Multiplication-stimulating activity (MSA) stimulates the uptake of xylose and alpha-aminoisobutyric acid (AIB) by intact rat soleus muscle. It is approximately 50 times less potent than insulin. A native insulin receptor species with apparent Mr = 350,000 in soleus muscle is revealed by affinity cross-linking to 125I-insulin with disuccinimidyl suberate. 125I-labeled insulin-like growth factor (IGF) I with the cross-linker affinity labels two native receptor types with Mr = 360,000 (type I) and Mr = 220,000 (type II). In order to distinguish which of the three insulin and IGF receptor systems identified mediate the rapid insulin-like effects of MSA, the biological actions of the unlabeled ligands at various concentrations were correlated with their ability to inhibit the affinity labeling of these receptor species. The stimulatory action of native insulin is closely related to its ability to inhibit the labeling of the insulin receptor by 125I-insulin such that the uptake of xylose and AIB is maximally stimulated when 80% of insulin receptor 125I-affinity labeling is inhibited. In contrast, MSA only displaced 16% of the 125I-labeling of the insulin receptor when it maximally stimulates the uptake of xylose and AIB, indicating that the insulin receptor is not primarily involved in mediating these effects. The affinity of MSA to the type II IGF receptor is 10 times higher than that to the type I IGF receptor. There is close to a 1:1 relationship between the stimulatory effects of MSA on xylose and AIB uptake and its inhibitory action on the affinity labeling of the type I receptor by 125I-IGF I. In contrast, MSA almost abolishes the labeling of the type II IGF receptor at concentrations which have no detectable effects on the uptake of xylose and AIB by soleus muscle. Thus, a marked dissociation can be observed between the rapid insulin-like action of MSA and its inhibitory effects on the affinity labeling of the type II receptor by 125I-IGF I. We conclude that MSA acts through the type I IGF receptor in soleus muscle to stimulate hexose and amino acid transport. The type II IGF receptor appears to be incapable of modulating these effects in this tissue.