Localization Of Insulin-like Growth factor-I Research Articles

Spatial distribution of insulin-like growth factor binding protein-2 following hypoxic-ischemic injury

BackgroundInsulin-like growth factor binding protein-2 (IGFBP-2) regulates the bioavailability, transportation, and localization of insulin-like growth factor-I (IGF-I), an effective neuroprotectant in animal stroke models especially when administered intranasally. Therefore, determining IGFBP-2′s endogenous distribution in the normal and ischemic brain is essential in maximizing the neuroprotective potential of the intranasal IGF-I treatment approach. However, current data on IGFBP-2 is limited to mRNA and in situ hybridization studies. The purpose of this study was to determine if there are any changes in IGFBP-2 protein levels and distribution in ischemic brain and also to determine if IGFBPs play a role in the transportation of intranasally administered IGF-I into the brain.ResultsUsing an in vitro approach, we show that ischemia causes changes in the distribution of IGFBP-2 in primary cortical neurons and astrocytes. In addition, we show using the transient middle cerebral artery occlusion (MCAO) model in mice that there is a significant increase in IGFBP-2 levels in the stroke penumbra and core after 72 h. This correlated with an overall increase in IGF-I after stroke, with the highest levels of IGF-I in the stroke core after 72 h. Brain sections from stroke mice indicate that neurons and astrocytes located in the penumbra both have increased expression of IGFBP-2, however, IGFBP-2 was not detected in microglia. We used binding competition studies to show that intranasally administered exogenous IGF-I uptake into the brain is not receptor mediated and is likely facilitated by IGFBPs.ConclusionsThe change in protein levels indicates that IGFBP-2 plays an IGF-I-dependent and -independent role in the brain’s acute (neuroprotection) and chronic (tissue remodeling) response to hypoxic-ischemic injury. Competition studies indicate that IGFBPs may have a role in rapid transportation of exogenous IGF-I from the nasal tissue to the site of injury.

Expression of Insulin-like Growth Factor-I in Uterus is Ovarian Steroid Dependent: An in situ Immunohistochemical Study in Rat

Objective To study the immunohistochemical localization of insulin-like growth factor-I (IGF-I) in rats' uteri to determine if expression of the growth factor is ovarian steroid dependent. Method The study was carried out in presence and absence of ovary in situ of non pregnant females and during early gestation (day 1 to day 5.5). Cyclic females were tested to observe the effect of native steroids on IGF-I expression. Adult females were ovariectomized (OVX) and injected (s.c) with estradiol-17β in a dose of 0.1 μg/ml per day for three consecutive days at interval of 24 h prior to the collection of uterine horns. During the early pregnancy, studies were carried out on day 3.5 and day 5.5 of geatation respectively to determine the steroids' effects during pre- and post-implantation period. Tamoxifen was administered (s.c) in a dose of 250 μg/ml per day from day 1 to day 3 of gestation while, the prostaglandin F 2α (PGF 2α ) was administered (s.c) from day 3 of gestation onward for three consecutive days at interval of 24 h in a dose of 150 μg/ml per day. Expression of IGF-I was immunohistochemically localised using IGF-I antibody in paraffin embedded sections. Results IGF-I was expressed in rat uterus during estrus phase as well as during pre and post-implantation period. The ovariectomized females' uteri lost the expression of IGF-I. Exogenous administration of tamoxifen and PGF 2α reduce the expression of the growth factor. Conclusion Expression of IGF-I in rat uterus during cyclic stage and early gestation depends upon the availability of circulating estrogen and progesterone. Uterine expression of IGF-I can be modulated by manipulating circulating ovarian steroid either during cyclic stage or during gestation.

Immunohistochemical detection of insulin-like growth factor-I, transforming growth factor-β2, basic fibroblast growth factor and epidermal growth factor-receptor expression in developing rat ovary

The aim of this study was to determine the immunohistochemical expression and localization of insulin-like growth factor-I (IGF-I), transforming growth factor-β2 (TGF-β2), basic fibroblast growth factor (bFGF) and epidermal growth factor-receptor (EGF-R) in developing rat ovaries.Eighteen female Wistar rats were enrolled in this study; newborn (n=6), one-month-old (n=6) and adult (n=6) rats. Formalin-fixed and parafin-embedded ovarian tissues were stained with antibodies against IGF-I, TGF-β2, bFGF and EGF-R, immunohistochemically. The ovarian cells were evaluated by semi-quantitative scoring system under light microscope.The staining of IGF-I, TGF-β2, bFGF and EGF-R were most intense in the oocytes and were heavily at one-month-old rats. A moderate immunostaining in theca cells and corpus luteii reacted with IGF-I in adult rats. Furthermore the staining intensity for IGF-I was moderate in granulosa cells of newborn rat ovaries. We detected also a moderate staining for TGF-β2 in corpus luteii of adult rats. In addition, we found a bFGF immunostaining mainly in oocytes of follicles of young and adult rats. Immunostaining for EGF-R was moderate in granulosa cells of one-month-old rats.In conclusion, this study suggests that growth factors play a pivotal role in ovarian function, especially in follicular development. The role of growth factor in controlling degeneration or growth (or both) of ovary follicles remain as explained.

Effects of Red Deer Antlers on Cutaneous Wound Healing in Full-thickness Rat Models

The process of wound repair involves an ordered sequence of events such as overlapping biochemical and cellular events that, in the best of circumstances, result in the restoration of both the structural and functional integrity of the damaged tissue. An important event during wound healing is the contraction of newly formed connective tissues by fibroblasts. The polypeptide growth factors, like transforming growth factor-β (TGF-β), insulin-like growth factor I (IGF-I) and epidermal growth factor (EGF), play very important mediator roles in the process of wound contraction. Deer antlers, as models of mammalian regeneration, are cranial appendages that develop after birth as extensions of a permanent protuberance (pedicle) on the frontal bone. Antlers contain various growth factors which stimulate dermal fibroblast growth. They are involved in digestion and respiration and are necessary for normal wound healing and skin health. In order to investigate and evaluate the effects of red deer antlers on skin wound site, the speed of full-thickness skin wound healing and the expression of IGF-I, TGF-β, and EGF in skin wounds, three groups of skin full-thickness rat models with a high concentration of antler ointment, a low concentration of antler ointment and without antler ointment were compared. At post-injury days 0, 2, 4, 8, 16, 20, 32, 40 and 60, the skin wound area was measured, the expressions of IGF-I, TGF-β, and EGF mRNA were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and collagen formation by sirius red dye and the localization of IGF-I, TGF-β, and EGF peptides were inspected by histological immunohistochemical techniques. Wound healing was significantly more rapid in antler treated skins. In addition, the wound treated with a high concentration antler ointment, a low concentration antler ointment, and the control closed completely at post-injury day 40, day 44 and day 60, respectively. Via RT-PCR, the expressions of IGF-I (day 8 and day 16), TGF-β (day 8, day 16 and day 20) and EGF (day 4, day 8, day 16, and day 32) were obviously up-regulated in high concentration antler-treated skins compared to control skins. Similar results could be seen in the histological detection of collagen dye and immunohistochemical methods using the corresponding polyclone antibodies of IGF-I, TGF-β, and EGF. These results illustrate that antlers stimulate and accelerate the repair of cutaneous wounds.

Real-time polymerase chain reaction, in situ hybridization and immunohistochemical localization of insulin-like growth factor-I and myostatin during development of Dicentrarchus labrax (Pisces: Osteichthyes)

The distribution of insulin-like growth factor-I (IGF-I) and myostatin (MSTN) was investigated in sea bass (Dicentrarchus labrax) by real-time polymerase chain reaction (PCR), in situ hybridization (ISH) and immunohistochemistry. Real-time PCR indicated that IGF-I mRNA increased from the second day post-hatching and that this trend became significant from day 4. ISH confirmed a strong IGF-I mRNA expression from the first week post-hatching, with the most abundant expression being detected in the liver of larvae and adults. Real-time PCR also showed that the level of MSTN mRNA increased significantly from day 25. The expression of MSTN mRNA was higher in muscle and almost absent in other anatomical regions in both larvae and adults. Interestingly, the lateral muscle showed a quantitative differential expression of IGF-I and MSTN mRNAs in red and white muscle, depending on the developmental stage examined. IGF-I immunoreactivity was detected in developing intestine at hatching and in skeletal muscle, skin and yolk sac. MSTN immunostaining was evident in several tissues and organs in both larvae and adults. Both IGF-I and MSTN proteins were detected in the liver from day 4 post-hatching and, subsequently, in the kidney and heart muscle from day 10. Our results suggest, on the basis of a combined methodological approach, that IGF-I and MSTN are involved in the regulation of somatic growth in the sea bass.

Expression and localization of insulin-like growth factor-I in four parts of the red deer antler

The expression and localization of insulin-like growth factor-I (IGF-I) in the four parts (tip, upper, mid and base) of the red deer antler has been extensively investigated. We used reverse transcriptase polymerase chain reaction (RT-PCR) and real-time reverse transcriptase polymerase chain reaction (real time RT-PCR), in situ hybridization, immunohistochemistry and Western blot techniques to localize IGF-I messenger ribonucleic acid (mRNA) and IGF-I peptide in the four parts of the antler. The specific sequence encoding IGF-I was detected by RT-PCR in all of the four specimens, and the 395 bp IGF-I sequence from the red deer antler was shown to have very high homology with human, goat and mouse IGF-I. In situ hybridization and immunohistochemistry results demonstrated that the expression of IGF-I occurred in chondrocytes and osteoblasts in the tip and upper parts of the antler. However, IGF-I was only detectable in osteoblasts around the bone in the mid and base parts. There were significant differences in the intensity of the signal obtained with the IGF-I probe in the tip, upper, mid and base tissues. The Western blot analysis also provided evidence that IGF-I expression was localized differentially in the four parts of the deer antler. This study indicates that antler tissue is an essential part of the IGF system, which is involved in the regulation of the growth of red deer antlers. The specific expression of IGF-I in the four parts of the deer antler suggests that the IGF-I molecule is present at significantly different levels throughout the deer antler development and regeneration processes. Localization of IGF-I in chondrocytes and osteoblasts suggests that IGF-I may play an important role in cartilage and bone formation. In addition, it may have a variety of biophysical effects that influence the rapid growth of deer antlers.

Localization of Insulin-Like Growth Factor I (IGF-I) in the Chicken Liver after Fasting and Refeeding: Demonstration by Using Antigen Retrieval Immunohistochemistry

Immunohistochemical localization of insulin-like growth factor-I (IGF-I) was investigated in the liver of fasted and refed chickens by using an antigen retrieval method. The present study is the first one showing the localization of IGF-I in the chicken liver. Immunoreactivity for IGF-I was detected on the paraffin sections of livers from the fed and refed chickens after the treatment with the antigen retrieval agent. A moderate number of cells showing IGF-I immunoreactivity were scattered in the parenchyma of the liver from fed chickens. These cells were relatively large and polygonal in shape and seemed to be hepatocytes. Reaction products were observed as a granular structure in the cytoplasm of IGF-I-immunoreactive hepatocytes. The number of immunoreactive hepatocytes was increased in the liver from refed chickens compared with fed chickens. Diffuse reaction products as well as granular ones were observed throughout the cytoplasm of IGF-I-immunoreactive hepatocytes of livers from refed chickens. There are, however, no regular patterns of the distribution of immunoreactive hepatocytes in the parenchyma of both fed and refed chickens. In the liver of the fasted chickens, clear immunoreactivity for the peptide was not observed. These data show that IGF-I is located in the chicken hepatocytes and influenced by the nutriture.

Localization of insulin-like growth factor-I in lung tissues of patients with fibroproliferative acute respiratory distress syndrome.

Insulin-like growth factor-I (IGF-I) is elevated in human fibrotic lung diseases and in animal models of pulmonary fibrosis, implicating IGF-I in the pathogenesis of fibrotic lung disease. We questioned whether IGF-I protein levels were enhanced in fibroproliferative acute respiratory distress syndrome (FP-ARDS). Serial lung tissue sections from a biopsy database were immunohistochemically stained for IGF-I, IGF-I receptor, CD68, alpha-smooth muscle actin, collagens I and III, and proliferating cell nuclear antigen. Our results show enhanced staining of IGF-I and IGF-I receptor, collagens I and III, smooth muscle actin, CD68, and proliferating cell nuclear antigen in FP-ARDS compared with control lung sections. In FP-ARDS specimens, prominent staining of IGF-I and IGF-I receptor was seen in alveolar and interstitial macrophages as well as in a variety of mesenchymal cells. There was a correlation between IGF-I staining and CD68-positive cells, suggesting macrophages as a potential source of the IGF-I protein present in lungs. IGF-I also correlated with enhanced collagen I, collagen III, and proliferating cell nuclear antigen immunoreactivity, suggesting that IGF-I may play a role in the extracellular matrix protein deposition and cellular proliferation seen in the lungs of individuals with FP-ARDS. Our results indicate that IGF-I is increased in FP-ARDS and may be an important mediator in the progression of acute lung injury to FP-ARDS.

IGF-I treatment attenuates renal abnormalities induced by neonatal ACE inhibition.

An intact renin-angiotensin system (RAS) during nephrogenesis is essential for normal renal development. We have shown previously that neonatal inhibition of the RAS, either with ANG II type 1-receptor blockade or angiotensin-converting enzyme (ACE) inhibition, induces irreversible renal abnormalities. The aim of the present study was to investigate whether an interrupted RAS can be compensated for by exogenous administration of another important renal growth-promoting factor, the insulin-like growth factor-I (IGF-I). Rats were treated daily with either the ACE inhibitor enalapril (10 mg/kg), recombinant human IGF-I (3 mg/kg), or the combination enalapril + IGF-I from perinatal day 3 to 13. Urinary concentrating ability, renal function, and renal morphology were assessed at adult age. The gene expression and localization of IGF-I, its receptor, and the growth hormone receptor (GHR) were investigated during ongoing ACE inhibition. The present study demonstrates normalized renal function and histology in enalapril + IGF-I-treated animals. Ongoing ACE inhibition suppressed the medullary IGF-I mRNA expression and altered the local distribution of both IGF-I and GHR. Thus the present study provides evidence for an interaction between the RAS and GH/IGF-I axis in renal development.

Pharmacological Characterization and Cellular Localization of Insulin-Like Growth Factor-I (IGF-I) Binding to Human Placenta during the Course of Pregnancy and at Term

Pharmacological Characterization and Cellular Localization of Insulin-Like Growth Factor-I (IGF-I) Binding to Human Placenta during the Course of Pregnancy and at Term

Immunohistochemical localization of insulin-like growth factor-I (IGF-I), its receptor and IGF-binding protein-1 (IGFBP-1) in the human placental membrane

The insulin-like growth factor (IGF) pathway plays a critical role in the growth and development of the uterus and is believed to function as a mediator of steroid hormone actions in the endometrium. The local expression of genes encoding IGFs and IGF-binding proteins (IGFBPs) is important in determining IGF bioactivity in the uterus. Genetic variation in key genes within the IGF pathway may influence the rate of cellular proliferation and differentiation in the uterus and ultimately affect the risk of endometrial cancer. Our hypothesis is that variant alleles in key genes involved in the IGF pathway will influence the development of endometrial cancer.We conducted a case-control study nested within the Nurses' Health Study (NHS) and the Women's Health Study (WHS) to investigate the association between forty-four polymorphisms within IGFI, IGFII, IGFBP-1, and IGFBP-3 with endometrial cancer risk using 692 invasive endometrial cancer cases and 1723 matched controls. We used conditional logistic regression to calculate odds ratios (ORs) and 95% confidence intervals (CIs) to assess the risk of endometrial cancer.We observed an inverse association with IGFII rs3741211 and endometrial cancer risk (OR = 0.79 (95% CI: 0.63, 0.99)) and IGFII rs1004446 and endometrial cancer risk (OR = 0.80 (95% CI: 0.68, 0.94)). We also observed an inverse association with IGFBP-3 rs2453839 and endometrial cancer risk (OR = 0.81 (95%CI: 0.67, 0.98). However, we did not observe any statistically significant associations with the polymorphisms in IGFI and IGFBP1 and endometrial cancer risk.Genetic variation with IGFII and IGFBP-3 may influence endometrial cancer risk in Caucasians. Polymorphisms in IGFI and IGFBP-1 were not associated with endometrial cancer risk, but further research is needed.

Immunohistorical localization of insulin-like growth factor-I (IGF-I), IGF-I receptor and IGF-binding protein-1 (IGFBP-1) in the human anchoring villi

During the first trimester of pregnancy, the human placenta is an actively dividing and highly invasive tumour-like tissue, while near term, it represents a fully developed, non-invasive unit. In order to understand the molecular basis of this marked difference in the placental phenotypes, an approach based on a differential display–reverse transcription–polymerase chain reaction (DD–RT–PCR) was adopted to analyse changes in gene expression, using total RNA isolated from first-trimester and term placental villi. Using this approach, T-plastin was initially identified as being differentially expressed in the human first-trimester placenta. T-plastin is an actin-bundling protein and is known to be highly expressed in actively dividing cells and up-regulated in several carcinomas. Using a homogenous population of cytotrophoblasts and syncytiotrophoblasts isolated from human placentae, the present authors demonstrate the differential expression of T-plastin in cytotrophoblasts compared with the terminally differentiated syncytiotrophoblasts. The down-regulation of T-plastin expression is further demonstrated in human trophoblastic BeWo cells induced to differentiate using transforming growth factor (TGF)β1, a growth factor known for its anti-proliferative and anti-invasive response in placental cells. These studies suggest that expression of T-plastin in the placental context may indeed be associated with the enhanced replicative potential of placental trophoblasts.

Insulin-like growth factor-I in the porcine endometrium and placenta: localization and concentration in relation to steroid influence during early pregnancy.

To initiate the establishment of an epitheliochorial placenta, the developing porcine conceptuses contact the maternal endometrium on its mesometrial side. The porcine conceptuses secrete estrogens which, together with circulating maternal hormones, govern variations in the structure as well as expression and levels of steroid receptors and growth factors during early pregnancy. Mesometrial samples of endometrium or placenta were collected from 15 early pregnant (8-30 days after the onset of estrus) and six cycling (days 1-14) gilts. The variations in tissue morphology and immunohistochemical localization of insulin-like growth factor-I (IGF-I) were related to the tissue levels (by enzyme immunoassay) of receptors for estrogen (ER) and progesterone (PR), as well as mRNA (by solution hybridization) concentrations for the two receptors and IGF-I. IGF-I immunoreactivity was present in samples from all animals, being principally located in maternal epithelium, trophoblast, endothelium and vascular smooth muscle; the latter showing the strongest labeling. The levels of receptor proteins, as well as mRNAs, were highest in the non-pregnant animals at estrus and metestrus. The pregnant animals showed decreasing concentrations to consistently low levels after day 14. Temporal changes in the studied parameters were clearly coincidental with the peak (days 13-14) in conceptus estrogen secretion, e.g. the more uniform IGF-I immunolabeling in the uterine glands (days 8-13) compared with the later stages studied; the conspicuous accumulation and release of secretory vesicles in the endometrial glands (days 8-13), marking the change in secretory quality and quantity, leading to a gradual shift from histotrophic to hemotrophic nutrition of the conceptuses, and finally, the peaking level of IGF-I mRNA in the pregnant endometrium (days 12-13) which decreased considerably thereafter. It is concluded that IGF-I activity in the porcine uterus changes with the early development of the placenta.

Insulin-like growth factor-I (IGF-I) mRNA in the small intestine of suckling and adult rats

The presence and cellular localization of insulin-like growth factor-I (IGF-I) mRNA in the small intestine of suckling and adult rats was studied. A sensitive reverse transcription (RT) competitive-polymerase chain reaction (PCR) revealed IGF-I gene expression in both age groups. Adult tissue contained 3-fold higher levels of IGF-I mRNA in comparison with sucklings. Using an in situ hybridization technique, IGF-I transcripts were localized mainly in enterocytes and goblet cells in the intestinal crypts of adult rats. By using this technique, IGF-I mRNA was not detected in jejunum of 12-day-old rats.

Localization of insulin-like growth factor-I (IGF-I) and -II messenger ribonucleic acid and type 1 IGF receptors in the ovine uterus during the estrous cycle and early pregnancy.

Insulin-like growth factor-I (IGF-I), IGF-II, and the type 1 IGF receptor (IGF-IR) gene transcripts were localized in sections of ovine uterus collected from 53 ewes at different stages during the estrous cycle and the first 3 weeks of pregnancy using in situ hybridization with synthetic oligonucleotides. Binding studies on cryostat sections using [125I]IGF-I or [125I]des(1-3)IGF-I were carried out to assess whether binding sites for IGF-I colocalized with sites of IGF-IR gene transcription. Peak IGF-I mRNA concentrations occurred at estrus in both the periepithelial endometrial stromal cells and the longitudinal and circular muscle cells of the myometrium. Concentrations of transcript returned to basal levels by 48 h after estrus. The variations in IGF-I mRNA concentration during the estrous cycle showed significant correlations with the estradiol receptor concentration in the same cell types, measured in the same animals by immunocytochemistry (P < 0.01). IGF-II mRNA was localized exclusively to the caruncular stroma (high transcription) and endometrial stroma (low transcription), with levels showing no positive correlation to the concentration of estradiol receptors, progesterone receptors, or plasma progesterone, but decreasing gradually between estrus and days 14-15 (P < 0.05). IGF-IR mRNA was localized mainly to the deep and superficial glandular epithelium, with lower levels of transcription in the caruncular stroma and myometrium. Transcription in the deep glands remained high throughout the cycle, but in the superficial glands and myometrium, a small peak occurred in the early luteal phase (days 1-2), 24-48 h after peak IGF-I gene transcription. [125I]IGF-I- and des(1-3)IGF-I-binding sites colocalized with sites of IGF-IR gene transcription. Additional binding sites were revealed with the iodinated ligands in the myometrium, superficial glands, deep stroma, and blood vessel walls, which were probably attributable to the presence of binding proteins (IGFBPs). The different affinities of des(1-3)IGF-I for IGFBP-3, -2, and -1 lead us to suggest that IGFBP-2 and/or -1 are present in the deep stroma and glands, whereas IGFBP-3 may be responsible for the additional binding sites in the myometrium and blood vessel walls. Comparisons between pregnant and nonpregnant ewes on days 2-15 after estrus did not reveal any clear pregnancy-associated differences in either the sites or levels of IGF-I, IGF-II, or IGF-IR gene transcription.

Localization of insulin-like growth factor-I (IGF-I) and -II messenger ribonucleic acid and type 1 IGF receptors in the ovine uterus during the estrous cycle and early pregnancy

Localization of insulin-like growth factor-I (IGF-I) and -II messenger ribonucleic acid and type 1 IGF receptors in the ovine uterus during the estrous cycle and early pregnancy

Insulinlike growth factor I and interleukin 1β messenger RNA in a rat model of granulomatous enterocolitis and hepatitis

Background: Insulinlike growth factor I (IGF-I) is mitogenic for fibroblasts and smooth muscle cells and stimulates collagen synthesis. The present study tested the hypothesis that IGF-I is important in the development of granulomatous inflammation and fibrosis. Methods: IGF-I messenger RNA (mRNA) was measured in bowel and liver of rats with peptidoglycan-polysaccharide-induced chronic granulomatous enterocolitis and hepatitis using RNase protection. Cellular sites of IGF-I mRNA and IGF-I peptide precursor were localized by in situ hybridization and immunohistochemistry, respectively. Sites of IGF-I synthesis were compared with sites of interleukin 1β mRNA expression. Results: IGF-I mRNA was increased 3.7-fold in cecal tissue from peptidoglycan-polysaccharide-injected rats compared with controls. IGF-I mRNA was up-regulated in fibroblastlike cells in the intensely fibrotic periphery of cecal and hepatic granulomas. This region also expressed IGF-I peptide precursor. Interleukin 1 mRNA localized to macrophage-like cells in the center of granulomas. Conclusions: IGF-I may be important in the development of fibrosis in this model of Crohn's disease. The localization of IGF-I and interleukin 1 mRNAs to distinct but adjacent sites is consistent with a paracrine interaction between cells expressing IGF-I and interleukin 1.

Ultrastructural localization of IGF-I in the rat kidney; an immunocytochemical study

We have shown recently by light microscopy that insulin-like growth factor I (IGF-I) immunoreactivity is localized in cells in the collecting ducts and in the thin loop of Henle in the normal rat kidney. In the present study, we have investigated the ultrastructural localisation of IGF-I using preembedding immunocytochemistry. The light microscopical findings were confirmed at the electronmicroscopical level. In collecting ducts as well as in the thin limb of Henle's loop a focal expression of IGF-I immunoreactivity was evident, i.e. distinctly IGF-I positive cells were intermingled with cells lacking IGF-I immunoreactivity. IGF-I immunoreactivity was found to have a diffuse cytoplasmatic distribution in both cell types. No specific association to organelles was found.

Receptor autoradiographic localization of insulin-like growth factor-I (IGF-I) binding sites in human fetal and adult adrenal glands

We report here the first evidence of insulin-like growth factor-I (IGF-I) binding sites in human fetal and adult adrenal glands, obtained at autopsy. Sections of tissue were incubated with 0.1 nM [ 125I]IGF-I and analyzed using [ 3H]Ultrofilm autoradiography with image analysis coupled to computerized microdensitometry. Specific binding sites of [ 125I]IGF-I were found to be localized in the definitive zone, fetal zone, and fetal medulla of the fetal adrenal glands. In the adult adrenal glands, the entire cortex and medulla were specifically labeled with [ 125I]IGF-I. Specific binding obtained at a concentration of 0.1 nM [ 125I]IGF-I to areas in the fetal and adult human adrenal glands was competitively displaced by unlabeled IGF-I, with an IC 50 value of 0.34 – 2.54 nM, and 0.38 – 0.73 nM, respectively, whereas insulin was much less potent in displacing the binding. Acquisition of this knowledge will aid in studies on cell growth and steroid — catecholamines biosynthesis of the human adrenal gland.