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Biomedical subjects

G Norstedt

Publications and source records attributed to G Norstedt.

At least 73 records · Page 4Linked to original sources
PubMed1990

Decreased expression of the growth hormone receptor and growth hormone binding protein in rat liver nodules.

The growth hormone receptor (GHR) and growth hormone-binding protein (GH-BP) expression were characterized in liver nodules and hepatomas from male Wistar rats. The mRNA levels of GHR and GH-BP, studied by northern blot analysis and solution hybridization, were 35-50% (in nodules) and 2-6% (in hepatomas) of the level found in liver from untreated, age-matched rats. The binding of 125I-labeled human growth hormone to a low-density membrane fraction (LDMF) containing Golgi and endosome membranes in the presence of excess ovine prolactin was 75-80% lower in nodules than in liver. When endogenous ligand was removed, the binding increased in nodules but not in liver. Affinity cross-linking experiments revealed identical specific receptor-binding protein complexes at Mr of 95,000, 55,000, and 43,000 in both nodules and liver, assuming stochiometric binding of ligand. The in vivo endocytosis of 125I-labeled bovine growth hormone correlated with the level of GH binding and was thus reduced 75-80% in nodules, compared with liver. The level of insulin-like growth factor-I (IGF-I) mRNA was reduced by 50% in nodules; however, GH administration resulted in a twofold induction of IGF-I mRNA in both nodules and liver. It is concluded that the greater proportion of occupied GHR in nodules could result from an impaired dissociation of endogenous GH, which might explain the reduced GHR mRNA expression. Furthermore the low GHR mRNA expression may reflect a de-differentiated phenotype in nodules.

Animals↗
PubMed1990

Introduction of exogenous growth hormone receptors augments growth hormone-responsive insulin biosynthesis in rat insulinoma cells.

The stimulation of insulin biosynthesis in the pancreatic insulinoma cell line RIN5-AH by growth hormone (GH) is initiated by GH binding to specific receptors. To determine whether the recently cloned rat hepatic GH receptor is able to mediate the insulinotropic effect of GH, we have transfected a GH receptor cDNA under the transcriptional control of the human metallothionein promoter into RIN5-AH cells. The transfected cells were found to exhibit an increased expression of GH receptors and to contain a specific GH receptor mRNA that was not expressed in the parent cell line. The expression of GH receptors in one clone (1.24) selected for detailed analysis was increased 2.6-fold compared to untransfected cells. The increased GH receptor expression was accompanied by an increased responsiveness to GH. Thus, the maximal GH-stimulated increase of insulin biosynthesis was 4.1-fold in 1.24 cells compared to 1.9-fold in the nontransfected RIN5-AH cells. The expression of the transfected receptor was stimulated 1.6- and 2.3-fold when cells were cultured in the presence of 25 or 50 microM Zn2+, respectively. The increased expression of the GH receptor by Zn2+ was associated with an increased magnitude of GH-stimulated insulin biosynthesis. A close stoichiometric relationship between the level of receptor expression and the level of GH-stimulated insulin biosynthesis was observed. We conclude from these results that the hepatic GH receptor is able to mediate the effect of GH on insulin biosynthesis in RIN5-AH cells.

Animals↗
PubMed1990

Regulation of the uterine expression of messenger ribonucleic acids encoding the oestrogen receptor and IGF-I peptides in the pig uterus.

The acute effects of oestradiol-17 beta on the expression of the oestrogen receptor (ER) and insulin-like growth factor I (IGF-I) in the endometrium of ovariectomized pigs were examined. The steroid receptor level was assayed by hormone binding techniques and specific mRNAs analyzed by solution hybridization using 35S-labelled RNA probes complementary to the ligand-binding domain of the ER receptor gene and a 160 bp PanI-Pvul fragment of the IGF-I gene. One hour after a single injection of oestradiol (1 micrograms/kg BW), the nuclear oestrogen receptor (ERn) mean level was increased 3-fold whereas the ER mRNA content had not changed significantly. After 3 hours the ERn mean concentration was still high; the mean ER mRNA level had decreased by 15% and the mean IGF-I mRNA had increased 3-fold above that in the samples collected prior to treatment from these ovariectomized animals. Six hours after the injection the ERn content had returned to the basal level and stayed there during the following six hours. The ER mRNA concentration continued to decline, reached its lowest value after six hours and had increased slightly by twelve hours. The IGF-I mRNA level increased steadily during the course of the experiment. At twelve hours after the injection it had increased 3-fold. From these data we conclude that in the pig uterus oestradiol down-regulates its own receptor and acts as a potent stimulator of endometrial growth by inducing IGF-I expression.

Animals↗
PubMed1990

Growth hormone (GH) stimulates protein synthesis in cells transfected with GH receptor complementary DNA.

An expression vector containing a rat GH receptor cDNA was transfected into Chinese hamster ovary (CHO) cells, and stable cell lines expressing GH receptors were established. In contrast to nontransfected CHO cells, expression of GH receptors in transfected cells resulted in the appearance of high affinity (Kd = 1.53 nM) specific binding of GH. Cross-linking of [125I]hGH to the receptors and subsequent sodium dodecyl sulfate (SDS)-electrophoresis gave an estimated receptor mol wt of 84,000. GH treatment stimulated protein synthesis 60% over basal levels in GH receptor-expressing CHO cells, but not in the receptor-negative parental cells. The effect was observed only under serum-free conditions and was time and dose dependent. These results show that heterologous expression of the rat GH receptor results in the appearance of specific binding of GH and the acquisition of a functional GH response.

Animals↗
PubMed1990

Regulation of insulin-like growth factor-I gene expression by growth factors in cultured vascular smooth muscle cells.

The aim of this investigation was to study the regulation of insulin-like growth factor-I (IGF-I) gene expression in cultured rat aortic smooth muscle cells. Near-confluent cells were deprived of serum for 24 h and then exposed to IGF-I, insulin, serum, basic fibroblast growth factor (basic FGF), platelet-derived growth factor (PDGF-BB; consisting of B-chain homodimer) or GH for 24 h. Levels of IGF-I mRNA were measured by solution hybridization. The level of IGF-I mRNA was markedly decreased by 10% (v/v) newborn calf serum (78 +/- 4 (S.E.M.) % decrease), 1 nmol basic FGF/l (53 +/- 8%), and 1 nmol PDGF-BB/l (40 +/- 3%) when measured after 24 h. The effect of PDGF-BB was significant after 6 h and became more marked after 24 h. GH (1 nmol/l or 0.1 mumol/l) or insulin (1 nmol/l) had no effect after 24 h, whereas IGF-I (1 nmol/l) and insulin (10 mumol/l) increased IGF-I mRNA 64 +/- 20% and 46 +/- 14% respectively. The increase caused by IGF-I was demonstrated after 3 h, and was most marked after 24 h. Using Northern blot analysis of cultured aortic smooth muscle cells, IGF-I transcripts of 7.4, 1.7 and 1.1-0.8 kilobases were observed. Exposure of the cells to 10% serum, 1 nmol basic FGF/l or 1 nmol PDGF-BB/l for 48 h increased the cell number by 104 +/- 7%, 64 +/- 3% and 61 +/- 22% respectively, while IGF-I, insulin and GH had little effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Regulation of rat growth hormone receptor gene expression.

A cDNA encoding the growth hormone (GH) receptor was cloned from rat liver. Both the nucleotide and translated amino acid sequence share greater than 70% similarity with the GH receptors from rabbit and human. An RNA probe was generated from this sequence for use in a solution hybridization assay to quantitate GH receptor mRNA expression in rat tissues. Expression was detected in 9/12 tissues examined, with the highest levels observed in the liver. Expression in liver, kidney, heart and muscle was developmentally regulated, being low at birth and rising to adult levels in 5 weeks. No difference was observed between hepatic expression in males and females, although livers from pregnant rats had elevated levels. Hypophysectomy and GH treatment did not affect hepatic GH receptor mRNA levels.

Aging↗
PubMed1989

Somatomedin C in the pancreas of young and adult, normal and obese, hyperinsulinemic mice.

Immunocytochemical, immunochemical and RNA-hybridization techniques were used to map the distribution of somatomedin C (Sm-C; insulin-like growth factor I; IGF-I) in the pancreas of young and adult lean and obese mice. The D cells in the islets of Langerhans showed intense cytoplasmic Sm-C immunoreactivity, extending into their processes. Only slight Sm-C immunoreactivity was seen in A and B cells, apparently confined to the plasma membranes. In the exocrine pancreas scattered duct cells were immunopositive. Starvation increased, while feeding decreased the Sm-C immunoreactivity in B cells. RNA-hybridization analyses revealed that roughly the same number of Sm-C mRNA molecules, as calculated per DNA amount in the pancreas, could be demonstrated in young and adult, lean and obese mice. Radioimmunoassay (RIA) determinations of total Sm-C showed that there were about equal concentrations in the pancreas of lean and obese mice. There were marked differences between the liver and the pancreas, in that the RIA Sm-C values for the former were twice those in the latter while, in contrast, the corresponding values for the Sm-C mRNA, i.e. the agent determining the synthesis of Sm-C, were about 100 times higher in the liver as compared to that in the pancreas. We interpret our results as follows: The D cells in the islets form and secrete Sm-C in both young and adult, lean and obese mice, while A and B cells bind, but do not necessarily synthesize this peptide.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Regulation of sexual differentiation in drug and steroid metabolism.

Certain members of the cytochrome P-450 family are expressed at different levels in the livers of male and female rats. Although little is known of the functional significance of these sex differences, progress has been made towards the understanding of the endocrine control of hepatic sex differences in cytochrome P-450 levels. Jan-Ake Gustafsson and colleagues describe a subpopulation of hepatic sexually differentiated P-450s that is regulated by sex differences in growth hormone (GH) secretory pattern. This secretory pattern is in turn regulated by gonadal steroids. These studies demonstrate a novel action of GH and suggest that the hormonal secretory rhythm is pivotal in determination of biological effects.

Animals↗
PubMed1989

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.

Animals↗
PubMed1989

Induction of insulin-like growth factor I messenger ribonucleic acid during regeneration of rat skeletal muscle.

Expression of insulin-like growth factor I (IGF-I) was studied in regenerating skeletal muscle. Irreversible damage to muscle cells was induced in the extensor digitorum longus muscle of adult rats by ischemia, preceded by glycogen depletion. IGF-I mRNA levels during the regeneration process were studied for periods up to 10 days after injury using a solution hybridization assay. Increased IGF-I mRNA levels could be demonstrated within 24 h after injury; maximum levels were achieved in 3 days and decreased to approximately normal levels by 10 days. Changes in IGF-I mRNA levels could not be seen in undamaged contralateral extensor digitorum longus muscles during the experimental period. An increase in IGF-I mRNA was also evident in injured muscles of hypophysectomized animals. In situ hybridization at the time of maximum induction showed the presence of IGF-I mRNA in proliferating myoblasts and in satellite cells. IGF-I, thus, may act as a locally produced non-GH dependent trophic factor during regeneration of skeletal muscle after injury.

Animals↗
PubMed1989

Hormonal regulation of estrogen receptor messenger ribonucleic acid in T47Dco and MCF-7 breast cancer cells.

Using a combination of hormone-binding assays, immunologic techniques, and mRNA hybridizations we have measured the estrogen receptor (ER) content and studied the hormonal regulation of ER mRNA in one estrogen responsive and one estrogen unresponsive breast cancer cell line, MCF-7 and T47Dco, respectively. Estradiol binding could be detected in cytosol from MCF-7 cells but not in T47Dco cells. However, when measured by an enzyme-linked immunosorbent assay, T47Dco cells were found to contain approximately 15 fmol ER/mg cytosolic protein or 10% of the ER content in MCF-7 cells. Immunologically reactive ER in T47Dco cells was indistinguishable in size (approximately equal to 68 KD) from the ER in MCF-7 cells, as shown by Western blotting using a monoclonal antihuman ER antibody. Quantification of ER mRNA in MCF-7 and T47Dco cells indicated that T47Dco cells contained approximately 50% of the ER mRNA levels found in MCF-7 cells. This basal level of ER mRNA in T47Dco cells was not decreased by estradiol treatment, as opposed to in MCF-7 cells where estradiol caused 40-60% decrease in the ER mRNA expression. Also, estradiol did not increase the progesterone receptor (PR) mRNA levels in T47Dco cells whereas in MCF-7 cells an approximately 5-fold increase of the PR mRNA levels occurred after estradiol treatment. However, incubation of the cells with the synthetic progestin R5020 decreased the ER mRNA levels to approximately the same degree in both cell lines. In conclusion, we have shown that estrogen down-regulates ER mRNA and up-regulates PR mRNA in MCF-7 cells. Neither of these estrogenic effects were seen in T47Dco cells. It appears that the steroid-resistance in T47Dco cells does not occur as a consequence of a complete absence of ER mRNA or protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Blotting, Northern↗
PubMed1989

Developmental and steroid hormonal regulation of insulin-like growth factor II expression.

We have investigated the influence of steroid hormones on insulin-like growth factor II (IGF-II) expression. Hepatic IGF-II mRNA decreased gradually during postnatal development, reaching adult levels at 3 weeks of age. Treatment of 1-day-old rats for 4 days with 10 micrograms/day of the glucocorticoid dexamethasone (DEX) reduced IGF-II mRNA levels 10-fold in liver and inhibited body weight gain. Estradiol and testosterone did not affect IGF-II expression. A dose-response relationship between IGF-II mRNA levels and the different amounts of DEX injected was seen. IGF-II levels remained low after withdrawal of DEX, indicating an irreversible effect. Albumin expression was increased in newborn rat livers after DEX treatment. Our results suggest that glucocorticoids play an important role in the regulation of IGF-II expression. The mechanism for glucocorticoid-induced reduction of IGF-II mRNA is still unclear; however, our findings indicate that DEX inhibits IGF-II by causing premature differentiation of the liver.

Aging↗
PubMed1989

Regulation of uterine insulin-like growth factor I mRNA and insulin-like growth factor II mRNA by estrogen in the rat.

IGF-I and IGF-II are peptides with mitogenic properties. In this study mRNA for IGF-I and IGF-II was analysed in rat uterine tissue after different endocrine manipulations and the possibility of an estrogenic regulation of IGF expression was investigated. Both IGF-I and IGF-II mRNA were present in uterine tissue. The level of IGF-I mRNA, but not IGF-II mRNA, was reduced following ovariectomy. Administration of estradiol (2.5 micrograms/day for 4 days) to ovariectomized rats increased IGF-I mRNA 8-fold to levels seen in intact animals. In adult animals hepatic IGF-I mRNA did not appear to be increased by estrogen treatment. Low levels of IGF-II mRNA were detected in the uterus, but showed no dependence on estrogen. The inductive effect of estrogen on uterine IGF-I mRNA could not be substituted for by growth hormone administration (0.5 mg/100 g, ip for 6 h). The present results suggest IGF-I as a potential candidate for a mediator of estrogen-induced growth. Both estrogen and GH induce IGF-I mRNA and a tissue specificity for these hormones is indicated where GH regulates hepatic and estrogen uterine IGF-I mRNA.

Animals↗
PubMed1989

Growth hormone regulates the rodent hepatic epidermal growth factor receptor at a pretranslational level.

Recent studies have implicated the involvement of pituitary factor(s) in the regulation of hepatic epidermal growth factor receptor (EGF-R) levels. In the present study the possible role of GH as a regulator of EGF-R has been examined by measuring hepatic EGF-R mRNA and EGF binding in intact and GH-deficient rats and mice before and after administration of GH. Using a human EGF-R probe, 10.5 and 6.0 kb transcripts were detected in mouse and rat liver by Northern gel analysis. EGF-R mRNA was quantified by solution hybridization, and EGF binding determined by incubation of 125I-labelled EGF with a low-density membrane fraction. Levels of hepatic EGF-R mRNA and binding of EGF in female rats were about two-thirds of those in male rats. GH-deficient (lit/lit) male and female mice had approximately 10 and 25% respectively of the levels of EGF-R mRNA and EGF binding of intact male and female mice. Furthermore, hypophysectomized rats exhibited a reduced level of EGF-R mRNA and EGF binding, corresponding to about 20% of the levels detected in intact male rats. When hypophysectomized male and female rats received recombinant human GH (hGH) either as intermittent injections or by continuous infusion using osmotic minipumps, the EGF-R mRNA and EGF binding levels increased to about half those of the intact male animals. No differences between intermittent or continuous administration of hGH on the induction of EGF-R mRNA or EGF binding could be seen.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Regulation of insulin-like growth factor-I and growth hormone receptor gene expression by diabetes and nutritional state in rat tissues.

Insulin-like growth factor-I (IGF-I) mRNA and GH receptor mRNA levels were analysed in different tissues from rats made diabetic with streptozotocin, fasted rats and rats fed with a protein-reduced diet. Diabetes decreased IGF-I mRNA levels in liver, heart, diaphragm, kidney and aorta, but not in brain. GH receptor mRNA levels were decreased in heart and diaphragm, but not in liver and kidney. Fasting decreased IGF-I mRNA in all tissues studied except brain, and decreased GH receptor mRNA in liver, heart and diaphragm, but not in kidney. A protein-reduced diet decreased hepatic IGF-I mRNA levels but did not significantly affect other tissues, while GH receptor mRNA levels were reduced in liver and diaphragm. In conclusion, both diabetes and limited nutrition affected IGF-I and GH receptor mRNA in different tissues, but the two mRNAs were not co-ordinately regulated in all tissues studied. While reduced GH receptor gene expression may thus be responsible for decreased IGF-I gene expression in some states and tissues, additional regulatory mechanisms may be of importance.

Animals↗
PubMed1988

Expression of insulin-like growth factor I (IGF-I) and IGF-II mRNA during hepatic development, proliferation and carcinogenesis in the rat.

Insulin-like growth factor I (IGF-I) and IGF-II are peptides that presumably are required for normal fetal and postpubertal growth. The production of IGFs is developmentally regulated and the liver appears to be a major site of production. By analysing mRNA levels for IGF-I and IGF-II in the rat liver we have attempted to further study the expression of these growth factors during development and regeneration as well as during the course of hepatic carcinogenesis. Fetal livers are characterized by a high level of IGF-II mRNA and a low level of IGF-I mRNA, while in adult livers the opposite situations occur, i.e. a high level of IGF-I mRNA and a non-measurable level of IGF-II mRNA. During the course of experimentally induced hepatic cancer, IGF-I mRNA was consistently reduced and in a majority of cancers analysed (6/9) IGF-II mRNA was increased, i.e. a fetal type of IGF expression can be switched on in some experimentally induced hepatocellular carcinomas. The onset of IGF-II production during hepatic carcinogenesis appears to be a late phenomenon since liver nodules, preceding the development of hepatocellular cancer, were found not to contain IGF-II mRNA. Furthermore, during hepatic regeneration following partial hepatectomy no marked change in IGF-I or IGF-II mRNA levels was noted. The above results suggest that the fetal growth factor IGF-II could have a role in hepatic cancer.

Aging↗
PubMed1988

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)

Age Factors↗