In autoradiographic studies in anesthetized rats, 125I-labeled amylin binding was associated with proximal convoluted tubules but not distal tubules, interstitium, or glomeruli in the renal cortex. Split-drop micropuncture experiments showed that perfusion of the peritubular capillaries with amylin (10(-9) M) stimulated proximal tubular fluid absorption by 28%. This effect was inhibited by luminal addition of ethylisopropylamiloride, indicating mediation by a brush-border Na+/H+ exchanger. Intravenous infusion of an amylin binding antagonist, AC-187, reduced proximal fluid reabsorption (22%) in anesthetized rats, indicating a role for endogenous amylin in salt homeostasis. In primary cultures of rat proximal tubule cells, amylin (10(-7) M) stimulated proliferation with a potency equal to epidermal growth factor. Peptide antagonists (AC-187, AC-413, and AC-512) of the amylin binding sites in the renal cortex blocked the mitogenic action of amylin. We conclude that amylin acts on renal proximal tubules to promote sodium and water reabsorption and cell proliferation. These novel actions may have implications for the development of hypertension for example in non-insulin-dependent diabetes mellitus and obesity in which hyperamylinemia has been observed.

Chorionic gonadotropin (CG), a pregnancy-specific heterodimeric hormone found in primates, is responsible for CL rescue with pregnancy maintenance. Of the primates, the human and baboon gene sequences are the only structures so far determined. In order to study the structure and function of CG in other primates, we have isolated and sequenced the coding regions for the two subunits of marmoset CG (mCG) by the reverse transcription/polymerase chain reaction method. Study of multiple clones confirmed a high degree of homology with the human sequences (88% and 80% for the alpha and beta nucleotide sequences, respectively). Marmoset CG alpha has an extra four amino acids compared to hCG alpha, whereas the mCG beta sequence has a 3-bp deletion that maintains the reading frame and C-terminal amino acid sequence. Most of the differences between hCG beta and mCG beta peptides occur in the C-terminal region, which includes the loss of two of the O-linked glycosylation consensus sequences and the presence of an N-linked glycosylation consensus sequence. When mCG alpha and beta were co-expressed in CHO cells, assembly of biologically active hormone was confirmed by induced steroid secretion by MA10 cells. Partially purified mCG beta was used to raise anti-mCG antibodies. To date, an antibody has been obtained that is capable of detecting recombinant mCG beta, recombinant mCG dimer, and mCG dimer secreted by cultured marmoset trophoblast. Marmoset CG alpha and beta were also detectable at the transcriptional level in cultured trophoblast by in situ hybridization. This suggests that the LH/CG bioactivity reported from marmoset placentae and embryos is due to a molecule with structural features common to hLH (glycosylation pattern) and hCG (CG beta C-terminal structure).

Insulin-like growth factor-I (IGF-I) is a peptide growth factor whose activity is modulated by interaction with the family of six IGF binding proteins (IGFBPs). IGF-I is detected in rat kidney and has metabolic and growth effects. We have used in situ hybridization to localize mRNAs for the IGFBPs in rat kidney. Messenger RNAs for all six IGFBPs were detected, each with a distinctive distribution. IGFBP-1 mRNA was expressed in the distal nephron, from the thick ascending limb of the loop of Henle to the cortical collecting ducts. IGFBP-2 mRNA expression was confined to epithelial cells of the glomeruli and the thin limbs of the loop of Henle. IGFBP-3 mRNA was localized to the cortical interstitium while IGFBP-4 was the only IGFBP mRNA found in the proximal tubule. IGFBP-5 mRNA, the most abundant and widely distributed of the IGFBP mRNAs in the kidney, occurred in the glomerular mesangium and the medullary interstitium as well as in the epithelial cells of the distal nephron. IGFBP-6 mRNA, the least abundant, was expressed mainly in fibroblasts associated with renal blood vessels and the ureter. This heterogeneous distribution of the IGFBPs may enable IGF action to be regulated by multiple factors in a site-specific manner.

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