Nn. Levina et al., THE ROLES OF CA2-MEMBRANE ION CHANNELS IN HYPHAL TIP GROWTH OF NEUROSPORA-CRASSA( AND PLASMA), Journal of Cell Science, 108, 1995, pp. 3405-3417
Growing hyphae of the ascomycete fungus Neurospora crassa contained a
tip-high gradient of cytoplasmic Ca2+, which was absent in non-growing
hyphae and was insensitive to Gd3+ in the medium. Patch clamp recordi
ngs in the cell-attached mode, from the plasma membrane of these hypha
e, showed two types of channel activities; spontaneous and stretch act
ivated. The spontaneous channels were identified as inward K+ channels
based on inhibition by tetraethylammonium. The stretch activated chan
nels had increased amplitudes in response to elevated Ca2+ in the pipe
tte solution, and thus are permeable to Ca2+ and mediate inward Ca2+ m
ovement. Gd3+ which is an inhibitor of some stretch activated channels
, incompletely inhibited stretch activated channel activity. Both tetr
aethylammonium and Gd3+ only transiently reduced the rates of tip grow
th without changing tip morphology, thus indicating that the channels
are not absolutely essential for tip growth. Furthermore, in contrast
to the hyphae of another tip growing organism, Saprolegnia ferax, tip-
high gradients of neither spontaneous nor stretch activated channels w
ere found. Voltage clamping of the apical plasma membrane potential in
the range from -300 to +150 mV did not affect the rates of hyphal elo
ngation. Collectively, these data suggest that ion transport across th
e plasma membrane at the growing tip in Neurospora is not obligatory f
or the maintainance of tip growth, but that a gradient of Ca2+, possib
ly generated from internal stores in an unknown way, is required.