Laser microsurgery permits fungal plasma membrane single-ion-channel resolution at the hyphal tip.
نویسندگان
چکیده
A method for formation of high-electrical-resistance seals on the Neurospora crassa plasma membrane, allowing resolution of single-ion-channel activity by patch clamp electrophysiology, is reported. Laser microsurgery permits access to the hyphal apex without enzymatic cell wall digestion and loss of morphological polarity. Cell wall reformation is delayed by brefeldin. This method can allow full characterization of apical plasma membrane channels, which are implicated in tip growth.
منابع مشابه
The roles of Ca2+ and plasma membrane ion channels in hyphal tip growth of Neurospora crassa.
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 recordings in the cell-attached mode, from the plasma membrane of these hyphae, showed two types of channel activities; spontaneous and stretch activated. The spontaneous channels were identifie...
متن کاملAn IP3-activated Ca2+ channel regulates fungal tip growth.
Hyphal extension in fungi requires a tip-high Ca(2+) gradient, which is generated and maintained internally by inositol (1,4,5)-trisphosphate (IP(3))-induced Ca(2+) release from tip-localized vesicles and subapical Ca(2+) sequestration. Using the planar bilayer method we demonstrated the presence of two types of IP(3)-activated Ca(2+) channels in Neurospora crassa membranes with different condu...
متن کاملMesA, a Novel Fungal Protein Required for the Stabilization of Polarity Axes in Aspergillus nidulans□D
The Aspergillus nidulans proteome possesses a single formin, SepA, which is required for actin ring formation at septation sites and also plays a role in polarized morphogenesis. Previous observations imply that complex regulatory mechanisms control the function of SepA and ensure its correct localization within hyphal tip cells. To characterize these mechanisms, we undertook a screen for mutat...
متن کاملMesA, a novel fungal protein required for the stabilization of polarity axes in Aspergillus nidulans.
The Aspergillus nidulans proteome possesses a single formin, SepA, which is required for actin ring formation at septation sites and also plays a role in polarized morphogenesis. Previous observations imply that complex regulatory mechanisms control the function of SepA and ensure its correct localization within hyphal tip cells. To characterize these mechanisms, we undertook a screen for mutat...
متن کاملThe Neurospora crassa exocyst complex tethers Spitzenkörper vesicles to the apical plasma membrane during polarized growth
Fungal hyphae are among the most highly polarized cells. Hyphal polarized growth is supported by tip-directed transport of secretory vesicles, which accumulate temporarily in a stratified manner in an apical vesicle cluster, the Spitzenkörper. The exocyst complex is required for tethering of secretory vesicles to the apical plasma membrane. We determined that the presence of an octameric exocys...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Applied and environmental microbiology
دوره 64 4 شماره
صفحات -
تاریخ انتشار 1998