scholarly journals Anticentromere antibody induced by immunization with centromere protein and Freund’s complete adjuvant may interfere with mouse early-stage embryo

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Hanyan Liu ◽  
Yufen Zhang ◽  
Haiying Liu ◽  
Qing Huang ◽  
Ying Ying
Keyword(s):  
Early Stage ◽  
Protein A ◽  
Immunofluorescence Assay ◽  
Immunofluorescence Test ◽  
Anticentromere Antibody ◽  
Indirect Immunofluorescence Test ◽  
Centromere Protein ◽  
Centromere Protein A ◽  
Stage Embryo ◽  
Early Stage Embryo

Abstract Background Anticentromere antibody (ACA) is a member of the antinuclear antibody spectrum (ANAs) which has been speculated to be associated with subfertility. Thus, the present study aimed to investigate the induction of ACA production and its potential interference with early-stage embryos. Methods Recombinant centromere protein-A (CENP-A) or centromere protein-B (CENP-B) and complete Freund’s adjuvant (CFA) were used to immunize mice. Serum ACA level was then evaluated by using an indirect immunofluorescence test. Immunofluorescence assay was performed to detect IgG in follicles in ovarian tissues and early-stage embryos. Results Following treatment, serum positive ACA was observed in mice treated with CENP and CFA. Furthermore, IgG were detected in follicular fluid and early-stage embryos from mice treated with CENP and CFA. Conclusions This study preliminarily indicated that ACA induced by CENP and CFA may penetrate into the living embryos of early-stage in mice.

scholarly journals Anticentromere antibody induced by immunization with centromere protein a and Freund’s complete adjuvant may interfere with mouse oocyte meiosis

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Ying Ying ◽  
Shuang Liu ◽  
Yixuan Wu ◽  
Sichen Li ◽  
Qing Huang
Keyword(s):  
Protein A ◽  
Underlying Mechanism ◽  
Saline Group ◽  
Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Groups ◽  
Anticentromere Antibody ◽  
Centromere Protein ◽  
Centromere Protein A ◽  
Oocyte Meiosis

Abstract Background Anticentromere antibody (ACA) is a member of the antinuclear antibody (ANA) family, and recent studies have found that ACA may be associated with oocyte maturation disorders; however, the possible mechanism behind this phenomenon remains unknown. We conducted this study to investigate whether ACA could penetrate into the living oocytes and interfere with oocyte meiosis in a mouse model. Methods We divided mice into three groups: human recombinant centromere protein-A (human CENP-A, HA) and complete Freund’s adjuvant (CFA) were used to immunize mice for the study group (HA + CFA), and mice injected with CFA (CFA group) or saline (Saline group), respectively, served as controls. After immunization, serum anti-CENP-A antibody was detected by indirect immunofluorescence assay (IIFT) and enzyme-linked immunosorbent assay (ELISA). Chromosome alignment and intracellular IgG localization in MI- and MII-stage oocytes were investigated by immunofluorescence analysis. Results Positive ACAs were successfully induced by immunization with CENP-A and CFA, and results showed that the serum level of anti-CENP-A antibody was significantly higher in the HA + CFA group compared with the control groups. There was marked increase of chromosome misalignments in MI and MII oocytes in the HA + CFA group compared to the control groups. However, no oocytes from any of the three groups showed intracellular antibody immunofluorescence. Conclusions The development and maturation of oocytes were impaired in peripheral ACA positive mice, which exhibited severe chromosomal misalignments in metaphase meiosis; however, no evidence of ACAs entering the oocytes was observed, thus the underlying mechanism needs further exploration.

scholarly journals An Exploration of the Impact of Anticentromere Antibody on Early-Stage Embryo

2017 ◽  
Vol 2017 ◽  
pp. 1-4
Author(s):  
Ying Ying ◽  
Xi Guo ◽  
Yiping Zhong ◽  
Canquan Zhou
Keyword(s):  
Early Stage ◽  
Protein A ◽  
Immunofluorescence Assay ◽  
Mouse Embryos ◽  
Anticentromere Antibody ◽  
Developmental Potential ◽  
Stage Embryo ◽  
Direct Target ◽  
The Impact

Background. Previously, we found women with positive anticentromere antibody showed impaired potential of oocyte maturation and embryo cleavage; the possible mechanism behind this phenomenon was still unknown. Objective. Thus, the present study aimed to preliminarily explore whether ACA could penetrate into the living embryos and impair their developmental potential via in vitro coculture with mouse embryos. Methods. Mouse embryos were collected and used for in vitro culture with polyclonal anticentromere protein A (CENP-A) antibody; then, immunofluorescence assay was performed to determine the penetration of antibody into embryos, and embryo development potential was observed. Results. All embryos cultured with anti-CENP-A antibody exhibited immunofluorescence on the nucleus, while none of the embryos from the control groups showed immunofluorescence. Additionally, embryos cultured with anti-CENP-A antibody experienced significant growth impairment compared with controls. Conclusion. Mouse embryos may be a direct target for ACA in vitro prior to implantation. However, the precise mechanism needs further clarification.

scholarly journals Fast Multi-Focus Fusion Based on Deep Learning for Early-Stage Embryo Image Enhancement

Sensors ◽  
2021 ◽  
Vol 21 (3) ◽  
pp. 863
Author(s):  
Vidas Raudonis ◽  
Agne Paulauskaite-Taraseviciene ◽  
Kristina Sutiene
Keyword(s):  
Deep Learning ◽  
Image Fusion ◽  
Early Stage ◽  
Image Data ◽  
Cell Detection ◽  
Processing Times ◽  
Fused Image ◽  
Stage Embryo ◽  
Early Stage Embryo

Background: Cell detection and counting is of essential importance in evaluating the quality of early-stage embryo. Full automation of this process remains a challenging task due to different cell size, shape, the presence of incomplete cell boundaries, partially or fully overlapping cells. Moreover, the algorithm to be developed should process a large number of image data of different quality in a reasonable amount of time. Methods: Multi-focus image fusion approach based on deep learning U-Net architecture is proposed in the paper, which allows reducing the amount of data up to 7 times without losing spectral information required for embryo enhancement in the microscopic image. Results: The experiment includes the visual and quantitative analysis by estimating the image similarity metrics and processing times, which is compared to the results achieved by two wellknown techniques—Inverse Laplacian Pyramid Transform and Enhanced Correlation Coefficient Maximization. Conclusion: Comparatively, the image fusion time is substantially improved for different image resolutions, whilst ensuring the high quality of the fused image.

scholarly journals NMR spectroscopy of a single mammalian early stage embryo

2021 ◽  
pp. 107142
Author(s):  
Giulia Sivelli ◽  
Gaurasundar M. Conley ◽  
Carolina Herrera ◽  
Kathryn Marable ◽  
Kyle J. Rodriguez ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Early Stage ◽  
Stage Embryo ◽  
Early Stage Embryo

Anti-centromere protein A antibodies in systemic sclerosis: Significance and origin

2016 ◽  
Vol 15 (1) ◽  
pp. 102-109 ◽  
Author(s):  
Federico Perosa ◽  
Marcella Prete ◽  
Giuseppe Di Lernia ◽  
Carmela Ostuni ◽  
Elvira Favoino ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Protein A ◽  
Centromere Protein ◽  
Centromere Protein A

scholarly journals Functional Complementation of Human Centromere Protein A (CENP-A) by Cse4p from Saccharomyces cerevisiae

2004 ◽  
Vol 24 (15) ◽  
pp. 6620-6630 ◽  
Author(s):  
Gerhard Wieland ◽  
Sandra Orthaus ◽  
Sabine Ohndorf ◽  
Stephan Diekmann ◽  
Peter Hemmerich
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Budding Yeast ◽  
Protein A ◽  
Human Cells ◽  
Cycle Arrest ◽  
Centromere Protein ◽  
Centromere Protein A

ABSTRACT We have employed a novel in vivo approach to study the structure and function of the eukaryotic kinetochore multiprotein complex. RNA interference (RNAi) was used to block the synthesis of centromere protein A (CENP-A) and Clip-170 in human cells. By coexpression, homologous kinetochore proteins from Saccharomyces cerevisiae were then tested for the ability to complement the RNAi-induced phenotypes. Cse4p, the budding yeast CENP-A homolog, was specifically incorporated into kinetochore nucleosomes and was able to complement RNAi-induced cell cycle arrest in CENP-A-depleted human cells. Thus, Cse4p can structurally and functionally substitute for CENP-A, strongly suggesting that the basic features of centromeric chromatin are conserved between yeast and mammals. Bik1p, the budding yeast homolog of human CLIP-170, also specifically localized to kinetochores during mitosis, but Bik1p did not rescue CLIP-170 depletion-induced cell cycle arrest. Generally, the newly developed in vivo complementation assay provides a powerful new tool for studying the function and evolutionary conservation of multiprotein complexes from yeast to humans.

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