Pitfalls in establishing mouse model of female infertility by immunization with human centromere protein

AbstractInfertility affects 10% - 15% of families worldwide. However, the pathogenesis of female infertility caused by abnormal early embryonic development is not clear. We constructed a mouse model (Pabpn1l -/-) simulating the splicing abnormality of human PABPN1L and found that the female was sterile and the male was fertile. The Pabpn1l -/- oocytes can be produced, ovulated and fertilized normally, but cannot develop beyond the 2-cell stage. Using RNA-Seq, we found a large-scale upregulation of RNA in Pabpn1l -/- MII oocytes. Of the 2401 transcripts upregulated in Pabpn1l-/- MII oocytes, 1523 transcripts (63.4%) were also upregulated in Btg4 -/- MII oocytes, while only 53 transcripts (2.2%) were upregulated in Ythdf2 -/- MII oocytes. We documented that transcripts in zygotes derived from Pabpn1l -/- oocytes have a longer poly(A) tail than the control group, a phenomenon similar to that in Btg4-/- mice. Surprisingly, the poly(A) tail of these mRNAs was significantly shorter in the Pabpn1l -/- MII oocytes than in the Pabpn1l +/+. These results suggest that PABPN1L is involved in BTG4-mediated maternal mRNA degradation, and may antagonize poly(A) tail shortening in oocytes independently of its involvement in maternal mRNA degradation. Thus, PABPN1L variants could be a genetic marker of female infertility.

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Mutational Analysis of the Central Centromere Targeting Domain of Human Centromere Protein C, (CENP-C)

Experimental Cell Research ◽

10.1006/excr.2002.5495 ◽

2002 ◽

Vol 275 (1) ◽

pp. 81-91 ◽

Cited By ~ 18

Author(s):

Kang Song ◽

Bobbi Gronemeyer ◽

Wei Lu ◽

Emily Eugster ◽

John E. Tomkiel

Keyword(s):

Protein C ◽

Mutational Analysis ◽

Human Centromere ◽

Centromere Protein

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Mapping of the Human Centromere Protein B Gene (CENPB) to Chromosome 20p13 by Fluorescence in Situ Hybridization

Genomics ◽

10.1006/geno.1994.1600 ◽

1994 ◽

Vol 24 (1) ◽

pp. 187-188 ◽

Cited By ~ 2

Author(s):

Naohiko Seki ◽

Toshiyuki Saito ◽

Katsumi Kitagawa ◽

Hiroshi Masumoto ◽

Tuneko Okazaki ◽

...

Keyword(s):

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Human Centromere ◽

Centromere Protein ◽

Chromosome 20P13 ◽

Protein B

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Native SAD Structure Solution from Merohedrally Twinned Data

Acta Crystallographica Section A Foundations and Advances ◽

10.1107/s2053273314093851 ◽

2014 ◽

Vol 70 (a1) ◽

pp. C614-C614

Author(s):

Tobias Weinert ◽

Sandro Waltersperger ◽

Vincent Olieric ◽

Federica Basilico ◽

Valentina Cecatiello ◽

...

Keyword(s):

Light Source ◽

Model Building ◽

Experimental Setup ◽

Difficult Case ◽

Space Group ◽

Human Centromere ◽

Centromere Protein ◽

Structure Solution ◽

Swiss Light Source ◽

Automatic Model Building

Up until now, comparatively few structures were solved by native SAD. Recent advances in multi crystal averaging [1] have shown that native SAD can be applied to an increasing number of cases. Though theoretically possible [2], successful structure solutions from twinned data have not been reported yet. Here, we report the structure solution of the human Centromere protein M from a merohedrally twinned crystal with a twinning fraction of 0.45 in the space group P3. The data were collected at the bending magnet beamline X06DA at the Swiss Light Source, which is equipped with the in-house developed multi-axis goniometer PRIGo and the PILATUS 2M detector. A highly redundant 2.2 Å dataset was collected in a number of different crystal orientations. A substructure solution could only be obtained after 50000 SHELXD [3] tries. Automatic model building after phasing and density modification resulted in a model with the majority of residues built correctly. We will present this particularly difficult case together with other more routine cases, all solved with the same experimental setup and at the beamline X06DA.

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PABPN1L assemble into “ring-like” aggregates in the cytoplasm of MII oocytes and is associated with female infertility†

Biology of Reproduction ◽

10.1093/biolre/ioab203 ◽

2021 ◽

Author(s):

Ying Wang ◽

Tianhao Feng ◽

Mingcong Zhu ◽

Xiaodan Shi ◽

Zerui Wang ◽

...

Keyword(s):

Embryonic Development ◽

Mouse Model ◽

Genetic Marker ◽

Large Scale ◽

Scale Up ◽

Mrna Degradation ◽

Female Infertility ◽

Rna Seq ◽

Maternal Mrna

Abstract Infertility affects 10% - 15% of families worldwide. However, the pathogenesis of female infertility caused by abnormal early embryonic development is not clear. A resent study showed that PABPN1L recruited BTG4 to mRNA 3′-poly(A) tails and was essential for maternal mRNA degradation. Here, we generated an PABPN1L-antibody and found “ring-like” PABPN1L aggregates in the cytoplasm of MII oocytes. PABPN1L-EGFP proteins spontaneously formed“ring-like” aggregates in vitro. This phenomenon is similar with CCR4–NOT catalytic subunit, CNOT7, when it starts deadenylation process in vitro. We constructed two mouse model (Pabpn1l −/− and Pabpn1l tm1a/tm1a) simulating the intron1-exon2 abnormality of human PABPN1L and found that the female was sterile and the male was fertile. Using RNA-Seq, we observed a large-scale up-regulation of RNA in zygotes derived from Pabpn1l−/− MII oocytes. We found that 9222 genes were up-regulated instead of being degraded in the Pabpn1l-♀/+♂zygote. Both the Btg4 and Cnot61 genes are necessary for the deadenylation process and Pabpn1l −/− resembled both the Btg4 and Cnot6l knockouts, where 71.2% genes stabilized in the Btg4-♀/+♂ zygote and 84.2% genes stabilized in the Cnot6l-♀/+♂zygote were also stabilized in Pabpn1l-♀/+♂ zygote. BTG4/CNOT7/CNOT6L was partially co-located with PABPN1L in MII oocytes. The above results suggest that PABPN1L is widely associated with CCR4–NOT-mediated maternal mRNA degradation and PABPN1L variants on intron1-exon2 could be a genetic marker of female infertility. Summary sentence. “Ring-like” PABPN1L aggregates was found in the cytoplasm of MII oocytes and in vitro; intron1-exon2 abnormality of Pabpn1l leads female sterile in mice.

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