IGF-I Immunoreactivity Research Articles

Distribution of Circulating Insulin-Like Growth Factor-I (IGF-I) into Tissues*

Intravenous infusions of amino terminal methionyl insulin-like growth factor-I (N-Met IGF-I; 8 micrograms/kg body wt x h; 24 h) were performed in lactating sheep and samples of mammary lymph, cerebrospinal fluid, and postinfusion tissues collected to examine distribution of the recombinant analog outside the vascular space. Samples were analyzed using an antibody specific for N-Met IGF-I and a second IGF-I antibody which recognized endogenous IGF-I and the N-Met variant equally. N-Met IGF-I infusion increased total plasma IGF-I immunoreactivity (ir) from 150 to 290 ng/ml. N-Met IGF-I was distributed into mammary lymph, increasing total lymph IGF-I from 60 to 130 ng/ml. By contrast iv N-Met IGF-I had no significant effect on IGF-I ir in cerebrospinal fluid. N-Met IGF-I was distributed on plasma and lymph IGF binding protein as endogenous IGF-I with binding to the 150,000 mol wt species predominant in plasma and the 40,000-50,000 mol wt pool of proteins predominant in lymph. N-Met IGF-I was also distributed into extra-vascular tissue accounting for 36% (kidney) to 62% (spleen) of total tissue IGF-I ir at the end of the infusion. The IGF-I antibodies were also used for the autoradiographical localization of IGF-I in postinfusion muscle and mammary tissue. No significant difference in antibody binding was observed to muscle fiber and mammary epithelium, but in marked contrast binding of the N-Met specific antibody to connective tissue of muscle and mammary was significantly less than the total IGF-I antibody (P less than 0.001; N-Met/total, 0.12). The data suggest that the contribution of blood-derived N-Met to total IGF-I varies markedly between tissues and provides evidence that blood-borne IGF-I may fill specific endocrine functions in selected tissues.

Secretion of Insulin-Like Growth Factor-I and Binding Proteins by Rat Liver Fat-Storing Cells: Regulatory Role of Platelet-Derived Growth Factor*

Insulin-like growth factor (IGF-I) is synthesized in multiple organs, including the liver, and may play a role in tissue growth and repair. We report that liver fat-storing cells (FSC) secrete IGF-I immunoreactivity in the culture medium. Secretion of IGF-I immunoreactivity was blocked in the presence of cycloheximide, suggesting de novo synthesis. Culture medium conditioned by FSC was concentrated and applied to a Sephadex G100 column equilibrated in a denaturing buffer. Two major species with apparent mol wts of 7.5 and greater than 25 k were identified by IGF-I RIA. Reverse phase HPLC of the 7.5 kilodalton species (the size of IGF-I) showed that it eluted in a single peak. To determine whether the higher mol wt species possessed IGF-I binding activity, appropriate fractions were desalted, incubated with [125I]IGF-I for 2 h at 30 C and applied to a Sephadex G100 column equilibrated in a nondissociating buffer. The major peak of radioactivity was confined to a high mol wt region. Western blot ligand analysis revealed the presence of two insulin-like growth factor binding proteins of approximately 28 and 31 kilodaltons. Platelet-derived growth factor, a potent mitogen for FSC, resulted in a 230% increase in release of IGF-I immunoreactivity that could be accounted for by an increase in IGF-I binding activity. In addition IGF-I increased DNA synthesis in FSC and this effect was additive to that of platelet-derived growth factor. IGF-I treatment also resulted in an increase in cell number. IGF-I and insulin-like growth factor binding proteins secreted by FSC may play a role in the hepatic tissue response to injury via autocrine and/or paracrine mechanisms.

Effect of GH status on IGF-I and IGF-II concentrations and serum IGF binding profiles in pigs.

The present study was conducted to determine 1) whether porcine growth hormone (pGH) affected insulin-like growth factor (IGF)-II concentrations in growing pigs and 2) the distribution of [125I]IGF-I and [125I]IGF-II binding activity and immunoreactive IGF-I and IGF-II in serum from control, pGH-treated and hypophysectomized (hypox) pigs. Seven weeks of pGH treatment had no effect on serum IGF-II concentrations, but after 11 wk serum IGF-II had increased by up to 50%. In normal pig serum, [125I]IGF-I bound to specific insulin-like growth factor binding proteins (IGF-BP) of 40 kDa (87.5%) and 150 kDa (12.5%). Treatment with pGH increased the proportion of IGF-I bound to the large IGF-BP (approximately 48%). In contrast, [125I]IGF-II partitioned to specific IGF-BP of 450 kDa (30%), 150 kDa (46%) and 40 kDa (23%). After treatment with pGH, binding of IGF-II to the 150-kDa IGF-BP increased to 57% of total specific binding. Binding of IGF-I and IGF-II to the 150-kDa IGF-BP was virtually abolished in hypox pig serum. IGF-I immunoreactivity was predominantly associated with the 150-kDa IGF-BP (67 to 83%), with the remainder bound to the small IGF-BP. There was no detectable free IGF-I in pig serum. Treatment with pGH did not affect the distribution of immunoreactive IGF-I between the two IGF-BP complexes, although there was a marked elevation in total serum IGF-I concentration. Immunoreactive IGF-II was detected in all three IGF-BP complexes. In control serum, the largest quantity (46%) of immunoreactive IGF-II was associated with the 40-kDa IGF-BP. Thirty-four percent was bound to the 150-kDa IGF-BP complex. Administration of pGH led to an increased association with the 150-kDa complex (50%) and a concomitant decrease in IGF-II associated with the 40-kDa species (35%). In summary, IGF-II concentrations in the pig are elevated by prolonged pGH treatment. Furthermore, there is a unique large IGF-BP in postnatal pig serum that specifically binds IGF-II but not IGF-I.

Sequential immunohistochemical expression of IGF-I and the transferrin receptor in regenerating rat muscle in vivo

The immunohistochemical expression of insulin-like growth factor I and the transferrin receptor was investigated in regenerating rat skeletal muscle. Muscle injury was induced in the extensor digitorum longus muscle in one hind limb by tourniquet ischemia. The regenerating muscles were investigated 3 to 7 days after the injury. Expression of IGF-I and the transferrin receptor was demonstrated in cryostat sections by a double-staining method. It was found that both markers were transiently expressed by the regenerating muscle cells. Expression of IGF-I immunoreactivity preceded that of the transferrin receptor and the staining for IGF-I disappeared 1 to 2 days before the staining for the transferrin receptor. The data are compatible with a regulatory role of IGF-I on the transferrin receptor.

Changes in the distribution of insulin-like growth factor I, thioredoxin, thioredoxin reductase and ribonucleotide reductase during the development of the retina

The postnatal development of the rat retina offers good opportunities to follow migration and differentiation of neuroectodermal cells. In this study we have analyzed, by immunohistochemical methods, the distribution of several protein antigens and the trophic peptide insulin-like growth factor I (IGF-I; somatomedin-C). The latter contains disulfide bonds, which in vitro are reduced by the thioredoxin system, i.e. thioredoxin, thioredoxin reductase and NADPH. Ribonucleotide reductase provides growing cells with deoxyribonucleotides, necessary for DNA synthesis, and thioredoxin is an in vitro hydrogen donor. By immunofluorescence IGF-I immunoreactivity was observed throughout the Müller neuroglial cells in the developing retina, but only to a very small extent in the mature retina. Nerve cells showed transient expression of IGF-I during their development. The IGF-I immunoreactivity is likely to be due to local synthesis, since we could demonstrate retinal IGF-I mRNA. Treatment with the transport-blocking agent colchicine caused the Müller glial cells and the retinal pigment epithelium cells to become IGF-I immunoreactive. Thioredoxin and thioredoxin reductase immunoreactivities are confined to neurons and photoreceptor cells in the developing retina. Ribonucleotide reductase subunit M1 immunoreactivity was only observed during the first postnatal week in proliferating neuroectodermal cells. In conclusion, IGF-I and ribonucleotide reductase subunit M1 immunoreactivities are transiently expressed by neuroectodermal cells in the developing rat retina. In contrast, the proteins of the thioredoxin system are demonstrable also in the mature retina.

Transient expression of insulin-like growth factor I immunoreactivity by vascular cells during angiogenesis

The present study was undertaken to investigate whether vascular cells show insulin-like growth factor I (IGF-I; somatomedin C) immunoreactivity under normal conditions and/or during angiogenesis in humans and animals, as the trophic peptide IGF-I is considered important for cell growth and differentiation. In adult animals normal blood vessels, i.e., arteries, veins, and capillaries, did not show any IGF-I immunoreactivity. In newborn animals every vascular cell showed IGF-I immunoreactivity; the frequency and intensity thereafter decreased and eventually vanished as the animals approached maturity. Injury of a tissue or organ rapidly induced extensive blood vessel formation and such new blood vessels transiently expressed IGF-I immunoreactivity. Endothelial cells in budding capillaries showed distinct cytoplasmic IGF-I immunoreactivity, as did endothelial cells, smooth muscle cells, and fibroblast in newly formed arteries and veins. In biopsies of human tissue, transient IGF-I immunoreactivity was evident in vascular cells during angiogenesis after injury, as it also was in granulation tissue, skin wounds, and scar capsules around implants. Increased IGF-I immunoreactivity was further demonstrated in vascular cells in biopsies from patients with other changes involving blood vessel formation, e.g., nasal polyps, and in specimens from patients with arteritis, tendonitis, synovitis, Wegener's granulomatosis, idiopathic midline destructive disease, neurofibromatosis (von Recklinghausen's disease), and muscular dystrophy. It is concluded that during angiogenesis, obviously irrespective of inducing factors and mechanisms, vascular wall cells transiently show IGF-I immunoreactivity.

Immunohistochemical Demonstration of Insulin-like Growth Factor I in Inflammatory Lesions in Wegener's Granulomatosis and Idiopathic Midline Destructive Disease

The immunoreactivity of the trophic peptide insulin-like growth factor I (IGF-I; somatomedin C) was mapped in nasal mucosa biopsies from three patients with Wegener's granulomatosis (WG) and one with idiopathic midline destructive disease (IMDD; idiopathic midline granuloma). Strongly increased IGF-I immuno-reactivity restricted to cells bordering and in vessel walls and in granulomas (WG) was demonstrated, while necrotic and noninflammatory areas were negative. Treatment with steroids and cyclophosphamide reduced the IGF-I immunoreactivity. The abnormally increased IGF-I immunoreactivities in WG and IMDD probably reflects the reactive growth processes in diseased tissue and is not thought to be the primary cause of either disease. IGF-I may be formed locally by cells in and close to the vascular walls in areas with active disease resulting in e.g. vascular growth, granuloma formation, and finally vessel obliteration and necrosis. IGF-I is likely to form, possibly in concert with other trophic factors, a link in the chain of events resulting in the tissue abnormalities in WG and IMDD.

Immunocytochemical and biochemical localization of insulin-like growth factor I in the kidney of rats before and after uninephrectomy.

The localization of insulin-like growth factor I (IGF-I, somatomedin C) was investigated in the kidney of adult rats during normal conditions and after nephrectomy, using immunocytochemical and biochemical methods. In the normal kidney, IGF-I immunoreactivity could be demonstrated mainly in cells in the medullary collecting ducts and in those parts of the thin limb of Henle's loop located in the outer medulla. During compensatory growth all parts of the collecting ducts, including those in the cortex, showed IGF-I immunoreactivity, as did cells in the entire thin limb of Henle's loop. No IGF-I immunoreactivity could be demonstrated in the proximal or distal tubules, either in the control kidney or during compensatory growth. Biochemical measurements showed a significantly higher content of IGF-I in the inner medulla than in the cortex in the normal kidney. Uninephrectomy resulted in significantly increased IGF-I content in the cortex. It is suggested that IGF-I is produced mainly in the collecting ducts and in the thin limb of Henle's loop and exerts its effect on other parts of the nephron by paracrine mechanisms.

Somatomedin C immunoreactivity in the Achilles tendon varies in a dynamic manner with the mechanical load.

Distribution of the trophic peptide somatomedin C (Sm-C; insulin-like growth factor I; IGF-I) immunoreactivity was mapped in normal Achilles and tibialis anterior tendons. The spindle-shaped tendon fibroblasts showed faint perinuclear staining. Fibroblasts in the paratenon mostly had a more intense IGF-I immunoreactivity, i.e. faint to moderate. When analysing either tendon in detail, areas with more intense IGF-I immunoreactivity could be recognized and seemed to correlate with areas of high mechanical stress. Increased mechanical load induced over 3 days elevated IGF-I immunoreactivity throughout the cytoplasm of tendon fibroblasts. Peak intensity was reached in 7 days, and thereafter the IGF-I immunoreactivity seemed to decrease irrespective of persistent high mechanical load. Training the animals on a treadmill for from 20 up to 60 min per day for 5 days induced after 3-5 days increased IGF-I immunoreactivity throughout the cytoplasm of the tendon and paratenon fibroblasts. Sudden curtailment of loading the Achilles tendon resulted in a marked reduction of the IGF-I immunoreactivity in most fibroblasts within 3 days. After a week only a small number of tendon fibroblasts showed any IGF-I immunoreactivity. The IGF-I immunoreactivity of tendon fibroblasts thus correlates to mechanical loading of the tendon. It is proposed that IGF-I may have a trophic influence on tendon and paratenon cells by autocrine and/or paracrine mechanisms.

Expression of insulin-like growth factor-I and -II genes in human smooth muscle tumours.

The insulin-like growth factors I and II (IGF-I and -II) are polypeptides which play an important role in growth and development of the organism. In the present report we describe the detection of human IGF-I RNAs (both type Ia and type Ib) and IGF-II RNAs in benign (leiomyoma) and malignant (leiomyosarcoma) tumours from smooth muscle origin, using Northern blot hybridization analysis. In normal smooth muscle tissue of the uterus we found low levels of IGF-I RNAs only. In the tumours the same IGF-I RNA species were detected as in adult non-tumour tissues (uterus, liver). For transcription of the IGF-II gene in these tumours, two promoters are used which are expressed in fetal liver, but not in adult liver. The presence of IGF-I and IGF-II RNAs was also established by nucleotide sequence analysis of recombinant DNA clones isolated from cDNA libraries derived from two leiomyosarcomas. The nucleotide sequences of these cDNA clones, together covering the entire coding regions of IGF-Ia and IGF-II var RNA, predict that IGFs encoded by the tumour RNAs do not differ in amino acid sequence from the corresponding polypeptides isolated from serum. In those tissues containing IGF-I RNAs, IGF-I immunoreactivity was also demonstrated.

Insulin‐like growth factor I in the pancreas of normal and diabetic adult rats

Insulin-like growth factor I (IGF-I, somatomedin C) was mapped by immunocytochemistry in the pancreas of normal and experimentally influenced rats. The polyclonal IGF-I antiserum K 37 was characterized and demonstrated to be specific. In the exocrine pancreas some duct cells showed IGF-I immunoreactivity, other components being negative. The three main endocrine cell types in the islets of Langerhans were IGF-I immunoreactive, most strikingly the D cells. Hypophysectomy resulted in loss of IGF-I immunoreactivity in all three endocrine cell types, i.e. D, A and B cells, while the levels of somatostatin, glucagon and insulin, respectively, remained unchanged. Starvation seemed to increase and feeding to decrease the IGF-I immunoreactivity in the B cells. Cysteamine pre-treatment reduced the normally intense IGF-I and somatostatin immunoreactivities in the D cells. In rats made diabetic with alloxan or streptozotocin, the B cells were irreversibly damaged and lost both their insulin and IGF-I immunoreactivities, while the IGF-I immunoreactivity was increased in A cells; the D cells remained unchanged. The concentrations of IGF-I mRNA in the pancreas were almost equal in normal and alloxan diabetic rats as were the concentrations of extractable IGF-I. We conclude that IGF-I immunoreactive material can be demonstrated in adult animals in all endocrine islet cells, most prominently in the D cells. The expression of IGF-I immunoreactivity is in part under pituitary control. In the adult rat only one islet cell type synthesizes IGF-I immunoreactive material, i.e. the D cells, while, in contrast, the B cells are likely to be a major IGF-I source in fetal and neonatal islets.

IGF-I immunoreactivity is expressed by regenerating renal tubular cells after ischaemic injury in the rat.

The immunohistochemical expression of the trophic peptide insulin-like growth factor I (IGF-I; somatomedin C) was investigated in the post-ischaemic regenerating rat kidney. The part of the nephron most severely injured by the ischaemic insult was the pars rectae or S3 region of the proximal tubules. In this part of the nephron no specific IGF-I immunoreactivity could be demonstrated under control conditions. Three days after injury, however, low regenerating tubular cells expressing IGF-I immunoreactivity could be demonstrated invading the damaged region. Five and 7 days post-ischaemia some of the regenerating cells showed IGF-I immunoreactivity while adjacent and similar-looking cells did not. In parallel with an increasing differentiation of the regenerating cells the IGF-I immunoreactivity vanished. Fourteen days after the injury the S3 cells had regained their normal morphology, and by this time the IGF-I immunoreactivity had returned to a normal undetectable level. It is concluded that there is a transient increase in the expression of IGF-I mainly confined to the early phase of post-ischaemic rat kidney regeneration. It is proposed that IGF-I exerts its trophic role by paracrine mechanisms.

Differing expression of insulin-like growth factor I in the developing and in the adult rat cerebellum.

Insulin-like growth factor I (IGF-I; somatomedin C) is a trophic peptide of importance for the development of several tissues and organs. In the present study we have mapped the cellular distribution and dynamic changes of IGF-I immunoreactivity in the rat cerebellum from its postnatal development to maturity. In vitro hybridization of IGF-I mRNA was used to demonstrate that the IGF-I immunoreactive material was synthesized in the cerebellum during a limited time period of cerebellar differentiation. IGF-I immunoreactivity was absent in primordial nerve cells but was present in neuroglial cells during the first two days after birth and then rapidly increased in intensity in the latter during the next few days. Proliferative nerve cells in the external granular layer did not express IGF-I immunoreactivity, while migrating and differentiating nerve cells as well as neuroglial cells showed intense labelling. Starting about 2 weeks postnatally, the IGF-I immunoreactivity declined, first in the neuroglial cells and eventually in the nerve cells. No IGF-I immunoreactivity could be demonstrated in the normal adult cerebellum. Colchicine pretreatment did, however, enable demonstration of IGF-I immunoreactivity in adult cerebellar nerve cells but not in neuroglial cells. In vitro hybridization revealed IGF-I mRNA in the developing cerebellum but only at very low levels in the adult cerebellum. It is concluded that IGF-I is likely to be a factor of importance for the development and maturation of nerve cells and neuroglial cells in the brain.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunohistochemical localization of insulin-like growth factor I in the adult rat.

Rabbit antisera against native human insulin-like growth factor I (IGF-I; somatomedin C) or a synthetic tetradecapeptide, representing the carboxyterminal amino acids 57-70 of human IGF-I, were used to map immunohistochemically the distribution of IGF-I immunoreactive material in adult rats. Both antisera were specific for IGF-I, as characterized by immunoabsorption, immunoblotting and radioimmunoassay. There was no cross-reactivity to IGF-II, relaxin or pro-insulin; substances having a high degree of structural homology with IGF-I. High IGF-I immunoreactivity was observed in spermatocytes of the testis; in oocytes, granulosa and theca interna cells of the ovary during early stages of follicle development; in some lymphocytes and in reticular cells of lymphoid and hematopoietic organs; in salivary gland duct cells; in the adrenal medulla, the parathyroid gland and the Langerhans' islets. Chondrocytes in the epiphyseal and rib growth plates and at articular surfaces showed strong IGF-I immunoreactivity. Brown but not white fat cells were stained. Nerve cells in the peripheral and autonomic nervous system showed faint to intense IGF-I immunoreactivity. In contrast, neurons and neuroglial cells in the central nervous system were generally negative; motor neurons being an exception. Erythropoietic, thrombocytopoietic and myeloic cells in the bone marrow showed IGF-I immunoreactivity, but only at defined developmental stages. Hepatocytes showed faint IGF-I immunoreactivity, but became more intensely stained after pretreatment with colchicine. The present results suggest that IGF-I is synthetized by cells in several tissues and organs in the adult rat. There was an apparent association between the localization of IGF-I and cell differentiation.(ABSTRACT TRUNCATED AT 250 WORDS)

Transiently increased insulin-like growth factor I immunoreactivity in tendons after vibration trauma. An immunohistochemical study on rats.

The hind limbs of anaesthetized rats were exposed to vibration trauma (81 Hz; amplitude peak to peak 0.50 mm) for 4 hours during 2 consecutive days. The animals were examined in groups of 4 immediately after the last exposure, and after 1, 2, 3, 5, 7, 10, 14 and 28 days. The Achilles tendons and the tendons of the anterior tibialis muscles were sampled and processed to demonstrate IGF-I immunoreactivity. In the normal Achilles tendon and in the tendon of the anterior tibial muscle, slight IGF-I immunoreactivity was seen in many of the long slender fibroblasts between the collagen bundles. A strong increase in the IGF-I immunoreactivity appeared in the anterior tibialis muscle tendon 3 days after the last vibration exposure. In addition, the tendon fibroblasts became hypertrophic. A similar but less striking increase in IGF-I immunoreactivity appeared in the Achilles tendon. The peak intensity and frequency of stained cells were achieved after 7 days for both tendons. The intensity then levelled off, and was normalized after 28 days. It is concluded that acute exposure to vibrations induces reactive changes in fibroblasts in tendons, which may reflect a change to a more active synthesising state, as a response to the vibration trauma. The transiently altered expression of IGF-I immunoreactivity forms a link in a chain of events regulating the functional activity level of fibroblasts in response to a trauma.

Transient expression of insulin‐like growth factor I immunoreactivity in skeletal muscle cells during postnatal development in the rat

The immunohistochemical expression of insulin-like growth factor I (IGF-I; somatomedin C) was investigated in hindlimb skeletal muscle of rats during postnatal development. IGF-I immunoreactivity could be demonstrated in satellite fibres and myotubes 1, 2 and 3 days after birth, while no IGF-I immunoreactivity could be demonstrated in the more mature primary fibres. Five days after birth only scattered cells showed IGF-I immunoreactivity and 10 days after birth no specific IGF-I immunoreactivity could be demonstrated in muscle cells, i.e. the adult pattern was established. It is concluded that IGF-I immunoreactivity is expressed during a limited period of postnatal skeletal muscle maturation in rats. IGF-I is probably synthesized by IGF-I immunoreactive muscle cells and contributes to the differentiation/maturation process by autocrine and/or paracrine mechanisms.

Regenerating skeletal muscle cells express insulin-like growth factor I.

The expression of the trophic peptide insulin-like growth factor I (IGF-I; somatomedin C) was investigated in the regenerating soleus muscle of mice after injury by the snake venom taipoxin. No specific IGF-I immunoreactivity was observed in muscle cells during control conditions. Within 2 days after taipoxin injection, IGF-I immunoreactivity could be demonstrated in activated satellite cells. Myoblasts and myotubes expressed high IGF-I immunoreactivity. The IGF-I immunoreactivity was strictly cytoplasmatic and obviously associated with polyribosomes. No vesicular or membraneous IGF-I immunoreactivity could be demonstrated. It is concluded that IGF-I is synthesized in myogenic cells during skeletal muscle regeneration. It is suggested that IGF-I exerts its effects on skeletal muscle mainly by autocrine mechanisms.

IGF-I expression in blood vessels varies with vascular load.

The present study was designed to investigate if the immunoreactivity of insulin-like growth factor I (IGF-I, somatomedin C) in blood vessels was changed in relation to the vascular load in adult rats. Occlusion of the right femoral artery resulted within 24 h in a strongly increased IGF-I immunoreactivity in the media of the contralateral, left, femoral artery and a significant decrease in the IGF-I expression in the right femoral artery distal to the occlusion. The increased labelling of the smooth muscle cells persisted for at least 21 days. Inactivity of the right hind limb, secondarily to an ischaemic insult, likewise induced a significant increase in IGF-I immunoreactivity, within 24 h, in the femoral vessels of the undamaged, contralateral left hind limb. It is concluded that IGF-I immunoreactivity varies with the dynamic load in the vascular walls. It is proposed that IGF-I may serve as a parahormone exerting its effect by paracrine and/or autocrine mechanisms, taking part in regulating the structural adaption of blood vessels in response to stress.

Satellite cells express the trophic factor IGF-I in regenerating skeletal muscle.

The expression of insulin-like growth factor I (IGF-I, somatomedin C) was studied in regenerating skeletal muscle. Irreversible damage to skeletal muscle cells was induced in the extensor digitorum longus muscle (EDL) of adult rats by ischaemia, preceded by glycogen depletion, and the regeneration process was studied for periods up to 14 days after injury. The IGF-I was demonstrated by indirect immunofluorescence. Immunoreactivity against ribonucleotide reductase (RR) was used as a marker for DNA synthesis, that is, cell proliferation. Increased IGF-I immunoreactivity could be demonstrated within 24 h after injury in satellite cells, intramuscular nerves and in blood vessels. The IGF-I immunoreactivity remained virtually unchanged in the contralateral, undamaged EDL. An increasing number of satellite cells, expressing high IGF-I immunoreactivity, could be demonstrated in the injured EDL, and within 72 h myoblasts, expressing high IGF-I and RR immunoreactivity, were formed. Small immature muscle cells, displaying high IGF-I immunoreactivity, were observed 4 days after injury. Increased IGF-I immunoreactivity was still obvious in the regenerated muscle cells 14 days after injury while RR immunoreactivity was seen only in scattered satellite cells. It is concluded that IGF-I may act as a trophic factor during regeneration of skeletal muscle after injury.

Rapid axoplasmic transport of insulin-like growth factor I in the sciatic nerve of adult rats

Somatomedin C (Sm-C; insulin-like growth factor I; IGF-I) is a polypeptide (Mr 7649), often dependent on growth hormone (GH), with trophic effects on several different tissues. Monospecific IGF-I antisera were used to investigate its localization in the sciatic nerve and corresponding nerve cells, as well as its possible axoplasmic transport in the adult rat. IGF-I-like immunoreactivity was demonstrated in anterior horn motor nerve cells in the spinal cord and in spinal- and autonomic ganglion nerve cells. Faint IGF-I immunoreactivity was under normal conditions observed in axons of the sciatic nerve and in the Schwann cells. Using crush technique, accumulation of IGF-I immunoreactivity was seen in dilated axons within 2 h, both proximal and distal to the crush. However, only a small fraction of the anterogradely transported IGF-I immunoreactive material could be demonstrated to be transported in retrograde direction. Colchicine injected proximal to a crush prevented accumulation of IGF-I immunoreactivity proximal to the crush, but not distal to it. IGF-I-immunoreactive material is synthesized in the cell bodies of peripheral sensory and motor nerve cells. It is transported at rapid rates in the axoplasm of the sciatic nerve of adult rats both in anterograde and retrograde directions. We propose that axonally transported IGF-I may be released and exert trophic influence on innervated cells, tissues and organs.