scholarly journals Rapid protocol for pre-conception genetic diagnosis of single gene mutations by first polar body analysis: a possible solution for the Italian patients

2007 ◽  
Vol 28 (1) ◽  
pp. 62-64 ◽  
Author(s):  
F. Fiorentino ◽  
A. Biricik ◽  
A. Nuccitelli ◽  
R. De Palma ◽  
S. Kahraman ◽  
...  
Keyword(s):  
Single Gene ◽  
Polar Body ◽  
Genetic Diagnosis ◽  
Gene Mutations ◽  
First Polar Body ◽  
Polar Body Analysis

Advantages of multiple markers and polar body analysis in preimplantation genetic diagnosis for Alagille disease

2007 ◽  
Vol 27 (4) ◽  
pp. 317-321 ◽  
Author(s):  
P. Renbaum ◽  
B. Brooks ◽  
Y. Kaplan ◽  
T. Eldar-Geva ◽  
E J. Margalioth ◽  
...  
Keyword(s):  
Preimplantation Genetic Diagnosis ◽  
Polar Body ◽  
Genetic Diagnosis ◽  
Multiple Markers ◽  
Polar Body Analysis

173 Assessment of the first polar body quality and viability in bovine

2019 ◽  
Vol 31 (1) ◽  
pp. 211
Author(s):  
O. Briski ◽  
M. Duque-Rodríguez ◽  
A. Gambini ◽  
N. P. Leopardo ◽  
E. A. Ynsaurralde ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Dna Fragmentation ◽  
Polar Body ◽  
Genetic Material ◽  
Membrane Integrity ◽  
Genetic Diagnosis ◽  
Mature Oocyte ◽  
Plasma Membrane Integrity ◽  
Apoptotic Process ◽  
First Polar Body

In female gametes, after the first asymmetric meiotic division, a mature oocyte in metaphase II and a first polar body (PB1) are generated. The PB1 contains one of each pair of homologous chromosomes present in the mature oocyte and its DNA can be used for preconception genetic diagnosis. The PB1 degenerates shortly after extrusion, possibly due to an apoptotic process; however, it has not yet been elucidated in bovine. Therefore, the objective of this study was to evaluate PB1 morphology changes, plasma membrane integrity, and the presence of DNA fragmentation during in vitro maturation (IVM). To this aim, cumulus-oocyte complexes collected from slaughterhouse ovaries were cultured in maturation medium in different groups according to IVM time: 16, 18, 20, 22, 24, and 26h. The PB1 were classified into 5 categories according to their morphology: grade (G)1, round PB1 with intact smooth membrane; G2, round or ovoid PB1 with intact membrane; G3, broken PB1 with a small PB1 fragment; G4, broken PB1 with a big PB1 fragment; and G5, completely damaged PB1. Grades 1 and 2 were considered good quality. Plasma membrane integrity was assessed by propidium iodide (PI) DNA staining; PI is a fluorescent intercalating agent that cannot cross the membrane of live cells. The presence of DNA fragmentation was detected at 16, 22, and 26h by TUNEL assay. Data were analysed by two-way ANOVA and Bonferroni post hoc test using GraphPad Prism 5.0 (GraphPad Inc., San Diego, CA, USA) and differences were considered significant at P < 0.05. Our results (mean%±s.e.m.) showed that significantly more oocytes assessed at 18, 20, 22, and 24h after onset of IVM presented high-quality (G1) PB1 (18 h: 61.5±13.4%, 20 h: 73.5±9.6%, 22 h: 61.0±9.5%, 24 h: 60±5.1%), compared with those assessed at 16 and 26h (43.0±4.7%, 22.3±3.4%, respectively). The percentage of G2 PB1 did not change throughout the period studied (16 h: 34.0±13.5%, 18 h: 29.9±14.1%, 20 h: 22.0±7.1%, 22 h: 26.5±4.2%, 24 h: 23.3±4.9%, 26 h: 21.33±9.9%), but was significantly lower than that of G1 PB1 at 20, 22, and 24h. The proportion of damaged (G5) PB1 started to increase at 24h (14.3±8.6%), being highest at 26h (30.0±10.5%), in parallel with positive PI staining (P<0.05). Moreover, there was a significant increase of PB1 with DNA fragmentation at 26h (82.0±18.0%) compared with 16h (13.9±9.0%) and 22h (2.5±2.5%). Altogether, these findings demonstrate that PB1 remains stable and of good quality between 18 and 24 h; however, after this time, plasma membrane integrity is compromised and the DNA is fragmented, suggesting the occurrence of an apoptotic process. Our results could be helpful to determine the optimal time for using PB1 as a potential donor of genetic material.

Review of the longevity of the second polar body in the mouse

Zygote ◽  
2003 ◽  
Vol 11 (1) ◽  
pp. 23-34 ◽  
Author(s):  
Roland Bartholomeusz
Keyword(s):  
Single Gene ◽  
Polar Body ◽  
Research Model ◽  
Polar Bodies ◽  
Single Gene Disorders ◽  
First Polar Body ◽  
Excellent Research ◽  
Second Polar Body ◽  
Normal Offspring ◽  
Potential Use

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.

Analysis of the first polar body: preconception genetic diagnosis

1990 ◽  
Vol 5 (7) ◽  
pp. 826-829 ◽  
Author(s):  
Yury Verlinsky ◽  
Norman Ginsberg ◽  
Aaron Lifchez ◽  
Jorge Valle ◽  
Jacob Moise ◽  
...  
Keyword(s):  
Polar Body ◽  
Genetic Diagnosis ◽  
First Polar Body

Prepregnancy Testing for Single-Gene Disorders by Polar Body Analysis

1999 ◽  
Vol 3 (2) ◽  
pp. 185-190 ◽  
Author(s):  
Y. VERLINSKY ◽  
S. RECHITSKY ◽  
O. VERLINSKY ◽  
V. IVACHNENKO ◽  
A. LIFCHEZ ◽  
...  
Keyword(s):  
Single Gene ◽  
Polar Body ◽  
Single Gene Disorders ◽  
Polar Body Analysis

scholarly journals Multiple aneuploidies in the oocytes of balanced translocation carriers: a preimplantation genetic diagnosis study using first polar body

Reproduction ◽  
2003 ◽  
pp. 701-711 ◽  
Author(s):  
A Pujol ◽  
M Durban ◽  
J Benet ◽  
I Boiso ◽  
JM Calafell ◽  
...  
Keyword(s):  
Preimplantation Genetic Diagnosis ◽  
Polar Body ◽  
Genetic Diagnosis ◽  
Single Embryo Transfer ◽  
Balanced Translocation ◽  
Aneuploidy Screening ◽  
Robertsonian Translocations ◽  
Polar Bodies ◽  
First Polar Body ◽  
Made In

Preimplantation genetic diagnosis (PGD) of first polar bodies (1PBs) has been used in carriers of balanced chromosomal reorganizations and also for aneuploidy screening. Although an acceptable number of normal or balanced embryos is usually obtained using PGD in translocation carriers, the pregnancy rate is disappointingly low. To determine whether aneuploidy of chromosomes not involved in the chromosome rearrangements could be the cause of the low pregnancy rates achieved, the present authors analysed the segregation products of three translocation carriers, t(8;13)(q24.1;q22) and two Robertsonian (Rob)(13;14), using 1PBs, and afterwards another eight chromosomes in the same 1PBs, for a total of 10 chromosomes in each 1PB, that is chromosomes 1, 8, 13, 14, 15, 16, 17, 18, 21, 22 and X. In the reciprocal translocation, chromosomes with different chromatids due to meiotic recombination were found. Only one out of nine 1PBs was normal for the reorganization products but no aneuploidies were found after PGD in this case. In the two balanced Rob(13;14), six out of 12 and four out of 11 1PBs were normal or balanced for the reorganization but only one oocyte was euploid for all the chromosomes analysed in each case; a single embryo transfer was made in both but no pregnancy was achieved. The incidence of aneuploidy for the chromosomes not involved in the Robertsonian translocations was extremely high (91.7% and 81.8%). Extra chromosomes were present in most of the aneuploid oocytes (81.8% and 90%). The reason for this increase could be the tendency to non-disjunction related to advanced maternal age combined with an interchromosomal effect resulting in the presence of synaptic errors in other chromosome pairs.

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