Estrogen effects on high-affinity choline uptake in primary cultures of rat basal forebrain.
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 muM HC-3, but not higher concentrations that inhibit LACU, could distinguish the two transport activities. 17-beta-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.
Bennett
Katie M
KM
Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center, School of Pharmacy, 1300 S Coulter, Amarillo, TX 79106, USA.
Hoelting
Courtney
C
Martin
Christopher P
CP
Stoll
James
J
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002794:Choline / Q000378:metabolism*; D017931:DNA Primers; D004967:Estrogens / Q000494:pharmacology*; D007150:Immunohistochemistry; D016548:Prosencephalon / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D004967:Estrogens; D002794:Choline