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Plant and Cell Physiology Advance Access originally published online on February 2, 2005
Plant and Cell Physiology 2005 46(4):598-608; doi:10.1093/pcp/pci063
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JSPP © 2005

Ca2+ Influx into Lily Pollen Grains Through a Hyperpolarization-activated Ca2+-permeable Channel Which Can be Regulated by Extracellular CaM

Zhong-lin Shang1,2, Li-geng Ma1, Hai-lin Zhang3, Rui-rong He4, Xue-chen Wang2, Su-juan Cui1 and Da-ye Sun1,5

1 Institute of Molecular and Cell Biology, Hebei Normal University, Shijiazhuang 050016, PR China
2 National Key Laboratory of Plant Physiology and Biochemistry, Beijing 100094, PR China
3 Department of Pharmacology, Hebei Medical University, Shijiazhuang 050016, PR China
4 Department of Physiology, Hebei Medical University, Shijiazhuang 050016, PR China

5 Corresponding author: E-mail, dysun{at}heinfo.net; Fax, +86-311-5820649.

Confocal laser scanning microscopy (CLSM) and whole-cell patch-clamp were used to investigate the role of Ca2+ influx in maintaining the cytosolic Ca2+ concentration ([Ca2+]c) and the features of the Ca2+ influx pathway in germinating pollen grains of Lilium davidii D. [Ca2+]c decreased when Ca2+ influx was inhibited by EGTA or Ca2+ channel blockers. A hyperpolarization-activated Ca2+-permeable channel, which can be suppressed by trivalent cations, verapamil, nifedipine or diltiazem, was identified on the plasma membrane of pollen protoplasts with whole-cell patch-clamp recording. Calmodulin (CaM) antiserum and W7-agarose, both of which are cell-impermeable CaM antagonists, lead to a [Ca2+]c decrease, while exogenous purified CaM triggers a transient increase of [Ca2+]c and also remarkably activated the hyperpolarization-activated Ca2+ conductance on plasma membrane of pollen protoplasts in a dose-dependent manner. Both the increase of [Ca2+]c and the activation of Ca2+ conductance which were induced by exogenous CaM were inhibited by EGTA or Ca2+ channel blockers. This primary evidence showed the presence of a voltage-dependent Ca2+-permeable channel, whose activity may be regulated by extracellular CaM, in pollen cells.

Received October 26, 2004; Accepted January 18, 2005
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