scholarly journals 276 THE EFFECT OF CUMULUS CELLS DURING MATURATION ON THE RISE IN THE CONCENTRATION OF INTRACELLULAR Ca2+ ([Ca2+]i) OF PORCINE OOCYTES INDUCEDBY INOSITOL 1,4,5-TRISPHOSPHATE

2005 ◽  
Vol 17 (2) ◽  
pp. 288
Author(s):  
T. Amano ◽  
T. Mori ◽  
K. Matsumoto ◽  
T. Watanabe ◽  
A. Iritani
Keyword(s):  
Cumulus Cells ◽  
Immature Oocytes ◽  
R Ratio ◽  
Transient Rise ◽  
Activation Rate ◽  
Sperm Penetration ◽  
Cytoplasmic Maturation ◽  
Student’S T ◽  
Rate Of Activation

Increase of inositol 1,4,5-triphosphate (IP3) in the cytoplasm of mammalian oocytes is said to be responsible for [Ca2+]i oscillation observed in the oocytes immediately after sperm penetration, and the [Ca2+]i oscillation is known to be essential for the development of embryos. On the other hand, cumulus cells have been reported to play an important role in cytoplasmic maturation of oocytes and affecting the embryonic development. To obtain more information about the role of cumulus cells in cytoplasmic maturation, the effects of cumulus cells during maturation on the rise in [Ca2+]i and on the rate of activation of porcine mature oocytes induced by IP3 injection were investigated. The immature porcine oocytes were divided into three groups: COCs (intact cumulus-oocyte complexes), DOs (oocytes denuded of their cumulus cells), Co-culture (DOs attached to separated cumulus cells). These groups of immature oocytes were cultured in NCSU23 46 h for maturation. To examine the function of cumulus cells, two groups of immature oocytes were also prepared: DOs + pyruvate (DOs put into NCSU23 with pyruvate) and COCs-glucose free (COCs put into NCSU23 without glucose). The mature oocytes from each group were loaded with Ca2+ indicator fluorescent dye Fura2-AM, and then were irradiated by 340 nm and 360 nm of ultraviolet immediately after the injection of IP3. The intensities of emission light caused by the irradiation of 340 nm and 360 nm ultraviolet were recorded as E340 and E360. Since coupling of Ca2+ and the dye intensifies E340, but does not change E360, the level of [Ca2+]i was shown as R (ratio = E340/E360) in this study. Activation rate was calculated by counting the number of the oocytes that formed pronuclei by injection of IP3. ANOVA and Student's t-test were used in this study. Transient rise in [Ca2+]i was observed in the mature oocytes from every group. The peak R of the rise in [Ca2+]i of the mature oocytes derived from COCs, Dos, and Co-culture and induced by IP3 were 7.2, 4.0, and 6.9, respectively. The R of DOs was significantly lower than those of the others (P < 0.05). Also, the activation rate of the mature oocytes from DOs was significantly lower than those from COCs and Co-culture (31, 66, and 66%). The mature oocytes from DOs + pyruvate showed the same level of peak R compared with those from COCs (7.4 and 6.3), but COCs-glucose free showed a slight but significantly lower peak R compared with the mature oocytes from COCs (6.0 and 7.4, P < 0.05). In conclusion, cumulus cells appeared to support the rise in [Ca2+]i of porcine oocytes induced by IP3 during maturation and the following activation. Moreover, a function of cumulus cells supposedly produces pyruvate by metabolizing glucose and provides it to oocytes during maturation for promoting the cytoplasmic maturation. A part of this study was supported by a Grant-in-Aid for the 21st Century COE Program of the Japan MEXT, and by a grant from the Wakayama Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence of the JST.

scholarly journals PACAP-mediated sperm–cumulus cell interaction promotes fertilization

Reproduction ◽  
2011 ◽  
Vol 141 (2) ◽  
pp. 163-171 ◽  
Author(s):  
Ichiro Tanii ◽  
Tadashi Aradate ◽  
Kouhei Matsuda ◽  
Akira Komiya ◽  
Hideki Fuse
Keyword(s):  
Sperm Motility ◽  
Acrosome Reaction ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Cell Interaction ◽  
Fertilization Rate ◽  
Type I ◽  
Dependent Manner ◽  
Sperm Penetration

The developing acrosome in spermatids contains pituitary adenylate cyclase-activating polypeptide (PACAP). However, the role of the acrosomal PACAP remains unclear because it has not been detected in mature spermatids and sperm. We reinvestigated whether the sperm acrosome contains PACAP. An antiserum produced against PACAP reacted to the anterior acrosome in epididymal sperm fixed under mild conditions, suggesting that PACAP acts on oocytes and/or cumulus cells at the site of fertilization. Immunolabeling and RT-PCR demonstrated the presence of PACAP type I receptor, a PACAP-specific receptor, in postovulatory cumulus cells. To investigate the role of PACAP in fertilization, we pretreated cumulus–oocyte complexes with the polypeptide. At a low concentration of sperm, the fertilization rate was significantly enhanced by PACAP in a dose-dependent manner. Sperm penetration through the oocyte investment, cumulus layer, and zona pellucida was also enhanced by PACAP. The enhancement was probably due to an enhancement in sperm motility and the zona-induced acrosome reaction, which were stimulated by a cumulus cell-releasing factor. Indeed, PACAP treatment increased the secretion of progesterone from the cumulus–oocyte complexes. These results strongly suggest that in response to PACAP, cumulus cells release a soluble factor that probably stimulates sperm motility and the acrosome reaction, thereby promoting fertilization.

The importance of manganese in the cytoplasmic maturation of cattle oocytes: blastocyst production improvement regardless of cumulus cells presence duringin vitromaturation

Zygote ◽  
2015 ◽  
Vol 24 (1) ◽  
pp. 139-148 ◽  
Author(s):  
Juan Patricio Anchordoquy ◽  
Juan Mateo Anchordoquy ◽  
Matias Angel Sirini ◽  
Juan Alberto Testa ◽  
Pilar Peral-García ◽  
...  
Keyword(s):  
Embryo Development ◽  
Embryo Quality ◽  
Cumulus Cells ◽  
Sod Activity ◽  
Cumulus Expansion ◽  
Production Improvement ◽  
Blastocyst Quality ◽  
Cytoplasmic Maturation

SummaryAdequate dietary intake of manganese (Mn) is required for normal reproductive performance in cattle. This study was carried out to investigate the effect of Mn duringin vitromaturation of bovine cumulus–oocyte complexes (COC) on apoptosis of cumulus cells, cumulus expansion, and superoxide dismutase (SOD) activity in the COC. The role of cumulus cells on Mn transport and subsequent embryo development was also evaluated. Early apoptosis decreased in cumulus cells matured with Mn compared with medium alone. Cumulus expansion did not show differences in COC matured with or without Mn supplementation. SOD activity was higher in COC matured with 6 ng/ml Mn than with 0 ng/ml Mn. Cleavage rates were higher in COC and denuded oocytes co-cultured with cumulus cells, either with or without Mn added toin vitromaturation (IVM) medium. Regardless of the presence of cumulus cells during IVM, the blastocyst rates were higher when 6 ng/ml Mn was supplemented into IVM medium compared with growth in medium alone. Blastocyst quality was enhanced when COC were matured in medium with Mn supplementation. The results of the present study indicated that Mn supplementation to IVM medium enhanced the ‘health’ of COC, and improved subsequent embryo development and embryo quality.

Lysophosphatidic acid stimulates nuclear and cytoplasmic maturation of golden hamster immature oocytes in vitro via cumulus cells

Life Sciences ◽  
2002 ◽  
Vol 70 (7) ◽  
pp. 759-767 ◽  
Author(s):  
Kenji Hinokio ◽  
Shuji Yamano ◽  
Koji Nakagawa ◽  
Minoru Irahara ◽  
Masaharu Kamada ◽  
...  
Keyword(s):  
Lysophosphatidic Acid ◽  
Golden Hamster ◽  
Cumulus Cells ◽  
Immature Oocytes ◽  
Cytoplasmic Maturation

The antioxidant dimethylthiourea improves IVF efficiency and decreases cumulus cell apoptosis in pigs

2019 ◽  
Vol 31 (10) ◽  
pp. 1607 ◽  
Author(s):  
M. S. Lorenzo ◽  
A. Maruri ◽  
P. R. Cruzans ◽  
G. M. Teplitz ◽  
M. F. Tello ◽  
...  
Keyword(s):  
Cumulus Cell ◽  
Cumulus Cells ◽  
Oocyte Quality ◽  
Control Group ◽  
Nuclear Maturation ◽  
Intracellular Ros ◽  
Sperm Penetration ◽  
The Media ◽  
Late Apoptosis ◽  
Cytoplasmic Maturation

Abattoir ovaries, which are the main source of oocytes for reproductive biotechnologies, arrive at the laboratory under ischaemic conditions. Reoxygenation generates reactive oxygen species (ROS) in ischaemic tissues, which could affect oocyte quality. The aim of this study was to evaluate the effect of supplementation of media with dimethylthiourea (DMTU) during the collection and washing of cumulus–oocyte complexes (COC) on ROS levels, COC apoptosis and oocyte nuclear and cytoplasmic maturation. Thus, the collection (TCM-199) and washing (TCM-199 with 10% porcine follicular fluid, sodium pyruvate and antibiotics) media were supplemented with 1 and 10mM DMTU. In the control group, the media were not supplemented with DMTU. Intracellular ROS levels decreased significantly in the DMTU-treated groups (P&lt;0.05). Although no effects on rate of nuclear maturation were observed, DMTU significantly increased sperm penetration rates without increasing polyspermy (P&lt;0.05). The addition of 10mM DMTU to the collection and washing media enhanced IVF efficiency. DMTU did not modify the early or late apoptosis of oocytes. Both concentrations of DMTU significantly increased viability and decreased the apoptosis of cumulus cells (P&lt;0.05). These results suggest that the addition of 1 or 10mM of DMTU to the media during the collection and washing of porcine COCs is useful for decreasing cumulus apoptosis mediated by ROS and for optimising the IVF of porcine oocytes.

scholarly journals Does the Apoptosis Value of Cumulus Cells Play a Role in Rescue Oocyte in Vitro Maturation?

2020 ◽  
pp. 1
Author(s):  
Tulay Irez ◽  
Sinem Ercan Dogan ◽  
Enver Ciraci ◽  
Saadet Busra Aksoyer ◽  
Muhammet Sait Toprak ◽  
...  
Keyword(s):  
Experimental Study ◽  
Luteinizing Hormone ◽  
Embryo Development ◽  
In Vitro Maturation ◽  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Immature Oocytes ◽  
Maturation Rate

<p><strong>OBJECTIVE:</strong> In this study, we aimed to investigate the role of the cumulus cell’s apoptosis parameter in the maturation of immature rescue oocytes. </p><p><strong>STUDY DESIGN:</strong> In this experimental study, donated immature germinal vesicle oocytes were cultured for, in vitro maturation, embryo development in matured germinal vesicle oocytes were compared with apoptotic properties of cumulus cells. </p><p><strong>RESULTS:</strong> In all of the immature oocytes after oocyte in vitro maturation, the maturation rate has been observed as 56.1% and 2PN rate as 63.0%. Afterin vitro maturation of germinal vesicle oocytes, there was no difference in apoptosis rates of the cumulus cells between mature and immature oocytes (p&gt; 0.05). The ratio of 2PN in matured germinal vesicle oocytes showing embryo development was 35.4%. A positive correlation was found between luteinizing hormone values on day 3 and E2 values during HCG days during oocyte maturation and embryo development (p=0.021, p=0.020). In addition, it has been observed that the germinal vesicle oocytes, which have completed their maturation and developed into embryos, have high E2 values during HCG days (p=0.020).</p><p><strong>CONCLUSION:</strong> In our study, it has been demonstrated that in vitro maturation in rescue oocytes from stimulated cycles, embryo development potential could not be explained by the apoptosis parameter.</p>

scholarly journals Routine examination of gallbladder specimens after cholecystectomy: a single-centre analysis of the incidence, clinical and histopathological aspects of incidental gallbladder carcinoma

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Davide Di Mauro ◽  
Amira Orabi ◽  
Aye Myintmo ◽  
Alex Reece-Smith ◽  
Shahjehan Wajed ◽  
...  
Keyword(s):  
Gallbladder Carcinoma ◽  
Common Finding ◽  
Adult Women ◽  
Advanced Stage ◽  
Continuous Variables ◽  
Single Centre ◽  
One Stage ◽  
Old Adult ◽  
Student’S T

Abstract Background Gallbladder carcinoma is often found incidentally on histopathologic examination after cholecystectomy—this is referred as incidental gallbladder carcinoma (IGC). Routine vs selective histopathological assessment of gallbladders is under debate and this study evaluates the role of regular specimens’ examination, based on a single-centre analysis of incidence, clinical and histopathological aspects of IGC. Methods Patients who underwent cholecystectomy, between July 2010 and January 2020, were considered. Exclusion criteria were age under 18 and preoperative diagnosis of GB carcinoma. Demographic, clinical and histopathological data were retrospectively collected, continuous variables with a normal distribution were evaluated with Student’s t-test and ANOVA. Results Some 5779 patients were included. The female/male ratio was 2.5:1. Chronic cholecystitis (CC) was the most common finding on specimens (99.3%), IGC was found in six cases (0.1%). In the latter group, there were 5 women and patients were older than those with benign disease—73.7 $$\pm$$ ±  5.38 years vs 55.8 $$\pm$$ ±  0.79 years (p < 0.05). In all the cases, the GB was abnormal on intraoperative inspection and beside cancer, histopathology showed associated CC and/or dysplasia. Upon diagnosis, disease was at advanced stage—one stage II, one stage IIIA, one stage IIIB, three stage IVA. Two patients are alive, three died of disease progression—median survival was 7 months (range 2–14). Conclusions In this series, ICG was rare, occurred most commonly in old adult women and was diagnosed at an advanced stage. In all the cases, the GB was abnormal intraoperatively, therefore macroscopic GB anomalies demand histopathological assessment of the specimen.

scholarly journals The kinase activity of human Rio1 is required for final steps of cytoplasmic maturation of 40S subunits

2012 ◽  
Vol 23 (1) ◽  
pp. 22-35 ◽  
Author(s):  
Barbara Widmann ◽  
Franziska Wandrey ◽  
Lukas Badertscher ◽  
Emanuel Wyler ◽  
Jens Pfannstiel ◽  
...  
Keyword(s):  
Rna Interference ◽  
Protein Kinases ◽  
18S Rrna ◽  
Kinase Activity ◽  
Ribosomal Biogenesis ◽  
Ribosomal Subunits ◽  
Binding Partner ◽  
Rrna Maturation ◽  
Cytoplasmic Maturation

RIO proteins form a conserved family of atypical protein kinases. Humans possess three distinct RIO kinases—hRio1, hRio2, and hRio3, of which only hRio2 has been characterized with respect to its role in ribosomal biogenesis. Here we show that both hRio1 and hRio3, like hRio2, are associated with precursors of 40S ribosomal subunits in human cells. Furthermore, we demonstrate that depletion of hRio1 by RNA interference affects the last step of 18S rRNA maturation and causes defects in the recycling of several trans-acting factors (hEnp1, hRio2, hLtv1, hDim2/PNO1, and hNob1) from pre-40S subunits in the cytoplasm. Although the effects of hRio1 and hRio2 depletion are similar, we show that the two kinases are not fully interchangeable. Moreover, rescue experiments with a kinase-dead mutant of hRio1 revealed that the kinase activity of hRio1 is essential for the recycling of the endonuclease hNob1 and its binding partner hDim2 from cytoplasmic pre-40S. Kinase-dead hRio1 is trapped on pre-40S particles containing hDim2 and hNob1 but devoid of hEnp1, hLtv1, and hRio2. These data reveal a role of hRio1 in the final stages of cytoplasmic pre-40S maturation.

Contrasting effects of the Toll-like receptor 4 in determining ovarian follicle endowment and fertility in female adult mice

Zygote ◽  
2021 ◽  
pp. 1-7
Author(s):  
Júlio Panzera Gonçalves ◽  
Breno Augusto Magalhães ◽  
Paulo Henrique Almeida Campos-Junior
Keyword(s):  
Ovarian Follicle ◽  
Cumulus Cells ◽  
Toll Like Receptor ◽  
Female Reproduction ◽  
Toll Like Receptor 4 ◽  
Genetic Ablation ◽  
Adult Mice ◽  
Cumulus Oocyte Complex ◽  
Estrous Cyclicity

Abstract Toll-like receptor 4 (TLR4) is best known for its role in bacteria-produced lipopolysaccharide recognition. Regarding female reproduction, TLR4 is expressed by murine cumulus cells and participates in ovulation and in cumulus–oocyte complex (COC) expansion, maternal–fetal interaction and preterm labour. Despite these facts, the role of TLR4 in ovarian physiology is not fully understood. Therefore, the aim of the present study was to investigate the effects of TLR4 genetic ablation on mice folliculogenesis and female fertility, through analysis of reproductive crosses, ovarian responsiveness and follicular quantification in TLR4−/− (n = 94) and C57BL/6 mice [wild type (WT), n = 102]. TLR4-deficient pairs showed a reduced number of pups per litter (P = 0.037) compared with WT. TLR4−/− mice presented more primordial, primary, secondary and antral follicles (P < 0.001), however there was no difference in estrous cyclicity (P > 0.05). A lower (P = 0.006) number of COC was recovered from TLR4−/− mice oviducts after superovulation, and in heterozygous pairs, TLR4−/− females also showed a reduction in the pregnancy rate and in the number of fetuses per uterus (P = 0.007) when compared with WT. Altogether, these data suggest that TLR4 plays a role in the regulation of murine folliculogenesis and in determining ovarian endowment. TLR4 deficiency may affect ovulation and pregnancy rates, potentially decreasing fertility, therefore the potential side effects of its blockade have to be carefully investigated.

Role of Cumulus Cells in Oocyte Maturation

1982 ◽  
pp. 175-188 ◽  
Author(s):  
Torbjörn Hillensjö ◽  
Claes Magnusson ◽  
Carl Ekholm ◽  
Håkan Billig ◽  
Lars Hedin
Keyword(s):  
Oocyte Maturation ◽  
Cumulus Cells

scholarly journals Sperm binding to ZP2-coated beads improve the efficiency of porcine in vitro fertilisation

Reproduction ◽  
2020 ◽  
Vol 160 (5) ◽  
pp. 725-735
Author(s):  
Julieta Gabriela Hamze ◽  
María Jiménez-Movilla ◽  
Raquel Romar
Keyword(s):  
Acrosome Reaction ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
In Vitro Fertilisation ◽  
Sperm Binding ◽  
Fertilisation Rate ◽  
Gamete Recognition ◽  
Boar Spermatozoa

The role of specific zona pellucida (ZP) glycoproteins in gamete interaction has not yet been elucidated in many species. A recently developed 3D model based on magnetic sepharose beads (B) conjugated to recombinant ZP glycoproteins (BZP) and cumulus cells (CBZP) allows the study of isolated ZP proteins in gamete recognition studies. The objective of this work was to study the role of porcine ZP2, ZP3 and ZP4 proteins in sperm binding, cumulus cell adhesion and acrosome reaction triggering. ZP protein-bound beads were incubated with fresh ejaculated boar spermatozoa and isolated cumulus cells for 24 h. The number of sperm bound to the beads, the acrosomal shrouds (presence of acrosomal content) on the bead’s surface, and the acrosome integrity (by means of PNA-FITC lectin) in bound and unbound sperm were studied. Finally, in vitro matured porcine oocytes mixed with BZP2 were inseminated in vitro using fresh sperm and fertilisation results evaluated. Over 60% of beads had at least one sperm bound after 2 h of coincubation. ZP2-beads (BZP2) and cumulus-ZP2-bead complexes (CBZP2) reached the highest number of sperm per bead, whereas BZP3 and BZP4 models showed the highest number of unbound reacted sperm cells and acrosomal shrouds. Fertilisation efficiency and monospermy rate increased when oocytes were fertilised in the presence of BZP2. We, therefore, conclude that in pigs, it is mainly ZP2 that is involved in sperm-ZP binding whereas ZP3 and ZP4 induce acrosome reaction. Using magnetic sepharose ZP2-bound beads might be a valuable tool to improve the fertilisation rate in pigs.

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