Subject(s)
Anti-HIV Agents/adverse effects , Chemical and Drug Induced Liver Injury/etiology , HIV Seropositivity/complications , Hydroxyurea/adverse effects , Adult , Anti-HIV Agents/therapeutic use , Female , HIV Seropositivity/drug therapy , Humans , Hydroxyurea/therapeutic use , Male , Middle AgedABSTRACT
We investigated the role of microtubules in the compartmentation of motility and endocytosis in the neurite shaft and growth cone of cultured chick sensory neurons. As reported previously by Letourneau and Ressler (J. Cell Biol., 98 (1984) 1355-1362), stimulating microtubule polymerization with taxol inhibits growth cone motility. In neurons that had grown for 18-30 h, taxol treatment caused growth cones to round up forming an obvious varicosity (taxol bulb) at the terminal. Removal of taxol allowed nearly immediate resumption of cortical motility in all 17 neurons observed by time-lapse videomicroscopy. However, only one of the 17 neurites was observed to elongate measurably even after 5 h of observation. In 14 cases taxol was rinsed out and the concentration of nerve growth factor was increased 10x, 11/14 neurites retracted within the next hour. Endocytic activity was investigated by incubating control and taxol treated neurons in either cationized ferritin or horseradish peroxidase for 30 min. The number and area of label-containing vesicles was measured along with the total area of the growth cone or taxol bulb. We found that taxol treatment caused a 7-fold decrease in the ratio of the area of the labeled vesicles to the area of growth cone or taxol bulb. Conversely, in neurite shafts, normally relatively quiescent with respect to endocytosis, those regions devoid of microtubules in both control and nocodazole-treated cells contained a high concentration of label-containing vesicles. We conclude that the presence of microtubules plays a role in regulating endocytic activity by the overlying cell cortex.(ABSTRACT TRUNCATED AT 250 WORDS)
Subject(s)
Cytoplasm/physiology , Endocytosis/drug effects , Microtubules/physiology , Neurons, Afferent/physiology , Alkaloids/pharmacology , Animals , Cells, Cultured , Chick Embryo , Cytoplasm/drug effects , Cytoplasm/ultrastructure , Dendrites/drug effects , Dendrites/physiology , Dendrites/ultrastructure , Ferritins/metabolism , Ganglia, Spinal/cytology , Ganglia, Spinal/physiology , Ganglia, Spinal/ultrastructure , Horseradish Peroxidase/metabolism , Microscopy, Electron , Microtubules/drug effects , Microtubules/ultrastructure , Neurons, Afferent/drug effects , Neurons, Afferent/ultrastructure , PaclitaxelABSTRACT
We have examined the role of microtubules (MTs) in the development and maintenance of the cytoplasmic compartmentation of ribosomes in cultured embryonic chick sensory neurons. Control neurons show a sharp cytoplasmic demarcation between the ribosome-rich/MT-deficient 'translational' cytoplasm of the perikaryon and the ribosome-deficient/MT-rich 'expressional' cytoplasm of the axon-like neurites. Neurites treated with 1.0 microgram/ml nocodazole for 15-20 min show in addition to the depolymerization of MTs a clear loss of the cytoplasmic demarcation between the soma and neurite. We found a decreasing density of ribosomes with increasing distance from the soma in the nocodazole treated neurites. Ribosomes, as confirmed by their diameter and sensitivity to ribonuclease treatment, populated the first 100 microns of neurites at roughly the same density as found in the soma (150-300 ribosomes per microns2 in 50 mm longitudinal thin sections). The second 100 microns contained a lower density of ribosomes than the first, 20-60 per microns2, but still higher than controls. Distances past 200 microns were devoid of unambiguous ribosomes. Neurites amputated between 50 and 150 microns from the soma then treated with nocodazole contained a dense population of ribosomes in the proximal segment still attached to the soma, but few or none in the distal segment. We interpret these findings as evidence that ribosomes are able to move from the soma down the neurite after MT disassembly.(ABSTRACT TRUNCATED AT 250 WORDS)
