Service dHistologie Embryologie Cytogenetique
Service dHistologie Embryologie Cytogenetique
Barraud-Lange V.,University of Paris Descartes |
Pont J.-C.,University of Paris Descartes |
Ziyyat A.,University of Paris Descartes |
Pocate K.,University of Paris Descartes |
And 5 more authors.
Fertility and Sterility | Year: 2011
Objective: To assess the effect of leukocytospermia on assisted reproductive technology outcomes. Design: Retrospective analysis. Setting: University laboratory. Patient(s): Couples attending the infertiliy clinic and involved in ART program for IVF or ICSI. Intervention(s): During a 7-year follow-up in an assisted reproductive technology program, leukocytospermia was routinely determined using the peroxidase technique. Donor sperm were excluded from the study. Main Outcome Measure(s): Egg retrievals (N = 3,508) were distributed in 3 groups according to the leukocyte levels in semen from which fertilizing sperm were extracted: group 1, absence of leukocytes (n = 3,026); group 2, moderate leukocytospermia (<10 6/mL) (n = 344); or group 3, high leukocytospermia (≥10 6/mL) (n = 138). They resulted in 1,463 IVF and 2,045 intracytoplasmic sperm injection procedures that gave 802 clinical pregnancies. Result(s): Surprisingly, the fertilization rate, cleavage rate, clinical pregnancy rate, gestational age, and mean infant weight were significantly improved when seminal leukocytes were present, regardless of the technique used. The only negative side effects associated with a high level of seminal leukocytes (group 3) were an elevated rate of early pregnancy loss (from 26.6% to 40.5%) and a 3-fold increase in the percentage of ectopic pregnancies. Conclusion(s): At moderate levels (<10 6/mL), leukocytospermia appears to be physiologic. It is associated with improved sperm fertilization ability and pregnancy outcome. At higher concentrations, leukocytospermia alters neither sperm fertilization ability nor the probability of clinical pregnancy when compared with nonleukocytic patients with infertility. However, the pregnancy outcome is reduced. Copyright © 2011 American Society for Reproductive Medicine.
PubMed | Howard Hughes Medical Institute, Genetique Medicale, Bordeaux University Hospital Center, University of Miami and 17 more.
Type: | Journal: American journal of human genetics | Year: 2017
Degradation of proteins by the ubiquitin-proteasome system (UPS) is an essential biological process in the development of eukaryotic organisms. Dysregulation of this mechanism leads to numerous human neurodegenerative or neurodevelopmental disorders. Through a multi-center collaboration, we identified six de novo genomic deletions and four de novo point mutations involving PSMD12, encoding the non-ATPase subunit PSMD12 (aka RPN5) of the 19S regulator of 26S proteasome complex, in unrelated individuals with intellectual disability, congenital malformations, ophthalmologic anomalies, feeding difficulties, deafness, and subtle dysmorphic facial features. We observed reduced PSMD12 levels and an accumulation of ubiquitinated proteins without any impairment of proteasome catalytic activity. Our PSMD12 loss-of-function zebrafish CRISPR/Cas9 model exhibited microcephaly, decreased convolution of the renal tubules, and abnormal craniofacial morphology. Our data support the biological importance of PSMD12 as a scaffolding subunit in proteasome function during development and neurogenesis in particular; they enable the definition of a neurodevelopmental disorder due to PSMD12 variants, expanding the phenotypic spectrum of UPS-dependent disorders.
Cederroth C.R.,University of Geneva |
Auger J.,Service dHistologie Embryologie |
Zimmermann C.,University of Geneva |
Eustache F.,Service dHistologie Embryologie Cytogenetique |
Nef S.,University of Geneva
International Journal of Andrology | Year: 2010
There is growing interest in the possible health threat posed by the effects of endocrine disruptors on reproduction. Soy and soy-derived products contain isoflavones that mimic the actions of oestrogens and may exert adverse effects on male fertility. The purpose of this review was to examine the evidence regarding the potential detrimental effects of soy and phyto-oestrogens on male reproductive function and fertility in humans and animals. Overall, there are some indications that phyto-oestrogens, alone or in combination with other endocrine disruptors, may alter reproductive hormones, spermatogenesis, sperm capacitation and fertility. However, these results must be interpreted with care, as a result of the paucity of human studies and as numerous reports did not reveal any adverse effects on male reproductive physiology. Further investigation is needed before a firm conclusion can be drawn. In the meantime, caution would suggest that perinatal phyto-oestrogen exposure, such as that found in infants feeding on soy-based formula, should be avoided. © 2009 European Academy of Andrology.
Kacem O.,University of Paris Descartes |
Sifer C.,Service dHistologie Embryologie Cytogenetique |
Barraud-Lange V.,University of Paris Descartes |
Ducot B.,French Institute of Health and Medical Research |
And 3 more authors.
Reproductive BioMedicine Online | Year: 2010
Microinjection of nuclear vacuole-free spermatozoa selected by motile sperm organellar morphological examination (MSOME) has been claimed to enhance assisted reproduction treatment outcome compared with intracytoplasmic sperm injection. However, the nature of these nuclear vacuoles is unclear, since their localization at the front of the sperm head suggests they might be of acrosomal origin. To study this hypothesis, acrosomal status was evaluated using Pisum sativum agglutinin staining on a smear, together with sperm organellar morphological examination using the same optics as for MSOME on 30 sperm samples from infertile patients, yielding >3200 spermatozoa. Vacuoles were present in 61% of spermatozoa when acrosomal material or intact acrosomes were observed, versus 29% when spermatozoa were acrosome reacted (P < 0.0001). Induction of the acrosomal reaction by ionophore A23587 from 17.4% to 36.1% significantly increased the percentage of vacuole-free spermatozoa from 41.2% to 63.8% (P < 0.001). These data suggest that most nuclear vacuoles are of acrosomal origin. Hence, the best spermatozoa selected by MSOME are mostly acrosome-reacted spermatozoa. As microinjection of spermatozoa with a persistent acrosome drastically hampers embryo development in animal models, this suggests that the improvement in pregnancy rates reported following intracytoplasmic injection of morphologically selected sperm might be due to the procedure allowing injection of acrosome-reacted spermatozoa. © 2009, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.
PubMed | Service de Pediatrie IIA, French National Center for Scientific Research, Institute National dHygiene and Service dHistologie Embryologie Cytogenetique
Type: | Journal: Journal of medical case reports | Year: 2015
Joubert syndrome is a rare congenital disorder characterized by brain malformation, developmental delay with hypotonia, ocular motor apraxia, and breathing abnormalities. Joubert syndrome is a genetically highly heterogeneous ciliopathy disorder with 23 identified causative genes. The diagnosis is based on brain imaging showing the molar tooth sign with cerebellar vermis agenesis. We describe a consanguineous Moroccan family with three affected siblings (18-year-old boy, 13-year-old girl, and 10-year-old boy) showing typical signs of Joubert syndrome, and attempt to identify the underlying genetic defect in this family.We performed genome-wide homozygosity mapping using a high-resolution array followed by targeted Sanger sequencing to identify the causative gene.This approach found three homozygous regions, one including the AHI1 gene. Direct sequencing of the 26 coding exons of AHI1 revealed a homozygous mutation (p.Thr304AsnfsX6) located in exon 7 present in the three Joubert syndrome-affected Moroccan siblings. Of more interest, this truncating mutation was previously reported in patients with compound heterozygous Joubert syndrome originating from Spain (one patient) and from the Netherlands (two patients), suggesting a possible founder effect or mutational hotspot.Combined homozygosity mapping and targeted sequencing allowed the rapid detection of the disease-causing mutation in the AHI1 gene in this family affected with a highly genetically heterogeneous disorder. Carriers of the same truncating mutation (p.Thr304AsnfsX6), originating from Spain and the Netherlands, presented variable clinical characteristics, thereby corroborating the extreme heterogeneity of Joubert syndrome.
Perrin A.,University of Western Brittany |
Perrin A.,French Institute of Health and Medical Research |
Nguyen M.H.,University of Western Brittany |
Nguyen M.H.,French Institute of Health and Medical Research |
And 13 more authors.
Andrology | Year: 2013
Summary: It has been previously shown that men with chromosomal structural abnormality had a higher rate of sperm DNA fragmentation. We studied 11 male carriers of a chromosomal structural abnormality (seven with a balanced reciprocal translocation, three with a Robertsonian translocation, one with a pericentric inversion) to determine whether spermatozoa with unbalanced chromosomes were more likely to have fragmented DNA. A sequential method combining analysis of DNA fragmentation using the TUNEL assay followed by analysis of meiotic segregation by fluorescent in situ hybridization was performed on the same spermatozoa. A statistically significant higher number of spermatozoa with unbalanced chromosomal content were found to have fragmented DNA for each man. The rate of spermatozoa with DNA fragmentation was higher than the rate of those without fragmented DNA in particular modes of segregation. Our findings provide a better understanding of the mechanisms involved in male infertility ascribable to chromosomal structural abnormality. © 2013 American Society of Andrology and European Academy of Andrology.
Poirier K.,University of Paris Descartes |
Poirier K.,French Institute of Health and Medical Research |
Saillour Y.,University of Paris Descartes |
Saillour Y.,French Institute of Health and Medical Research |
And 23 more authors.
Human Molecular Genetics | Year: 2010
Mutations in the TUBB3 gene, encoding β-tubulin isotype III, were recently shown to be associated with various neurological syndromes which all have in common the ocular motility disorder, congenital fibrosis of the extraocular muscle type 3 (CFEOM3). Surprisingly and in contrast to previously described TUBA1A and TUBB2B phenotypes, no evidence of dysfunctional neuronal migration and cortical organization was reported. In our study, we report the discovery of six novel missense mutations in the TUBB3 gene, including one fetal case and one homozygous variation, in nine patients that all share cortical disorganization, axonal abnormalities associated with pontocerebellar hypoplasia, but with no ocular motility defects, CFEOM3. These new findings demonstrate that the spectrum of TUBB3-related phenotype is broader than previously described and includes malformations of cortical development (MCD) associated with neuronal migration and differentiation defects, axonal guidance and tract organization impairment. Complementary functional studies revealed that the mutated βIII-tubulin causing the MCD phenotype results in a reduction of heterodimer formation, yet produce correctly formed microtubules (MTs) in mammalian cells. Further to this, we investigated the properties of the MT network in patients' fibroblasts and revealed that MCD mutations can alter the resistance of MTs to depolymerization. Interestingly, this finding contrasts with the increased MT stability observed in the case of CFEOM3-related mutations. These results led us to hypothesize that either MT dynamics or their interactions with various MT-interacting proteins could be differently affected by TUBB3 variations, thus resulting in distinct alteration of downstream processes and therefore explaining the phenotypic diversity of the TUBB3-related spectrum. © The Author 2010. Published by Oxford University Press. All rights reserved.
Sifer C.,Service dHistologie embryologie cytogenetique |
Sermondade N.,Service dHistologie embryologie cytogenetique |
Dupont C.,Service dHistologie embryologie cytogenetique |
Poncelet C.,Service de Gynecologie obstetrique |
And 4 more authors.
Gynecologie Obstetrique Fertilite | Year: 2012
Objectives: Since the end of 2010, France by "l'Agence de Biomédecine" has validated the embryo vitrification procedure as an improvement of the slow freezing method. We presented here data concerning biological and clinical outcomes from a prospective observational study where early cleavage stage good quality embryos were vitrified and warmed. We compared these results to those of a retrospective series where embryos were thawed after a slow freezing procedure (SF). We report also the first French live birth following embryo vitrification. Patients and methods: In all, 58 cycles of frozen-thawed embryo transfers (FET) following vitrification were prospectively included and compared with 189 FET from SF method. Primary end points were the (i) survival rate (SR) (% of embryos with ≥50% post-thaw intact blastomeres), (ii) intact survival rate (ISR) (% of embryos with 100% post-thaw intact blastomeres) and (iii) survival blastomeres index (SBI) (% of post thaw intact blastomeres per survival embryo). Secondary end point was the clinical pregnancy rate (CPR) defined as the presence of an intra-uterine gestational sac with positive foetal heart beat. We report here the first French live birth following embryo vitrification. Results: In all, 87 and 412 embryos have been thawed following vitrification and SF, respectively. We observed a highly significant increase of SR, ISR et SBI respectively when thawing concerned vitrified embryos rather than those from SF method (98.3 ± 13.1% vs. 77.3 ± 32.0%, P < 10 -4; 88.2 ± 28.3% vs. 47.7 ± 41.4%, P < 10 -4; 97.7 ± 6.1% vs. 87.3 ± 14.4%, P < 10 -4). Furthermore, CPR were of 32.7% (19/58) and of 18.5% (35/189) following FET performed after vitrification or SF and thawing (P = 0.03), respectively. The live birth of two healthy girls occurred following a caesarean section after 38 weeks of amenorrhea the 8th of August 2011. Discussion and conclusion: We experienced in our study that the post-thaw survival of vitrified embryos was significantly better than those of embryos resulting from SF. Then, a better CPR per thawed embryo cycle was observed following vitrification. © 2011 Elsevier Masson SAS. All rights reserved.
Malan V.,Service dHistologie Embryologie Cytogenetique
Cytogenetic and Genome Research | Year: 2016
Cytogenetic microarray analysis is now the first-tier genetic test used in a postnatal clinical setting to explore genomic imbalances in individuals with developmental disability and/or birth defects. However, in a prenatal setting, this technique is not widely implemented, largely because the clinical impact of some copy number variants (CNVs) remains difficult to assess. This limitation is especially true in France where termination of pregnancy for medical reasons may be performed at any stage of gestation. During a period of 15 months, we investigated 382 fetuses presenting with ultrasound anomalies, using a customized microarray designed to avoid the detection of CNVs raising challenges for genetic counseling. After excluding common aneuploidies, 20/374 (5.3%) fetuses had a pathogenic CNV, among which 12/374 (3.2%) could have been detected by karyotyping, whereas 8/374 (2.1%) were cryptic. Within these 374 cases, 300 were ongoing pregnancies at the time of array comparative genomic hybridization (aCGH) testing. For these pregnancies, we detected 18/300 (6%) pathogenic CNVs, among which 6/300 (2%) were cryptic. Using this approach, only 2/300 (0.6%) of the detected CNVs raised difficulties for genetic counseling. This study confirms the added value of this strategy in a prenatal clinical setting to minimize ethical issues for genetic counseling while enhancing the detection of genomic imbalances. © 2016 S. Karger AG, Basel Copyright © 2016, S. Karger AG. All rights reserved.