Enteral absorption of erythropoietin in the suckling rat.

Milk contains biologically relevant concentrations of erythropoietin (Epo), the primary hormone responsible for erythrocyte production. In animals, milk-borne Epo stimulates erythropoiesis. Epo receptors have been found in nonerythropoietic tissues including gastrointestinal tract. We hypothesized that milk-borne Epo is distributed to local gastrointestinal tissues, absorbed intact, and then distributed peripherally via the systemic circulation. Rat milk protected recombinant human Epo (rhEpo) from degradation in the suckling rat gastrointestinal tract. Simulated digestion of (125)I-rhEpo in suckling rat gastrointestinal juices was performed. When measured by acid precipitation and immunoassay, rat milk protected rhEpo from gastrointestinal juices better than saline (p < 0.0001). The fate of enterally administered milk-borne (125)I-rhEpo was examined in 10-d-old rats. RhEpo fed in rat milk was better protected from in vivo proteolytic degradation than rhEpo in saline (p < 0.05). After enteral (125)I-rhEpo dosing, radiolabeled protein from gastric tissue comigrated on SDS-PAGE with intact rhEpo at 36.5 kD. To determine the local and systemic distribution of physiologic intakes of rhEpo, suckling rats were fed (125)I-rhEpo in rat milk, and tissues were harvested 1, 2, and 4 h later. Intact (125)I-rhEpo was found in gastric and small intestinal walls and lumens. Five percent of total administered dose was found intact in the plasma, whereas another 8 to 10% of total administered dose was localized to bone marrow, percentages comparable to those seen after parenteral administration. Radiolabel was also localized to liver and peripheral solid tissues. These patterns of localization and degradation of rhEpo after acute administration support both systemic absorption and gastrointestinal cellular processing.

Transforming growth factor-alpha delays gastric emptying and small intestinal transit in suckling rats.

Transforming growth factor-alpha (TGF-alpha) is a biologically potent polypeptide detected in the gastrointestinal tract in suckling rats. The major goal of the present study was to test the hypothesis that the administration of TGF-alpha affects gastric emptying and small intestinal transit in suckling rats. Suckling (12-day-old) rats fasted 16 h received rat TGF-alpha subcutaneously (s.c.) or orogastrically in varying doses (0, 0.5, 1.0 microg/rat in 0.1% BSA). Control animals received 0.1% BSA only. Poly R-478 dye was used as a motility marker. Rats were decapitated 45 min after marker administration and the amount of dye in the stomach and the small intestine was measured by spectrophotometry. Subcutaneous administration of TGF-alpha significantly delayed stomach evacuation. In controls, the stomach contained 21.4 +/- 1.4% (mean +/- s(x)) of the Poly R-478 marker, whereas in TGF-alpha treated rats the stomach contained 37.2 +/- 2.8% of the total Poly R-478 dye given to animals. The delaying effect of TGF-alpha was time- and dose-dependent. Small intestinal transit was also significantly delayed. The proximal jejunum of TGF-alpha treated rats contained a 1.4-fold higher amount of marker in comparison with control rats. Orogastrically administered rTGF-alpha did not affect gastric emptying or intestinal transit. In conclusion, s.c. administration of rat TGF-alpha significantly delayed the gastrointestinal motility in vivo in suckling rats.

Secretion of insulin-like growth factor-II into bile of rats of different ages.

Bile from rats of different ages (suckling 10-12 days; weanling 30-33 days, and adult 60-70 days) was collected and studied for the presence of immuno- and receptor-assayable insulin-like growth factor-II (IGF-II) concentrations. Concentrations of RIA IGF-II in bile were highest in suckling rats (230 +/- 38 ng/ml) and lowest in adults (47 +/- 7 ng/ml). These concentrations were approximately twice those of the bile IGF-I concentration in sucklings, as measured in a previous study. Selected bile samples were also assayed using a competitive binding assay with a crude preparation of adult rat liver membranes bearing the IGF-II receptor. These studies confirmed the presence of receptor- (as well as immuno-) active IGF-II in bile. Since bile flow rates increased dramatically after the suckling period, bile delivery rates of IGF-II were normalized as picograms per gram body weight per hour. When such calculations were done, bile IGF-II delivery rates to the small intestine were highest in sucklings and weanlings in comparison to adult rats. Thus non-enterically derived (milk- and bile-borne) IGF-II delivery to the suckling small intestine can be approximated at roughly 1 microg/day. Unlike IGF-I, intravenously injected IGF-II could not be detected in suckling bile, suggesting a predominantly hepatic origin. From this study we conclude that there exists a significant delivery of receptor-active IGF-II to the gastrointestinal tract of rats of all ages.

Absorption of milk-borne insulin-like growth factor-I into portal blood of suckling rats.

BACKGROUND: Insulin-like growth factors (IGFs) are potent mitogens that have been implicated in control of growth and development during the perinatal period. These hormones are also present in biologically significant quantities in mammalian milks. Although one site of action of these IGFs may be at the intestinal level, current information about whether they pass intact into the circulation is conflicting. METHODS: To test the hypothesis that milk-borne IGFs are absorbed into blood in receptor-active form, suckling rats were given either recombinant human (rh)125I-IGF-I or -II (4 x 10(6) counts per minute [cpm]), and the activity present in portal and cardiac blood was examined at 5, 10, 20, and 30 minutes after ingestion for presence of appropriate molecular weight peptides in these samples. In selected samples, purified radioactive samples were tested for their ability to bind competitively to crude membranes bearing IGF receptors. RESULTS: The results of these studies indicate that rh125I-IGF-I is absorbed in receptor-active form into the portal circulation and that maximal amounts are present 20 to 30 minutes after ingestion. Estimation of the presence of intact hormone was made on the basis of the elution profile of samples when run on gel chromatography as well as reversed-phase high-performance liquid chromatography. Isolated samples from portal blood also bound competitively to placental membranes bearing IGF receptors. In contrast, rh125I-IGF-II could not be demonstrated in receptor-active form in portal blood. Chromatography showed appropriate sized peaks with greater activity in portal than cardiac samples, but competitive binding was not appreciated. CONCLUSIONS: It is likely that at least milk-borne IGF-I is absorbed intact and may exert effects on liver and other peripheral tissues. In addition, this study lends further credence to the possibility of an enterohepatic circulation for IGF-I.

Milk-borne epidermal growth factor modulates intestinal transforming growth factor-alpha levels in neonatal rats.

Epidermal growth factor (EGF) is present in milk from various mammalian species, but its physiologic function in neonatal development remains unclear. Transforming growth factor-alpha (TGF-alpha) is a peptide structurally related to EGF, and its presence is detected in the developing small intestine of rats. The purpose of the present study was to examine the effect of milk-borne EGF on endogenous production of EGF and TGF-alpha in the small intestine of suckling rats. Neonatal rats were fed via gastrostomy either growth factor-free rat milk substitute (RMS) or RMS supplemented with EGF (100 ng/mL of RMS) from 8 to 12 d of age. Artificially reared rats were then compared with their dam-fed littermates. Animals fed the EGF-deficient diet RMS had markedly increased EGF and TGF-alpha mRNA levels in duodenum and ileum compared with dam-fed controls and significantly elevated total intestinal content of TGF-alpha peptide. Intestinal EGF content and EGF serum levels were significantly decreased in the RMS group compared with controls. The addition of EGF to the RMS diet normalized TGF-alpha mRNA levels in the duodenum and ileum, EGF mRNA levels in the ileum, and total intestinal TGF-alpha content and EGF serum levels to the levels measured in dam-fed littermates. Motility studies showed that enteral administration of EGF did not affect stomach emptying and intestinal transit. These studies indicate that exogenous milk-borne EGF modulates endogenous production of TGF-alpha in developing small intestine. It is likely that neither TGF-alpha nor EGF are solely responsible for small intestinal overgrowth of artificially reared neonatal rats.

Effect of milk on somatostatin degradation in suckling rat jejunum in vivo.

BACKGROUND: Somatostatin-14 is present in breast milk, and intact somatostatin-14 has been recovered from gastric lumen of infants. Studies have shown that somatostatin-14 is metabolized in the intestinal luminal contents in vitro, which could be prevented by the presence of breast milk. In this study, the effect of milk on stability of somatostatin-14 in suckling rat jejunum in vivo was examined. METHODS: 125I-Somatostatin-14[Tyr 11] was administered to the isolated jejunal loops in anesthetized suckling rats in the absence or presence of milk, fractions of milk, or known protease-peptidase inhibitors. Structural integrity of 125I-somatostatin-14[Tyr 11] recovered from tissues at different intervals was analyzed by gel filtration and high-performance liquid chromatography. RESULTS: Radioactivity rapidly disappeared from the jejunal lumen with a 50% clearance achieved by 1.2 minutes. Gel filtration and high-performance liquid chromatography analyses showed that 125I-somatostatin- 14[Tyr 11] was rapidly degraded into smaller fragments. At 1 minute, jejunal luminal radioactivity was eluted in a major peak with retention time of 42.4 minutes, along with other minor peaks (retention time, 5.6, 8.0, 10.4, and 14.4 minutes); only a trace amount of intact 125I-somatostatin-14[Tyr 11] (retention time, 44.8 minutes) was present. Coadministration of rat's milk or its soluble fraction increased the level of intact 125I-somatostatin-14[Tyr 11] in the jejunal lumen and jejunal tissue. Presence of rat's milk-casein or peptidase inhibitors (bestatin, phosphoramidon, or Bowman-Birk inhibitor), however, failed to increase the level of intact 125I-somatostatin-14[Tyr 11]. CONCLUSION: These results suggest that somatostatin-14 is rapidly degraded in the jejunal lumen of suckling rats, and that milk-borne peptidase inhibitors prevent this somatostatin-14 degradation.

The expression of epidermal growth factor and transforming growth factor-alpha mRNA in the small intestine of suckling rats: organ culture study.

Epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) are associated with regulation of various gastrointestinal functions. In order to better understand their role in developing small intestine EGF, TGF-alpha and EGF-R steady-state mRNA levels and transcript stability were determined. Reverse transcription (RT) competitive-polymerase chain reaction (PCR) revealed that intestinal TGF-alpha mRNA levels were 10-fold higher in comparison with EGF mRNA. The primary intestinal culture technique was used to evaluate mRNA stability. The stability of TGF-alpha mRNA was remarkably lower than the stability of EGF mRNA. High levels of TGF-alpha mRNA accompanied by high degradation rate of this mRNA suggested a rapid turnover of intestinal TGF-alpha mRNA.

Luminal stability of insulin-like growth factors I and II in developing rat gastrointestinal tract.

BACKGROUND: Insulin-like growth factors (IGF)-I and -II are present in milk of a number of mammalian species. The stability of IGF-I and -II in the intestinal lumen was investigated by measuring the proteolytic degradation of 125I-labeled IGF-I and IGF-II by rat (suckling and adult) intestinal luminal flushings in vitro. METHODS: Degradation of 125I-labeled IGF-I and IGF-II was assessed by measuring the generation of acid-soluble radioactivity and the reduction of the amounts of peak activity (gel filtration). Degradation was confirmed by measuring the loss of immunoreactivity and receptor activity. RESULTS: Incubation of 125I-IGF-I with midjejunal luminal flushings from 12-day-old suckling rats generated acid-soluble radioactivity in a time- and dose-(flushing) dependent manner, whereas incubation of 125I-IGF-II produced only minor amounts of acid-soluble radioactivity. Degradation activity in luminal flushings from adult rat intestine was several times greater than that in luminal flushings from suckling rats. Degradation of 125I-IGF-II was several times lower than that of 125I-IGF-I in the intestinal luminal flushings from suckling and adult rats. The rate of decrease in immunoprecipitable 125I-IGF-I was considerably lower than the rate of decrease in receptor-active radioactivity. Except for immunoreactivity, analyses of acid-precipitable, peak-A, and receptor-active radioactivities demonstrate that IGF-II is relatively more stable than IGF-I in luminal flushings of suckling rat duodenum, jejunum, and midjejunum. CONCLUSIONS: These results show that the stability of IGF in the gastrointestinal lumen depends on the age of the animal and the segment of the gastrointestinal tract, as well as on the peptide isoform.

Human milk as a potential enteral source of erythropoietin.

In addition to its content of traditional nutrients, milk is a rich source of hormones and peptides, which survive digestion in the neonatal gastrointestinal tract secondary to lower proteolytic activity and increased protein permeability. Previous studies have shown accelerated erythropoiesis or elevated serum erythropoietin (Epo) levels in neonatal (suckling) animals after maternal phlebotomy or maternal hypoxia exposure. We sought to determine whether significant quantities of Epo are present in human milk and whether Epo remains intact under physiologic digestion conditions. Immunoreactive Epo concentrations were determined in 409 human milk samples obtained from mothers of term and premature infants. Samples collected between birth and postpartum d 134 were divided into 11 postpartum day groups. Mean milk-borne Epo concentrations were within the normal range for plasma Epo concentrations and rose with postpartum day (F10,398 = 5.82, p < 0.0001). No differences were observed between milk collected from mothers of premature versus term infants. Estimated weekly human milk-borne Epo intakes approximated the lower range of published parenteral therapeutic doses. In simulated digestion at physiologic pH levels of 3.2, 5.8, and 7.4, milk-borne Epo resisted degradation at 1 and 2 h, compared with baseline. Therefore, we conclude that human milk contains considerable amounts of Epo which resist degradation after exposure to gastric juices at physiologic pH levels. These results support continued investigation into the fate and developmental roles of Epo in human milk.

Growth of artificially fed infant rats: effect of supplementation with insulin-like growth factor I.

Insulin-like growth factor I (IGF-I), a potent mitogenic peptide, is present in considerable quantities in most mammalian milks, but its importance for the neonate is unknown. To test the hypothesis that milk-borne IGF-I is an important factor in the regulation of neonatal growth, as well as that of the gastrointestinal tract, rat pups were fed a rat milk substitute (RMS) devoid of growth factors via gastrostomy. These animals were compared with those given RMS supplemented with recombinant human IGF-I added at a concentration of 500 ng/ml. Animals given RMS + IGF-I gained mere weight than controls, although skeletal growth as represented by elongation of the tail was no different. Animals fed RMS + IGF-I had increased brain and liver wet weights as well as increased liver and small intestine protein contents. Serum IGF-I concentrations in the IGF-I-supplemented group were more than twofold above RMS controls and were similar to dam-fed rat pups. Semiquantification of serum IGF-binding proteins (IGFBP) in these animals documented that in IGF-I-supplemented pups the amount of 38- to 40-kDa molecular mass IGFBP species was also greater than in RMS controls. The rate of migration of enterocytes from crypts in duodenum and proximal jejunum was greater in IGF-I-supplemented animals than in rats fed RMS alone. These studies suggest that milk-borne IGF-I is important in modulation of somatic and gastrointestinal tract growth in the neonatal rat.

Effect of milk-borne epidermal growth factor on the hepatic microcirculation and Kupffer cell function in suckling rats.

Epidermal growth factor (EGF) is contained in breast milk, is transported intact across the gastrointestinal mucosa during the suckling period, retains its biological activity, and affects hepatic growth. Whether or not EGF also influences postnatal hepatic development by affecting nonparenchymal cells or by modifying intrahepatic blood is not clear. As a result, the effect of EGF on the hepatic microvasculature was studied in suckling rats fed rat milk substitute (RMS) with and without EGF (100 ng/ml, i.e. twice the normal intake in breast milk) between days 11 and 14 and compared to pups breast-fed for 14 days. The livers of anesthetized pups were examined by in vivo microscopy to determine the numbers of sinusoids with flow (SCF) in each of 10 microscopic fields and the numbers of phagocytic Kupffer cells (KC) in the same fields following an intraportal injection of fluorescent 1-micron latex particles. Phagocytic activity was expressed as the ratio KC/SCF. In pups fed RMS without EGF, SCF and KC/SCF was 75 and 45%, respectively, of that in breast-fed animals. The addition of EGF to the RMS restored SCF and KC/SCF nearly to the levels measured in breast-fed pups. Thus, the results suggest that milk-borne EGF plays a role in the development of KC phagocytic function and affects the amount of blood that perfuses the sinusoidal bed in the suckling.

Organ distribution and biliary excretion of intravenously injected insulin-like growth factor-I in suckling rats.

The organ distribution of intravenously injected 125I-IGF-I at a dose of 2-4 x 10(6) cpm or 5-10 ng/animal was studied in 10- to 12-day-old Sprague-Dawley rats at 5 and 30 min after injection. Results of the study suggest that, although the main portion of intravenous IGF-I remains in the circulation, significant amounts are also found in the carcass, liver and kidney. Blood radioactivity fell by 50% 30 min after injection, but concentrations in the carcass, liver, kidney and skin either remained stable or increased. Gel chromatography demonstrated that significant portions of radioactivity recovered from serum, liver and kidney coeluted in a position identical to the injected IGF-I. In addition, the extracted peptide bound competitively to a membrane IGF-I receptor preparation. Studies performed on liver and kidney from these animals 5 min after injection showed that on a per gram wet weight basis, these organs contained equivalent amounts of 125I-IGF-I. However, although by 30 min, 65% of the intact labelled IGF-I has been removed from the liver, the amount remaining in kidney tissue was equal to that noted 5 min after injection. Bile was collected over a 2-hour period and contained approximately 2% of the injected radioactivity and a significant portion (30%) of this radioactivity coeluted with native IGF-I. This material also bound competitively in a radioreceptor assay, suggesting 'intactness' of this peptide. From this study, we conclude that (a) IGF-I, when administered intravenously, remains for at least 30 min in a receptor-active form in blood and several organs; (b) IGF-I derived from the circulation is cleared from the liver more quickly than from the kidney of suckling rats, and (c) that IGF-I is transferred from blood to bile.

The potential physiological significance of milk-borne hormonally active substances for the neonate.

This article reviews the presence and potential physiological significance of hormones and hormonally active substances (including growth factors) in human milk. Human milk has been found to contain several nonpeptide hormones and many peptide hormones and growth factors. In contrast to human breast milk, infant formulae lack some hormonally active peptides. There is little data concerning the effects of these agents on human neonates. Studies in immature experimental animals showing effects of orogastically administered hormones are summarized. The problems of supplementation of infant formula are discussed. Since hormones are present in the milk as a "cocktail" of potentially agonistic and antagonistic substances, one question is whether supplementation with a single agent would disturb this balance.

Milk carnitine affects organ carnitine concentration in newborn rats.

Previous studies suggest that exogenous milk carnitine may be necessary during the suckling period to maintain normal fat metabolism. To characterize the relationship between milk carnitine and carnitine in body organs, newborn rats were fed from birth a rat milk substitute with or without 300 micromol/L L-carnitine, corresponding to the concentration present in rat milk, for either 2 or 4 d. Carnitine concentrations in heart, skeletal muscle, liver and small intestine were compared with levels in rat pups that were never fed (d 0) and those that were nursed by their mothers for 4 d. Carnitine supplementation resulted in significantly higher concentrations of carnitine in all organs studied after 4 d compared with nursed controls. Relative intestinal carnitine pool size was 38.1 +/- 3.0, 22.6 +/- 1.0, 7.9 +/- 0.5 and 2.3 +/- 0.7 micromol/g body wt in supplemented, nursed, unsupplemented and never fed pups, respectively (P < 0.05, compared with one another). These results indicate that carnitine organ concentrations are related to dietary intake during the early suckling period and that the small intestine is a considerable and previously unrecognized proportion of the carnitine pool of suckling animals.

Nutrient absorption in the preterm neonate.

The majority of what we know about the development of the absorptive process is derived from animal studies, studies in human fetal or stillborn tissues, and epidemiologic investigations derived from clinical experience. One can readily ascertain from this review that the absorption of nutrients in the intestine of the premature infant relates to a dynamic developmental process in which the consecutive stages are pre-programmed but can also be regulated by environmental factors. An understanding of these factors may lead to therapeutic intervention in premature infants, as has been the case for the developing lung and respiratory distress syndrome. Application of this knowledge to the critically ill premature infant in the intensive care unit will need to proceed cautiously, but it is likely to yield major benefits in terms of decreased short- and long-term morbidity in these highly vulnerable patients.

Epidermal growth factor delays gastric emptying and small intestinal transit in suckling rats.

Suckling (12-d-old) rats that were fasted for 15 h received epidermal growth factor (EGF) s.c. (0.5 and 1.0 microgram per rat, i.e. approximately 2 and 4 micrograms/100 g of body weight), together with motility markers 51Cr-EDTA or Poly R-478, and were killed 45 min later. Counts were measured separately in the stomach and the small intestine, which was divided into 12 segments. Administration of EGF delayed gastric emptying. In controls, the stomach contained 26.1 +/- 1.6% (mean +/- SEM); in EGF-treated rats the stomach contained 75.9 +/- 10.2% and 75.7 +/- 2.5% of the total 51Cr-EDTA counts given. EGF had the maximum effect (1.0 microgram) when given simultaneously with 51Cr-EDTA. Significant, but lower, effects of EGF were seen with the administration of EGF preceded by 10 min or followed by 10 and 20 min with the administration of 51Cr-EDTA (65.8 +/- 5.8%, 60.0 +/- 6.4%, and 54.1 +/- 4.2%, respectively). Small intestinal transit was also delayed. Administration of anti-EGF antiserum did not affect gastric emptying, but accelerated small intestinal transit as determined 30 min after administration of 51Cr-EDTA. These studies are the first to demonstrate the effect of EGF on gastrointestinal motility in vivo in suckling mammals.

Diet-induced changes in gene expression of lactase in rat jejunum.

To explore the mechanisms by which jejunal lactase activity is modified by carbohydrate and/or fat intake, mRNA levels and the absolute synthesis rate of lactase-phlorizin hydrolase (LPH) were determined in 6-wk-old rats that were fed either low-starch diets containing long-chain triacylglycerol (LCT, 73% energy as corn oil) or medium-chain triacylglycerol (MCT, 66% energy as MCT, 7% energy as corn oil), or a high-starch diet (70% energy as cornstarch) for 7 days. LPH mRNA levels in the jejunum were similar between LCT-fed and MCT-fed rats, but animals fed the high-starch diet exhibited a greater (2x) LPH mRNA level than other groups. The absolute synthesis rate of LPH, estimated by the flooding dose technique using [3H]phenylalanine, was greater (2.4x) in rats fed the high-starch diet than in other groups. A short-term force-feeding experiment revealed that sucrose was able to evoke LPH mRNA levels within 12 h but that a nonmetabolizable sugar (alpha-methylglucoside) was unable to enhance it. By contrast, animals fed the high-LCT diet showed a lower (by 30%) lactase activity than rats fed the low-starch, high-MCT diet, which was accompanied by not only a reduction of immunoreactive LPH in brush-border membranes but also a reduction in lactase activity per unit weight of immunoreactive LPH. These results suggest that both gene expression and posttranslational events of LPH might be influenced by dietary manipulations; carbohydrate intake primarily increases LPH mRNA levels, and LCT accelerates inactivation and/or degradation of lactase.

Fate of insulin-like growth factors I and II administered orogastrically to suckling rats.

Milk-borne insulin-like growth factors I and II (IGF-I and -II) may be of importance in the differentiation of the gastrointestinal tract of the suckling. To test this hypothesis, 10- to 11-d-old suckling rats were given via an orogastric tube 125I-IGF-I (n = 6) or 125I-IGF-II (n = 6) in rat milk and killed 30 min later. The results of this study demonstrated that approximately 40% of the radioactivity administered was detected in the gastrointestinal tract for both 125I-IGF-I and 125I-IGF-II experiments. Gel chromatography of acid extracts of homogenates of gastrointestinal tissues and luminal contents demonstrated that a significant fraction of recovered radioactivity eluted in a position identical to "native" IGF. These findings were confirmed by subjecting similarly treated samples to high performance liquid chromatography. In addition, radioactive material recovered from M(r) 7,500 fractions bound specifically to crude membrane IGF-I and -II receptor preparations, further suggesting the preservation of biologic activity of the recovered peptides. Although skin homogenates contained large peptide fragments of 125I-IGF-I, no "intact" IGF was found in the blood or other tissues. These findings suggest that milk-borne IGFs are stable in the neonatal gastrointestinal tract and remain biologically active for as long as 30 min postingestion.

Presence of insulin-like growth factor I but absence of the binding proteins in the bile of rats.

Whereas insulin-like growth factor I (IGF-I) has been found in various body fluids from different species, the presence or absence of IGF and associated binding proteins (IGFBPs) in bile has not been clearly defined. Bile concentration of IGF-I was measured in this study and found to be highest in the neonate and lowest in adult rats [133 +/- 15.9, 79.4 +/- 10.5, 45.3 +/- 12.7 ng/ml (mean +/- SE) in 12-day-old, 33-day-old, and adult rats, respectively]. When bile delivery rates of IGF-I (i.e., the product of IGF-I concentration in bile and the biliary flow rate) were calculated, IGF-I delivery was highest in weanling rats (469 pg.h-1.g body wt-1). When expressed as amount of IGF-I in bile delivered per day, however, delivery rates rose from 0.2 micrograms/day in the suckling and remained constant at 1.6-1.7 micrograms/day in both weanling and adult animals. Bile samples exposed to a placental membrane IGF receptor preparation showed significant dose-dependent inhibition of binding of native IGF-I. Because no IGF binding proteins were identified by Western ligand blot or by Sephadex gel chromatography, the results suggest the presence of biologically significant quantities of bioactive IGF-I in bile. We speculate that IGF-I in bile may play an important role in the growth of the gastrointestinal tract, both in the suckling as well as later in life.