Showing papers in "Development Growth & Differentiation in 1976"

Crystal property of the larval sea urchin spicule

Abstract: A pair of pluteus skeletal spicules arises from a pair of calcareous granules via the triradiate form. In polarized light, each spicule behaves as though carved out of a single crystal of magnesian calcite. The optic axis lies perpendicular to the plane of the triradiate and parallel to the body rod of the pluteus. However, in the scanning electron microscope, the spicule surface appeared smooth or somewhat spongy and manifested no crystal faces. Neither etching nor fracturing revealed underlying crystalline texture. Nevertheless, rhombohedral calcite crystals could be grown epitaxially onto isolated spicules immersed in a medium containing CaCl2 and NaHCO3. The optic axes of all crystals coincided with the optic axis of the spicule on which they were grown. Corresponding faces of the crystals were all aligned parallel to each other despite the complex shape of each spicule. Where the left and right spicules joined, two mutually tilted sets of crystals were observed but not crystals of intermediate orientation. Thus, the sea urchin larval spicule is built from a stack of molecularly contiguous microcrystals but its overall shape is generated by the mesenchyme cells independent of the magnesian calcite crystal habit.

Growth of lens and ocular environment: role of neural retina in the growth of mouse lens as revealed by an implantation experiment

Abstract: The lens of 6-day-old normal mouse was implanted into the lentectomized eye of adult mouse to examine the effect of retina upon the growth of the implanted lens in vivo. The implanted lens grew normally and its transparency was kept for more than 5 months after implantation. The connection between the implanted lens and the ciliary part of the recipient iris was well established with the regeneration of zonular fibers from the recipient. In young lenses implanted reversely into adult eyes, the epithelial cells facing the retina elongated and a new epithelium was formed on the corneal side of the lens within 5 days. Young lenses implanted either in normal or reverse orientation into eyes from which the retina was previously removed did not grow. The cells of the original lens epithelium of these lenses were randomly accumulated beneath the posterior lens capsule, while the anterior portion of the implanted lenses became an epithelial structure without cell elongation. These results suggest that the growth of the implanted lens may be dependent on the retina of the adult eye.

Induction of chromosome motion in the glycerol‐isolated mitotic apparatus: nucleotide specificity and effects of antidynein and myosin sera on the motion

Abstract: Chromosome motion in glycerol-isolated mitotic apparatus (MA) of sea urchin and starfish eggs was investigated with respect to nucleotide specificity and the effects of antisera against tryptic fragment (Fragment A) of flagellar dynein and starfish egg myosin. The motion was highly specific for ATP. GTP, ITP, CTP, UTP, and ADP caused no displacement of the chromosomes towards the poles. The anti-Fragment A serum completely inhibited chromosome motion in the MA of the sea urchin egg, while antiserum against starfish egg myosin as well as its γ-globulin fraction did not inhibit the motion in the isolated MA of the starfish egg, suggesting that chromosome motion depends upon dynein-microtubule but not upon myosin-actin interaction. In addition, colchicine completely suppressed the chromosome motion in vitro.

Effects of the surfactants on the cleavage and further development of the sea urchin embryos 1. the inhibition of micromere formation at the fourth cleavage

Abstract: Eggs of the sea urchins, Hemicentrotus pulcherrimus, Temnopleurus toreumaticus and Pseudocentrotus depressus were used as materials. Embryos were exposed to the surfactants such as SLS, CTAB, digitonin, Tween 80, sodium deoxycholate and Lubrol P. If embryos are kept in the solutions of SLS, CTAB and digitonin, 4 vegetal cells of the 8-cell stage divide equally at the fourth cleavage and consequently 16 equal-sized blastomeres are formed at the 16-cell stage. In this case, micromere formation is inhibited by the equal cleavage. The minimum effective concentration of the surfactants for the equal cleavage gradually increases as the time performing the treatment is postponed. The continuous exposure to the surfactant is unnecessary for the inhibition of micromere formation. In the egg temporarily exposed during the earlier stage, the equal cleavage occurs at the fourth division in natural seawater. Micromere formation is strongly affected by the surfactant (SLS) at the mid 4-cell stage.

Localization of dynein in sea urchin eggs during cleavage

Abstract: Detection and localization of dynein in cleaving sea urchin eggs were attempted using antidynein serum (prepared against a tryptic fragment of dynein, Fragment A, of sea urchin sperm flagella) and fluorescein conjugated goat antiserum to rabbit γ-globulin. In both unfertilized and newly fertilized eggs, fluorescence was distributed rather uniformly within the cells but was absent from the nuclei. At prophase, intense fluorescence was observed on both sides of nucleus, suggesting accumulation of dynein in developing asters. From metaphase to anaphase, the whole mitotic apparatus (MA) was stained with the exceptions of the chromosomes and pole areas. Fluorescence then again became dispersed within the eggs. Throughout the mitotic process and cytokinesis, the egg cortex including the cleavage furrow was stained intensely, presumably reflecting the presence of dynein in this region. Similar distributions of fluorescence were obtained with the isolated MAs. Neither non-immune serum nor the antiserum to which Fragment A was absorbed stained the eggs. Little staining was obtained with the antiserum against starfish egg myosin. The results, together with the finding that the chromosome motion in the isolated MAs was completely inhibited by anti-dynein serum, but not with the anti-myosin serum, suggest an active role played by a tubulin-dynein system in mitosis.

Microinjection of toad sperm into oocytes undergoing maturation division

Abstract: Spermatozoa of Bufo bufo japonicus were briefly treated with Triton X-100 to remove their plasma membrane, and were injected into oocytes at various stages of maturation division. All the sperm injected into mature coelomic eggs transformed into pronuclei and synthesized DNA, as a normally fertilizing sperm does. The sperm injected into oocytes at the germinal vesicle (GV) stage did not show any change as long as the GV remained intact. In the oocytes which were induced to mature by progesterone, the injected sperm displayed characteristic features in synchrony with those of the resident female nucleus. These included the formation of several sperm-derived chromosomes in association with multipolar spindles in the oocytes from the stage of the germinal vesicle breakdown to the first polar spindle; the appearance of swollen, vesicular nuclei without concomitant DNA synthesis in those at the stage of the first polar body emission; and the reappearance of the condensed chromosomes with giant spindles in those at the stage of the second meiotic metaphase. Pricking of these last oocytes induced the formation of several male pronuclei and DNA synthesis. These results prove that the injection of detergent-treated sperm employed here provides an excellent means of studying the cytoplasmic state that regulates nuclear behavior.

Electron microscopic observations of guinea pig spermatozoa penetrating eggs in vitro

Abstract: Guinea pig ovarian oocytes matured in vitro were inseminated in vitro with capacitated, acrosome-reacted spermatozoa and sperm penetration through the zona pellucida and into the egg cytoplasm were examined. Sperm heads passing through the zona pellucida had already lost all their acrosomal elements except for the inner acrosomal membrane and the equatorial segment. It was often observed that the texture of the zona material around the sperm head was distorted, giving the impression that the zona pellucida was parted, at least partially, by a shearing force produced by the sperm head advancing through the zona. When eggs were freed from their zonae pellucidae and inseminated, the acrosome-reacted spermatozoa immediately bound to the egg surfaces and began to fuse with the eggs; whereas the spermatozoa with intact acrosomes failed to do so. Fusion began between the egg plasma membrane and the sperm plasma membrane at the central region of the sperm head. The anterior half of the sperm head was engulfed by the egg in a phagocytic fashion, while its posterior half was incorporated into the egg by a fussion between egg and sperm plasma membranes. Incorporation of the sperm tail into the egg was achieved by fusion between the sperm and egg plasma membranes.

On the role of calcium ions in oocyte maturation in the polychaete chaetopterus pergamentaceus

Abstract: The germinal vesicle of mechanically released Chaetopterus oocytes disintegrates in natural sea water (NSW), but not in artificial sea water of normal composition (ASW), calcium-free sea water (CaFSW), magnesium-free sea water (MgFSW) or calcium and magnesium-free sea water (CaMgFSW). Several methods of inducing oocyte maturation using chemically well-defined medium have been established. (1) Germinal vesicle breakdown was induced by the treatment of immature oocytes with KCl (60 mM) in ASW or MgFSW. The presence of Ca2+ is necessary for inducing oocyte maturation with high potassium concentration. “Differentiation without cleavage” was observed after this treatment. (2) Trypsin (0.3%) induced oocyte maturation in ASW, but not in CaFSW. Oocytes matured in this manner developed to trochophores upon insemination. (3) Immature oocytes, treated with isotonic CaCl2 for less than 1 min and then transferred to ASW, underwent germinal vesicle breakdown. The oocytes were arrested at the first meiotic metaphase and upon insemination developed to trochophore larvae. (4) Tetracaine (0.4 mM) induced oocyte maturation in the absence of Ca2+ in the medium. In ASW, CaFSW or CaMgFSW containing the drug, oocytes were arrested at the first meiotic metaphase, while in MgFSW with tetracaine they developed parthenogenetically up to the 4- and 8-cell stages. The role of calcium in oocyte maturation was established and its importance was discussed based on the results obtained with the different ways of inducing oocyte maturation.

Role of glucocorticoids in terminal differentiation and tissue‐specific function (a review)

Abstract: Glucocorticoids (GCs) are required by many kinds of organs, tissues and cells for initiation or maintenance of their specific functions in vivo and in vitro. It is noticeable that most of these GC actions can be induced at much lower levels of dosage or concentration than the well-known actions of the steroids in gluconeogenesis, immune suppression and anti-inflammation. Such “differentiation-stimulating” actions of GC are regarded as main physiological roles of the steroid.

A “normal” development of denuded eggs of the starfish, asterina pectinifera*

Abstract: Experimental conditions that allow “normal” development of starfish eggs stripped of the fertilization membrane are reported in this paper. Four kinds of intercellular relation are distinguished during the pre-hatching stages of these eggs. Cells from 2- to 8-cell stages are hardly related to each other, while those from 16- to 128-cell stages are bound loosely together. After the 8th division (about 5.5 hr after insemination at 21°C) cells adhere closely and cooperate with each other to perform morphogenetic movement of “blastulation”. This relation is taken over by that of a true multicellular system at about 10 hr after insemination. Closely after this, the function of cilia carries the embryo away from the substratum.

Motility of dissociated embryonic cells in xenopus laevis: its significance to morphogenetic movements *

Abstract: Motilities of dissociated embryonic cells of Xenopus laevis were investigated in order to elucidate the role of cell motilities in gastrulation. Various shapes and motile forms of the cells were classified into six types, i.e., globular cells with large lobopodia, locomotive vermiform cells with a hyaline cap, globular cells with many bulbous processes, oval cells with filiform pseudopodia, flattened cells with membraneous processes or lamellipodia which attached to glass, and attached cells with hyaline lobopodia. Cells dissociated from the ectodermal region began to exhibit pseudopodial activity at stage 11, while isolated mesoderm cells did so at stage 10 1/2. The pseudopodial activity of cells from these two regions increased coincidently with gastrulation. Approximately 10% of the cells isolated from the dorsal part of the neurula transformed into vermiform cells. Cells dissociated from the endodermal region were less motile during gastrulation. Invaginating cells of the presumptive pharyngeal endoderm were also immotile, when they were isolated.

Mechanical properties of sea urchin eggs iii. visco‐elasticity of the cell surface

Abstract: When a sea urchin egg was compressed between two parallel plates, the force required to keep the distance between the plates constant gradually decreased with time. The contours of the compressed egg were different from the contours expected from the assumption that the surface forces are uniform over the entire surface. The surface forces of the egg without deformation computed from the area of the cell surface in contact with the substratum, the density of the egg and its size were 0.02–0.04 dynes/cm in Hemicentrotus pulcherrimus. Larger values were obtained in eggs during compression. Surface forces, which were computed from measurements of the form of the egg and the applied force when the egg was deformed by a rod and a plate supporting the egg, increased as the deformation increased. From these results, it was concluded that the cell surface is visco-elastic in sea urchin eggs.

Molecular properties and differentiation of acetylcholinesterase in sea urchin embryos

Abstract: The two molecular forms of acethylcholinesterase (EC 3.1.1.7) in sea urchin embryos were characterized by several physical methods. The sedimentation coefficients determined by sucrose gradient centrifugation are 7.6S and 10.6S. The Stokes radii determined by gel filtration are 65 A and 91 A. From these parameters, molecular weights were estimated as 190,000 and 380,000; the one is twice as large as the other. Both forms have similar electric property and buoyant density in a CsCl gradient. When the enzyme solution was concentrated, the 10.6S form became predominant. These results suggest that the two forms are monomer and dimer. The sea urchin enzymes resemble globular forms of acetylcholinesterase of the electric organ of fishes. The activity of the enzyme abruptly increases in post-gastrulation embryos. Inhibition of concomitant protein synthesis by a specific inhibitor, emetine, does not affect the increase in enzyme activity. The result suggests that post-translational processes may be involved in the differentiation of this enzyme in sea urchin development. The following sea urchins were used in the study: Strongylocentrotus purpuratus, Strongylocentrotus franciscanus, and Dendraster excentricus.

Periodic changes in the content of protein-bound sulfhydryl groups and tension at the surface of starfish oocytes in correlation with the meiotic division cycle*

Abstract: The sulfhydryl content of protein and the tension at the surface were measured for starfish oocytes from the first meiotic division to the cleavage stage. A cyclic change in both the protein-SH and the tension at the surface was found to accompany the division cycle, including the first and second meiotic divisions. It is concluded that the unequal meiotic divisions share the same character with the equal divisions of cleavage, with respect to changes both in the protein-SH and the tension at the surface.

Effect of ecdysone on the ovarian development of bombyx silkworm

Abstract: From the ligation experiments, it is indicated that the secretion from prothoracic glands during early pupal period induces the ovarian development in female pupa of the silkworm, Bombyx mori. The exogenous β-ecdysone injected into isolated pupal abdomen also induces the initiation of the ovarian development.

Mechanical properties of starfish oocytes

Abstract: The stiffness of the starfish oocyte was determined from the degree of deformation when it was compressed by a definite force between a pair of parallel plates. The deformation of the oocyte increases during continued application of a constant force, indicating visco-elasticity of the cell. A cyclic change in stiffness of the oocyte accompanying meiotic divisions was found: the stiffness of the oocyte decreases during early stage of meiotic division, increases before the onset of the first polar body formation, then decreases, increases again before the onset of the second polar body formation, and decreases thereafter. Deuteration causes increase in stiffness of the oocyte.

Enzymatic dissociation of nascent macrocysts and partition of the liberated cytophagic giant cells in dictyostelium mucoroides

Abstract: Nascent macrocysts of the cellular slime mold Dictyostelium mucoroides were dissociated enzymatically and the liberated cytophagic giant cells were partitioned by dextrin density gradient centrifugation. Enzymatic and cytochemical studies revealed that the primary wall is composed mainly of cellulose (β-1,4-glucan) associated with polysaccharides including hemicellulose, pectic substances and a-1,4-glucan. The buoyant density of the liberated cytophagic giant cells and peripheral cells was determined by density gradient centrifugation, and partitioning of the cells was possible due to the difference in this property. The process of macrocyst reconstitution was investigated using dissociated cells. The isolated cytophagic giant cell has a specific affinity for other cytophagic giant cells and predominantly ingests them by phagocytosis, while it retains the ability to ingest peripheral cells. The present study provides a clue for investigating the differentiation and development of sexual cells, since only the cytophagic giant cell gives rise to a zygote in macrocyst formation.

The influence of environmental lighting on the growth and prometamorphic development of larval rana pipiens

Abstract: This study was designed to investigate whether the larval development of an anuran amphibian could be modified by raising the animals in continuous light or darkness instead of under conditions of diurnal illumination, and to quantify the effects of these treatments at various intervals during this period of development. Larvae of the frog, Rana pipiens, were raised through metamorphosis under conditions of constant light, constant darkness, or diurnal lighting. As measured by stages of development, body weight, tail length and body length at 20-day intervals, no significant differences in growth rate or metamorphic change were observed until near the middle of the prometamorphic period, which began at approximately the 50th day of development. After midmetamorphosis, a significant acceleration in the measured parameters was seen for the animals raised in conditions of constant light in comparison with those in constant darkness. Those with diurnal lighting were intermediate. These results suggested that light, or its absence, can respectively stimulate or retard amphibian metamorphosis in late larval stages after the hypothalamo-hypophyseal-thyroid axis has matured. Neither continuous light nor continuous darkness during larval development prevented the transformation from tadpole to frog.

Methionine‐activating enzyme in starfish follicle cells *

Abstract: In starfish follicle cells 1-methyladenine is produced under the influence of a gonad-stimulating hormonal peptide (GSS) Since such production of the substance is enhanced by the addition of L-methionine or S-adenosylmethionine in vitro, the presence of methionine-activating enzyme in the follicle cells of the starfish, Asterina pectinifera, was investigated To detect enzyme activity, the enzyme was partially purified from the supernatant of the follicle-cell homogenate by precipitation with ammonium sulfate followed by gel-filtration on a Sephadex G-150 column Using such a preparation of the enzyme, the production of S-adenosylmethionine from L-methionine and adenosine triphosphate was clearly demonstrated by thin-layer chromatography GSS was found to exert no effect on the activity of the methionine-activating enzyme The hormonal peptide, GSS, is therefore considered to take part in some reaction other than this step in the formation of 1-methyladenine

Biochemical and cytological examination on the initiation of ribosomal rna synthesis during gastrulation of xenopus laevis

Abstract: Embryos of Xenopus laevis at stage 6 were labeled with 14CO2 for 4 hr and then allowed to develop under nonradioactive conditions until they reached stage 9, 10, 11 or 12. RNA was extracted and electrophoresed on a polyacrylamide-agarose gel. From the pattern of newly synthesized RNAs, the incorporation into 18S and 28S ribosomal RNAs was measured. At the same time, the specific radioactivity of nucleoside triphosphates in the acid-soluble fraction was determined. On the basis of the results obtained, the absolute amounts of the RNAs synthesized were calculated. The results show that the synthesis of the ribosomal RNAs begins, or is at least markedly activated, around stage 10. Moreover, cytological examination has shown that cells with nucleolated nuclei appeared between stages 9 and 10 and increased thereafter. Thus, from the results of these studies along two different lines, it can safely be concluded that the initiation of 18S and 28S RNA synthesis takes place around stage 10.

Analysis of cell proliferation during early embryogenesis

Abstract: Cell proliferation was examined during early embryogenesis of the newt (Triturus pyrrhogaster) by various methods. After the two-cell stage, at 23°C, the blastomere (cell) number per whole embryo increased logarithmically until the mid-blastula stage (for about 19 hr) and the rate of increase slowed down in and after the late blastula stage. On the other hand, the synchronous cleavage of the blastomeres at the animal pole continued for 18 hr until the twelfth cleavage (mid-blastula) and the transition from synchronous to asynchronous division occurred abruptly at and after the thirteenth cell division (late blastula). The study also showed that the presumptive neuro-ectoderm consisted mainly of cells of the fifteenth generation (G-15) at the onset of gastrulation (pigment stage). The present study suggested that the number of ectodermal cells of the early gastrula (stage 12a) nearly doubled during gastrulation at the presumptive neuro-ectoderm. This means that most of the ectodermal cells are in G-16 at the end of gastrulation. On the other hand, both mitotic activity and the rate of cell increase gradually diminished during gastrulation in the ectoderms of both the presumptive neural and epidermal regions, and there are evidently significant differences in both activities between the neuro-ectoderm and the epidermal ectoderm after stage 13b: the epidermal ectoderm showed greater decrease in the rate of both mitotic activity and cell proliferation than the neuro-ectoderm. These facts suggested that, whether the ectodermal cells will differentiate into neural cells or epidermal cells is determined during G-15 or G-16 in normal primary induction.

Mitotic activity of vaginal epithelial cells following neonatal injections of different doses of estrogen in mice

Abstract: In C57BlackfTw mice given injections of 1 pg estradiol-170 (E) for 5 days beginning on the day of birth, and killed a few days after the treatment, the vaginal epithelium showed estrogen-dependent proliferation and parakeratosis. In contrast, in the mice treated neonatally with 30 pg E for 5 days, the vaginal epithelium exhibited estrogen-independent proliferation and cornification or parakeratosis. Two peaks occurred in the mitotic rate in vaginal epithelial cells in the proximal and middle vaginae of the 1 pgE-treated mice, at 1 and 5 days of age, respectively, while the first peak was lacking in the distal vagina. The mitotic activity in 1 pgE-treated mice declined to the control level at 60 days. In the 30 pgE-treated animals also, 2 peaks were found in the mitotic rate at 1 and 7 days in both the proximal and middle vaginae. In contrast to the 1 pgE-treated mice, although the rate dropped once at 10 days, it increased again at 20 days and remained high thereafter. The second peak at 7 days of age coincided with the active proliferation of nodules appearing in the 30 pgE-treated mice. In the distal vagina, a peak occurred in the mitotic rate at 7 days without a preceding peak like that observed in the other parts of the vagina following the first injection of E on the day of birth. It is well established that treatment of female mice with estrogen during neonatal life induces two different types of persistent proliferation and cornification or parakeratosis of the vaginal epithelium in adulthood. One type of persistent vaginal change, brought about by low doses of estrogen, is ascribable to a permanent alteration of the hypothalamo-hypophysial system and is directly related to a continued secretion of ovarian estrogen; while the other, elicited by high doses of estrogen, is due to permanent changes in the vaginal epithelium itself and is estrogenindependent (TAKASUGI, 1963; KIMURA et al., 1967a, b; MORI, 1967; KOHRMAN and GREENBERG, 1968; for review see TAKASUGI et al., 1970). From the estrogenindependent vaginal changes, cancerous or precancerous lesions frequently develop

Electron microscopic demonstration of tyrosinase in pterinosomes of the frog xanthophore, and the origin of pterinosomes*

Abstract: In the frog, Rana japonica, the successive appearance of types I, II and III pterinosomes, which were defined according to the degree of lamellar structure, is in keeping with the xanthophore differentiation at the larval stage, but these three types coexist in a single xanthophore in the adult. An intense tyrosinase reaction was found in type I–II intermediate form in the larval and adult xanthophores, but it was rarely observed in types I and III. A tyrosinase reaction was always found in the GERL (Golgi-associated Endoplasmic Reticulum) of larval and adult xanthophores, and it was similarly evident in small Golgi vesicles which were separated from the GERL and dispersed in the cytoplasm. The above findings suggest that tyrosinase and pterinosome originate from different parts of the cytoplasm. The hypothesis that small Golgi vesicles are transported to the tyrosinase-negative premelanosomes involved in the origin of the melanosome is also applicable to the origin of pterinosomes.

Effect of actinomycin d on the differentiation of hatching gland cell and cilia cell in the frog embryo

Abstract: The forehead epidermis of the stage 18–20 R. japonica embryo includes the hatching gland cell (HGC) which contains cell-specific secretory granules. The cilia cell (CC) and common epidermal cell (CEC) constitute the epidermis of the entire body surface, in addition to the forehead region. Culture of superficial epidermal explants from various embryonic portions at various developmental stages revealed that HGCs are derived from cells localized on the neural crest in the stage 13a (early neural plate) embryo. When explants from the presumptive HGC area were treated with 1 ug/ml actinomycin D (AMD), the formation of secretory granules in HGCs was inhibited either by continuous treatment from stage 13 or by an 8-hr treatment at stage 13b. Similarly, the ciliogenesis in CCs was inhibited. The differentiation of CECs was entirely unaffected by any of the AMD treatment. After release from AMD, mucous vesicles, characteristic of the CEC, were formed in cells whose differentiation into HGC and CC had been suppressed by the antibiotic. Thread complexes and clumps of coiled strings were found in the nuclei of AMD-affected cells. It is concluded that the DNA-dependent RNA syntheses which direct secretory granule formation in the HGC and ciliogenesis in the CC occur during a limited period at stage 13b, viz., 20 hr before their cytodifferentiation becomes appreciable.

Ultrastructural observations on the discharge of two kinds of granules in the fertilized eggs of cyprinus carpio and carassius auratus

Abstract: This paper describes mainly ultrastructural evidence on the discharge of two kinds of granules, CA and CB, as well as the cortical alveoli in an egg of the common carp and the goldfish. The cortical alveoli (about 2–28 μ in diameter) and CA-granules (about 0.4–2 μ in diameter) are located in the cortical cytoplasm of the mature egg, and the latter is distinguishable in size and texture from the former which contains an eccentric core at the very least as a basal internal structure. The CB-granules (about 100–950 mμ in diameter) appear in small or large clusters in the cortical cytoplasm after fertilization, being formed in connection with constriction or pinching-off of dilated tubular elements. After fertilization the cortical alveoli, CA- and CB-granules are discharged at different times. The new plasmalemma of a fertilized egg appears to be a joint production of the original plasmalemma and the limiting membranes of the cortical alveoli, CA- and CB-granules.

Prefurrow behavior of the equatorial surface in arbacia lixula eggs

Abstract: Study of equatorial surface activity occurring immediately before furrowing in Arbacia lixula (=pustulosa) eggs was undertaken to learn more about the establishment of the cleavage mechanism. Behavior of echinochrome granules in the egg surface, observed and recorded with a Nikon AFM camera, was used as the indicator of surface events. An hour after fertilization A. lixula eggs were slightly flattened and periodically photographed until the furrow appeared. By measuring regional changes in the concentration of echinochrome granules, we found that a band of equatorial surface approximately 22 μm wide, which comprises about 32% of the uncleaved egg surface, shrinks about 34% and forms a densely pigmented band averaging 15 μm wide. This contraction in the equatorial zone is accompanied by expansion or stretching in the subequatorial surfaces. The possible relation between these events and formation of the microfilamentous contractile ring is discussed.

Spermiogenesis of the teleost, oryzias latipes, with special reference to the formation of flagellar membrane *

Abstract: In the early stage of Oryzias spermiogenesis, an axonemal bud appears at the distal end of a centriole characterized by its electron dense accessories. When the axoneme begins to grow in the cytoplasm, small vesicles come to surround it. These vesicles are similar to those produced by the Golgi apparatus which lies close to the growing axoneme. At this stage, the spermatid cell membranes disappear, causing transformation of the mononuclear spermatids into a multinucleated syncytium. As each axoneme elongates in the syncytium, it is enveloped by a cylindrical array of vesicles which are most likely derived from the Golgi apparatus. Shortly after this stage, the syncytium is again partitioned by cell membranes, restoring the existence of mononuclear spermatids. The arrayed vesicles fuse with each other to form two concentric membranes surrounding the axoneme. The inner membrane becomes the flagellar membrane and the outer one, the membrane of a flagellar sheath. These observations lead to the conclusion that the formation of the flagellar membrane is due to the fusion of vesicles surrounding the axoneme which are derived from the Golgi apparatus. In the course of spermiogenesis, no indication of an acrosomal structure is observed.

Effects of two protease inhibitors on sea urchin and amphibian egg development

Abstract: After a review of our present knowledge about the effects of proteases and protease inhibitors on cell growth and egg fertilization, the results of experiments where sea urchin and amphibian eggs were treated with two protease inhibitors (TPCK, TLCK) are described. Cleavage was hardly affected, but gastrulation quickly stopped or was incomplete. The morphogenetic abnormalities which follow can be explained by abnormal gastrulation and other factors: persistence of remnants of the fertilization membrane in sea urchin larvae and dissociation of the ectoderm cells in Xenopus embryos. Copyright © 1976, Wiley Blackwell. All rights reserved

Analysis of a hertwig effect induced by the irradiation of rana pipiens sperm with ultraviolet

Abstract: Some of the underlying parameters of the classical Hertwig Effect were delineated in this study. It was shown that the irradiation of Rana pipiens sperm with an incremental regime of ultraviolet is accompanied by an initial decrease in viability followed by its recrudescence at higher doses. The most impaired survival is concomittant with aneuploid chromosomal conditions. The types of growth abnormalities are also seen to be somewhat dose-specific particularly at 8 sec where acephalic embryos seem to characterize the various aneuploid states observed. This paves the way for a detailed study of both the relationship between aneuploidy, growth anomalies and the important question of the mechanism of chromosomal loss (or removal). That more fundamental, perhaps molecular, events may underlie the Hertwig Effect is further demonstrated by the severe depression in mitotic activity that accompanies the aneuploid conditions.

In vivo study on the process of solid tumor formation from embryoid bodies of mouse teratocarcinoma

Abstract: Embryoid bodies of mouse teratocarcinoma OTT 6050 were studied with special reference to their growth and differentiation in vivo. They were grown either in the peritoneal cavity or in the lung. When injected intraperitoneally, embryoid bodies doubled their number every three days. Some of them attached to a small intra-peritoneal fat body and were soon surrounded by mesenchymal cells of host origin. They grew, fused with each other and became large solid tumors which contained many differentiated tissues. When injected intravenously, almost all the embryoid bodies lodged in the lungs and individually grew into discrete solid tumors which doubled in volume every 2.9 days. After about thirty days, some tumors were composed of only one type of tissue while others contained several types of tissues. The possible mechanisms for the uni- and multi-tissue types of differentiation are discussed.