GnRH agonist versus GnRH antagonist in assisted reproduction cycles: oocyte morphology

Background: The selection of developmentally competent human gametes may increase the efficiency of assisted reproduction. Spermatozoa and oocytes are usually assessed according to morphological criteria. Oocyte morphology can be affected by the age, genetic characteristics, and factors related to controlled ovarian stimulation. However, there is a lack of evidence in the literature concerning the effect of gonadotropin-releasing hormone (GnRH) analogues, either agonists or antagonists, on oocyte morphology. The aim of this randomized study was to investigate whether the prevalence of oocyte dysmorphism is influenced by the type of pituitary suppression used in ovarian stimulation.Methods: A total of 64 patients in the first intracytoplasmic sperm injection (ICSI) cycle were prospectively randomized to receive treatment with either a GnRH agonist with a long-term protocol (n: 32) or a GnRH antagonist with a multi-dose protocol (n: 32). Before being subjected to ICSI, the oocytes at metaphase II from both groups were morphologically analyzed under an inverted light microscope at 400x magnification. The oocytes were classified as follows: normal or with cytoplasmic dysmorphism, extracytoplasmic dysmorphism, or both. The number of dysmorphic oocytes per total number of oocytes was analyzed.Results: Out of a total of 681 oocytes, 189 (27.8 %) were morphologically normal, 220 (32.3 %) showed cytoplasmic dysmorphism, 124 (18.2%) showed extracytoplasmic alterations, and 148 (21.7%) exhibited both types of dysmorphism. No significant difference in oocyte dysmorphism was observed between the agonist- and antagonist- treated groups (P > 0.05). Analysis for each dysmorphism revealed that the most common conditions were alterations in polar body shape (31.3%) and the presence of diffuse cytoplasmic granulations (22.8%), refractile bodies (18.5%) and central cytoplasmic granulations (13.6%). There was no significant difference among individual oocyte dysmorphisms in the agonist- and antagonist-treated groups (P > 0.05).Conclusions: Our randomized data indicate that in terms of the quality of oocyte morphology, there is no difference between the antagonist multi-dose protocol and the long-term agonist protocol. If a GnRH analogue used for pituitary suppression in IVF cycles influences the prevalence of oocyte dysmorphisms, there does not appear to be a difference between the use of an agonist as opposed to an antagonist.

Relationship between first polar body morphology before intracytoplasmic sperm injection and fertilization rate, cleavage rate, and embryo quality

Objectives: To evaluate the influence of the morphology of the first polar body (PB) on intracytoplasmic sperm injection (ICSI) outcomes. Methods: The morphology of the first PB was assessed in 3177 metaphase 11 oocytes and classified as: intact and normal size, fragmented, or enlarged size. The rates of fertilization, cleavage, and embryo quality were evaluated on day 2. Results: The rates of fertilization, cleavage, and formation of good quality embryos resulting from the insemination of oocytes with an enlarged first PB (20.7%, 18.7%, and 5.0%, respectively) were significantly lower than those for oocytes with an intact first PB of normal size (70.8%, 62.5%, and 19%, respectively) or a fragmented first PB (69.7%, 60.5%, and 17.1%, respectively). Rates did not differ significantly between oocytes with an intact first PB of normal size and oocytes with a fragmented first PB (P>0.05). Conclusions: The presence of an enlarged PB is related to poorer rates of fertilization, cleavage, and top quality embryos. However, identification of first PB fragmentation does not seem to interfere with ICSI outcomes. (C) 2008 International Federation ofGynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.

Collection Rates and Morphology of Equine Oocytes Obtained from Immature Follicles after Treatment with Equine Pituitary Extract (EPE) and Human Chorionic Gonadotropin

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Correlação entre os dismorfismos oocitários e os resultados da fertilização in vitro

In assisted reproduction, the selection of gametes to achieve better clinical outcomes is a crucial task of embryologists. The quality of the oocyte is a key factor in female fertility, reflecting the intrinsic potential of gamete development, and has a vital role not only in conception but also in subsequent embryonic development. Oocyte dysmorphisms are classified into two types: cytoplasmic, including the presence of granules and/or cytoplasmic inclusions (vacuoles, refractive bodies, and aggregates of the endoplasmic reticulum), and extracytoplasmic (changes in the shape of the oocyte, the zona pellucida, the space perivitelline changes and the polar body). Variations in oocyte morphology may occur due to factors such as the age of women, genetic problems and changes in the hormonal environment to which the oocyte is exposed in ovarian hyperstimulation. The classification of oocyte morphology and its correlation with embryo development and pregnancy rates are controversial in the literature. Several studies show no association between oocyte dysmorphisms and the results of in vitro fertilization, while others report an association between oocyte morphology and embryo development. These differences in the results can be explained by the use of different morphological criteria due to a lack of standardization of oocyte evaluation. © Todos os direitos reservados a SBRA - Sociedade Brasileira de Reprodução Assistida.

Agonista versus antagonista do GnRH em ciclos de reprodução assistida: morfologia oocitária

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Maturação in vitro de oócitos caninos: influência da suplementação hormonal seriada sobre a ultraestrutura oocitária

Pós-graduação em Medicina Veterinária - FCAV

Aspectos reprodutivos, morfologia dos gametas e desenvolvimento embrionário de Astyanax altiparanae Garutti & Britski (Teleostei, Characidae)

Pós-graduação em Aquicultura - FCAV

Histochemical and morphological features of biopsied and stripped oocytes from the Brazilian endangered teleost pirapitinga, Brycon nattereri (Characiformes)

The aim of the present study was to characterize biopsied and stripped oocytes from the Brazilian endangered teleost pirapitinga (Brycon nattereri) using histochemical and morphological analyses. Biopsied oocytes had a mean diameter of 2.225 mm (modal diameter: 2.312 mm), complete vitellogenesis and a central or slightly eccentric nucleus. Neutral polysaccharides were detected in the follicular cells, zona radiata and yolk globules, while acidic polysaccharides were detected in the follicular cells and cortical alveoli. Ten out of the 19 females treated with two doses of carp pituitary extract (cPE) released oocytes, which were also analysed. Stripping occurred 292 +/- 39 degree-hours after the second dose of cPE and led to a mean spawning weight of 36.2 g, 10% spawning index, 241 oocytes/g of ova, 8222 oocytes/female and 23 oocytes/g of body weight. Stripped oocytes had a mean diameter of 2.33 mm and a mode at 2.375 mm, were weakly adhesive and coloration ranged from wine to brown. Under scanning electron microscopy, stripped oocytes exhibited a single funnel-shaped micropyle located at the animal pole and a zona radiata that measured 7.7 m in thickness with eight pore canals/m(2). Oocyte morphology in Brycon nattereri is similar to that found in other species of the genus, except for the larger size and weaker adhesiveness. These findings provide essential information for a better understanding of the reproductive biology of B. nattereri and the establishment of conservation measures for this threatened species.

Efeito da adição de vesículas extracelulares intrafoliculares de vacas holandesas submetidas ao estresse térmico em meio de maturação oocitária in vitro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of lineage of oocyte donor on the yield and morphology of oocytes recovered by ultrasound-guided follicular aspiration in nellore cows

The objective of this study was to investigate the influence of lineage of oocytes donors on the number and quality of oocytes obtained through ultrasound-guided follicular aspiration in Nellore breed cows derived from two lineages of bulls (Karvadi; K and Taj-Mahal; T). Both maternal (Km and Tm) and paternal (Kp and Tp) lineages, as well as their combinations were investigated. Oocyte aspirations were repeatedly performed with an aspiration interval of 15 days in 56 donor females. Recovered cumulus oocyte-complexes (COCs) were counted, morphologically examined and classified into seven categories (grades from I to VII) according to the number of layers of the cumulus oocyte and cytoplasm appearance. The mean number of oocytes retrieved from donors of lineage Tp-Tm was significantly higher (28.23±1.92, P<0.05) than those obtained from lineages Kp-Tm, Kp-Km, and Tp-Km (21.34±1.32, 21.28±1.73, and 16.72±1.31, respectively). There was no significant difference in the mean number of recovered oocytes between donors of lineages Kp-Km and Kp-Tm, whereas animals of lineage Tp-Km yielded the lowest number of oocytes. Higher mean number of grade III oocytes was recovered from donors of lineage Kp than lineage Tp (10.11±0.66 versus 8.79±0.58, respectively), with more grade III oocytes being obtained in both lineages as compared to the others. Paternal lineage did not influence the quality of recovered oocytes in any other category, but both Kp and Tp yielded a great mean number of oocytes graded as I, II, and III (3.14±0.21; 4.93±0.33, and 10.11±0.66 versus 3.19±0.21, 5.59±0.44, and 8.79±0.58, respectively) than those classified as IV, V, VI, and VII. However, when considering the data from the maternal lineage significantly more oocytes (P<0.05) of grade I, II and III were obtained from Taj-Mahal (11.67 ± 0.67, 5.9±0.42 and 3.64±0.25, respectively) than for lineage Karvadi, with similar results for oocytes of grades IV, V, VI, and VII. Similarly to the paternal lineage, the number of oocytes of grade III was superior (P<0.05) when compared to other categories for both lineages. In conclusion, we demonstrate here a direct influence of lineage of oocyte donor on the production and quality of oocytes obtained through ultrasound-guided follicular aspiration in Nellore cows.

Influence of vitrification on mouse metaphase II oocyte spindle dynamics and chromatin alignment

Objective: To evaluate influences of vitrification and warming of metaphase II (MII) mouse oocytes on survival, spindle dynamics. spindle morphology, and chromatin alignment on metaphase plates. Design: Experimental animal Study. Setting: University animal laboratory. Animal(s): Eight-week-old B6D2F1 mice. Intervention(s): Denuded MII oocytes were used fresh (control), exposed to vitrification/warming solutions (Sol Expos), or vitrified and warmed (Vitr). Main Outcome Measure(s): Oocyte recovery and survival after warming and the influence of solution exposure and cryopreservation on spindle dynamics and chromatin alignment. Result(s): Cryopreservation of two or 10 oocytes per straw resulted in recovery (100% +/- 0% and 95% +/- 4%, respectively; mean SE) and survival (95% 2% and 98% 2%, respectively). Immediately after warming (Vitr), significantly fewer oocytes assessed with immunocytochemistry contained spindles, compared with control and Sol Expos. When oocytes were placed into a 3 degrees 7C environment for 2 hours after exposure or warming, the ability to recognize spindles by immunocytochemistry was not significantly different between groups. Using live-cell time-lapse imaging with LC-Polscope, similar time-dependent spindle formation dynamics were observed. At 2 hours after collection or treatment, spindle morphology and length were not significantly different between the groups, nor was the incidence of aberrant alignment of chromatin on metaphase plates. Conclusion(s): Immediately after warming of vitrified MII oocytes, beta-tubulin is depolymerized and chromatin remains condensed on the metaphase plate. Within a 2-hour period, beta-tubulin repolymerizes, forming morphologically normal metaphase spindles with properly aligned chromatin.

Prospective randomized comparison of human oocyte cryopreservation with slow-rate freezing or vitrification

Objective: To compare cryopreservation of mature human oocytes with slow-rate freezing and vitrification and determine which is most efficient at establishing a pregnancy. Design: Prospective randomized. Setting: Academically affiliated, private fertility center. Patient(s): Consenting patients with concerns about embryo cryopreservation and more than nine mature oocytes at retrieval were randomized to slow-rate freezing or vitrification of supernumerary (more than nine) oocytes. Intervention(s): Oocytes were frozen or vitrified, and upon request oocytes were thawed or warmed, respectively. Main Outcome Measure(s): Oocyte survival, fertilization, embryo development, and clinical pregnancy. Result(s): Patient use has resulted in 30 thaws and 48 warmings. Women`s age at time of cryopreservation was similar. Oocyte survival was significantly higher following vitrification/warming (81%) compared with freezing/thawing (67%). Fertilization was more successful in oocytes vitrified/warmed compared with frozen/thawed. Fertilized oocytes from vitrification/warming had significantly better cleavage rates (84%) compared with freezing/thawing (71%) and resulted in embryos with significantly better morphology. Although similar numbers of embryos were transferred, embryos resulting from vitrified oocytes had significantly enhanced clinical (38%) pregnancy rates compared with embryos resulting from frozen oocyte (13%). Miscarriage and/or spontaneous abortion rates were similar. Conclusion(s): Our results suggest that vitrification/warming is currently the most efficient means of oocyte cryopreservation in relation to subsequent success in establishing pregnancy. (Fertil Steril (R) 2010; 94: 2088-95. (C) 2010 by American Society for Reproductive Medicine.)

Morphology and Histology of the Female Reproductive System of Abedus ovatus Stal (Belostomatidae: Hemiptera: Insecta)

In Abedus ovatus Stal, the female reproductive system consists of a pair of ovaries leading to oviduct and opens out by genital aperture. Each ovary is composed of five telotrophic ovarioles, with four different regions, terminal filament, germarium, vitellarium and pedicel. Germarium and vitellarium are the germ tubes in which the development of oocyte occurs. In the germarium, the differentiation of trophocytes into young oocytes was observed in three zones, ZI, ZII and ZIII. Further development of oocytes in the vitellarium could be divided into seven stages, STI STVII. The developed eggs are lodged in the pedicel

A retrospective model of oocyte competence: global mRNA and housekeeping transcripts are not associated with in vitro developmental outcome

Oocyte developmental competence depends on maternal stores that support development throughout a transcriptionally silent period during early embryogenesis. Previous attempts to investigate transcripts associated with oocyte competence have relied on prospective models, which are mostly based on morphological. criteria. Using a retrospective model, we quantitatively compared mRNA among oocytes with different embryo development competence. A cytoplasm biopsy was removed from in vitro matured oocytes to perform comparative analysis of amounts of global polyadenylated (polyA) mRNA and housekeeping gene transcripts. After parthenogenetic activation of biopsied oocytes, presumptive zygotes were cultured individually in vitro and oocytes were classified according to embryo development: (i) blocked before the 8-cell stage; (ii) blocked between the 8-cell and morulae stages; or (iii) developed to the blastocyst stage. Sham-manipulated controls confirmed that biopsies did not alter development outcome. Total polyA mRNA amounts correlate with oocyte diameter but not with the ability to develop to the 8-cell and blastocyst stages. The last was also confirmed by relative quantification of GAPDH, H2A and Hprt1 transcripts. In conclusion, we describe a novel retrospective model to identify putative markers of development competence in single oocytes and demonstrate that global mRNA amounts at the metaphase II stage do not correlate with embryo development in vitro.