Male mice lacking ADAMTS-16 are fertile but exhibit testes of reduced weight

Catherine Livermore,Nicolas Chalon,Joffrey Mianné,Andy Greenfield,Sara Wells,Pam Siggers,Lydia Teboul,Gemma Codner,Nick Warr

doi:10.1038/s41598-019-53900-0

Abstract

Adamts16 encodes a disintegrin-like and metalloproteinase with thrombospondin motifs, 16, a member of a family of multi-domain, zinc-binding proteinases. ADAMTS-16 is implicated in a number of pathological conditions, including hypertension, cancer and osteoarthritis. A large number of observations, including a recent report of human ADAMTS16 variants in cases of 46,XY disorders/differences of sex development (DSD), also implicate this gene in human testis determination. We used CRISPR/Cas9 genome editing to generate a loss-of-function allele in the mouse in order to examine whether ADAMTS-16 functions in mouse testis determination or testicular function. Male mice lacking Adamts16 on the C57BL/6N background undergo normal testis determination in the fetal period. However, adult homozygotes have an average testis weight that is around 10% lower than age-matched controls. Cohorts of mutant males tested at 3-months and 6-months of age were fertile. We conclude that ADAMTS-16 is not required for testis determination or male fertility in mice. We discuss these phenotypic data and their significance for our understanding of ADAMTS-16 function.

Highlights

Adamts[16] encodes a disintegrin-like and metalloproteinase with thrombospondin motifs, 16, a member of a family of multi-domain, zinc-binding proteinases
A recent report describes the identification of three human ADAMTS16 variants, resulting in amino acid substitutions predicted to be damaging, in cases of 46,XY disorders/differences of sex development (DSD)[7]
Expression was detected by wholemount in situ hybridisation (WMISH) in XY gonads at all stages from 11.5 dpc, and between 12.5 and 14.5 dpc this was prominent in the cords of the developing testis in a profile consistent with Sertoli cell expression (Fig. 1A–D)

Summary

Introduction

Adamts[16] encodes a disintegrin-like and metalloproteinase with thrombospondin motifs, 16, a member of a family of multi-domain, zinc-binding proteinases. A large number of observations, including a recent report of human ADAMTS16 variants in cases of 46,XY disorders/ differences of sex development (DSD), implicate this gene in human testis determination. ADAMTS (a disintegrin-like and metalloproteinase with thrombospondin motifs) proteins comprise a 19-member family of extracellular metalloproteinases implicated in a number of molecular processes, including collagen processing, proteoglycan cleavage and inhibition of angiogenesis[1] Their control of the structure and function of the extracellular matrix (ECM) is thought to underpin diverse roles in tissue morphogenesis and patho-physiological remodelling[2]. In order to determine whether ADAMTS-16 plays a role in XY mouse reproductive biology, in particular sex determination and testicular function, we used CRISPR/Cas[9] genome editing to introduce a 906 bp deletion into the Adamts[16] gene that removes exon 5 This deletion event removes part of the highly conserved protease domain of the enzyme and results in a predicted premature stop codon and nonsense-mediated decay. We discuss the significance of these data for our understanding of ADAMTS-16 function in reproduction

Methods

Results

Conclusion