scholarly journals THE EFFECTS OF NICOTINE ON FERTILIZATION IN THE SEA URCHIN, ARBACIA PUNCTULATA

1970 ◽  
Vol 46 (2) ◽  
pp. 308-325 ◽  
Author(s):  
Frank J. Longo ◽  
Everett Anderson
Keyword(s):  
Sea Urchin ◽  
Cortical Response ◽  
Large Nucleus ◽  
Zygote Nucleus ◽  
Arbacia Punctulata ◽  
Male Pronucleus ◽  
Initial Series ◽  
Female Pronucleus

The number of sperm incorporated into eggs made polyspermic with varying concentrations of nicotine (0.025–0.25%, v/v) appears to be directly related to the concentrations employed. The cortical response is morphologically equivalent to that observed in control preparations. Shortly after their incorporation all of the spermatozoa undergo structural events normally associated with the development of the male pronucleus in monospermic eggs. During the reorganization of the spermatozoa, sperm asters are formed. The number of male pronuclei that initially migrate to and encounter the female pronucleus is usually one to three. When pronuclei come into proximity to one another the surface of the female pronucleus proximal to the advancing male pronuclei flattens and becomes highly convoluted. Subsequently, the pronuclei contact each other and the outer and inner membranes of the pronuclear envelopes fuse, thereby producing the zygote nucleus. The male pronuclei remaining in the zygote after this initial series of pronuclear fusions continue to differentiate, i.e. they enlarge, form nucleolus-like bodies, and undergo further chromatin dispersion. In approximately 90% of the zygotes, all of the remaining male pronuclei progressively migrate to the zygote nucleus and fuse to form one large nucleus by 80 min postinsemination. Mitosis and cleavage of the polyspermic zygote occurs later than in monospermic eggs.

scholarly journals A CYTOLOGICAL STUDY OF THE RELATION OF THE CORTICAL REACTION TO SUBSEQUENT EVENTS OF FERTILIZATION IN URETHANE-TREATED EGGS OF THE SEA URCHIN, ARBACIA PUNCTULATA

1970 ◽  
Vol 47 (3) ◽  
pp. 646-665 ◽  
Author(s):  
Frank J. Longo ◽  
Everett Anderson
Keyword(s):  
Sea Urchin ◽  
Cortical Granule ◽  
Cortical Granules ◽  
Cell Stage ◽  
Zygote Nucleus ◽  
Arbacia Punctulata ◽  
Cortical Reaction ◽  
Female Pronucleus ◽  
Almost All ◽  
Granule Release

Eggs of the sea urchin, Arbacia punctulata, treated with 3% urethane for 30 sec followed by 0.3% urethane and inseminated are polyspermic and fail to undergo a typical cortical reaction. Upon insemination the vitelline layer of urethane-treated eggs either does not separate or is raised only a short distance from the oolemma. 1–6 min after insemination, almost all of the cortical granules remain intact and are dislodged from the plasmalemma. Later (6 min to the two-cell stage) some cortical granules are released randomly along the surface of the zygote. Not all zygotes show the same degree of cortical granule dehiscence; most of them experience little if any granule release whereas others demonstrate considerably more. The thickness of the hyaline layer appears to be directly related to the number of cortical granules released. Subsequent to pronuclear migration, several male pronuclei become associated with the female pronucleus. Later the male and female pronuclear envelopes contact and the outer and the inner laminae fuse, thereby forming the zygote nucleus. The male pronuclei remaining in the cytoplasm increase in size and progressively migrate to, and fuse with, the zygote nucleus. By 60 min some zygotes appear to contain only one large zygote nucleus which subsequently enters mitosis. Other zygotes possess a number of male pronuclei which remain unfused, and later these pronuclei along with the zygote nucleus undergo mitosis. There does not appear to be a direct relation between the number of cortical granules a zygote possesses and the above mentioned dichotomy.

scholarly journals THE FINE STRUCTURE OF PRONUCLEAR DEVELOPMENT AND FUSION IN THE SEA URCHIN, ARBACIA PUNCTULATA

1968 ◽  
Vol 39 (2) ◽  
pp. 339-368 ◽  
Author(s):  
Frank J. Longo ◽  
Everett Anderson
Keyword(s):  
Nuclear Envelope ◽  
Sea Urchin ◽  
Plasma Membranes ◽  
Light And Electron Microscopy ◽  
Sperm Nucleus ◽  
Sperm Chromatin ◽  
Zygote Nucleus ◽  
Arbacia Punctulata ◽  
Male Pronucleus ◽  
Sperm Aster

Fertilization events following coalescence of the gamete plasma membranes and culminating in the formation of the zygote nucleus were investigated by light and electron microscopy in the sea urchin, Arbacia punctulata. Shortly after the spermatozoon passes through the fertilization cone, it rotates approximately 180° and comes to rest lateral to its point of entrance. Concomitantly, the nonperforated nuclear envelope of the sperm nucleus undergoes degeneration followed by dispersal of the sperm chromatin and development of the pronuclear envelope. During this reorganization of the sperm nucleus, the sperm aster is formed. The latter is composed of ooplasmic lamellar structures and fasciles of microtubules. The male pronucleus, sperm mitochondrion, and flagellum accompany the sperm aster during its migration. As the pronuclei encounter one another, the surface of the female pronucleus proximal to the advancing male pronucleus becomes highly convoluted. Subsequently, the formation of the zygote nucleus commences with the fusion of the outer and the inner membranes of the pronuclear envelopes, thereby producing a small internuclear bridge and one continuous, perforated zygote nuclear envelope.

scholarly journals An ultrastructural study of cross-fertilization (Arbacia female x Mytilus male).

1977 ◽  
Vol 73 (1) ◽  
pp. 14-26 ◽  
Author(s):  
F J Longo
Keyword(s):  
Sea Urchin ◽  
Ultrastructural Study ◽  
Cortical Granule ◽  
Male Pronucleus ◽  
Female Pronucleus ◽  
Sperm Nuclei ◽  
Cross Fertilization ◽  
Sperm Aster

Insemination of sea urchin (Arbacia) ova with mussel (Mytilus) sperm has been accomplished by treating eggs with trypsin and suspending the gametes in seawater made alkaline with NaOH. Not all inseminated eggs undergo a cortical granule reaction. Some eggs either elevate what remains of their vitelline layer or demonstrate no cortical modification whatsoever. After its incorporation into the egg, the nucleus of Mytilus sperm undergoes changes which eventually give rise to the formation of a male pronucleus. Concomitant with these transformations, a sperm aster may develop in association with the centrioles brought into the egg with the spermatozoon. Both the male pronucleus and the sperm aster may then migrate centrad to the female pronucleus. Evidence is presented which suggests that fusion of the male pronuclei from Mytilus sperm with female pronuclei from Arbacia eggs may occur, although this was not directly observed. These results demonstrate that Mytilus sperm nuclei are able to react to conditions within Arbacia eggs and differentiate into male pronuclei.

scholarly journals Nucleo-cytoplasmic interactions that control nuclear envelope breakdown and entry into mitosis in the sea urchin zygote

1999 ◽  
Vol 112 (8) ◽  
pp. 1139-1148 ◽  
Author(s):  
E.H. Hinchcliffe ◽  
E.A. Thompson ◽  
F.J. Miller ◽  
J. Yang ◽  
G. Sluder
Keyword(s):  
Nuclear Envelope ◽  
Dna Synthesis ◽  
Sea Urchin ◽  
Mammalian Cells ◽  
Cyclin B1 ◽  
Nuclear Accumulation ◽  
Dna Breaks ◽  
Male Pronucleus ◽  
Nuclear Envelope Breakdown ◽  
Female Pronucleus

In sea urchin zygotes and mammalian cells nuclear envelope breakdown (NEB) is not driven simply by a rise in cytoplasmic cyclin dependent kinase 1-cyclin B (Cdk1-B) activity; the checkpoint monitoring DNA synthesis can prevent NEB in the face of mitotic levels of Cdk1-B. Using sea urchin zygotes we investigated whether this checkpoint prevents NEB by restricting import of regulatory proteins into the nucleus. We find that cyclin B1-GFP accumulates in nuclei that cannot complete DNA synthesis and do not break down. Thus, this checkpoint limits NEB downstream of both the cytoplasmic activation and nuclear accumulation of Cdk1-B1. In separate experiments we fertilize sea urchin eggs with sperm whose DNA has been covalently cross-linked to inhibit replication. When the pronuclei fuse, the resulting zygote nucleus does not break down for >180 minutes (equivalent to three cell cycles), even though Cdk1-B activity rises to greater than mitotic levels. If pronuclear fusion is prevented, then the female pronucleus breaks down at the normal time (average 68 minutes) and the male pronucleus with cross-linked DNA breaks down 16 minutes later. This male pronucleus has a functional checkpoint because it does not break down for >120 minutes if the female pronucleus is removed just prior to NEB. These results reveal the existence of an activity released by the female pronucleus upon its breakdown, that overrides the checkpoint in the male pronucleus and induces NEB. Microinjecting wheat germ agglutinin into binucleate zygotes reveals that this activity involves molecules that must be actively translocated into the male pronucleus.

C-Microtubules in Isolated Mitotic Spindles

1971 ◽  
Vol 9 (3) ◽  
pp. 603-619
Author(s):  
W. D. COHEN ◽  
T. GOTTLIEB
Keyword(s):  
Cross Section ◽  
Sea Urchin ◽  
Cross Sections ◽  
Metaphase Chromosome ◽  
Inverse Relationship ◽  
Mitotic Spindles ◽  
Cylindrical Structure ◽  
Arbacia Punctulata ◽  
Alternate Possibility

Microtubules with incomplete cylindrical structure are present in isolated mitotic spindles of the sea urchin, Arbacia punctulata. In cross-section they appear C-shaped, and are thus similar to the ‘C-microtubules’ or ‘C-filaments’ observed previously in other systems. The C-microtubules are not uniformly distributed within isolated spindles, but are typically numerous in the interzonal region of anaphase spindles and in the metaphase chromosome ‘plate’. In chromosome-to-pole regions they are seen much less frequently, and microtubules with the usual O-configuration predominate. Counts of C- and O-microtubules in anaphase spindle cross-sections of known location show an inverse relationship between the number of C-microtubules present and the total number of microtubules present. The observations suggest that the C-microtubules are not simple artifacts of fixation or isolation, but rather may represent a stage of microtubule disassembly which occurs in the interzone during isolation or during anaphase in vivo. The alternate possibility of assembly is not excluded, however. The significance of C-microtubules is further discussed with respect to their occurrence in other systems, and to potential differences between mitotic microtubules.

scholarly journals THE EXTRACELLULAR RELEASE OF ECHINOCHROME

1946 ◽  
Vol 29 (5) ◽  
pp. 267-275 ◽  
Author(s):  
Herbert Shapiro
Keyword(s):  
Sea Urchin ◽  
Sea Water ◽  
Potassium Content ◽  
Red Pigment ◽  
Anaerobic Conditions ◽  
Outward Diffusion ◽  
Arbacia Punctulata ◽  
Extracellular Release

A study was made of the diffusion of the red pigment echinochrome from the eggs of the sea urchin, Arbacia punctulata, into sea water. Unfertilized eggs retained their pigment, over periods of hours. Outward diffusion of pigment from unfertilized eggs normally is entirely negligible, or does not occur at all. Enchancing the calcium or potassium content of the artificial sea water (while retaining isosmotic conditions) did not induce pigment release. Under anaerobic conditions, unfertilized eggs release pigment in small quantities. Fertilization alone brings about echinochrome release. Fertilized eggs invariably released pigment, whether in normal sea water, or sea water with increased calcium or potassium. This diffusion of the pigment began during the first cleavage, possibly soon after fertilization. The pigment release is not a consequence solely of the cell's permeability to echinochrome (or chromoprotein, or other pigment combination) but is preceded by events leading to a release of echinochrome from the granules in which it is concentrated within the cell. These events may be initiated by activation or by anaerobiosis. The phenomenon was not due to cytolysis.

Pinocytosis in fertilized sea urchin (Arbacia punctulata) eggs

1984 ◽  
Vol 231 (3) ◽  
pp. 413-422 ◽  
Author(s):  
Christopher P. Carron ◽  
Frank J. Longo
Keyword(s):  
Sea Urchin ◽  
Arbacia Punctulata

scholarly journals OOCYTE DIFFERENTIATION IN THE SEA URCHIN, ARBACIA PUNCTULATA, WITH PARTICULAR REFERENCE TO THE ORIGIN OF CORTICAL GRANULES AND THEIR PARTICIPATION IN THE CORTICAL REACTION

1968 ◽  
Vol 37 (2) ◽  
pp. 514-539 ◽  
Author(s):  
Everett Anderson
Keyword(s):  
Sea Urchin ◽  
Golgi Complex ◽  
Morphological Evidence ◽  
Cortical Granule ◽  
Perivitelline Space ◽  
Unit Membrane ◽  
Cortical Granules ◽  
Arbacia Punctulata ◽  
Cortical Reaction ◽  
Oocyte Differentiation

This paper presents morphological evidence on the origin of cortical granules in the oocytes of Arbacia punctulata and other echinoderms. During oocyte differentiation, those Golgi complexes associated with the production of cortical granules are composed of numerous saccules with companion vesicles. Each element of the Golgi complex contains a rather dense homogeneous substance. The vesicular component of the Golgi complex is thought to be derived from the saccular member by a pinching-off process. The pinched-off vesicles are viewed as containers of the precursor(s) of the cortical granules. In time, they coalesce and form a mature cortical granule whose content is bounded by a unit membrane. Thus, it is asserted that the Golgi complex is involved in both the synthesis and concentration of precursors utilized in the construction of the cortical granule. Immediately after the egg is activated by the sperm the primary envelope becomes detached from the oolemma, thereby forming what we have called the activation calyx (see Discussion). Subsequent to the elaboration of the activation calyx, the contents of cortical granules are released (cortical reaction) into the perivitelline space. The discharge of the constituents of a cortical granule is accomplished by the union of its encompassing unit membrane, in several places, with the oolemma.

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