Effects of di(n-butyl) phthalate exposure on foetal rat germ-cell number and differentiation: identification of age-specific windows of vulnerability

M S Jobling,G R Hutchison,R M Sharpe,S Van Den Driesche

doi:10.1111/j.1365-2605.2010.01140.x

M S Jobling, G R Hutchison + Show 2 more

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https://doi.org/10.1111/j.1365-2605.2010.01140.x

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Abstract

Environmental factors are implicated in increased incidence of human testicular germ-cell cancer (TGCC). TGCC has foetal origins and may be one component of a testicular dysgenesis syndrome (TDS). Certain phthalates induce TDS in rats, including effects on foetal germ cells (GC). As humans are widely exposed to phthalates, study of the effects of phthalates on foetal rat GC could provide an insight into the vulnerability of foetal GC to disruption by environmental factors, and thus to origins of TGCC. This study has therefore characterized foetal GC development in rats after in utero exposure to di(n-butyl) phthalate (DBP) with emphasis on GC numbers/proliferation, differentiation and time course for inducing effects. Pregnant rats were treated orally from embryonic day 13.5 (e13.5) with 500 mg/kg/day DBP for varying periods. GC number, proliferation, apoptosis, differentiation (loss of OCT4, DMRT1 expression, DMRT1 re-expression, GC migration) and aggregation were evaluated at various foetal and postnatal ages. DBP exposure reduced foetal GC number by ∼60% by e15.5 and prolonged GC proliferation, OCT4 and DMRT1 immunoexpression; these effects were induced in the period immediately after testis differentiation (e13.5–e15.5). In contrast, DBP-induced GC aggregation stemmed from late gestation effects (beyond e19.5). Foetal DBP exposure delayed postnatal resumption of GC proliferation, leading to bigger deficits in numbers, and delayed re-expression of DMRT1 and radial GC migration. Therefore, DBP differentially affects foetal GC in rats according to stage of gestation, effects that may be relevant to the human because of their nature (OCT4, DMRT1 effects) or because similar effects are demonstrable in vitro on human foetal testes (GC number). Identification of the mechanisms underlying these effects could give a new insight into environment-sensitive mechanisms in early foetal GC development that could potentially be relevant to TGCC origins.

Highlights

Testicular germ-cell cancer (TGCC) is the most common cancer of young Caucasian men with a peak incidence occurring at around 25–30 years (Skakkebaek et al, 2001)
Environmental factors are implicated in increased incidence of human testicular germ-cell cancer (TGCC)
As humans are widely exposed to phthalates, study of the effects of phthalates on foetal rat germ cells (GCs) could provide an insight into the vulnerability of foetal GC to disruption by environmental factors, and to origins of TGCC

Summary

Introduction

Testicular germ-cell cancer (TGCC) is the most common cancer of young Caucasian men with a peak incidence occurring at around 25–30 years (Skakkebaek et al, 2001). In most Western countries, TGCC has increased progressively in incidence over the last 40–70 years, indicative of lifestyle ⁄ environmental causes (Bray et al, 2006). If CIS cells are identified in a testis, it predicts that TGCC will eventually develop (Skakkebaek et al, 2001). Two major unresolved issues about CIS are: (1) why does differentiation fail in some foetal GC, and (2) what lifestyle ⁄ environmental factors can influence this, and explain the progressive increase in TGCC incidence.

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