Responses of growth cones to changes in osmolality of the surrounding medium.
The possible involvement of osmotically generated hydrostatic pressure in driving actin-rich extensions of the cell surface was examined using cultures of chick neurons. Estimation of the excess internal osmotic pressure of chick neural tissue by vapor pressure deficit osmometry, and of the excess internal hydrostatic pressure in cultured chick neurons using a calibrated pressure pipette, gave upper limits of 10 mosM and 0.1 atmosphere (1 atmosphere = 101325 Pa), respectively. Increases in the osmolality of the medium surrounding cultured neurons by addition of sucrose, mannitol or polyethylene glycol by amounts that should eliminate any internal pressure not only failed to arrest the growth of filopodia but caused them to increase in length up to twofold in 3-5 min. Lamellipodia remained unchanged following hyperosmotic shifts of 20 mosM, but higher levels caused a small decrease in area. Reduction of osmolality by the addition of water to the culture fluid down to 50% of its normal value failed to show any detectable change in either filopodial length or lamellipodia area. These observations argue against an osmotic mechanism for growth cone extension and show that the growth of filopodia, in particular, is unlikely to be driven by osmotically generated hydrostatic pressure. In contrast to the short-term effects on growth cone morphology, the slower elongation of the neuritic cylinder showed a consistent osmotic response. Growth rates were reduced following addition of osmolytes and increased in rate (as much as sixfold) following addition of water to the culture medium.(ABSTRACT TRUNCATED AT 250 WORDS)
Bray
D
D
Department of Biochemistry, Colorado State University, Fort Collins 80523.
Money
N P
NP
Harold
F M
FM
Bamburg
J R
JR
D000199:Actins / Q000032:analysis; D000818:Animals; D001369:Axons / Q000187:drug effects* / Q000648:ultrastructure; D002462:Cell Membrane / Q000187:drug effects / Q000648:ultrastructure; D002642:Chick Embryo; D003470:Culture Media / Q000737:chemistry* / Q000494:pharmacology; D046508:Culture Techniques; D003593:Cytoplasm / Q000737:chemistry / Q000187:drug effects; D005724:Ganglia / Q000187:drug effects* / Q000196:embryology / Q000648:ultrastructure; D006874:Hydrostatic Pressure; D007091:Image Processing, Computer-Assisted; D008353:Mannitol; D009994:Osmolar Concentration; D009997:Osmotic Pressure; D011092:Polyethylene Glycols; D013997:Time Factors
D000199:Actins; D003470:Culture Media; D008353:Mannitol; D011092:Polyethylene Glycols