RECEPTOR-MEDIATED ENDOCYTOSIS OF INSULIN IN LOWER-VERTEBRATES - INTERNALIZATION AND INTRACELLULAR PROCESSING OF I-125 INSULIN IN ISOLATED HEPATOCYTES OF LAMPREY AND FROG
Yl. Lappova et Bn. Leibush, RECEPTOR-MEDIATED ENDOCYTOSIS OF INSULIN IN LOWER-VERTEBRATES - INTERNALIZATION AND INTRACELLULAR PROCESSING OF I-125 INSULIN IN ISOLATED HEPATOCYTES OF LAMPREY AND FROG, General and comparative endocrinology, 100(1), 1995, pp. 1-9
The binding of I-125-insulin to cellular insulin receptors and the int
ernalization of insulin-receptor complexes have been studied in isolat
ed hepatocytes of frog and lamprey. Two classes of binding sites (K-d
10(-9) and 10(-8) M) were found in cells of both species. The molecula
r weight of the insulin receptor a-subunit was 130 kDa in both species
. Internalization of bound I-125-insulin in both species was found in
the temperature range 0 to 20 degrees Cells ''loaded'' with I-125-insu
lin were used to estimate the fate of the internalized ligand. Release
of internalized ligand from frog cells increased at temperatures rang
ing from 0 to 20 degrees. At 0 degrees the degraded I-125-insulin was
5%, at 5 degrees 7%, and at 20 degrees 17% of total radioactivity accu
mulated in the medium. In lamprey hepatocytes there was neither radioa
ctivity accumulation in the incubation medium nor release from cells a
t all temperatures studied. The intracellular degradation of internali
zed I-125-insulin in frog hepatocytes was much lower than that in lamp
rey cells. In frog hepatocytes the specific binding of I-125-insulin w
as increased twofold in the presence of the lysosomal inhibitor chloro
quine. In contrast no increase was found in lamprey hepatocytes. In co
nclusion, the processing pathways of internalized insulin in the cells
of ectothermal and endothermal vertebrates are generally similar but
in ectothermal animals all events take place at lower temperatures and
at lower rates. The peculiarities of insulin processing in lamprey he
patocytes most likely result from the transformation of hepatocytes du
ring the nonfeeding prespawning period. (C) 1995 Academic Press, Inc.