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© The Rockefeller University Press, 0021-9525/1998//473 $5.00
The Journal of Cell Biology, Volume 142, Number 2, , 1998 473-484

Articles

Cyclic Nucleotide-gated Channels on the Flagellum Control Ca2+ Entry into Sperm



Burkhard Wiesner*, Jocelyn Weiner{ddagger}, Ralf Middendorff§, Volker Hagen*, U. Benjamin Kaupp{ddagger}, and Ingo Weyand{ddagger}

* Forschungsinstitut für Molekulare Pharmakologie, D-10315 Berlin; {ddagger} Institut für Biologische Informationsverarbeitung, Forschungszentrum Jülich, D-52425 Jülich; and § Anatomisches Institut, Universitätskrankenhaus Eppendorf, D-20246 Hamburg

Cyclic nucleotide-gated (CNG) channels are key elements of cGMP- and cAMP-signaling pathways in vertebrate photoreceptor cells and in olfactory sensory neurons, respectively. These channels form heterooligomeric complexes composed of at least two distinct subunits ({alpha} and β). The {alpha} subunit of cone photoreceptors is also present in mammalian sperm. Here we identify one short and several long less abundant transcripts of β subunits in testis. The {alpha} and β subunits are expressed in a characteristic temporal and spatial pattern in sperm and precursor cells. In mature sperm, the {alpha} subunit is observed along the entire flagellum, whereas the short β subunit is restricted to the principal piece of the flagellum. These findings suggest that different forms of CNG channels coexist in the flagellum. Confocal microscopy in conjunction with the Ca2+ indicator Fluo-3 shows that the CNG channels serve as a Ca2+ entry pathway that responds more sensitively to cGMP than to cAMP. Assuming that CNG channel subtypes differ in their Ca2+ permeability, dissimilar localization of {alpha} and β subunits may give rise to a pattern of Ca2+ microdomains along the flagellum, thereby providing the structural basis for control of flagellar bending waves.

Key Words: signal transduction • cGMP • fertilization • chemotaxis • caged compounds

Abbreviations used in this paper: aa, amino acid residue; CNC, cyclic nucleotide-gated channel; CNG, cyclic nucleotide-gated; DMNB, 4,5-dimethoxy-2-nitrobenzyl; GARP, glutamic acid-rich protein; MCM, (7-methoxy-coumarin-4-yl)methyl.

Burkhard Wiesner and Jocelyn Weiner contributed equally to this work.

Address all correspondence to Dr. I. Weyand, IBI, Forschungszentrum Jülich, 52425 Jülich, Germany. Tel.: 49-2461-614040; Fax: 49-2461-614216; E-mail: i.weyand{at}fz-juelich.de


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