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Estrogen Effects on High-Affinity Choline Uptake in Primary Cultures of Rat Basal Forebrain

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Abstract

Basal forebrain cholinergic neurons (BFCNs) degenerate in aging and Alzheimer’s disease. It has been proposed that estrogen can affect the survival and function of BFCNs. This study characterized primary rat BFCN cultures and investigated the effect of estrogen on high-affinity choline uptake (HACU). BFCNs were identified by immunoreactivity to the vesicular acetylcholine transporter (VAChT) and represented up to 5% of total cells. HACU was measured in living BFCN cultures and differentiated from low-affinity choline uptake by hemicholinium-3 (HC-3) inhibition. A HC-3 concentration curve showed that 0.3 μM HC-3, but not higher concentrations that inhibit LACU, could distinguish the two transport activities. 17-β-Estradiol treatment increased HACU in some culture preparations that contained non-neuronal cells. Elimination of dividing cells using antimitotic treatments resulted in a lack of estrogen effects on HACU. These results suggest that estrogen may have indirect effects on BFCNs that are mediated through non-neuronal cells.

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Abbreviations

AD:

Alzheimer’s disease

BFCNs:

Basal forebrain cholinergic neurons

ChAT:

Choline acetyltransferase

HACU:

High-affinity choline uptake

HC-3:

Hemicholinium-3

LACU:

Low-affinity choline uptake

NGF:

Nerve growth factor

VAChT:

Vesicular acetylcholine transporter

VAChT-IR:

VAChT-immunoreactive

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Acknowledgments

This research was supported by NIH RO3 AG21735 (J.S.), the Laura W. Bush Institute for Women’s Health (J.S.), the Achievement Rewards for College Scientists scholarship foundation (K.M.B.), and the Merck/AFAR Summer Research Scholarship in Geriatric Pharmacology (C.H. and C.M.). We would like to thank Dr. Joachim Hartmann for his expertise and Dr. Jochen Klein for providing access to instruments and for critical reading of the manuscript.

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  1. Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center, School of Pharmacy, 1300 S Coulter, Amarillo, TX, 79106, USA

    Katie M. Bennett, Courtney Hoelting, Christopher P. Martin & James Stoll

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  1. Katie M. Bennett
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Correspondence to James Stoll.

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Bennett, K.M., Hoelting, C., Martin, C.P. et al. Estrogen Effects on High-Affinity Choline Uptake in Primary Cultures of Rat Basal Forebrain. Neurochem Res 34, 205–214 (2009). https://doi.org/10.1007/s11064-008-9746-1

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  • DOI: https://doi.org/10.1007/s11064-008-9746-1

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