Biomedical subjects

A Gertler

Publications and source records attributed to A Gertler.

At least 19 recordsLinked to original sources

PubMed2006

Large-scale preparation and in vitro characterization of biologically active human placental (20 and 22K) and pituitary (20K) growth hormones: placental growth hormones have no lactogenic activity in humans.

Expression plasmids containing DNA sequences optimized for expression in Escherichia coli were prepared encoding human pituitary (hGH-N 20K) and placental (hGH-V 20 and 22K) growth hormones. The proteins were expressed in bacteria, refolded and purified to homogeneity by anion-exchange chromatography on Q-Sepharose according to a unique protocol developed for each protein. The yields from 5l of fermentation culture varied between 400 and 700mg of electrophoretically pure, over 95% monomeric protein. Circular dichroism (CD) analysis revealed similarity of the purified hGHs' secondary structure to that of the pituitary hGH-N 22K, except for hGH-V 20K, in which the alpha-helix content was lower. The purified proteins were stable as a 0.1% sterile solution held at pH 10-11 at 4 degrees C for at least one month. All three purified hGH molecules formed a 1:2 complex with hGH receptor extracellular domain (hGHR-ECD), similar to hGH-N 22K. Binding experiments using hGHR-ECD revealed that the differences between the two 22K variants or between the two 20K variants were not significant, except that hGH-V 20K exhibited slightly lower affinity. Somatogenic activity was tested in vitro using FDC-P1 cell lines. Whereas the bioactivity of 22K hGHs and hGH-N 20K in FDC-P1-9D11 cells stably transfected with hGHR was almost equal and two to threefold higher than that of hGH-V 20K, in FDC-P1 3B9 cells stably transfected with rabbit (rb) GHR, the bioactivity of both 20K analogues was significantly (five to ninefold) lower than that of the 22K hormones. The lactogenic activity measured in heterologous assays (Nb2-11C cells and Baf/3 cells stably transfected with the long form of rabbit prolactin receptor) revealed that the activity of hGH-N 20K was close to that of hGH-N 22K in the Baf/3 cells, but 4.5-fold lower in the Nb2 cells. The activity of hGH-V 22K was ninefold less in Nb2 cells and 55-fold less in Baf/3 cells, whereas hGH-V 20K had no lactogenic activity in either bioassay. In contrast, in a homologous lactogenic assay using Baf/3 LP cells stably transfected with hPRLR, the activity of both placental hGHs was nil and the activity of hGH-N 20K was 4.3-fold lower than that of hGH-N 22K. The latter finding raises the question of whether the lack of intrinsic lactogenic activity in the placental hGHs that dominate during pregnancy has any physiological relevance.

Animals↗

PubMed2006

Active immunization against leptin fails to affect reproduction and exerts only marginal effects on glucose metabolism in young female goats.

Approximately 150 days before expected breeding time, 12 female goats (3 months of age) were actively immunized against ovine leptin. Booster injections were given throughout the following year. Control animals (n = 6) were sham-immunized. After the first observed oestrus, a buck was introduced and goats were mated. Blood samples were collected twice weekly and frequent blood sampling series were performed on days -15, 76, 153 and 286 relative to the first immunization. Nine of the immunized goats developed titres within 3 months and had elevated serum concentrations of leptin compared with controls (p < 0.0001). Hematological parameters and blood chemistry were not affected by the immunization. No differences were detectable in all reproductive parameters recorded. Serum insulin was higher in immunized goats during the frequent blood sampling series of day 287 after the first immunization. Glucose metabolism was investigated during pregnancy using hyperglycaemic and euglycaemic/hyperinsulinaemic clamps. None of the parameters derived from the clamp studies was different (p > 0.05) between the two groups. During the hyperglycaemic clamp there was a trend (p < 0.15) towards increased insulin concentrations in immunized animals whereas glucose infusion rates were not different between the groups. This indicates decreased insulin sensitivity in immunized goats. Our study describes the ontogenesis of serum concentrations of leptin during growth, puberty and first pregnancy and parturition for the caprine species. The effects of the immunization were not detectable or only marginal and the approach aimed at therefore not effective to investigate leptin action in detail.

Animals↗

PubMed2005

Mapping leptin-interacting sites in recombinant leptin-binding domain (LBD) subcloned from chicken leptin receptor.

The binding domain of the chicken leptin receptor [chLBD (chicken leptin-binding domain)], subcloned from the full-size chicken leptin receptor and prepared in an Escherichia coli system, was subjected to site-directed mutagenesis to identify the amino acids involved in leptin binding. A total of 22 electrophoretically pure, >90% monomer-containing mutants were expressed, refolded and purified. The effects of the mutations were tested by the ability to form complexes with ovine leptin, and the kinetic parameters of interaction were determined by surface plasmon resonance. Six mutants were used to determine whether mutations of several amino acids that differ between chLBD and mammalian LBDs will affect affinity: none showed any such effect, except the mutant A105D (Ala(105)-->Asp), which exhibited some decrease in affinity. Surface plasmon resonance analysis identified six mutants in which binding activity was totally abolished (F73A, Y14A/F73A, V76A/F77A, L78A/L79A, V76A/F77A/L78A/L79A and A105D/D106V) and six mutants (Y14A, R41A, R41A/S42A/K43A, V103A, V135A/F136A and F136A) in which affinity for the hormone was reduced, mainly by increased dissociation rates. Gel-filtration experiments indicated the formation of a 1:1 ovine or human leptin-chLBD complex with a molecular mass of approx. 41 kDa. Gel-filtration experiments yielded 1:1 complexes with those mutants in which affinity had decreased, but not with the six mutants, which had totally lost their binding capacity. Modelling the leptin-chLBD complex indicated that the binding domain of the latter is located mainly in the L3 loop, which contributes nine amino acid residues interacting with leptin. Contact-surface analysis identified the residues having the highest contribution to the recognition site to be Phe73, Phe77 and Leu79.

Amino Acid Sequence↗

PubMed2005

Neonatal leptin treatment reverses developmental programming.

An adverse prenatal environment may induce long-term metabolic consequences, in particular obesity and insulin resistance. Although the mechanisms are unclear, this programming has generally been considered an irreversible change in developmental trajectory. Adult offspring of rats subjected to undernutrition during pregnancy develop obesity, hyperinsulinemia, and hyperleptinemia, especially in the presence of a high-fat diet. Reduced locomotor activity and hyperphagia contribute to the increased fat mass. Using this model of maternal undernutrition, we investigated the effects of neonatal leptin treatment on the metabolic phenotype of adult female offspring. Leptin treatment (rec-rat leptin, 2.5 microg/g.d, sc) from postnatal d 3-13 resulted in a transient slowing of neonatal weight gain, particularly in programmed offspring, and normalized caloric intake, locomotor activity, body weight, fat mass, and fasting plasma glucose, insulin, and leptin concentrations in programmed offspring in adult life in contrast to saline-treated offspring of undernourished mothers who developed all these features on a high-fat diet. Neonatal leptin had no demonstrable effects on the adult offspring of normally fed mothers. This study suggests that developmental metabolic programming is potentially reversible by an intervention late in the phase of developmental plasticity. The complete normalization of the programmed phenotype by neonatal leptin treatment implies that leptin has effects that reverse the prenatal adaptations resulting from relative fetal undernutrition.

Absorptiometry, Photon↗

PubMed2004

Leptin attenuates the acute effects of centrally administered neuropeptide Y on somatotropin but not gonadotropin secretion in ovariectomized cows.

We tested the hypothesis that recombinant ovine leptin would attenuate the acute effects of neuropeptide Y (NPY) on secretion of GH and gonadotropins (LH and FSH) in cows. Ovariectomized cows (n=6) fitted with third ventricle guide cannulas were assigned randomly to each of three groups in a Latin square arrangement: (1) control; saline treatment only, (2) NPY; saline followed by NPY, and (3) L-NPY; leptin pretreatment followed by NPY. Treatments were: s.c. injection of saline or leptin (30 microg/kg BW) at time 0, i.v. injection of saline or leptin (30 microg/kg BW) at 70 min, and intracerebroventricular (i.c.v.) injection of saline or NPY (500 microg) at 90 min. Plasma leptin was elevated (P<0.01) at least four-fold throughout the experiment in the L-NPY group. Mean plasma concentrations of LH declined within 1 h and were lower (P<0.03) than controls in both the NPY and L-NPY groups beginning 2 h after NPY injection. An acute increase in plasma concentrations of GH was observed within 1 h after NPY in the NPY group and mean values were greater (P<0.01) than controls. However, in the L-NPY group, leptin pretreatment attenuated the NPY effect on GH. Treatments had no effect on FSH secretion. Results confirm suppressive and stimulatory effects of NPY on LH and GH secretion, respectively, and indicate that leptin can attenuate the acute effects of NPY on GH secretion in cattle.

Animals↗

PubMed2004

Intercellular signaling between adipose tissue and muscle tissue.

Adipose and muscle tissues undergo regulated growth and differentiation processes that are modulated by a wide range of factors. The interactions between myogenic cells and adipocytes play a significant role in growth and development, including the rate and extent of myogenesis, muscle growth, adipogenesis, lipogenesis/lipolysis, and in the utilization of energy substrates. Important hormones and growth factors involved in the regulation of these processes include glucocorticoids, insulin-like growth factors, various cytokines, insulin, and leptin. Interactions among these axes have important implications in their influence on relative fat and lean deposition and the efficiency of energy utilization in growth and development. As research progresses to better clarify the interactions among adipose tissue depots and muscle of different fiber types, pathways will become better understood, ultimately leading to the optimized management of fat and lean growth in domestic livestock species. This review will focus on elements of intercellular signaling, using data from cell culture studies to illustrate specific examples of signaling pathways between cells.

Adipocytes↗

PubMed2003

Study of the alteration of gene expression in adipose tissue of diet-induced obese mice by microarray and reverse transcription-polymerase chain reaction analyses.

In the present study we developed a model of diet-induced obesity (DIO) in male C57 BL/6J mice using an 8-wk high fat diet. This model should better reflect the physiology of the majority of the human obese patients than mouse genetic models of obesity with defects in leptin or leptin signaling. At the end of the diet, DIO mice displayed an increased weight (20%) and higher leptin, insulin, glucose, and corticosterone plasma levels compared with mice fed a standard diet during the same period. Moreover, they became resistant to the central effect of peripheral administration of leptin. Oligonucleotide microarray studies were conducted in adipose tissue. They showed that a great number of genes are differentially expressed. The majority of these genes (69%) are down-regulated in DIO mice. Among those are genes encoding enzymes of the lipid metabolism or markers of adipocyte differentiation, enzymes involved in detoxification processes, as well as structural components of the cytoskeleton. Some other groups of genes displayed increased expression, such as those encoding inflammatory markers. The results of the microarray analysis were confirmed by semiquantitative RT-PCR studies run on a selected number of genes that were differentially expressed or not modified.

Adipose Tissue↗

PubMed2003

Attenuation by leptin of the effects of fasting on ovarian function in hens (Gallus domesticus).

Thirty-four-week-old laying hens received injections of recombinant chicken leptin to assess the role of leptin in avian ovarian function. In the first experiment, the hens (n=60) were divided into three groups: (i). fed ad libitum; (ii). fasted; and (iii). fasted + leptin. Hens were fasted for 5 days and those treated with leptin received 250 microg leptin kg-1 body weight twice a day, i.p. In the second experiment, the hens (n=72) were divided into four groups: (i). fed ad libitum; (ii). fasted; (iii). fasted + leptin given only during fasting (5 days); or (iv). fasted and leptin given during both fasting and 5 days of re-feeding (10 days). LH was measured in blood plasma, and progesterone and oestradiol were measured in blood plasma and the ovary by radioimmunoassay. Apoptosis was examined in the walls of the three largest yellow hierarchical follicles (F3-F1; F3 8-12 mm), and the granulosa layer of F3 follicles. The expression of leptin receptor in the granulosa layer of F2 and F1 follicles was barely detectable. This was in contrast to a much higher expression of leptin receptor maintained in the theca layer of F3-F1 follicles. The present results indicate that in chickens leptin might be involved in the adaptation to starvation due to attenuation of follicular apoptosis. The presence of leptin receptors in the ovary indicates the possibility of a peripheral effect of the hormone.

Adaptation, Physiological↗

PubMed2002

The late gestation increase in circulating ACTH and cortisol in the fetal sheep is suppressed by intracerebroventricular infusion of recombinant ovine leptin.

The obese gene product leptin, originally characterised as an adipocyte hormone coordinating the behavioural and neuroendocrine responses to starvation, is expressed in fetal adipocytes and placental trophoblast cells and is present in the fetal circulation. Concentrations of leptin in fetal blood correlate with fetal bodyweight and fat mass. In post-natal life, leptin conveys information about calorie intake and the state of adipose tissue energy stores, and plasma leptin levels are generally inversely correlated with hypothalamo-pituitary adrenal (HPA) activity. Late fetal life is characterised by increasing HPA activity that prepares the fetus for extrauterine life and initiates the endocrine cascade leading to parturition. We have investigated the hypothesis that leptin in the fetal circulation can inhibit the fetal HPA axis, thereby providing a mechanism by which the fetus can determine the fine timing of parturition as long as it is adequately nourished and growing appropriately. Here we show that a 5-day intracerebroventricular infusion of leptin to the sheep fetus in late gestation inhibits the pre-parturient rise in ACTH and cortisol concentrations, and that this seems to be a centrally mediated effect.

Adrenocorticotropic Hormone↗

PubMed2001

Leptin attenuates follicular apoptosis and accelerates the onset of puberty in immature rats.

Human and rat granulosa cells express receptors to leptin which synergies with glucocorticoid hormones in stimulation of ovarian steroidogenesis. To examine whether leptin affects follicular development and maturation, we injected recombinant ovine leptin (300 ng-10 microg/animal) daily to immature 21 day-old female rats. Non-treated rats reached puberty at 44.5+/-1.6 (n=9) days. In contrast, in leptin treated animals, puberty was reached at 34.5+/-1.6 (n=9) days. Ovarian sections revealed hypertrophy of granulosa cells in leptin treated animals. Moreover, the number of ovulations was 2-fold higher in the treated animals compared to controls (3-4 ovulations versus 7-8 on the first three estrous cycles, P<0.001). Leptin dramatically reduced incidence of follicular apoptosis measured by TUNEL, and was already evident after 7 days of leptin injection (12% of apoptosis in leptin treated group compared to 52% in controls, P<0.001). Maximal protection against apoptosis was achieved at 1-3 microg leptin/animal. The levels of FSH, LH, progesterone and the steroidogenic factors ADX and STAR were elevated earlier in development in the leptin treated animals compared to control animals which is in line with the achievement of early puberty in the leptin treated animals compared to non treated ones. To reveal whether modulation of death and survival genes is involved in leptin attenuation of follicular apoptosis, we examined the expression of the survival gene Bcl-2 and the death gene Bax in Western blots of ovarian homogenates. There was a pronounced elevation in Bcl-2 expression during 7-14 days of leptin injections up to 16.3-fold (P<0.001) compared to Bcl-2 expression in controls. Bax expression was elevated only 3.4 fold (P<0.001), leading to an increase in the Bcl-2/Bax ratio of 4.7 fold (P<0.001). Expression of the tumor suppressor gene p 53 and the oncogene Mdm2 did not change significantly. Our data suggests that leptin may be involved in accelerating follicular maturation by attenuating follicular atresia and increasing the ratio of Bcl-2/Bax.

Adrenodoxin↗

PubMed2001

Evidence of rainbow trout prolactin interaction with its receptor through unstable homodimerisation.

This study aims to characterise Prolactin receptor (PRLR) in rainbow trout for which no information is available despite the availability of Salmonid PRL preparations. By screening a freshwater rainbow trout intestine cDNA library with a probe corresponding to the extracellular domain (ECD) of tilapia PRLR, we have cloned a 2.5 kb insert coding for the PRLR. The mature protein of 614 amino acid residues is similar to PRLR isolated in tilapia and also the long form of mammalian PRLR. Analysis of PRLR gene expression in osmoregulatory organs revealed the presence of a unique transcript, thus confirming the involvement of this hormone in the control of osmoregulation in this fish species. By using surface plasmon resonance (SPR) technology, kinetic measurement of interaction between trout PRL and its receptor ECD was studied. This approach allowed us to demonstrate the formation of a transient, unstable homodimeric complex. This unstability could explain the inability to perform binding experiments using homologous PRL. In contrast, heterologous lactogenic ligands were able to interact through a more stable complex. Whether these characteristics of PRL-receptor interaction in rainbow trout are different to what occurs in tilapia where a homologous radioreceptor assay was developed would require further studies.

Animals↗

PubMed2001

Large-scale preparation of recombinant ovine prolactin and determination of its in vitro and in vivo activity.

Recombinant bovine Ala-prolactin (PRL) (GenBank Accession No. V00112) in prokaryotic expression plasmid pMON3401 was mutated using a mutagenesis kit, to prepare plasmid encoding ovine PRL (oPRL) (GenBank Accession No. M27057) Escherichia coli cells transformed with this latter plasmid overexpressed large amounts of oPRL upon induction with nalidixic acid. The expressed protein, found in inclusion bodies, was refolded and purified to homogeneity on a Q-Sepharose column, yielding an electrophoretically pure fraction composed of over 98% monomeric protein of the expected molecular mass of approximately 23 kDa. The biological activity of the recombinant oPRL after proper renaturation was evidenced in vitro by its ability to stimulate proliferation of rat lymphoma Nb(2) cells possessing PRL receptors, to stimulate luciferase activity in HEK 293 cells transiently transfected with oPRL receptors, and to induce progesterone secretion in primary cultures of luteal cells obtained from midpregnant ewes. In contrast to ovine growth hormone or ovine placental lactogen, recombinant oPRL had no galactopoietic effect in lactating ewes.

Animals↗

PubMed2001

Chicken leptin: properties and actions.

Chicken leptin cDNA shows a high homology to mammalian homologous, with an expression localized in the liver and adipose tissue. It is noteworthy, that the hepatic expression is most likely associated with the primary role that this organ plays in lipogenic activity in avian species. As in mammals, chicken leptin expression is regulated by hormonal and nutritional status. This regulation is tissue-specific and with a high sensitivity in the liver compared to adipose tissue. The blood leptin levels are regulated by the nutritional state with high levels in the fed state compared to the fasted state. The recombinant chicken leptin markedly inhibits food intake as reported in mammals, suggesting the presence of an hypothalamic leptin receptor. The chicken leptin receptor has been identified and all functional motifs are highly conserved compared to mammalian homologous. Chicken leptin receptor is expressed in the hypothalamus but also in other tissues such as pancreas, where leptin inhibits insulin secretion and thus may have a key role in regulating nutrient utilization in this species.

Adipose Tissue↗

PubMed2001

Direct modification of somatotrope function by long-term leptin treatment of primary cultured ovine pituitary cells.

Leptin is produced primarily in adipocytes and regulates body energy balance. A close link between leptin and pituitary hormones, including GH, has been reported. The mechanisms employed by leptin to influence somatotropes are not clear, however. Here we report a direct action of recombinant ovine leptin on primary cultured ovine somatotropes by analyzing the levels of mRNA encoding for GH or the receptors for GHRH (GHRH-R) and GH-releasing peptides (GHRP). Treatment of ovine somatotropes with leptin (10(-7)-10(-9) M) for 1-3 d reduced the mRNA levels encoding GH and GHRH-R, but increased GHRP receptor mRNA levels in a time- and dose-dependent manner. Three-day treatment of cells with leptin decreased the GH response to GHRH stimulation, but the GH response to GHRP-2 stimulation was increased. The combined effect of GHRH and GHRP-2 on GH secretion was not altered by treatment of the cells with leptin. These results demonstrated a direct action of leptin on ovine pituitary cells, leading to a reduced sensitivity of somatotropes to GHRH. It is also suggested that GHRP may be useful to correct the decrease in GHRH-induced GH secretion by leptin.

Animals↗

PubMed2001

The in vitro effect of leptin on growth hormone secretion from primary cultured ovine somatotrophs.

Although existing data suggest an influence of leptin on circulating levels of growth hormone (GH), the action site and properties of leptin are still controversial. Using primary cultured ovine pituitary cells, we studied the direct effect of leptin on the secretion of GH. Pituitary cells were dissociated by collagenase and subjected to Percoll gradient centrifugation to enrich the somatotroph population to 60-80% of cells. Treatment of primary cultured ovine somatotrophs with leptin (10(-9)-10(-7) M) for 30 min did not affect basal, GH-releasing hormone (GHRH) (10(-7) M)- or GH-releasing peptide-2 (GHRP-2)(10(-7) M)-stimulated GH secretion. Following treatment of cells for 1-3 d with leptin, GHRH-stimulated GH secretion was reduced and GHRP-2-stimulated GH secretion increased. The combined effect of GHRH and GHRP-2 on GH secretion was not altered by the treatment of cells with leptin for 3 d. GHRH receptor mRNA levels in cultured somatotrophs were decreased but GHRP receptor mRNA levels were increased by 3-d leptin treatment. These results suggest that leptin has a long-term effect on somatotrophs to reduce GHRH receptor synthesis leading to a decrease in GHRH-stimulated GH secretion. Leptin appears, however, to have an opposite effect on GHRP receptor synthesis leading to an increase in GHRP-stimulated GH secretion.

Animals↗

PubMed2001

Central infusion of leptin into well-fed and undernourished ewe lambs: effects on feed intake and serum concentrations of growth hormone and luteinizing hormone.

Leptin has been implicated in the regulation of feed intake, growth, and reproduction. The objective of this study was to determine if centrally administered leptin would affect feed intake and the secretion of growth hormone (GH) and luteinizing hormone (LH) in ewe lambs. Eighteen ewe lambs were ovariectomized and fitted with intracerebroventricular (i.c.v.) cannulae. Lambs were randomly assigned to receive either a maintenance diet (fed), or a diet that provided 38% of maintenance requirements (diet-restricted) for 14 weeks. Subsequently, recombinant ovine leptin or vehicle was continuously infused, via i.c.v. cannulae, in a linearly increasing dose for 8 days, reaching a maximum of 1.25 microg/kg per h. Feed intake was recorded on days -1 to 7. Blood was collected via jugular cannulae every 10 min for 4 h on days 0, 2, 4, 6 and 8 for the determination of serum leptin, insulin, LH and GH. Leptin suppressed feed intake in fed lambs on days 4 to 7 (P<0.001), but had no effect on feed intake in diet-restricted lambs (P>0.25). Fed lambs had greater serum concentrations of leptin than diet-restricted lambs (P=0.007). Also, although not different on day 0 (pretreatment), on day 8 serum leptin concentrations were greater in leptin-treated lambs than in saline-treated lambs (P=0.003). Insulin was lower in diet-restricted than in fed lambs (P=0.003), but was not affected by leptin treatment (P=0.82). LH pulse frequencies were lower in diet-restricted lambs than in fed lambs (P=0.038), but were not affected by leptin treatment (P=0.85). Mean serum GH was greater in diet-restricted than in fed lambs (P<0.01). In diet-restricted lambs treated with leptin or saline, mean GH did not differ on day 0, but increased in response to leptin treatment (P<0.006). Treatment of fed lambs with leptin did not affect serum GH (P>0.32). From this work, we propose that leptin represents an important functional link between adipose stores and hypothalamic function in ruminants. We demonstrate that leptin concentrations change in response to reduced nutritional status, and that leptin has the ability to regulate multiple physiological processes in lambs, including both feed intake and secretion of GH.

Animal Nutritional Physiological Phenomena↗

PubMed2001

Mutations of ovine and bovine placental lactogens change, in different ways, the biological activity mediated through homologous and heterologous lactogenic receptors.

The biological activities of ovine (o) and bovine (b) placental lactogens (PLs) and their mutated analogues were compared using several binding and in vitro bioassays. In almost all cases, the biological activities of these analogues mediated through rat (r) prolactin receptor (PRLR) showed little or no change, despite a remarkable decrease in their capacity to bind to the extracellular domain of rPRLR and despite compromised stability of the 2:1 complexes. These results indicate that mutations impairing the ability of oPL or bPL to form stable complexes with lactogenic receptors do not necessarily lead to a decrease in the biological activity, because the transient existence of the homodimeric complex is still sufficient to initiate the signal transduction. In contrast, oPL and bPL analogues completely, or almost completely, lost their ability to activate homologous PRLRs, and some of them even acted as site-2 antagonists. To explain the difference between the activity transduced through homologous and that transduced through heterologous PRLRs, we propose the novel term 'minimal time of homodimer persistence'. This concept assumes that in order to initiate the signal transduction, the associated kinase JAK2 has to be transphosphorylated and this requires a 'minimal time' of homodimer existence. In the case of homologous interaction between ruminant PLs and homologous PRLRs, this 'minimal time' is met, though the interaction with homologous PRLRs has a shorter half-life than that with heterologous PRLRs. Therefore oPL or bPL are active in cells possessing both homologous and heterologous PRLRs. Mutations of oPL or bPL lead to reduced affinity and, consequently, the 'time of homodimer persistence' is shortened. Although in the case of heterologous interaction the 'minimal time' is still sufficient to initiate the biological activity, in homologous interactions, which are already weaker than heterologous interactions, further destabilization of the complex shortens its persistence to below the 'minimal time', leading to full or partial loss of biological activity.

Animals↗

PubMed2001

Effects of recombinant ovine leptin on in vitro lipolysis and lipogenesis in subcutaneous adipose tissue from lactating and nonlactating sheep.

Direct effects of recombinant ovine leptin on adipose metabolism in sheep were investigated. Lipolytic and lipogenic rates were assessed following preincubation of subcutaneous adipose tissue explants with recombinant ovine leptin. Leptin had no consistent effect on the basal (unstimulated) lipolytic rate in adipose tissue from wethers. Lipolytic rate measured in the presence of combinations of adenosine deaminase, isoprenaline, and N6-phenylisopropyl adenosine was unaffected by pretreatment with leptin. In lactating ewes, there was no relationship between increasing concentrations of leptin and basal lipolytic rate. Leptin had no effect on basal (unstimulated) lipogenesis, or on insulin-stimulation or growth hormone inhibition of lipogenesis in adipose tissue from wethers. Lipogenesis in adipose tissue from lactating ewes was also unaffected by preincubation with leptin; however, at supraphysiological concentrations of leptin, there was a small reduction in the rate of insulin-stimulated lipogenesis. Leptin failed to induce phosphorylation of the signal transducers and activators of transcription, STAT 3 and STAT 5, in sheep adipocytes. These results suggest that leptin does not have a direct physiological effect on subcutaneous adipose tissue metabolism in sheep.

Adenosine Deaminase↗