sobald die zentripetalen Invaginations-Versuche der Urgeschlechtszellen — von der sechsten bis zur achten Teilung — einsetzen, dauern die Teilungsschritte länger als vorher; bis zur achten Teilung nimmt die Zeit erheblich zu; nach der neunten verlaufen die Teilungen nicht mehr synchron.13. Die große Aktivität der vier Urgeschlechtszellen bleibt auch noch nach ihrer Einordnung in das Entoderm des Urdarmdaches erhalten, offenbar im Rhythmus der nunmehr lokalisierten Teilungen im Entoderm.14. Nach Verlagerung der Urgeschlechtszellen in das Lumen des Archenteron, unter Beibehaltung des Kontaktes mit dem Entoderm, ist das Blastocoel bis auf den schmalen spaltartigen Raum zwischen Ekto- und Entoderm verdrängt. Das Auswandern der vier Zellen erfolgt offenbar durch aktive, mit starker Metabolie verbundenen Bewegungen. Das Archenteron weist unter Z.R. rhythmische Erweiterungen und Verengungen auf.15. Der Verschluß des Prostoma (Deuterostomia) verläuft synchron mit dem Auswandern der Urgeschlechtszellen.

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The dynamics of the early development ofSagitta setosa. Film-Projection and single-frame analysis of time lapse movies

The dynamics of the early development of the chaetognathSagitta setosa Joh. Müll. has been studied up to the completed gastrulation, the leaving of the primordial germ cells and the closure of the prostoma. Comparative studies have been conducted onSagitta elegans arctica Aurv. The analysis was performed on the basis of micro-time-lapse movies and exact investigation of single frames (Teilbild-Analyse). A detailed account is presented on rhythmic behaviour of the blastomeres, such as the expansion during the cell division and the contraction during interphase, which is combined with an almost complete rounding off of the embryo and the production of fluid drops in the contact areas between the blastomeres. These drops fuse with the minute blastocoel and enlarge it. The Urgeschlechtszellen (primordial germ cells) are very active; during the sixth to eight division of the blastomeres they show an active centripetal movement, depress the blastocoel to some extent and move backward until they project in part into the region of the vegetative pole (during interphase) beyond the surface of the rounded embryo. Gastrulation by means of invagination becomes only possible after three advancing attempts of the primordial cells. During invagination active-passive movements of blastomeres occur in the Umbiegungszone between ectoderm and entoderm. Duration of cell division increases considerably after the sixth division. After the ninth division the blastomeres no longer divide synchronically. Within the entoderm the 4 primordial germ cells remain very motile. Closure of the prostoma occurs synchronically with the dislocation of the primordial germ cells into the archenteron lumen. They stay in further contact with the entoderm of the Urdarmdach. The cinematic diagrams produced by employing the method of single frame-analysis of time lapse series allow an exact survey of the cytodynamic processes during the embryonic development.

     

Organization of the nematocyst battery in the tentacle of hydra: Arrangement of the complex anchoring junctions between nematocytes,epithelial cells,and basement membrane

Summary Nematocytes (stinging cells) of hydra tentacles are anchored to the basement membrane by peculiar complex junctions in which a flattened tongue of an epithelial cell is interposed between the nematocyte and the basement membrane. In this paper we describe the arrangement of these junctions with emphasis on how they are related to the architecture of the epithelial cell. Each epithelial cell, called a battery cell, harbors 10–20 nematocytes and bears muscle processes that extend along the basement membrane. The epithelial cell component of the complex junction is usually a lateral extension of a muscle process. All nematocytes within a battery cell make junctions with muscle processes of the same (resident) epithelial battery cell despite the presence of numerous muscle processes from adjacent (foreign) cells. Some nematocytes make junctions with several resident processes, spanning the foreign processes to do so. Most junctions reside near the proximal ends of the muscle processes. New findings are reported on the substructure of the junctions. They are composed of aggregates of smaller elements, and the cytoskeleton within the complexes has a pronounced longitudinal organization. These observations are consistent with a suggestion that the complex junctions develop by aggregation of smaller junctional units originating elsewhere on the cells.     

Intercellular communication and tissue growth: IX. Junctional membrane structure of hybrids between communication-competent and communication-incompetent cells

Summary The structure of the membrane junctions of the hybrid cell system, examined in the companion paper in respect to competence for communication through cell-to-cell membrane channels, is here examined by freeze-fracture electron microscopy. The junctions of the channel-competent parent cell and of the channel-competent hybrid cells present aggregates of intramembranous particles typical of gap junction; those of the channel-incompetent parent cell and channel-incompetent segregant hybrid cells do not. Competence for junctional communication and for gap junction formation are genetically related. The junctions of the intermediate hybrid cells with incomplete channel-competence (characterized by cell-to-cell transfer of small inorganic ions but not of fluorescein), present special intramembranous fibrillar structures instead of discrete gap-junctional particles. The possibility that these structures may constitute coupling elements with subnormal permeability is discussed in terms of incomplete dominance of the genetic determinants of gap junction.     

Gap junction formation between normal and reaggregated endoderm cells ofXenopus laevis neurulae

Summary Using freeze-fracture electron microscopy and fluorescent dye injection we have analysed the contacts between cells of the deeper endoderm taken from neurulae ofXenopus laevis. Endodermal cells in situ have large 1.5 m diameter gap junctions composed of 8 nm P-face particles and corresponding E-face pits. Beside gap junctions, particle aggregates typical of desmosomal plaques are present but there are no tight junctions. The dissociation of endoderm into single cells involves profound structural alterations in the surface membrane including the complete disappearance of junctional structures among them gap junctions. The reaggregation of endoderm cells leads to the restoration of the surface membrane IMP (Intra Membrane Particle) pattern and, after ca. 30 min, to the establishment of functional pathways allowing for the intercellular transfer of fluorescent dye. Concomitantly gap junctions reappear. The observation that the dissociation and reaggregation of endodermal cells involves IMP alterations which go beyond the cell junctions themselves is discussed as an adaptation of the plasma membrane to changing environmental conditions.     

Starch hydrolysing enzymes in the developing barley grain

Summary The enzymes -amylase (-1, 4-glucan 4-glucanohydrolase, 3.2.1.1), -amylase (-1,4-glucan maltohydrolase, 3.2.1.2) and phosphorylase (-1,4-glucan: orthophosphate glucosyltransferase, 2.4.1.1) were assayed in whole grains of barley throughout the maturation period. -amylase and phosphorylase had peaks of activity between 25 and 30 days after anthesis. On the other hand the activity of -amylase in both the available and latent forms reached a maximum value at 35 days after anthesis which did not decrease thereafter. -amylase activity was also assayed throughout development in the endosperm, aleurone, testa pericarp and embryo. Latent -amylase reached a constant maximum value in endosperm at 35 days but available -amylase reached a peak of activity at 25 days and then declined to zero at 45 days. Only latent -amylase was associated with the aleurone layer and activity rose to a maximum value at 35 days. The testa pericarp had mainly latent -amylase whose activity fell from an early maximum at 21 days to zero at 35 days. No hydrolytic activity was associated with the embryo. The phosphorylase activity was low and mainly associated with the endosperm fraction.     

Ultrastructure of the oocyte and embryo of the calcified sponge,Petrobiona massiliana (Porifera,Calcarea)

Summary In the spongePetrobiona massiliana, a Calcarea related to pharetronid fossils, the oocyte and the embryo both receive an unusual amount of maternal nurse cells. Yolk granules are large and display a lamellar structure throughout the entire growth period, which allows them to be used as markers of the oocyte reserves. The cruciform cells (cellules en croix) of the embryo appear to degenerate at an early stage. These features are compared to those found in other Calcarea.     

Some observations on the fine structure of the myotendinous junction in myotomal muscles of the tadpole tail

Summary Myotendinous junctions in the myotomal tail muscles of the tadpole of Rana rugosa were examined by electron microscopy. At the site of the myotendinous junction, the sarcolemma is covered on its sarcoplasmic aspect by the connecting filament layer and the attachment layer, and on the extracellular aspect by the intermediary layer and the external lamina, with associated collagen fibrils. The intermediary layer consists of filamentous structures which closely resemble microfibrils (Hanak and Böck, 1971), spine-like or thread-like profiles (Korneliussen, 1973) and intermediary layer (Nakao, 1975a, b) in the myotendinous junctions of other vertebrate skeletal muscles.Particularly interesting is the fact that all the coverings and linings of the sarcolemma, including the external lamina, are completely absent in the terminal segment of the finger-like sarcolemmal invagination characteristic of the myotendinous junction. Furthermore, special types of coupling between a sac of sarcoplasmic reticulum and a part of the sarcolemmal invagination are frequently observed. These couplings always occur along the region of the sarcolemma where the external lamina is absent. The couplings show features similar to those of the triad, such as SR feet , scalloped SR membranes and granular content of the SR sac, suggesting that they are analogous and functionally similar to the triad and other equivalent structures.     

Role of histamine and cyclic nucleotides in implantation in the rabbit

Summary The effect of histamine on cAMP and cGMP levels in day 6 (144 h post coitum) rabbit blastocysts was determined. Histamine at 200 M and 1000 M concentrations stimulated the increased formation of cAMP in vitro, whereas stimulation of cGMP occurred only in the presence of 1000 M histamine. Furthermore, intrauterine injection of RMI-12330A (50 g or 500 g/uterine horn), an inhibitor of adenylyl cyclase, on day 5 of pregnancy interrupted embryro development and implantation of the embryo. The drug was also effective in reducing the cAMP level in the endometrial cells in vitro. A relationship between histamine and cyclic nucleotide changes in embryo development and implantation is suggested.