In vitro fertilization and early embryogenesis: a cytological analysis

Anderson, Everett; Hoppe, Peter C.; Whitten, W. K.; Lee, Gloria S.

doi:10.1016/s0022-5320(75)80054-2

Cited by 49 publications

(6 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Detectable lamins are vastly reduced in sperm, and the formation of the male pronucleus in both systems, involving dramatic biochemical (reviewed in refs. 25 and 26) and ultrastructural rearrangements (27,28), is coupled with the appearance of lamins associated with the male pronuclear envelope. In mouse oocytes, lamins appear on the female pronucleus as it develops after the completion of meiosis.…”

Section: Results Mouse Fertilization: Lamins Appear On Pronuclel But mentioning

confidence: 98%

Nuclear lamins and peripheral nuclear antigens during fertilization and embryogenesis in mice and sea urchins.

Schatten¹,

Maul²,

Schatten³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

128

View full text Add to dashboard Cite

Nuclear structural changes during fertilization and embryogenesis in mice and in sea urchins have been followed by using antibodies against the nuclear lamins A/C and B and against antigens at the periphery of nuclei and chromosomes. Lamins are found on all pronuclei and nuclei during mouse fertilization, but with a diminished intensity on the second polar body nucleus. On sperm in both systems, lamins are reduced and detected only at the acrosomal and centriolar fossae. In sea urchin eggs, lamins are found on both pronuclei. Unlike in other dividing cells, the mitotic chromosomes of sea urchin eggs and embryos retain an association with lamins. The peripheral antibodies delineate each chromosome and nucleus except the mature mouse sperm nucleus. A dramatic change from the expected lamin distribution occurs during early development. In mouse morulae or blastocysts, lamins A/C are no longer recognized, although lamin B remains. In sea urchins both lamins A/C and lamin B, as detected with polyclonal antibodies, are lost after the blastula stage, although a different lamin A/C epitope emerges as recognized by a monoclonal antibody. These results demonstrate that pronucleus formation in both systems involves a new association or exposure of lamins, that the polar body nucleus is largely restricted from the cytoplasmic pool of lamins, and that mitotic chromosomes in the rapidly proliferating sea urchin egg retain associated lamins. They also suggest that changes in the expression or exposure of different lamins are a common feature of embryogenesis.Fertilization requires several dramatic changes in nuclear organization. The architecture of the nuclear surface (reviewed in refs. 1-4) involves the nuclear lamins, which are typically three proteins subjacent to the inner nuclear membrane (5, 6), and nuclear peripheral proteins referred to as "P1" (7) or "Perichromin" (8), which probably reside between the chromatin and the nuclear lamins. During mitosis in somatic cells, the lamins dissociate from the nuclear envelope at prophase and reappear with the reconstituting envelope at telophase (9). Unlike the behavior of the lamins at mitosis, the peripheral antigens separate from the nuclear periphery and ensheathe the condensing chromosomes before nuclear envelope breakdown and dissolution of the lamins during mitosis (7,8). During spermatogenesis the nuclear lamins are lost or vastly reduced (10-12), whereas during oogenesis the lamina may be comprised of only a single lamin (13-16), which differs from somatic lamins (17).In this study the presence and distribution of nuclear lamins and of the nuclear and chromosomal P1 peripheral antigens are traced during fertilization and embryogenesis in mice and in sea urchins. These two systems represent

show abstract

Section: Results Mouse Fertilization: Lamins Appear On Pronuclel But mentioning

confidence: 98%

Nuclear lamins and peripheral nuclear antigens during fertilization and embryogenesis in mice and sea urchins.

Schatten¹,

Maul²,

Schatten³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

128

View full text Add to dashboard Cite

show abstract

“…This protuberance has been termed the incorporation cone (Stefanini et al, 1969;Zamboni, 1971) or, by analogy with invertebrates, the fertilization cone (Longo, 1973). Although a small cone has been reported in rabbit (Pincus, 1930), rat (Austin and Braden, 1956) and mouse (Stefanini e t al, 1969;Zamboni, 1971, andAnderson et al, 1975), there has been no detailed study of its structure or functions. In the persent study we have used time lapse cinematography, electron microscopy and a pharmaceutical agent to investigate the sequence of appearance and structure of the cone.…”

Section: Ntrod Uctl Onmentioning

confidence: 99%

Observation on the incorporation cone in the rat

1978

View full text Add to dashboard Cite

At the time of in vivo sperm-egg fusion in the rat, a small region of the oolemma under the head of the fertilizing sperm is observed to be free of microvilli. The microvilli-free region increases in area, and by one hour after sperm-egg contact extends over an area 20-30 p in circumference and bulges out t o form an "incorporation cone" visible by light microscopy. The microvilli-free incorporation cone reaches its maximum size at about two hours after sperm-egg interaction. It soon becomes smaller and has disappeared three to four hours after sperm-oocyte fusion. The cone cytoplasm is characterized by a 0.1 p zone of thin filaments below the plasma membrane. Cytochalasin-B, 2.5 pg/ml, prevents formation of the cone or destroys the intact cone. It is suggested that microfilaments may be involved in the formation of the incorporation cone.

show abstract

“…1). The participation of the egg microtubules during mammalian fertilization is less well understood, though microtubule inhibitors (2)(3)(4) prevent the completion of meiosis, resulting in polyploidy; microtubules have also been found within fertilized mammalian eggs with electron microscopy (5)(6)(7)(8) and during oogenesis with immunofluorescence microscopy (9).…”

mentioning

confidence: 99%

Microtubule configurations during fertilization, mitosis, and early development in the mouse and the requirement for egg microtubule-mediated motility during mammalian fertilization.

Schatten¹,

Simerly²,

Schatten³

1985

Proc. Natl. Acad. Sci. U.S.A.

335

205

View full text Add to dashboard Cite

Microtubules forming within the mouse egg during fertilization are required for the movements leading to the union of the sperm and egg nuclei (male and female pronuclei, respectively). In the unfertilized oocyte, microtubules are predominantly found in the arrested meiotic spindle. At the time for sperm incorporation, a dozen cytoplasmic asters assemble, often associated with the pronuclei. As the pronuclei move to the egg center, these asters enlarge into a dense array. At the end of first interphase, the dense array disassembles and is replaced by sheaths of microtubules surrounding the adjacent pronuclei. Syngamy (pronuclear fusion) is not observed; rather the adjacent paternal and maternal chromosome sets first meet at metaphase. The mitotic apparatus emerges from these perinuclear microtubules and is barrelshaped and anastral, reminiscent of plant cell spindles; the sperm centriole does not nucleate mitotic microtubules. After cleavage, monasters extend from each blastomere nucleus. The second division mitotic spindles also have broad poles, though by third and later diisions the spindles are typical for higher animals, with narrow mitotic poles and fusiform shapes. Colcemid, griseofulvin, and nocodazole inhibit the microtubule formation and prevent the movements leading to pronuclear union; the meiotic spindle is disassembled, and the maternal chromosomes are scattered throughout the oocyte cortex. These results indicate that microtubules forming within fertilized mouse oocytes are required for the union of the sperm and egg nuclei and raise questions about the paternal inheritance of centrioles in mammals.Fertilization results in the union of the parental genomes, and in most animals a microtubule-containing cytoskeleton forming within the activated egg participates in the motility necessary for the cytoplasmic migrations of the sperm and egg nuclei (reviewed in ref. 1). The participation of the egg microtubules during mammalian fertilization is less well understood, though microtubule inhibitors (2-4) prevent the completion of meiosis, resulting in polyploidy; microtubules have also been found within fertilized mammalian eggs with electron microscopy (5-8) and during oogenesis with immunofluorescence microscopy (9).To explore the participation of egg cytoplasmic microtubules during mammalian fertilization and early development, we have performed anti-tubulin immunofluorescence and transmission electron microscopy on mouse oocytes and zygotes* throughout fertilization and have studied the effects of microtubule inhibitors. These results indicate that the egg cytoplasmic microtubules, organized by sources other than the sperm centriole, are required during mammalian fertilization. MATERIALS AND METHODSVirgin CD-1 mice (Charles River Breeding Laboratories) were superovulated with 10 international units of pregnant mare serum followed 48 hr later with 10 international units of human chorionic gonadotropin (10) and introduced to experienced males. After mating, fertilized oocytes were collected (11, ...

show abstract

In vitro fertilization and early embryogenesis: a cytological analysis

Cited by 49 publications

References 34 publications

Nuclear lamins and peripheral nuclear antigens during fertilization and embryogenesis in mice and sea urchins.

Nuclear lamins and peripheral nuclear antigens during fertilization and embryogenesis in mice and sea urchins.

Observation on the incorporation cone in the rat

Microtubule configurations during fertilization, mitosis, and early development in the mouse and the requirement for egg microtubule-mediated motility during mammalian fertilization.

Contact Info

Product

Resources

About