Through the smoke: Use of in vivo and in vitro cigarette smoking models to elucidate its effect on female fertility

https://doi.org/10.1016/j.taap.2014.10.010Get rights and content

Highlights

  • In vivo exposure to individual cigarette smoke chemicals alters female fertility.

  • The use of in vitro models in determining molecular mechanisms

  • Whole cigarette smoke inhalation animal models negatively affect ovarian function.

Abstract

A finite number of oocytes are established within the mammalian ovary prior to birth to form a precious ovarian reserve. Damage to this limited pool of gametes by environmental factors such as cigarette smoke and its constituents therefore represents a significant risk to a woman's reproductive capacity. Although evidence from human studies to date implicates a detrimental effect of cigarette smoking on female fertility, these retrospective studies are limited and present conflicting results. In an effort to more clearly understand the effect of cigarette smoke, and its chemical constituents, on female fertility, a variety of in vivo and in vitro animal models have been developed. This article represents a systematic review of the literature regarding four of experimental model types: 1) direct exposure of ovarian cells and follicles to smoking constituents’ in vitro, 2) direct exposure of whole ovarian tissue with smoking constituents in vitro, 3) whole body exposure of animals to smoking constituents and 4) whole body exposure of animals to cigarette smoke. We summarise key findings and highlight the strengths and weaknesses of each model system, and link these to the molecular mechanisms identified in smoke-induced fertility changes.

Section snippets

Introduction: smoking and human fertility

Females are born with a finite number of oocytes, meiotically arrested at meiosis I in primordial follicles (Baker, 1963) (Fig. 1). Once puberty begins, a number of these follicles are recruited each cycle to mature into primary, secondary and finally antral follicles, ready for ovulation (Jones et al., 2013, Skinner, 2005). This recruitment and growth process continues until the follicular pool is exhausted and menopause occurs (Richardson et al., 1987). Accelerated depletion of this reserve

Oocyte and follicle maturation

Mammalian folliculogenesis and oocyte maturation are interlinked processes that are required for the production of oocytes capable of fertilisation. Quiescent primordial follicles consist of an immature oocyte meiotically arrested at prophase I surrounded by a single layer of granulosa cells (Skinner, 2005). As the follicle begins to grow through primary, secondary and antral stages, granulosa cells multiply and theca cells develop at the basement membrane, while cumulus cells — a granulosa

Major constituents of cigarette smoke implicated in ovotoxicity

Cigarettes smoke contains over 4000 chemicals from a wide variety of chemical classes including hydrocarbons, alcohols, phenols, aldehydes, ketones, alkaloids, acids and heavy metals (Rodgman and Perfetti, 2013). Of these classes, the most widely studied with regard to female fertility are the hydrocarbons, specifically polycyclic aromatic hydrocarbons (PAHs) (Ganesan et al., 2013, Ganesan and Keating, 2014, Igawa et al., 2009, Neal et al., 2007, Neal et al., 2010, Sobinoff et al., 2011,

Studies on the effect of smoking on human fertility

Most data regarding the effect of smoking on female fecundity are derived from studies of assisted reproductive technology (ART) patients, and reports present conflicting results. Several studies have found smoking decreases the number of oocytes retrieved for ART (El-Nemr et al., 1998, Fuentes et al., 2010, Zenzes et al., 1995). Those that are collected have a reduced ability to be fertilised, and subsequent embryo quality is decreased (El-Nemr et al., 1998, Gruber et al., 2008, Neal et al.,

Oocyte, follicle and ovarian cell cultures: in vitro testing of ovarian cytotoxicity

In vitro culture of follicles, oocytes and isolated ovarian somatic cells, represent simple methods used to examine the direct effects of cigarette smoke constituents on fertility. These culture systems are well defined in rodent models and allow for toxicant exposure at very specific developmental time points. Additionally, ovarian cell and follicle cultures allow the mechanism of action to be more easily pinpointed in relation to the follicle/oocyte stage affected and the defined outcomes

Ovarian explant culture: effects of smoking constituents

Ovarian explant culture has advantages over isolated follicle culture, as it mimics the effects of the metabolism of the constituents by all ovarian cells. Additionally it allows normal interaction between the follicle and the ovarian stromal cells. Mouse and rat neonatal ovaries have been cultured in the PAH B[a]P, 3-methylcholanthrene (3-MC), DMBA, and its metabolite DMBA-3,4-diol (Ganesan et al., 2013, Ganesan and Keating, 2014, Igawa et al., 2009, Sobinoff et al., 2011, Sobinoff et al.,

Direct systemic exposure to individual cigarette toxicants

Treatment of whole animal models with the individual constituents of cigarette smoke is another method commonly used to elucidate their effects on ovarian health. The advantage of these models is their ability to incorporate metabolism of the chemicals by organs other than the ovary, such as the liver, against the backdrop of the hypothalamic–pituitary axis. Whole animal models also avoid potential artifactual changes that may occur as a result of the in vitro culturing process, such as

Inhalation models of cigarette smoke exposure

Recently mouse inhalation models of cigarette smoke exposure have been used to fully elucidate the negative impact smoking has on ovarian health (Gannon et al., 2012, Gannon et al., 2013, Jennings et al., 2011, Sobinoff et al., 2013, Tuttle et al., 2009). These models have distinct advantages over those previously discussed as they allow exposure to all cigarette smoke components simultaneously via the nasal route, thus closely mimicking human exposure. Two models have been developed: whole

Conclusions

It is widely known that cigarette smoking is linked to a number of diseases and pathologies, including decreased fertility. Human studies have shed some light on this phenomenon, however as the bulk of these studies are performed retrospectively or on ART patients, these findings can be compromised by numerous confounding factors. In an effort to truly understand how smoking affects female fertility, a variety of in vivo and in vitro animal models have been developed. Each of these models has

Conflict of interest

The authors declare that they have no conflict of interest.

Acknowledgments

This work was supported by the Australian Postgraduate award to NJC and Australian Research Council DECRA Award to JEH (DE120101242) and National Health and Medical Research Council to EAM (510735).

Abbreviations

2-EP
2-ethylpyridine
3-EP
3-ethylpyridine
3-MC
3-methylcholanthrene
DMBA
7,12 dimethylbenz[a]anthracene
AMH
anti-Müllerian hormone
AHR
aryl hydrocarbon receptor
ART
assisted reproductive technology
BaA
benzo[a]anthracene
B[a]P
benzo[a]pyrene
BkF
benzo[k]fluoranthene
CSC
cigarette smoke condensate
COCs
cumulus–oocyte-complexes
E2
estradiol
FSH
follicle stimulating hormone
Lc3
light chain protein 3
LH
luteinising hormone
NO-S
nose-only Smoking
PAHs
polycyclic aromatic hydrocarbons
SOD2
superoxide dismutase 2
WB-S
whole body smoking

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