Mayu Fukutomi1, Chiharu Uedono1, Aki Fujii1, Youichi Sato2. 1. Department of Pharmaceutical Information Science, Tokushima University Graduate School of Biomedical Sciences, 1-78-1 Sho-machi, Tokushima, Tokushima, 770-8505, Japan. 2. Department of Pharmaceutical Information Science, Tokushima University Graduate School of Biomedical Sciences, 1-78-1 Sho-machi, Tokushima, Tokushima, 770-8505, Japan. youichi.sato@tokushima-u.ac.jp.
Abstract
PURPOSE: Leucine-rich repeats and IQ motif containing 1 (LRRIQ1) gene is reportedly associated with plasma inhibin B levels. However, the function of LRRIQ1 remains unknown. In this study, we generated Lrriq1 knockout mice (Lrriq1-/- mice) and examined the effects of LRRIQ1 on inhibin B and fertility. METHODS: Lrriq1-/- mice were generated using CRISPR/Cas9 genome editing technology. The expression of Inhibin B was examined by Western blotting using a protein extracted from the testis of a 3-month-old male mouse. Mating experiments were conducted using 7-week-old Lrriq1-/- mice and wild-type (WT) mice to examine fertility. Sperm concentration and sperm motility were measured using 3-month-old male mice. RESULTS: Expression analysis of inhibin B revealed that Lrriq1-/- mice exhibited reduced mRNA and protein levels of inhibin alpha (Inha), which constitutes the α subunit. In the mating experiment, the litter size of Lrriq1-/- male mice was 4.3 ± 2.9, which was significantly lower than that of WT male mice (8.3 ± 1.3) (p < 0.001). No difference in sperm count was observed between Lrriq1-/- and WT male mice; however, sperm motility (%) was significantly reduced in Lrriq1-/- mice (48.4 ± 4.9) when compared with WT mice (70.2 ± 4.7) (p < 0.001). Based on TUNEL staining, the testes and epididymal sperm of Lrriq1-/- mice showed high numbers of apoptosis-positive cells. CONCLUSION: Lrriq1 knockout reduced sperm motility and litter size by inducing apoptosis of testicular germ cells and epididymal sperm.
PURPOSE: Leucine-rich repeats and IQ motif containing 1 (LRRIQ1) gene is reportedly associated with plasma inhibin B levels. However, the function of LRRIQ1 remains unknown. In this study, we generated Lrriq1 knockout mice (Lrriq1-/- mice) and examined the effects of LRRIQ1 on inhibin B and fertility. METHODS: Lrriq1-/- mice were generated using CRISPR/Cas9 genome editing technology. The expression of Inhibin B was examined by Western blotting using a protein extracted from the testis of a 3-month-old male mouse. Mating experiments were conducted using 7-week-old Lrriq1-/- mice and wild-type (WT) mice to examine fertility. Sperm concentration and sperm motility were measured using 3-month-old male mice. RESULTS: Expression analysis of inhibin B revealed that Lrriq1-/- mice exhibited reduced mRNA and protein levels of inhibin alpha (Inha), which constitutes the α subunit. In the mating experiment, the litter size of Lrriq1-/- male mice was 4.3 ± 2.9, which was significantly lower than that of WT male mice (8.3 ± 1.3) (p < 0.001). No difference in sperm count was observed between Lrriq1-/- and WT male mice; however, sperm motility (%) was significantly reduced in Lrriq1-/- mice (48.4 ± 4.9) when compared with WT mice (70.2 ± 4.7) (p < 0.001). Based on TUNEL staining, the testes and epididymal sperm of Lrriq1-/- mice showed high numbers of apoptosis-positive cells. CONCLUSION: Lrriq1 knockout reduced sperm motility and litter size by inducing apoptosis of testicular germ cells and epididymal sperm.
Authors: E A M Kuijper; C B Lambalk; D I Boomsma; S van der Sluis; M A Blankenstein; E J C de Geus; D Posthuma Journal: Hum Reprod Date: 2007-06-13 Impact factor: 6.918
Authors: Claes Ohlsson; Henri Wallaschofski; Kathryn L Lunetta; Lisette Stolk; John R B Perry; Annemarie Koster; Ann-Kristin Petersen; Joel Eriksson; Terho Lehtimäki; Ilpo T Huhtaniemi; Geoffrey L Hammond; Marcello Maggio; Andrea D Coviello; Luigi Ferrucci; Margit Heier; Albert Hofman; Kate L Holliday; John-Olov Jansson; Mika Kähönen; David Karasik; Magnus K Karlsson; Douglas P Kiel; Yongmei Liu; Osten Ljunggren; Mattias Lorentzon; Leo-Pekka Lyytikäinen; Thomas Meitinger; Dan Mellström; David Melzer; Iva Miljkovic; Matthias Nauck; Maria Nilsson; Brenda Penninx; Stephen R Pye; Ramachandran S Vasan; Martin Reincke; Fernando Rivadeneira; Abdelouahid Tajar; Alexander Teumer; André G Uitterlinden; Jagadish Ulloor; Jorma Viikari; Uwe Völker; Henry Völzke; H Erich Wichmann; Tsung-Sheng Wu; Wei Vivian Zhuang; Elad Ziv; Frederick C W Wu; Olli Raitakari; Anna Eriksson; Martin Bidlingmaier; Tamara B Harris; Anna Murray; Frank H de Jong; Joanne M Murabito; Shalender Bhasin; Liesbeth Vandenput; Robin Haring Journal: PLoS Genet Date: 2011-10-06 Impact factor: 5.917