We have investigated the hypothesis that there is local regulation of insulin-like growth factor (IGF) gene expression during skeletal muscle growth. Compensatory hypertrophy was induced in the soleus, a predominantly slow-twitch muscle, and plantaris, a fast-twitch muscle, in 11- to 12-wk-old female Wistar rats by unilateral cutting of the distal gastrocnemius tendon. Animals were killed 2, 4, or 8 days later, and muscles of the nonoperated leg served as controls. Muscle weight increased throughout the experimental period, reaching 127% (soleus) or 122% (plantaris) of control values by day 8. In both growing muscles, IGF-I mRNA, quantitated by a solution-hybridization nuclease-protection assay, rose by nearly threefold on day 2 and remained elevated throughout the experimental period. IGF-II mRNA levels also increased over controls. A more dramatic response was seen in hypophysectomized rats, where IGF-I mRNA levels rose by 8- to 13-fold, IGF-II values by 3- to 7-fold, and muscle mass increased on day 8 to 149% (soleus) or 133% (plantaris) of the control contralateral limb. These results indicate that signals propagated during muscle hypertrophy enhance the expression of both IGF genes, that modulation of IGF-I mRNA levels can occur in the absence of growth hormone, and that locally produced IGF-I and IGF-II may play a role in skeletal muscle growth.

The role of polypeptide growth factors in promoting muscle differentiation is uncharacterized. We have used a fusing skeletal muscle cell line, C2, to examine the endogenous expression of one peptide, insulin-like growth factor II (IGF-II), and its receptor during differentiation. The synthesis of IGF-II is low during proliferation of myoblasts; IGF-ll mRNA can be detected only through use of a highly sensitive solution-hybridization assay. Competition binding studies reveal that the IGF-II receptor is similarly nonabundant in myoblasts. During differentiation IGF-ll mRNA rises rapidly. A nearly 4-fold increase is seen within 16 hr of onset of the differentiation process, and levels are 25 times higher than those in myoblasts by 96 hr, when myotubes have formed and muscle-specific a-actin mRNAs are synthesized. IGF-H accumulates in conditioned culture medium with similar kinetics. The expression of IGF-II receptors on the cell surface increases almost 6-fold 24 hr after the onset of differentiation and remains high. These studies suggest that IGF-II and its receptor are coordinately regulated during myogenic differentiation in C2 cells and that IGF-ll may be an autocrine factor for skeletal muscle.The formation of skeletal muscle involves proliferation of a committed stem cell, the myoblast, and subsequent differentiation and fusion into myotubes. Recent studies have suggested (1, 2) that commitment to the myoblast lineage is a consequence of induction of a limited number of genes whose expression is required for the later ordered activation of the differentiation program. Although the phenomenology of myoblast differentiation is well described, the mechanisms involved at the onset of myogenesis in withdrawal from the cell cycle or terminally in expression of muscle-specific proteins (3-6) are not understood. In this context, it has been shown that specific mitogens and activated oncogenes inhibit myogenesis (7)(8)(9)(10)(11)(12)(13). Whether other growth factors are capable of promoting muscle differentiation or whether specific differentiation agents are produced by skeletal muscle cells has not been established.The insulin-like growth factors (IGFs) I and II are structurally related polypeptides with several actions on muscle. IGFs appear able to stimulate both proliferation and differentiation of myoblasts at subnanomolar concentrations and are able to promote nutrient uptake and inhibit proteolysis in muscle cell lines (14-17). In some of these actions, IGF-I is nearly an order of magnitude more potent than IGF-II. It has been suggested that both growth factors exert their effects on myoblasts through the IGF-I receptor (15, 18), an IGFsensitive tyrosine-specific protein kinase structurally similar to the insulin receptor (19). In contrast, the role of the IGF-II receptor in IGF action is less clear, although this protein appears to be identical to the cation-independent mannose-6-phosphate receptor involved in lysosomal enzyme targeting (20)(21)(22)(23)(24).Several studies have suggested that IGFs ...

Proteinuria and tubulointerstitial inflammation (TII) correlate with progression to renal failure in human glomerulonephritis. Various forms of experimental nephrotic syndrome are associated with TII. To study the genesis of TII, we utilized the model of albumin overload. Rats received intraperitoneal bovine serum albumin (BSA) for 1 to 14 days, developing heavy proteinuria. A predominantly macrophage interstitial infiltrate was present at days 3, 7 and 14. The urine of the rats contained a factor chemotactic for macrophages which partitioned into the organic phase with ethyl acetate extraction. TLC and HPLC characteristics were those of a novel, non-polar lipid. Supernatant from the culture of proximal tubule (PT) segments after in vivo or in vitro exposure to high concentrations of lipid-replete BSA showed chemotactic activity with similar chromatographic characteristics. PT cultured with delipidated BSA produced little activity. Thus, the generation of this inflammatory factor occurs as a consequence of tubular metabolism of albumin-borne fatty acids and may contribute to the development of proteinuria-associated TII.

We examined the regulation of insulin-like growth factor I (IGF-I) in kidney during the renal hypertrophy produced by two different experimental models: growth hormone treatment of hypophysectomized rats and compensatory hypertrophy subsequent to unilateral nephrectomy. Immunostaining for IGF-I in collecting ducts was enhanced in kidneys from growth hormone-repleted hypophysectomized rats, and the levels of IGF-I mRNAs were increased. In compensatory hypertrophy, no enhancement of the intensity of immunostaining was observed in kidneys of nephrectomized rats until 5 days postnephrectomy, at which time immunostainable IGF-I was increased markedly in medullary collecting ducts of hypertrophied kidneys compared with kidneys from sham-operated animals. No difference in steady-state levels of any IGF-I mRNA species was detected in whole kidneys or in collecting ducts from nephrectomized or sham-operated rats at any time postnephrectomy. Our findings demonstrate an increase in both IGF-I mRNA and in immunostainable IGF-I in collecting duct in the setting of growth hormone-induced renal hypertrophy but suggest that other, possibly translational, mechanisms underlie the induction of IGF-I synthesis during compensatory hypertrophy.

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