Developmental changes in the half-life of acetylcholine receptors in the myotomal muscle of Xenopus laevis.
1. Tail preparations, containing myotomal muscle and associated spinal cord, were isolated from embryos and tadpoles of Xenopus laevis between stages 25 and 49 (1.15-12 days) and were pulse-labelled with 125I-alpha-bungarotoxin (125I alpha BT) so that the half-life (T1/2) of their acetylcholine receptors (AChRs) could be estimated in organ culture. 2. For the entire population of AChRs, estimates of T1/2 based on a single exponential decline in radioactivity (but see item 4 below) increased from 53-55 h at stages 25-31 (1.15-1.56 days) to approximately 135 h at stage 47 (5.5 days). Beyond stage 47 T1/2 increased only slightly. 3. Radioautographic estimates of the T1/2 of extrajunctional AChRs at stages 47-48 (5.5-7.5 days) were 41-50 h. It follows that the developmental change in the T1/2 of the entire population of AChRs was due to the junctional AChRs. 4. At stages 47-49 (5.5-12 days) the decline in radioactivity for the entire population of AChRs was fitted well by a double exponential. Assuming a T1/2 of 50 h for the extrajunctional AChRs and 210 h for the junctional AChRs, the correlation coefficient (r) was 0.9947 +/- 0.0014 (mean +/- S.E.M.; n = 14) and junctional AChRs were estimated to comprise 80 +/- 3% of the entire population. Similar analysis, as well as experiments in which the degradation of junctional AChRs was assessed by pulse-labelling with fluorescent alpha-bungarotoxin, suggested that at earlier stages of development the junctional AChRs have a shorter T1/2 and comprise a smaller fraction of the entire population. 5. The developmental increase in T1/2 occurred even when animals were raised in the anaesthetic tricaine or in tetrodotoxin, conditions which abolished all motor activity. 6. Developmental increases in T1/2 also occurred in culture but were smaller than those in vivo. The increases in culture did not occur amongst those AChRs which were pre-labelled with 125I alpha BT. 7. It is concluded that in Xenopus myotomal muscle the T1/2 of junctional AChRs begins to increase within a day after the onset of innervation and that the increase does not require nerve or muscle impulse activity. We suggest, among other possibilities, that it may depend upon incorporation of a different molecular species of AChR into the postsynaptic membrane.
Cohen
M W
MW
Department of Physiology, McGill University, Montreal, Quebec, Canada.
Frair
P F
PF
Cantin
C
C
Hébert
G
G
D062365:Aminobenzoates / Q000494:pharmacology; D000818:Animals; D002038:Bungarotoxins / Q000378:metabolism; D006207:Half-Life; D007814:Larva; D009043:Motor Activity / Q000187:drug effects; D009132:Muscles / Q000378:metabolism*; D009924:Organ Culture Techniques; D011950:Receptors, Cholinergic / Q000378:metabolism*; D013779:Tetrodotoxin / Q000494:pharmacology; D013997:Time Factors; D014982:Xenopus laevis
D062365:Aminobenzoates; D002038:Bungarotoxins; D011950:Receptors, Cholinergic; C003636:tricaine; D013779:Tetrodotoxin