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Plant and Cell Physiology Advance Access published online on February 2, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci063
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received October 26, 2004
Accepted January 18, 2005

Regular Paper

Ca2+ Influx into Lily Pollen Grains through a Hyperpolarization-Activated Ca2+ Permeable Channel, Which Can Be Regulated by Extracellular CaM

Zhong-lin Shang 1, Li-geng Ma 2, Hai-lin Zhang 3, Rui-rong He 4, Xue-chen Wang 5, Su-juan Cui 2, and Da-ye Sun 2*

1 Institute of Molecular & Cell Biology, Hebei Normal University, Shijiazhuang 050016, PR China; National Key Laboratory of Plant Physiology & Biochemistry, Beijing 100094, PR China
2 Institute of Molecular & Cell Biology, Hebei Normal University, Shijiazhuang 050016, 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 National Key Laboratory of Plant Physiology & Biochemistry, Beijing 100094, PR China

* To whom correspondence should be addressed.
Da-ye Sun, E-mail: dysun{at}heinfo.net


  Abstract

Confocal laser scanning microscopy (CLSM) and whole-cell patch clamp was used to investigate the role of Ca2+ influx in maintaining cytosolic Ca2+ concentration ([Ca2+]c) and the feature of Ca2+ influx pathway in germinating pollen grains of Lilium davidiiD. [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 plasma membrane of pollen protoplasts with whole-cell patch clamp recording. CaM antiserum and W7-agarose, both of which are cell-impermeable CaM antagonist, lead to [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 activation of Ca2+ conductance which induced by exogenous CaM were inhibited by EGTA or Ca2+ channel blockers. These primary evidences firstly showed the presence of a voltage-dependent Ca2+ permeable channel, whose activity may be regulated by extracellular CaM, in pollen cells.

Keywords: cytosolic calcium; Lilium davidii; pollen grain; hyperpolarization-activated Ca2+ permeable channel; extracellular CaM.
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