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doi: 10.1242/10.1242/dev.00353
1 Department of Physiology and Biophysics, Box 357290, University of Washington,
Seattle, WA 98195-7290, USA
2 Department of Anatomy and Cell Biology, Philipps University Marburg, 35037
Marburg, Germany
3 Department of Obstetrics and Gynecology, Box 356460, University of Washington,
Seattle, WA 98195-6460, USA
* Author for correspondence (e-mail: donner{at}u.washington.edu)
Accepted 16 December 2002
At mating, mammalian sperm are diluted in the male and female reproductive
fluids, which brings contact with HCO3- and initiates
several cellular responses. We have identified and studied two of the most
rapid of these responses. Stop-motion imaging and flagellar waveform analysis
show that for mouse epididymal sperm in vitro, the resting flagellar beat
frequency is 2-3 Hz at 22-25°C. Local perfusion with
HCO3- produces a robust, reversible acceleration to 7 Hz
or more. At 15 mM the action of HCO3- begins within 5
seconds and is near-maximal by 30 seconds. The half-times of response are
8.8±0.2 seconds at 15 mM HCO3- and
17.5±0.4 seconds at 1 mM HCO3-. Removal of
external HCO3- allows a slow return to basal beat
frequency over 
10 minutes. Increases in beat symmetry accompany the
accelerating action of HCO3-. As in our past work,
HCO3- also facilitates opening of voltagegated
Ca2+ channels, increasing the depolarization-evoked rate of rise of
intracellular Ca2+ concentration by more than fivefold. This action
also is detectable at 1 mM HCO3- and occurs with an
apparent halftime of 60 seconds at 15 mM HCO3-. The
dual actions of HCO3- respond similarly to
pharmacological intervention. Thus, the phosphodiesterase inhibitor IBMX
promotes the actions of HCO3- on flagellar and channel
function, and the protein kinase A inhibitor H89 blocks these actions. In
addition, a 30 minute incubation with 60 µM cAMP acetoxylmethyl ester
increases flagellar beat frequency to nearly 7 Hz and increases the evoked
rates of rise of intracellular Ca2+ concentration from 17±4
to 41±6 nM second-1. However, treatment with several other
analogs of cAMP produces only scant evidence of the expected mimicry or
blockade of the actions of HCO3-, perhaps as a
consequence of limited permeation. Our findings indicate a requirement for
cAMP-mediated protein phosphorylation in the enhancement of flagellar and
channel functions that HCO3- produces during sperm
activation.
Key words: Capacitation, sAC, cAMP-AM, PKA, Motility, Shear angle, Tan angle, Asymmetry
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