Elsevier

Neuroscience

Volume 105, Issue 3, 15 August 2001, Pages 527-534
Neuroscience

Nicotine-induced alterations in the expression of nicotinic receptors in primary cultures from human prenatal brain

https://doi.org/10.1016/S0306-4522(01)00209-3Get rights and content

Abstract

The nicotinic receptor proteins and gene transcripts for the different nicotinic receptor subunits exist in human prenatal brain already at 4–5 weeks of gestation. The early presence of nicotinic receptors suggests an important role for these receptors in modulating dendritic outgrowth, establishment of neuronal connections and synaptogenesis during development. When measurements of nicotinic receptors using [3H]epibatidine (labelling both the α3 and α4 subtype) and [3H]cytisine (labelling the α4 subtype) were performed in intact cells from the cortex, subcortical forebrain and mesencephalon (7.5–11 weeks of gestation), the highest specific binding for both ligands was detected in cells from mesencephalon, followed by subcortical forebrain and cortex. The effects of nicotine exposure were studied in primary cultures of prenatal brain (7.5–11 weeks of gestation). Treatment with nicotine (1–100 μM) for 3 days significantly increased the specific binding of [3H]epibatidine and [3H]cytisine in cortical cells but not in cells from subcortical forebrain and mesencephalon brain regions, indicating region-specific differences in the sensitivity to nicotine exposure. Relative quantification of mRNA showed that the expression of the nicotinic receptor subunits α3 and α7, but not α4, was increased in cortical cells after nicotine treatment.

These findings support the assumption of a potential risk of disturbance in the functional role of nicotinic receptors during brain development as a consequence of maternal smoking during pregnancy.

Section snippets

Brain tissues

First trimester brain tissue from humans (7.5–11 weeks of gestation) was obtained following routine abortions by vacuum aspiration after the pregnant women’s consent. Brain regions were dissected under sterile conditions in Ringer’s solution within 1–1.5 h of surgical tissue retrieval and separated on the basis of their morphological appearance according to Seiger (1989). Subcortical forebrain is defined as all CNS tissue rostral to mesencephalon, with the exception of the neocortical anlage,

[3H]Epibatidine and [3H]cytisine binding to intact primary neuronal cells from human fetal brain

Specific binding of [3H]epibatidine and [3H]cytisine to intact primary cell cultures from cortex, subcortical forebrain and mesencephalon of 7.5–11 weeks gestational age was measured using a single concentration (0.2 and 2.0 nM, respectively). The specific binding of both ligands was highest in cultured cells from the mesencephalon (approximately 30 fmol/mg protein) followed by the subcortical forebrain (15–18 fmol/mg protein) and cortical cells (11 fmol/mg protein) (Table 1). In saturation

Discussion

Developmental animal models exposing the fetus to nicotine by itself and without involvement of other confounding factors of smoking have demonstrated nicotine being neuroteratogenic (Slotkin, 1998). Direct specific actions of nicotine on the developing brain are quite plausible during the major part of prenatal life, since the nAChRs are already detectable in brain and spinal cord at 4–5 weeks gestational age (Hellström-Lindahl et al., 1998). It is believed that nicotine exposure of prenatal

Conclusion

This study shows that it is possible to detect and characterize nAChRs in primary cell cultures from human fetal brain and that nicotine exposure to these cells at relevant concentrations up-regulates nAChR binding sites and increases the expression of their transcripts. These findings suggest that a potential risk exists that the normal functional role of nAChRs during brain development might be disturbed as a consequence of maternal smoking or treatment with transdermal nicotine patches

Acknowledgements

This work was supported by grants from the Swedish Medical Research Council (Project numbers 13426, 05817 and 06555), Council for Medical Tobacco Research, Swedish Match, Magnus Bergwall Foundation, Alcohol Research Council for the Swedish Alcohol Retailing Monopoly, Stiftelsen Gamla Tjänarinnor, Åke Wiberg’s Foundation and KI Foundations.

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