title: Cholesterol depletion with (2-hydroxypropyl)- β-cyclodextrin modifies the gating of membrane electroporation-induced inward current in pituitary tumor GH3 cells: experimental and analytical studies.
abstract: The effects of (2-hydroxypropyl)-β-cyclodextrin (HPβCD), a cyclic oligomer, on membrane electroporation-induced inward current (I(MEP)) in pituitary tumor (GH(3)) cells were experimentally and analytically characterized. Depletion of membrane cholesterol by exposing cells to HPβCD (2 mM) increased the activation time constant of delayed rectifier K(+) current. Such maneuver resulted in a significant reduction of I(MEP) density. 2,2'-Azo-bis(2-amidinopropane) dihydrochloride (AAPH), an initiator of free radicals, increased the magnitude of I(MEP). AAPH-stimulated I(MEP) was not reversed by the blockers of Ca(2+)-activated K(+) channels, but by LaCl(3) or MnCl(2). However, in HPβCD-treated cells, the ability of AAPH to enhance I(MEP) was abolished. Under such maneuver, the gating charge of I(MEP) activation was increased by 2 fold, along with a hyperpolarized shift of the activation curve by 30 mV. No change in single-channel conductance of MEP-induced channels during cell exposure to HPβCD was demonstrated. The energy change of I(MEP) in untreated and HPβCD-treated cells was estimated to be -17.7 and -44.8 kJ/mol, respectively, and the perturbation of free energy following HPβCD treatment was -27.1 kJ/mol. Based on an MEP model, cell exposure to HPβCD increased the edge energy of the electropore size. By use of a two barrier-one site barrier model, HPβCD treatment can increase both the peak height and well depth of the barrier profile. Taken together, depletion of membrane cholesterol by HPβCD can elevate the edge energy of pore formation, thereby decreasing the I(MEP) magnitude. The channel-suppressing properties during membrane cholesterol depletion with HPβCD might thus contribute to the underlying mechanisms by which such maneuver alters neuronal or neuroendocrine function.
authors.affiliation: Department of Physiology, National Cheng Kung University Medical College, Tainan City, Taiwan. snwu@mail.ncku.edu.tw
authors.forename: Sheng-Nan
authors.initials: SN
authors.lastname: Wu
authors.forename: Chia-Chen
authors.initials: CC
authors.lastname: Yeh
authors.forename: Hsien-Ching
authors.initials: HC
authors.lastname: Huang
authors.forename: Wei-Hsin
authors.initials: WH
authors.lastname: Yang
mesh_terms: D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000578:Amidines / Q000494:pharmacology; D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects* / Q000502:physiology; D002784:Cholesterol / Q000378:metabolism* / Q000502:physiology; D018274:Electroporation; D015640:Ion Channel Gating / Q000187:drug effects; D008564:Membrane Potentials / Q000187:drug effects; D010911:Pituitary Neoplasms / Q000503:physiopathology; D051381:Rats; D047392:beta-Cyclodextrins / Q000494:pharmacology*
chemical_list: D000578:Amidines; D000924:Anticholesteremic Agents; D047392:beta-Cyclodextrins; D000073738:2-Hydroxypropyl-beta-cyclodextrin; C046728:2,2'-azobis(2-amidinopropane); D002784:Cholesterol