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Normal spermatogenesis in Fank1 (fibronectin type 3 and ankyrin repeat domains 1) mutant mice

, , , , ,
1 Department of Histology and Embryology, Nanjing Medical University, Nanjing, China
2 Animal Core Facility of Nanjing Medical University, Nanjing, China
DOI
10.7287/peerj.preprints.27356v1
Published
Accepted
Subject Areas
Andrology, Histology
Keywords
Fank1, male infertility, gene knockout, spermatogenesis
Copyright
© 2018 Zhang et al.
Licence
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
Cite this article
Zhang J, Zhang X, Zhang Y, Zeng W, Zhao S, Liu M. 2018. Normal spermatogenesis in Fank1 (fibronectin type 3 and ankyrin repeat domains 1) mutant mice. PeerJ Preprints 6:e27356v1

Abstract

Background. The fibronectin type 3 and ankyrin repeat domains 1 gene, Fank1, is an ancient, evolutionarily conserved gene present in vertebrates. Fank1-knockdown mice have oligospermia caused by an increase in apoptotic germ cells. In this study, we investigated the in vivo function of Fank1.

Methods. In this study, we generated Fank1-knockout mice using the CRISPR/Cas9 system. We then investigated the phenotype and in vivo function of Fank1. Testes and epididymis tissues were analyzed by histological and immunofluorescence staining. Apoptotic cells were analyzed in TUNEL assays. Fertility and sperm counts were also evaluated. The GTEx database were used to assess gene expression quantitative trait loci (eQTL) and mRNA expression of candidate genes and genes neighboring single nucleotide polymorphisms was analyzed by quantitative RT-PCR.

Results. In contrast to the Fank1-knockdown model, no significant changes in epididymal sperm content and the number of apoptotic cells were observed in Fank1-/- homozygotes. In addition, a different pattern of Dusp1, Klk1b21 and Klk1b27 mRNA expression was detected in Fank1-knockout testis. These results reveal differences in the molecular changes between Fank1-knockdown mice and Fank1 -knockout mice and provide a basic resource for population genetics studies.

Author Comment

This is a peer-review submission to PeerJ.

Supplemental Information

Table S1

SNPs associated with expression of the mRNAs of Fank1

DOI: 10.7287/peerj.preprints.27356v1/supp-1

Table S2

List of Antibodies

DOI: 10.7287/peerj.preprints.27356v1/supp-2

Supplemental File.1

Rawdata of Figure.2B

DOI: 10.7287/peerj.preprints.27356v1/supp-5

Supplemental File.2

Rawdata of Figure.2D

DOI: 10.7287/peerj.preprints.27356v1/supp-6

Supplemental File.3

Rawdata of Figure.5C, D and E

DOI: 10.7287/peerj.preprints.27356v1/supp-7

Supplemental File.4

Rawdata of Figure.5F

DOI: 10.7287/peerj.preprints.27356v1/supp-8

Supplemental File.5

Rawdata of Figure.6

DOI: 10.7287/peerj.preprints.27356v1/supp-9

Supplemental File.6

Rawdata of Figure.7

DOI: 10.7287/peerj.preprints.27356v1/supp-10

Additional Information

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Jintao Zhang performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, prepared figures and/or tables, authored or reviewed drafts of the paper.

Xin Zhang performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, prepared figures and/or tables.

Yue Zhang performed the experiments, analyzed the data.

Wentao Zeng performed the experiments.

Shuqin Zhao performed the experiments.

Mingxi Liu conceived and designed the experiments, prepared figures and/or tables, authored or reviewed drafts of the paper, approved the final draft.

Animal Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

Institutional Animal Care and Use Committee of Nanjing Medical University (China)

Data Deposition

The following information was supplied regarding data availability:

The raw measurements are provided in the supplementary files. The raw data shows no significant changes in epididymal sperm content and the number of apoptotic cells were observed in Fank1-/- homozygotes. In addition, a different pattern of Dusp1, Klk1b21 and Klk1b27 mRNA expression was detected in Fank1-knockout testis.

Funding

This work was funded by the Natural Science Foundation of China (31571536 and 31771654), The Natural Science Foundation of the Jiangsu Higher Education Institutions of China (16KJA310003), Natural Science Foundation of Jiangsu Province (No. BK20150990) and Qing Lan Project. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Normal spermatogenesis in Fank1 (fibronectin type 3 and ankyrin repeat domains 1) mutant mice
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