Human oocyte showing first polar body and metaphase stage in formation of second polar body

1927 ◽  
Vol 2 (4) ◽  
pp. 149-151 ◽  
Author(s):  
A. Francis Dixon
Development ◽  
1996 ◽  
Vol 122 (7) ◽  
pp. 1995-2003 ◽  
Author(s):  
G.L. Russo ◽  
K. Kyozuka ◽  
L. Antonazzo ◽  
E. Tosti ◽  
B. Dale

Using the fluorescent dye Calcium Green-dextran, we measured intracellular Ca2+ in oocytes of the ascidian Ciona intestinalis at fertilization and during progression through meiosis. The relative fluorescence intensity increased shortly after insemination in a single transient, the activation peak, and this was followed by several smaller oscillations that lasted for approximately 5 minutes (phase 1). The first polar body was extruded after the completion of the phase 1 transients, about 9 minutes after insemination, and then the intracellular calcium level remained at baseline for a period of 5 minutes (phase 2). At 14 minutes postinsemination a second series of oscillations was initiated that lasted 11 minutes (phase 3) and terminated at the time of second polar body extrusion. Phases 1 and 3 were inhibited by preloading oocytes with 5 mM heparin. Simultaneous measurements of membrane currents, in the whole-cell clamp configuration, showed that the 1–2 nA inward fertilization current correlated temporally with the activation peak, while a series of smaller oscillations of 0.1-0.3 nA amplitude were generated at the time of the phase 3 oscillations. Biochemical characterization of Maturation Promoting Factor (MPF) in ascidian oocytes led to the identification of a Cdc2-like kinase activity. Using p13suc1-sepharose as a reagent to precipitate the MPF complex, a 67 kDa (67 × 10(3) Mr) protein was identified as cyclin B. Histone H1 kinase activity was high at metaphase I and decreased within 5 minutes of insemination reaching a minimum level during phase 2, corresponding to telophase I. During phase 3, H1 kinase activity increased and then decayed again during telophase II. Oocytes preloaded with BAPTA and subsequently inseminated did not generate any calcium transients, nonetheless H1 kinase activity decreased 5 minutes after insemination, as in the controls, and remained low for at least 30 minutes. Injection of BAPTA during phase 2 suppressed the phase 3 calcium transients, and inhibited both the increase in H1 kinase activity normally encountered at metaphase II and second polar body extrusion.


Zygote ◽  
2003 ◽  
Vol 11 (1) ◽  
pp. 23-34 ◽  
Author(s):  
Roland Bartholomeusz

The polar bodies are derived from meiotic divisions during oogenesis and are contained together with the oocyte within the zona pellucida. Fertilisation triggers the second meiotic division, at which time the second polar body (PB2) is formed (Hogan et al., 1986; Schatten et al., 1988; Johnson & Everitt, 1995) There is no clear evidence on the fate of the polar bodies in any mammal including the mouse, which is the commonly used research model. However, the polar bodies are generally considered as waste material, and therefore not essential to embryo development. In recent years the polar bodies have gained prominence as they have been used in humans for pre-implantation genetic diagnostic purposes (PGD), of single gene disorders, such as determining whether an embryo may have inherited the cystic fibrosis allele from its mother (Munne et al., 1995; Strom et al., 1998; Rechitsky et al., 2000). PB2 also has a potential use in cloning, for the harvesting of stem cells. Wakayama et al. (1997) have shown that PB2 has the same genetic potential as the female pronuclei and can be used for the production of normal offspring in mice. The successful use of PB2 for these purposes is dependent on its age, for its longevity, rate and nature of degeneration has yet to be determined. While there is little doubt that the first polar body (PB1) experiences a necrotic fate, the same cannot be said for PB2, which may experience an apoptotic fate. Furthermore if PB2 experiences an apoptotic fate rather than a necrotic one, it would not only be the earliest evidence of apoptosis in a mammal but also provide an excellent research model for the study of apoptosis.


1913 ◽  
Vol s2-59 (233) ◽  
pp. 133-174
Author(s):  
J. P. HILL ◽  
CHAS. H. O'DONOGHUE

Dasyurus is monœstrous and has one breeding season a year, which begins at the end of May or early in June and lasts into the first fortnight in August (i.e. it extends over the winter months). The male does not appear to experience a marked rutting season. Copulation is similar to that of Didelphys (Selenka), and the sperms can remain alive in the Fallopian tubes for at least two weeks. Anœtrus.--The anœstral period lasts more than half the year. Pro-œstrus.--Pro-œstrus appears to extend over a varying period of from four to twelve days. During this time, the lips of the cloaca become swollen, the pouch enlarges somewhat and becomes slightly tumid and moist, and the Graafian follicles increase in size and become vesicular. The uterine mucosa increases in thickness and becomes more vascular, its glands lengthen and become convoluted and the uterine epithelium also tends to thicken. Œstrus.--Œstrus lasts usually for one or two days and is the period during which copulation occurs. The changes already initiated during pro-oestrus in the various parts of the reproductive system are continued without interruption. Post-œstrus.--Post-œstrus, which term we employ to designate the period following œstrus and terminated by ovulation, occupies as a rule about five or six days. The tumidity of the cloacal lips disappears, but the changes in the pouch and uterus still continue, not, however, very actively. In the ovary (1) the ova give off the first polar body and the spindle for the second meiotic division is formed. (2) The follicles attain maturity and ultimately rupture, setting free the ova. Ovulation.--Ovulation marks the end of this period and occurs generally about five or six days after cestrus. It is spontaneous and independent of copulation and is remarkable because of the large number of ova liberated. Ovulation is succeeded (a) by pregnancy or (b) by pseudo-pregnancy. Pregnancy.--Fertilisation is effected in the upper part of the Fallopian tube and the second polar body is there given off. As a rule more young are born than can possibly survive owing to the limited accommodation in the pouch. The gestation period is not less than eight and not more than fourteen days, but the interval between copulation and birth is usually considerably longer. Corpora lutea are formed and persist throughout the greater part of the time that the animal is lactating. Nursing Period.--This period may be divided into two phases. (1) Period of Fixation.--A period, lasting for seven or eight weeks, during which the young are firmly attached to the teats. (2) Free Period.--A period of eight or nine weeks when the young are free in the pouch but dependent on the mother for food. After this time the various organs gradually return to a state of rest. Pseudo-pregnancy.--We have applied this term to the period following ovulation iu cases where the ova have failed to develop, because of the occurrence in it of a series of changes in the reproductive organs essentially identical with those met with in pregnant females. Corpora lutea are formed in the ovaries which are identical with those in the pregnant female. The pouch enlarges, and its sweat and sebaceous glands reach a state of hypertrophy and functional activity comparable to that in the pregnant female. The mammary glands also enlarge and reach a state of development equal to that in a female thirty-six hours after parturition. The uteri enlarge and become vascular, often to a marked degree. The uterine mucosa undergoes a series of changes, progressive, regressive and regenerative. Metœstrus.--This is an indefinite period during which the whole of the reproductive organs return to a state of rest.


Reproduction ◽  
2005 ◽  
Vol 129 (1) ◽  
pp. 27-38 ◽  
Author(s):  
Elena Ibáñez ◽  
David F Albertini ◽  
Eric W Overström

With the aim of investigating the effects of oocyte genotype and activating stimulus on the timing of nuclear events after activation, oocytes collected from hybrid B6D2F1, inbred C57BL/6 and outbred CF-1 and immunodeficient nude (NU/+) females were activated using ethanol or strontium and fixed at various time-points. Meiotic status, spindle rotation and second polar body (PB2) extrusion were monitored by fluorescence microscopy using DNA-, microtubule- and microfilament-selective probes. Although activation efficiency was similar in all groups of oocytes, a significant percentage of CF-1 and NU/+ oocytes treated with ethanol and of C57BL/6 oocytes treated either with ethanol or strontium failed to complete activation and became arrested at a new metaphase stage (MIII) after PB2 extrusion. C57BL/6 oocytes also showed slower release from MII arrest but faster progression to telophase (TII) after ethanol exposure, and they exhibited the most rapid exit from TII under both activation treatments. Strontium caused delayed meiotic resumption, spindle rotation and PB2 extrusion, but rapid TII exit, in B6D2F1, CF-1 and NU/+ oocytes when compared with ethanol. Compared with all other strains, NU/+ oocytes were significantly slower in completing spindle rotation and PB2 extrusion, irrespective of the activating stimulus, and a significant decrease in activation rates and pace of meiotic progression was observed after strontium exposure. Thus, our findings demonstrated that the kinetics of meiosis resumption and completion, spindle rotation and PB2 extrusion following parthenogenetic activation depends on both genotype-specific factors and on the activation treatment applied.


2004 ◽  
Vol 16 (6) ◽  
pp. 599 ◽  
Author(s):  
Marek Maleszewski ◽  
Lynne Selwood

Maturation of marsupial oocytes in vitro, an important step in the analysis of early developmental events, has a low success rate and results from the artificial activation of oocytes, which may not include nuclear maturation. In Sminthopsis macroura, 24-h culture of advanced antral follicles in medium containing 10 μg mL−1 porcine pituitary luteinising hormone (LH) yielded 60% of mature polarised oocytes with the first polar body; follicles cultured in medium without LH yielded only immature oocytes. Parthenogenetic activation of follicular, oviducal or uterine oocytes occurred when a two-step protocol was used. Sixty-one oocytes, exposed to 10 μm calcium ionophore A23187 for 10 min followed by 10 μg mL−1 cycloheximide (protein synthesis inhibitor) for 5 h and then cultured for 20–24 h, were scored for signs of activation, namely extrusion of the second polar body and formation of the pronucleus. In each of 43 oocytes (70%), the extruded second polar body was present. Sixteen oocytes were analysed on slides after fixation and staining and, in 13 oocytes (81%) in this group, the female pronucleus was visible. No activation occurred following incubation of oocytes in medium containing Sr2+ for 5 h (n = 14), 8% ethyl alcohol solution for 8 or 12 min (n = 13) or 10 μm calcium ionophore A23187 (n = 13) for 10–20 min followed by culture for 20–24 h.


Zygote ◽  
1995 ◽  
Vol 3 (2) ◽  
pp. 157-161 ◽  
Author(s):  
Jacob Levron ◽  
Jacques Cohen ◽  
Steen Willadsen

SummaryFresh and aged unfertilised human oocytes were activated by electroporation and by exposure to isotonic solution of mannitol supplemented with low concentrations of calcium magnesium and chloride. Over 95% of the fresh oocytes were activated, all showing formation of one pronucleus and extrusion of the second polar body. Oocytes activated 1 and 2 days post-collection showed activation rates of 66.6% and 64.1%, respectively; however, the proportion of one-pronucleate oocytes in these groups was significantly lower (61.6% and 23.5%, respectively). There was no difference in the activation efficiency between the two activation modes. Twelve activated oocytes from the freshly collected group cleaved when left in culture. It is concluded that, in the human, a brief exposure to isotonic solution of mannitol with low concentrations of calcium, magnesium and chloride is a very effective activation stimulus.


1946 ◽  
Vol 7 (1) ◽  
pp. 91-121 ◽  
Author(s):  
Chr P. Raven

AbstractThe structure and physico-chemical composition of the egg of Limnaea stagnalis and the changes occurring during the uncleaved stage of the egg were studied by a variety of methods. A. Composition of egg: I. The cytoplasm of the egg consists of 2 parts: ectoplasm and endoplasm,which differ in their staining reactions. Immediately after oviposition, the ectoplasm occupies a sector at the vegetative pole of the egg, the endoplasm the rest of the egg. 2. Three sorts of granules can be distinguished: α-granules (probably mitochondria), β-granules and γ-granules. Moreover, fat droplets and Golgi bodies are present in the egg. 3. The α-granules, which are small, are accumulated especially in the endoplasm. Probably, glycogen, phenolases and peroxydases are bound in some way or other to these granules. 4. The β-granules, of medium size, form a major part of the ectoplasm. They consist of albumen and contain pentosenucleic acids. 5. The γ-granules are coarse, probably albuminous in nature; they lie mostly in the endoplasm. 6. Both fat droplets and Golgi bodies are distributed rather evenly in the cytoplasm, leaving free only the spindle and asters. "Praesubstances" and "Golgi systems" can be distinguished. 7. The hyaloplasm contains pentosenucleic acids in small quantity and, probably, bound sulfhydril components, especially in its central part. 8. The freshly laid egg contains the first maturation spindle in metaphase. The spindle area is free of fat droplets and Golgi bodies, but is characterized by the presence of free glutathion in reduced form. The chromosomes contain thymonucleic acid. B. Changes during uncleaved stage: 9. The course of the maturation divisions and the formation of the polar bodies are described. The egg shows amoeboid movements shortly after the extrusion of either of the polar bodies. This is accompanied with a distinct drop of the tension at the surface. 10. The α- and γ-granules of the endoplasm are attracted by the maturation spindle and asters, forming a halo surrounding the amphiaster. 11. The ectoplasm spreads beneath the egg cortex to the animal side. At first, a gap remains at the animal pole; after the completion of the maturation divisions, the ectoplasm surrounds the whole egg. 12. A fine chorion is formed, which lies inside the first, but outside the second polar body. 13. The spermastcr makes its appearance shortly before the extrusion of the first polar body; during the maturation divisions, it grows slowly. This is accompanied with a gradual rise of the viscosity of the protoplasm. No division of the spermaster with formation of an amphiaster takes place. 14. The sperm-head remains in a subcortical position till shortly after the extrusion of the second polar body; then, it migrates to the spermaster and develops into the male pronucleus. 15. The chromosomes left in the egg after the maturation divisions swell into karyomeres which fuse to the female pro-nucleus. 16. The copulation of the pronuclei takes place immediately beneath the egg cortex at the animal pole. 17. About one hour before cleavage, the animal pole plasm is formed by a local accumulation of substances attracted, probably, by the egg cortex at the animal pole. The centripetal flow of protoplasm in the dilating maturation aster may aid in this localisation process. Many α-granules are transported by it to the animal pole plasm. 18. From the time of oviposition till first cleavage the egg swells considerably, probably by the intake of water. This is accompanied with a decrease in density. At the same time, the γ-granules of the egg give rise to the formation of vacuoles, by an attraction of water from the neighbourhood. Eventually, the egg protoplasm has a vacuolated appearance throughout, with the exception of the animal pole plasm. 19. The viscosity, which is high one hour before cleavage, has a minimum 30 minutes later, then it rises again. The tension at the surface reaches a minimum immediately before the beginning of cleavage.


Zygote ◽  
2013 ◽  
Vol 23 (1) ◽  
pp. 145-157 ◽  
Author(s):  
M. Sousa ◽  
J. Teixeira da Silva ◽  
J. Silva ◽  
M. Cunha ◽  
P. Viana ◽  
...  

SummaryHuman oocyte dysmorphisms attain a large proportion of retrieved oocytes from assisted reproductive technology (ART) treatment cycles. Extracytoplasmic defects involve abnormal morphology of the zona pellucida (ZP), perivitelline space and first polar body. The aim of the present study was to describe a novel dysmorphism affecting the ZP, indented ZP. We also evaluated the clinical, embryological and ultrastructural features of these cases. We evaluated all ART treatment cycles during 7 consecutive years and found 13 treatment cycles (six patients) with all oocytes presenting an indented ZP. In addition, these oocytes presented total or partial absence of the perivitelline space, absence of resistance to ZP and oolemma penetration during microinjection, and low ooplasm viscosity during aspiration. This novel described dysmorphism was recurrent and attained all oocytes in three cases that had more than one treatment cycle. When compared with controls, data showed significant low oocyte maturity (42% versus 81.6%) and high cycle cancellation (30.8% versus 8.5%) rates, normal degeneration (3.4% versus 6.3%) and fertilization rates (69% versus 69.5%), and low pregnancy (15.4% versus 33.3%) and live-birth delivery (7.7% versus 27.7%) rates per cycle. Ultrastructure analysis revealed a zona pellucida structure with large empty electrolucent regions, an outer ZP layer with an indented surface with protuberances and a thick inner ZP that obliterated the perivitelline space. There was evidence of exocytosis of ZP material by the oocyte. In conclusion, oocytes with this novel described dysmorphism (indented ZP) are associated with low maturity, pregnancy and live-birth delivery rates.


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