Intracytoplasmic sperm injection induces transgenerational abnormalities in mice
Mito Kanatsu-Shinohara, … , Atsuo Ogura, Takashi Shinohara
Mito Kanatsu-Shinohara, … , Atsuo Ogura, Takashi Shinohara
Published November 15, 2023
Citation Information: J Clin Invest. 2023;133(22):e170140. https://doi.org/10.1172/JCI170140.
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Research Article Reproductive biology Stem cells

Intracytoplasmic sperm injection induces transgenerational abnormalities in mice

Abstract

In vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) are 2 major assisted reproductive techniques (ARTs) used widely to treat infertility. Recently, spermatogonial transplantation emerged as a new ART to restore fertility to young patients with cancer after cancer therapy. To examine the influence of germ cell manipulation on behavior of offspring, we produced F1 offspring by a combination of two ARTs, spermatogonial transplantation and ICSI. When these animals were compared with F1 offspring produced by ICSI using fresh wild-type sperm, not only spermatogonial transplantation–ICSI mice but also ICSI-only control mice exhibited behavioral abnormalities, which persisted in the F2 generation. Furthermore, although these F1 offspring appeared normal, F2 offspring produced by IVF using F1 sperm and wild-type oocytes showed various types of congenital abnormalities, including anophthalmia, hydrocephalus, and missing limbs. Therefore, ARTs can induce morphological and functional defects in mice, some of which become evident only after germline transmission.

Authors

Mito Kanatsu-Shinohara, Yusuke Shiromoto, Narumi Ogonuki, Kimiko Inoue, Satoko Hattori, Kento Miura, Naomi Watanabe, Ayumi Hasegawa, Keiji Mochida, Takuya Yamamoto, Tsuyoshi Miyakawa, Atsuo Ogura, Takashi Shinohara

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Figure 4

Analysis of spermatogenesis and GS cells derived from F1 mice.

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Analysis of spermatogenesis and GS cells derived from F1 mice. (A) Immun...
(A) Immunostaining of F1 testes using antibodies against H3K4me2, H3K9me2, H3K27me2, H3K27me3, H3K36me2, and H3K79me2. One hundred cells in 5 tubules of 3 mice were analyzed per group. Each antigen was assessed using a single antibody. Signal intensity in PNA+ cells was measured. H3K9me2 was omitted for quantification because PNA+ cells did not show H3K9me2 signals. Scale bar: 30 μm. (B) A scatter plot with a list of genes, showing correlation of the DNA methylation data at individual CpG sites in gene promoters (n = 4). Methylation statuses at 237,680 CpG sites were covered. The numbers of identified hypermethylated sites and hypomethylated sites in ICSI-F1 compared with control-F1 GS cells are shown in red and blue, respectively, along with the percentage of commonly covered sites. Red or blue lines indicate 20% increased methylation levels or 20% decreased methylation levels in ICSI-F1 GS cells, respectively. The dashed line indicates the linear regression line. Up, upregulation; Down, downregulation. (C) A scatter plot of gene expression by RNA-Seq (n = 4). (D) Real-time PCR analysis of F1 GS cells (n = 3). See Supplemental Tables 6 and 7 for details.