PLAG1 deficiency impairs spermatogenesis and sperm motility in mice

Almas R Juma,Moira K O’Bryan,Adam H Hart,Anne E O’Connor,Wim J M Van De Ven,Daniel Barriga,Sylvia V H Grommen,Nathan E Hall,Stephen R Doyle,Bert De Groef,Pauliina E Damdimopoulou,D Jo Merriner

doi:10.1038/s41598-017-05676-4

Abstract

Deficiency in pleomorphic adenoma gene 1 (PLAG1) leads to reduced fertility in male mice, but the mechanism by which PLAG1 contributes to reproduction is unknown. To investigate the involvement of PLAG1 in testicular function, we determined (i) the spatial distribution of PLAG1 in the testis using X-gal staining; (ii) transcriptomic consequences of PLAG1 deficiency in knock-out and heterozygous mice compared to wild-type mice using RNA-seq; and (iii) morphological and functional consequences of PLAG1 deficiency by determining testicular histology, daily sperm production and sperm motility in knock-out and wild-type mice. PLAG1 was sparsely expressed in germ cells and in Sertoli cells. Genes known to be involved in spermatogenesis were downregulated in the testes of knock-out mice, as well as Hsd17b3, which encodes a key enzyme in androgen biosynthesis. In the absence of Plag1, a number of genes involved in immune processes and epididymis-specific genes were upregulated in the testes. Finally, loss of PLAG1 resulted in significantly lowered daily sperm production, in reduced sperm motility, and in several animals, in sloughing of the germinal epithelium. Our results demonstrate that the subfertility seen in male PLAG1-deficient mice is, at least in part, the result of significantly reduced sperm output and sperm motility.

Highlights

Infertility is a major health problem worldwide: the prevalence of infertility in developed and developing countries ranges from 3.5% to 16.7%1
We present data demonstrating the spatial distribution of pleomorphic adenoma gene 1 (PLAG1) in the testes, the transcriptomic changes associated with the loss of one or two alleles of Plag[1] relative to WT mice, and the functional consequences of PLAG1 deficiency via histology of the testis, daily sperm production and sperm motility in male KO mice and age-matched WT mice, in order to characterise the reproductive abnormalities of PLAG1-deficient mice
While X-gal staining was abundantly present in the pars distalis of the pituitary gland (Supplementary Fig. S1), known to express ample PLAG113, sparse X-gal staining was observed across the seminiferous tubules in testes from KO mice (Fig. 1a,b)

Summary

Introduction

Infertility is a major health problem worldwide: the prevalence of infertility in developed and developing countries ranges from 3.5% to 16.7%1. Undifferentiated spermatogonia proliferate to produce type A and B spermatogonia via mitosis; type B spermatogonia differentiate to primary spermatocytes, which undergo meiosis to round spermatids This involves coordinated regulation by thousands of genes[5, 6]. Mating of both male and female KO mice resulted in infrequent conception and smaller litter sizes[13] The cause of these fertility issues in Plag[1] KO mice is not known. We present data demonstrating the spatial distribution of PLAG1 in the testes, the transcriptomic changes associated with the loss of one or two alleles of Plag[1] relative to WT mice, and the functional consequences of PLAG1 deficiency via histology of the testis, daily sperm production and sperm motility in male KO mice and age-matched WT mice, in order to characterise the reproductive abnormalities of PLAG1-deficient mice

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Conclusion