Supplementary Material for: Overexpression of Lis1 in Different Stages of Spermatogenesis Does Not Result in an Aberrant Phenotype

Previous studies showed that in the mouse mutant Lis1^GT/GT gene trap integration in intron 2 of Lis1 gene leads to male infertility in homozygous Lis1^GT/GT mice. We further analyzed this line and could confirm the suggested downregulation of a testis-specific Lis1 transcript in mutant animals in a quantitative manner. Moreover, we analyzed the gene trap mutation on different genetic backgrounds in incipient congenic animals and could exclude a genetic background effect. To gain further insights into the role and requirement of LIS1 in spermatogenesis, 3 transgenic lines were generated, that overexpress Lis1 under control of the testis-specific promoters hEF-1α, which is exclusively active in spermatogonial cells, PGK2, which is active in pachytene spermatocytes and following stages of spermatogenesis, and Tnp2 which is active in round spermatids and following stages of spermatogenesis, respectively. All 3 transgenic lines remained fertile and testis sections displayed no abnormalities. To overcome the infertility of Lis1^GT/GT males, these transgenic Lis1-overexpressing animals were mated with Lis1^GT/GT mice to generate ‘rescued’ Lis1^GT/GT/Lis1^Tpos males. ‘Rescued’ animals from all transgenic lines remained infertile, thus overexpression of Lis1 in different stages of spermatogenesis could not rescue the infertility phenotype of homozygous gene trap males.