Núria Baños

Insulin, insulin-like growth factor-I (IGF-I) and glucagon: the evolution of their receptors

Insulin and glucagon, two of the most studied pancreatic hormones bind to specific membrane receptors to exert their biological actions. Insulin-like growth factors IGF-I and IGF-II are structurally related to insulin, although they are expressed ubiquitously. The biological functions of the IGFs are mediated by different transmembrane receptors, which includes the insulin, IGF-I and IGF-II receptors. The interaction of insulin, insulin related peptides and glucagon with the corresponding receptors has been studied extensively in mammals and continues to be so. At the same time, research on ectothermic animals has made enormous progress in the recent years. This paper summarizes current knowledge on insulin, IGF-I and glucagon receptors, from a comparative point of view with special attention to non-mammalian vertebrates. The review covers adult and mostly typical target tissues, and with very few exceptions, developmental aspects are not considered. Binding characteristics, tissue distribution and structure of insulin and IGF-I receptors will be considered first, because both ligands and receptors are structurally related and have overlapping functions. These sections will be followed by similar distribution of information on glucagon receptors. Readers interested in either structure or functions of insulin, IGFs and glucagon in nonmammalian vertebrates are referred to other reviews (Mommsen TP, Plisetskaya EM. Insulin in fishes and agnathans: history, structure and metabolic regulation. Rev Aquat Sci 1991;4:225-259; Mommsen TP, Plisetskaya EM. Metabolic and endocrine functions of glucagon-like peptides: evolutionary and biochemical perspectives. Fish Physiol Biochem 1993;11:429-438; Duguay SJ, Mommsen TP. Molecular aspects of pancreatic peptides. In: Sherwood NM, Hew CL, editors, Fish Physiology. vol 13. 1994:225-271; Plisetskaya EM, Mommsen TP. Glucagon and glucagon-like peptides in fishes. Int Rev Citol 1996;168:187-257.).

Influence of high-carbohydrate enriched diets on plasma insulin levels and insulin and IGF-I receptors in trout

Insulin and insulin growth factor-I (IGF-I) binding to skeletal muscle semipurified receptors were assessed in rainbow trout (Oncorhynchus mykiss) fed with different enriched carbohydrate diets. The animals were fed for 2 months, either in spring, summer or autumn with a control diet (C, commercial diet containing 21% raw carbohydrates) or with two diets supplied with highly digestible carbohydrates (E1, 22% expanded wheat; and E2, 37% expanded wheat). Insulin and IGF-I receptors were semipurified by affinity chromatography (WGA-agarose). Fish fed with a carbohydrate enriched diet did not show lower growth rates than those fed with the control diet. Independently of the season, rainbow trout fed E1 and E2 presented higher insulin and glucose plasma levels as well as higher tissue glycogen reserves than fish fed C. An increase in the number of insulin receptors during the diet adaptation was observed especially in fish fed with E2. No differences in the affinity of receptors were observed. IGF-I specific binding in skeletal muscle was higher than that of insulin in all groups and in all seasons. Furthermore, IGF-I receptors showed the same tendency as insulin receptors, with increases in their number in experimentally fed fish, especially those fed with E2. Insulin and IGF-I receptors TKA increased only slightly, as a consequence of E1 and E2 diet adaptation. In conclusion, rainbow trout can be fed high-carbohydrate levels and show good rates of growth. This adaptation determines increases in circulating glucose and insulin, and muscle insulin receptors, which indicate an adaptation of the fish to higher levels of glucose supply. The response of IGF-I receptors also suggests a possible role in the regulation of metabolism.

Regulation of plasma insulin-like growth factor-I levels in brown trout (Salmo trutta).

In this study we report that the use of a heterologous radioimmunoassay (RIA) is valid for the detection of insulin-like growth factor-I (IGF-I) levels in plasma of a variety of fish species. Parallelism between standard curves and plasma dilutions were observed and the standard curve obtained with mammalian IGF-I presented the same characteristics as that obtained with coho salmon recombinant IGF-I. The RIA was biologically validated since total plasma IGF-I values were significantly modified by different experimental conditions. Hyperinsulinemia induced either by arginine or insulin injection was accompanied by increases in IGF-I plasma levels in brown trout (Salmo trutta). In contrast, parallel decreases in insulin and IGF-I circulating levels were observed after 45 days of fasting and 20 days after a single streptozotocin injection. Administration of arginine in fasted fish led to a relative increase in insulin and IGF-I plasma concentrations, while arginine injection in fish previously treated with streptozotocin increased IGF-I levels only. The above data suggest that insulin, together with other factors, may act to increase the levels of IGF-I in plasma.

Up-regulation of insulin binding in fish skeletal muscle by high insulin levels.

The effects of insulin titres on regulation of receptor binding were studied in several fish species. Insulin receptors were semi-purified by affinity chromatography (WGA-agarose) from skeletal muscle of carp, brown trout and rainbow trout that had been subjected to increases in insulinemia produced either by arginine injection, food administration, or adaptation to an experimental diet (extruded diet with high-digestibility carbohydrates). Arginine injection provoked acute hyperinsulinemia in both carp and trout. Specific binding of insulin to the skeletal muscle was significantly increased 3 h after injection (from 5.8 +/- 0.3 to 9.6 +/- 0.9%/10 micrograms protein in carp and from 0.8 +/- 0.2 to 1.5 +/- 0.4%/10 micrograms in trout). The same effect was observed in carp liver preparations (from 6.0 +/- 0.75 to 9.9 +/- 1.25%/10 micrograms). No alterations in tyrosine kinase activity of the receptors were detected in either carp or trout preparations: basal activities of the receptors were maintained (3100 +/- 200 fmol P/fmol receptors/30 min and 3700 +/- 400 fmol P/fmol receptors/30 min, in carp and trout, respectively), as were the percentage of stimulation over basal levels obtained by incubation with insulin (227 +/- 25% and 160 +/- 10% respectively). Food ingestion raised plasma insulin levels more steadily. Specific binding also increased in skeletal muscle preparations, especially in carp (from 5.7 +/- 0.3 to 11 +/- 1.7%/10 micrograms at 4 h and 10 +/- 0.7%/10 micrograms at 8 h). Tyrosine kinase activity was maintained without significant changes. Rainbow trout adapted for 2 months to an extruded diet presented higher insulin titres and higher glycogen reserves in liver and muscle. Insulin binding to skeletal muscle preparations was also significantly increased (from 0.36 +/- 0.02 to 0.77 +/- 0.1%/10 micrograms), as was tyrosine kinase activity (from 132 +/- 4% to 156 +/- 6%, without alterations in the basal activity). Results showed that fish can respond to both acute and maintained increases in insulinemia by increasing the number of insulin receptors. Tyrosine kinase activity, in contrast, is only modified after long-term adaptation.

Fish Insulin, IGF-I and IGF-II Receptors: A Phylogenetic Approach1

SYNOPSIS. In fish, the structural and functional characteristics of insulin and IGFI receptors have been well studied. Current evidence indicates that all gnatostome animals, from fish to mammals, contain separate insulin and IGF-I molecules and specific receptors for insulin and IGF-I. However, qualitative differences in the functional aspects of insulin and IGF-I receptors among vertebrate species can account for variations in the biological activity of insulin and IGF-I. In this paper we will focus on the functional evolution of the insulin and IGF-I receptors in vertebrates and on the appearance of the unrelated IGF-II receptors.

Insulin and insulin-like growth factor-I (IGF-I) binding in fish red muscle : regulation by high insulin levels

Insulin and IGF-I binding to semi-purified red muscle receptors was characterized in brown trout, Salmo trutta and the common carp. Cyprinus carpio. The yield of glycoprotein obtained after semipurification of receptors with WGA-agarose affinity chromatography in microgram g-1 initial tissue was 210.6 +/- 21 micrograms g-1 in trout and 108.5 +/- 2.5 micrograms g-1 in carp. IGF-I specific binding (4.72 +/- 0.64%/10 micrograms glycoprotein) was 4-5-times higher than insulin binding (1.04 +/- 0.12%/10 micrograms glycoprotein) in trout red muscle. This difference in binding was due to a higher number and a greater affinity of the IGF-I (Kd, 0.21 +/- 0.03 nM) compared with the insulin (Kd, 0.67 +/- 0.06 nM) receptors in this tissue. Carp red muscle IGF-I binding (9.14 +/- 0.55%/10 micrograms glycoprotein) surpassed insulin binding (2.59 +/- 0.094%/10 micrograms glycoprotein) mainly because of a greater affinity of the IGF-I (Kd, 0.092 +/- 0.027 nM) compared with the insulin (Kd, 0.1515 +/- 0.0285 nM) receptor. IGF-I and insulin binding in carp red muscle were higher than in trout, as a consequence of a higher affinity of carp red muscle receptors. Arginine injection provoked acute hyperinsulinemia in both trout (23.3 +/- 1.01 ng ml-1) and carp (24.3 +/- 1.34 ng ml-1. Specific binding of insulin and IGF-I to the red muscle decreased 4 h after injection. In trout, a decrease of insulin and IGF-I binding of 47.0% and 63.3%, respectively was observed compared with controls, in carp, these values were 44.0% and 45.0%. The number of insulin and IGF-I receptors decreased (42-55%) but affinities did not change suggesting that receptor down-regulation is a consequence of high insulin levels.

Effects of fasting and feeding on plasma amino acid levels in brown trout

Plasma amino acid (AA) levels were analysed at various intervals during fasting and after feeding in juvenile brown trout, Salmo trutta. After 3 days of food deprivation, total plasma AAs decreased significantly (–26% vs. control) due to a fall in the most abundant essential AAs (EAA): branched-chain amino acid (BCAA) and Thr, and there was also a decrease in non-essential AA (NEAA) levels, especially Ala and Asn. After 15 days of fasting, BCAA increased, possibly related to muscle protein mobilization. After 50 days of food deprivation, a significant increase in Gln levels (40% vs. 15 days of fasting) and a decrease in BCAA (–25% vs. 15 days of fasting) was noted. Feeding a commercial trout pellet caused a clear and progressive increase in plasma AA levels. Both EAA and NEAA concentrations peaked at 11 h post-feeding (4068 ± 160 and 4261 ± 239 µM NEAA, respectively) with a greater increase in EAA (+62%) than NEAA (40%) with respect to control values. These data show that changes in the plasma AA pool clearly reflect the nutritional state of fish and help us to understand the complex AA metabolism.

Appearance of insulin and insulin-like growth factor-I (IGF-I) receptors throughout the ontogeny of brown trout (Salmo trutta fario).

Insulin and IGF-I receptors were characterized in glycoprotein fractions prepared by affinity chromatography from different developmental stages of brown trout. The specificity of insulin and IGF-I binding was demonstrated by crossed-competition assays: unlabelled insulin displaced bound radiolabelled insulin at concentrations 45-fold lower than unlabelled IGF-I, whilst unlabelled IGF-I displaced bound radiolabelled IGF-I at concentrations 2,000-fold lower than unlabelled insulin. The affinity of these receptors did not change significantly during trout development. Insulin-specific binding was detectable 3 weeks after spawning, after which it increased to a maximum in fry weighing 0.4 g, and decreased progressively to adult levels. IGF-I specific binding was detectable in newly laid eggs and increased to a maximum during organogenesis in eyed eggs. It then decreased progressively during subsequent stages of development to adult levels. The apparent molecular weight (Mr) of the alpha-subunit of brown trout insulin and IGF-I receptors was smaller than that of the alpha-subunit of the rat insulin receptor. Receptor tyrosine kinase activity was stimulated in a dose-dependent manner by insulin and IGF-I. Insulin and IGF-I stimulated tyrosine kinase activity and reached a maximum of 201 +/- 17.6 and 240 +/- 29.6% of basal phosphorylation, respectively.

Definition of Failed Induction of Labor and Its Predictive Factors: Two Unsolved Issues of an Everyday Clinical Situation.

Objective: The objectives of this review were to identify the predictive factors of induction of labor (IOL) failure or success as well as to highlight the current heterogeneity regarding the definition and diagnosis of failed IOL. Materials and Methods: Only studies in which the main or secondary outcome was failed IOL, defined as not entering the active phase of labor after 24 h of prostaglandin administration ± 12 h of oxytocin infusion, were included in the review. The data collected were: study design, definition of failed IOL, induction method, IOL indications, failed IOL rate, cesarean section because of failed IOL and predictors of failed IOL. Results: The database search detected 507 publications. The main reason for exclusion was that the primary or secondary outcomes were not the predetermined definition of failed IOL (not achieving active phase of labor). Finally, 7 studies were eligible. The main predictive factors identified in the review were cervical status, evaluated by the Bishop score or cervical length. Discussion: Failed IOL should be defined as the inability to achieve the active phase of labor, considering that the definition of IOL is to enter the active phase of labor. A universal definition of failed IOL is an essential requisite to analyze and obtain solid results and conclusions on this issue. An important finding of this review is that only 7 of all the studies reviewed assessed achieving the active phase of labor as a primary or secondary IOL outcome. Another conclusion is that cervical status remains the most important predictor of IOL outcome, although the value of the parameters explored up to now is limited. To find or develop predictive tools to identify those women exposed to IOL who may not reach the active phase of labor is crucial to minimize the risks and costs associated with IOL failure while opening a great opportunity for investigation. Therefore, other predictive tools should be studied in order to improve IOL outcome in terms of health and economic burden.

Quantitative Analysis of the Cervical Texture by Ultrasound and Correlation with Gestational Age

Objectives: Quantitative texture analysis has been proposed to extract robust features from the ultrasound image to detect subtle changes in the textures of the images. The aim of this study was to evaluate the feasibility of quantitative cervical texture analysis to assess cervical tissue changes throughout pregnancy. Methods: This was a cross-sectional study including singleton pregnancies between 20.0 and 41.6 weeks of gestation from women who delivered at term. Cervical length was measured, and a selected region of interest in the cervix was delineated. A model to predict gestational age based on features extracted from cervical images was developed following three steps: data splitting, feature transformation, and regression model computation. Results: Seven hundred images, 30 per gestational week, were included for analysis. There was a strong correlation between the gestational age at which the images were obtained and the estimated gestational age by quantitative analysis of the cervical texture (R = 0.88). Discussion: This study provides evidence that quantitative analysis of cervical texture can extract features from cervical ultrasound images which correlate with gestational age. Further research is needed to evaluate its applicability as a biomarker of the risk of spontaneous preterm birth, as well as its role in cervical assessment in other clinical situations in which cervical evaluation might be relevant.