Insulin binding to liver plasma membranes in salmonids with modified plasma insulin levels

1991 ◽  
Vol 69 (11) ◽  
pp. 2745-2750 ◽  
Author(s):  
Joaquim Gutiérrez ◽  
Erika M. Plisetskaya

Insulin binding to the liver plasma membranes was assessed in coho salmon (Oncorhynchus kisutch) injected intraperitoneally with 6.6 μmol arginine/g body weight, and in rainbow trout (Oncorhynchus mykiss) fasted for 40 days and then refed for 15 days. Corresponding control groups of fish were injected with saline (coho salmon) or fed regularly (trout). Arginine injection was followed by a substantial elevation of plasma insulin titres from 12.5 to 76.8 ng/mL, an increase in the binding capacity and, consequently, an increase in specific binding of insulin to the liver plasma membranes from 3.4 to 5.5%. The binding affinity remained unchanged. Food deprivation lowered plasma insulin titres from 13.2 to 3.0 ng/mL, increased the binding capacity, but decreased the binding affinity, so the specific binding remained essentially unchanged (5.6% in fed versus 5.1% in fasted fish). Refeeding of fasted fish resulted in restoration of insulin levels and an increase in binding affinity relative to both control and fasted groups of fish. This led to a substantial elevation of the specific binding of insulin up to 7.1%. The binding of insulin to liver plasma membranes in salmonids depends both on the number of binding sites (binding capactity) and on the binding affinity. Long-term treatment, such as food deprivation, resulted in altered affinity and capacity of binding, whereas short-term treatment, such as arginine injection, affected mostly the binding capacity. Modulation of the number of binding sites in liver plasma membrane according to the insulin level was observed only in experiments on trout conducted at a higher (15 °C) water temperature.

1993 ◽  
Vol 40 (2) ◽  
pp. 231-236
Author(s):  
K W Nowak ◽  
P Maćkowiak ◽  
L Nogowski

The comparative studies were performed on insulin receptors of erythrocytes and liver plasma membranes in fish (tench and carp) and bird (duck). The Scatchard plots indicated the presence of two pools of binding sites both in fish and duck. These pools show inter-species differences in binding ability and the number of receptors. Specific binding of insulin and the binding affinity are higher in duck than in fish.


1979 ◽  
Vol 92 (2) ◽  
pp. 228-241 ◽  
Author(s):  
Rudolf Baumann ◽  
Herbert Kuhl

ABSTRACT The specific binding of [125I]LH-RH to isolated plasma membranes of rat pituitaries was investigated. The binding process was found to be highly specific, temperature-dependent and saturable. The dissociation constant as calculated by three different methods was approximately 1.3 · 10−8 m, indicating a single type of binding sites. Maximal binding capacity was 1 · 10−12 moles/mg protein (=2 ng LH-RH/pituitary gland), and the number of binding sites was calculated to be 6 · 1011 per mg membrane protein (= 1 · 1010 binding sites/pituitary gland). When diluted with icecold buffer the dissociation of specifically bound LH-RH occurred very rapidly (half-life 3.17 min) with a rate constant of 0.219 min−1. The dissociation process followed first-order kinetics. Specificity of binding was demonstrated by dose-dependent competition of unlabelled LH-RH, the highly potent analogue D-glutamine-(cyclohexyl)6-LH-RH-nonapeptide-ethylamide and the fragment of an analogue (6-D-Ser(TBu))-LH-RH-(3-9)-heptapeptide-ethylamide with the binding [125I]LH-RH, while angiotensin I, II, oxytocin and bacitracin did not compete. The affinities of LH-RH and the analogue to the binding sites of the pituitary plasma membranes were not consistent with the respective biological activities.


1991 ◽  
Vol 260 (2) ◽  
pp. G290-G298 ◽  
Author(s):  
B. K. De ◽  
T. L. Brown ◽  
F. J. Suchy

The binding of epidermal growth factor (EGF) to its receptor and the activity of the receptor intrinsic protein-tyrosine kinase were studied during the ontogeny of rat liver. The number of EGF receptors during pre- and postnatal development was first compared in crude liver plasma membranes using 1) specific binding of 125I-labeled EGF and 2) immunoblot analysis using any antireceptor polyclonal rabbit antibody. Both methods detected the expression of the EGF receptor in fetal rat liver on day 17 of gestation, but in an amount markedly less than the adult. Within 24 h, there was a more than twofold increase in EGF binding to plasma membranes as well as a marked increase in receptor immunoreactivity. However, after birth, there was a precipitous drop in receptor number to less than 20% of the adult level by the end of the first postnatal day (P less than 0.001). Next, the presence of EGF-stimulated tyrosine kinase activity (autophosphorylation) was determined during the same stages of development. Electrophoresis of membranes phosphorylated in the presence or absence of EGF followed by autoradiography demonstrated autokinase activity stimulated by EGF in day 18 and 19 fetal liver plasma membranes, but not in membranes on day 17 of gestation. Similar to the pattern observed with EGF binding, there was a decrease in autokinase activity in early neonatal plasma membranes followed by an increase to near adult levels by 7 days postnatally. Quantitation of the amount of 32P radioactivity associated with the EGF receptor bands in each age group, correlated with the degree of autophosphorylation assessed by autoradiography.(ABSTRACT TRUNCATED AT 250 WORDS)


1975 ◽  
Vol 80 (1) ◽  
pp. 126-137 ◽  
Author(s):  
Tokuko Saito ◽  
Brij B. Saxena

ABSTRACT A role of prolactin (PRL) in ovarian function has been suggested in several species, but not unequivocally established except in the rat. We, therefore, examined the presence of specific receptor for PRL in ovaries of rat, cow, and human. Human PRL (hPRL) labelled with 125I by the lactoperoxidase method was shown to be capable of specific binding to rat mammary tissue homogenate. Human, cow, and rat ovarian homogenates and/or partially purified plasma membranes were also shown to specifically bind 125I-hPRL. Binding was a saturable phenomenon and was dependent on receptor protein concentration. Optimal binding was observed at pH 7.0 and at 37°C. Binding was reversibly inhibited by exposure of membranes to pH 10.0 and irreversibly destroyed by exposure to pH 3.0. Bound 125I-hPRL was displaceable by unlabelled human, ovine, and bovine PRL but not by FSH or LH. However, human chorionic somatomammotrophin (hCS) and hGH showed some competition with 125I-hPRL. Number of binding sites/mg protein was lowest (0.8 × 10−12 m) during metoestrus and increased during dioestrus (11 × 10−12 m) reaching the maximum number at pro-oestrus (24.6 × 10−12 m). These results demonstrate that presence of specific PRL receptor in the ovaries and are consistent with a role of PRL at the ovarian level.


Diabetes ◽  
1982 ◽  
Vol 31 (10) ◽  
pp. 867-873 ◽  
Author(s):  
J. B. Clark ◽  
S. Keen ◽  
C. M. Clark

1989 ◽  
Vol 108 (2) ◽  
pp. 115-124 ◽  
Author(s):  
Miguel Trueba ◽  
Ana I. Vallejo ◽  
Isabel Rodriguez ◽  
Iñaki Ibarrola ◽  
María J. Sancho ◽  
...  

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