Sonic hedgehog promotes porcine oocyte maturation and early embryo development

2009 ◽  
Vol 21 (6) ◽  
pp. 805 ◽  
Author(s):  
Ngoc Tan Nguyen ◽  
David Pei-Cheng Lin ◽  
Shih-Ying Yen ◽  
Jung-Kai Tseng ◽  
Jui-Fen Chuang ◽  
...  

In the present study, we investigated the effects of the Sonic hedgehog (Shh) protein on porcine oocyte maturation and early embryo development. Immunohistochemistry showed activation of Shh signalling in cumulus–oocyte complexes (COCs), as reflected by Patched (Ptc), Smoothened (Smo) and Gli1 expression in oocytes, cumulus cells and granulosa cells, particularly those of small follicles (<2 mm in diameter). Western blot analysis showed Smo expression in COCs and in denuded oocytes derived from small and medium (3–7 mm)-sized follicles. Small follicles contained the highest concentration of Shh in follicular fluid compared with medium-sized and large (>7 mm in diameter) follicles. Supplementation with Shh (0.5 or 1 μg mL–1) enhanced oocyte maturation compared with the control group (92.4% and 90.4% v. 81.9%, respectively; P < 0.05). This effect was reversed by the simultaneous addition of cyclopamine (1–2 μm), an Shh inhibitor. Similar to intact COCs, denuded COCs showed enhanced maturation following Shh supplementation. Furthermore, cyclin B1 content, extracellular signal-regulated kinase 1/2 phosphorylation, intracellular calcium release, blastocyst rate and total cell numbers were greater (P < 0.05) in oocytes matured in the presence of 0.5 and 1 μg mL–1 Shh compared with control oocytes. The findings of the present study provide the first evidence that the Shh signalling pathway is active, or at least partially activated, in the porcine ovary and is likely to promote oocyte cytoplasmic and nuclear maturation, as well as subsequent in vitro development, although the underlying mechanisms remain to be elucidated.

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 108-108
Author(s):  
Malavika K Adur ◽  
Yunsheng Li ◽  
Jason W Ross

Abstract MicroRNA are small non-coding RNA involved in post-transcriptional gene regulation impacting oocyte maturation and embryo development. MIR574-3p abundance decreases during oocyte in vitro maturation (IVM) and blastocyst development. The study objective was to evaluate the role of MIR574-3p during porcine oocyte maturation and early embryo development. To assess the function of MIR574-3p during these processes, denuded GV stage oocytes injected with MIR574-3p mimic (MIR574-3p-M), MIR574-3p inhibitor (MIR574-3p-I) or negative control oligo (NC) underwent IVM for 42 hours. The number of MII arrested oocytes was decreased (P = 0.06) in the MIR574-3p-M group (67.7 ± 1.4%) as compared to the NC group (76.1 ± 1.3%), whereas maturation was not affected by MIR574-3p-I (75.6 ± 1.5%) as compared to the NC group (73.1 ± 3.6%). MII arrested oocytes were parthenogenetically activated and cultured for 7 days. Neither mimic nor inhibitor affected the cleavage or blastocyst rate. Using LC-MS/MS we evaluated changes in global protein abundance in injected oocytes after 42 hours of IVM. We identified 971 proteins in MIR574-3p-M injected oocytes, of which 57 were differentially abundant as compared to the control group. In MIR574-3p-I injected oocytes, 1007 proteins were identified, of which 107 were differentially abundant as compared to the control group. Overall, MIR574-3p-M upregulated proteins critical to membrane binding mediating sperm receptors on the zona pellucida, while it downregulated intranuclear activity such as nucleotide biosynthesis, mitotic spindle assembly and orientation; whereas MIR574-3p-I induced upregulation of proteins involved in the processes between and including protein biosynthesis and metabolism, while downregulating proteins critical to ATP, RNA, DNA and protein binding. These data suggest artificially increasing MIR574-3p abundance during IVM alters the oocyte proteome and influences meiotic progression to MII. Project was supported by Agriculture and Food Research Initiative Competitive Grant no. 2017-67015-26459 from the USDA National Institute of Food and Agriculture.


2008 ◽  
Vol 78 (Suppl_1) ◽  
pp. 94-94
Author(s):  
Nguyen Ngoc Tan ◽  
Tzu-An Lin ◽  
Shi-Ing Yen ◽  
Jung-Kai Tseng ◽  
Rui-Fen Zhuang ◽  
...  

2020 ◽  
Vol 21 (10) ◽  
pp. 3581
Author(s):  
Anthony Estienne ◽  
Adeline Brossaud ◽  
Maxime Reverchon ◽  
Christelle Ramé ◽  
Pascal Froment ◽  
...  

Some evidence shows that body mass index in humans and extreme weights in animal models, including avian species, are associated with low in vitro fertilization, bad oocyte quality, and embryo development failures. Adipokines are hormones mainly produced and released by white adipose tissue. They play a key role in the regulation of energy metabolism. However, they are also involved in many other physiological processes including reproductive functions. Indeed, leptin and adiponectin, the most studied adipokines, but also novel adipokines including visfatin and chemerin, are expressed within the reproductive tract and modulate female fertility. Much of the literature has focused on the physiological and pathological roles of these adipokines in ovary, placenta, and uterine functions. The purpose of this review is to summarize the current knowledge regarding the involvement of leptin, adiponectin, visfatin, and chemerin in the oocyte maturation, fertilization, and embryo development in both mammals and birds.


2015 ◽  
Vol 27 (1) ◽  
pp. 203
Author(s):  
I. Lindgren ◽  
P. Humblot ◽  
D. Laskowski ◽  
Y. Sjunnesson

Dairy cow fertility has decreased during the last decades, and much evidence indicates that metabolic disorders are an important part of this decline. Insulin is a key factor in the metabolic challenge during the transition period that coincides with the oocyte maturation and may therefore have an impact on the early embryo development. The aim of this study was to test the effect of insulin during oocyte maturation on early embryo development by adding insulin during the oocyte maturation in vitro. In this study, abattoir-derived bovine ovaries were used and cumulus-oocyte complexes (n = 991) were in vitro matured for 22 h according to standard protocols. Insulin was added during maturation in vitro as follows: H (10 µg mL–1 of insulin), L (0.1 µg mL–1 of insulin), or Z (0 µg mL–1 of insulin). After maturation, oocytes were removed and fixed in paraformaldehyde before staining. Click-it TUNEL assay (Invitrogen, Stockholm, Sweden) was used for apoptotic staining and DRAQ5 (BioNordika, Stockholm, Sweden) for nuclear staining (n = 132). Cumulus-oocyte complexes were evaluated using laser scanning confocal microscope (Zeiss LSM 510, Zeiss, Oberkochen, Germany). Five levels of scans were used to assess oocyte maturation (MII stage) and apoptosis. Because of incomplete penetration of the TUNEL stain (3–5 layers of cumulus cells), only the outer 2 layers of the cumulus complex were investigated regarding apoptosis. Apoptotic index was calculated as apoptotic cells/total cells visualised. Remaining oocytes were fertilized and cultured in vitro until Day 8. Day 7 and Day 8 blastocyst formation was assessed as well as blastocyst stage and grade. Effect of insulin treatment on variables was analysed by ANOVA following arc sin √p transformation. Post-ANOVA comparisons between H+L group v. Z were performed by using the contrast option under GLM (Scheffé test). Results are presented as least squares means ± s.e. P-values ≤ 0.05 were considered as statistically significant. Insulin treatment during oocyte maturation in vitro had no significant effect on oocyte nuclear maturation or apoptotic index of the cumulus cells (Z: 0.052 ± 0.025, L: 0.039 ± 0.016, H: 0.077 ± 0.044, P > 0.05). No effect was seen on cleavage rates (Z: 0.85 ± 0.02, L: 0.85 ± 0.02, H: 0.89 ± 0.03, P > 0.05), but insulin treatment significantly decreased Day 7 rates from fertilized oocytes (Z: 0.19 ± 0.02, L: 0.14 ± 0.02, H: 0.12 ± 0.02, P < 0.05). This study also showed a significantly retarded developmental stage and decreased grade of blastocysts in insulin-treated groups taken together when compared with the control group (P < 0.05). In this study, no effect of insulin supplementation during in vitro maturation was seen on bovine oocyte maturation and apoptosis of cumulus cells, but blastocyst formation and development were negatively affected. Further studies are needed for understanding the relationship between the addition of insulin during maturation in vitro and impaired blastocyst formation. Insulin is a common supplement in the first phase of the first in vitro maturation medium for pig oocytes and is believed to have a beneficial effect on this species.Funding was received from Stiftelsen Nils Lagerlöfs Fond H12–0051-NLA.


Cell Cycle ◽  
2014 ◽  
Vol 13 (21) ◽  
pp. 3390-3403 ◽  
Author(s):  
Yu Zhang ◽  
Xing Duan ◽  
Rui Cao ◽  
Hong-Lin Liu ◽  
Xiang-Shun Cui ◽  
...  

2018 ◽  
Vol 30 (2) ◽  
pp. 391 ◽  
Author(s):  
C. Liu ◽  
Y. Liu ◽  
K. Larsen ◽  
Y. P. Hou ◽  
H. Callesen

It has been demonstrated that extracellular calcium is necessary in fertilisation and embryo development but the mechanism is still not well understood. The present study mainly focussed on the extracellular calcium effector called the calcium-sensing receptor (CASR) and examined its expression in porcine gametes and embryos and its function during fertilisation and early embryo development. By using reverse transcription polymerase chain reaction, CASR was found to be expressed in porcine oocytes, spermatozoa and embryos at different developmental stages. Functionally, medium supplementation with a CASR agonist or an antagonist during in vitro fertilisation (IVF) and in vitro culture (IVC) was tested. During fertilisation, the presence of a CASR agonist increased sperm penetration rate and decreased polyspermy rate leading to an increased normal fertilisation rate. During embryo development, for the IVF embryos, agonist treatment during IVC significantly increased cleavage rate and blastocyst formation rate compared with the control group. Furthermore, parthenogenetically activated embryos showed similar results with lower cleavage and blastocyst formation rates in the antagonist group than in the other groups. It was concluded that CASR, as the effector of extracellular calcium, modulates porcine fertilisation and early embryo development.


2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Si-Jing Song ◽  
Qiao-Chu Wang ◽  
Ru-Xia Jia ◽  
Xiang-Shun Cui ◽  
Nam-Hyung Kim ◽  
...  

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