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Biomedical subjects

R G Vernon

Publications and source records attributed to R G Vernon.

At least 37 records · Page 2Linked to original sources
PubMed1998

Stearoyl-CoA desaturase mRNA is transcribed from a single gene in the ovine genome.

Clones corresponding to ovine stearoyl-CoA desaturase (SCD) cDNA were isolated from an adipose tissue cDNA library. All of these clones represented a single mRNA species as judged by restriction fragment and DNA sequence analysis. RNase protection analysis demonstrated that this SCD transcript is highly expressed in adipose tissue and liver, and in the mammary gland of lactating animals. A lower level of expression was detectable in a variety of other tissues including brain. Levels of the SCD transcript were decreased in adipose tissue during lactation, and this appears to be related to a marked decline in serum insulin and insulin-responsiveness of the tissue. Southern analysis of ovine and mouse genomic DNA demonstrated that the ovine SCD cDNA hybridised in a manner consistent with a single gene for SCD in ovine DNA; mouse genomic DNA produced a pattern of hybridisation consistent with the previously characterised mouse SCD-1 and SCD-2 genes. Three ovine cosmids were isolated that comprised the restriction fragments predicted by the genomic Southern analysis. The ovine SCD gene was predicted to be encompassed within a 23 kbp region that was present in all three cosmids. These results demonstrate that SCD is transcribed from a single gene in the ovine genome and this gene is insulin-responsive in ovine adipose tissue.

Adipose Tissue↗
PubMed1998

Effects of food restriction on the responses of the mammary gland and adipose tissue to prolactin and growth hormone in the lactating rat.

Exogenous GH is used extensively in the USA to stimulate milk production in dairy cattle but its effectiveness is reduced in undernourished animals. It has been proposed that GH increases milk yield by stimulating IGF-I secretion and that this IGF-I-response is nutritionally sensitive and thus acts as a 'sensor' of energy balance. To investigate this possibility, we placed lactating rats on three planes of nutrition, ad libitum, 50% or 25% of ad libitum for 48 h. Subgroups of these animals were treated for 48 h with bromocriptine, to suppress prolactin secretion, and anti-rat GH, to neutralize GH action. From 24 to 48 h some of the treated animals were assessed for their milk yield response to prolactin or GH. Food restriction reduced milk yield in control rats by approximately 50% and was accompanied by a catabolic state, as judged by lipid mobilization from adipose tissue and by low concentrations of serum insulin, IGF-I, triiodothyronine and thyroxine, and increased serum nonesterified fatty acid concentrations. In animals fed ad libitum, anti-rat GH plus bromocriptine treatment produced an 80% decrease in milk yield and a dramatic fall in the activity of acetyl-CoA carboxylase in mammary tissue. GH was able to stimulate milk yield when given from 24 to 48 h; however, its effectiveness decreased progressively as food intake was reduced. The milk yield response to GH was accompanied by an increase in serum IGF-I concentrations and this response also decreased progressively with reduction of food intake, consistent with the hypothesis that IGF-I determines the milk yield response to GH and thus regulates GH action on the mammary gland in a nutritionally dependent fashion. However, the milk yield response to prolactin and the milk yield of control rats decreased in line with food intake without any changes in serum IGF-I concentrations. This clearly indicates that factors other than IGF-I are responsible for restricting milk yield. In order to assess other possible candidates for this role, we monitored serum glucose, non-esterified fatty acids, insulin triiodothyronine and thyroxine concentrations, but found no evidence for any simple relationship between these parameters and the milk yield response to prolactin and GH. Surprisingly we found that the ability of GH or prolactin to prevent epithelial cell loss in in the mammary gland was completely insensitive to nutrient intake, despite the fact that IGF-I is considered to be an important survival factor for mammary epithelial cells. Finally, we also demonstrated that, at least during short-term food restriction, the lactating rat is capable of mobilizing significant amounts of lipid from adipose tissue, such that it could provide the total output of triglyceride in milk, which is much greater than has previously been proposed.

Acetyl-CoA Hydrolase↗
PubMed1998

Regulation of the GTP-binding protein-based antilipolytic system of sheep adipocytes by growth hormone.

Chronic exposure of sheep adipose tissue to growth hormone (GH) in vitro decreases the ability of the adenosine analogue, N6-phenylisopropyladenosine (PIA), to inhibit isoprenaline-stimulated lipolysis by a mechanism which is dependent on both gene transcription and protein serine/threonine phosphorylation. The inhibition is not due to a change in ligand binding to the adenosine receptor, the amounts of the three isoforms of the inhibitory GTP-binding protein, Gi, or the maximum (forskolin-stimulated) adenylate cyclase activity. The ability of GH to modulate the PIA-activated adenosine receptor to stimulate dissociation of heterotrimeric Gi was assessed by measurement of pertussis toxin-catalysed ADP-ribosylation of Gi; GH does not appear to alter the interaction between the activated receptor and Gi. The ability of GH to alter the ability of activated Gi to inhibit adenylate cyclase activity was assessed by measuring the ability of a GTP analogue, guanosine 5'-[beta gamma-imido]triphosphate (p[NH]ppG), to inhibit forskolin-stimulated adenylate cyclase activity; chronic exposure to GH prevented this effect of p[NH]ppG. Thus the attenuation of the inhibition of lipolysis by PIA by chronic exposure of adipocytes to GH appears to be due to an impairment in the interaction between adenylate cyclase and the alpha subunit of one or more isoforms of Gi.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1997

Adaptations of maternal adipose tissue to lactation.

The ability to store substantial amounts of energy as lipid in adipose tissue has allowed development of a variety of strategies in wild animals to meet the considerable metabolic challenge of lactation. The ability to use adipose tissue energy has also been critical for development of the exceptional rates of milk production achieved in the dairy cow. Lactation thus results in profound changes in adipose tissue metabolism, the molecular bases of which are beginning to be resolved in domestic ruminants and laboratory rodents. In addition to its role as an energy store, adipose tissue has a variety of other functions (e.g., modulation of mammary development, appetite, immune system function), some of which are important for lactation.

Adaptation, Physiological↗
PubMed1997

Repression of the acetyl-CoA carboxylase gene in ovine adipose tissue during lactation: the role of insulin responsiveness.

We have investigated the mechanisms whereby lipogenesis is markedly suppressed in adipose tissue depots of lactating sheep. Expression of the gene encoding acetyl-CoA carboxylase (ACC), the flux-determining enzyme of the lipogenic pathway, is reduced approximately threefold in both omental and subcutaneous adipose tissue depots during late pregnancy and remains so into lactation when compared with non-pregnant, non-lactating animals. By comparison, total ACC enzyme activity in these adipose depots is suppressed approximately 25- to 30-fold in lactation. Culture of explants from the subcutaneous depot of lactating sheep with insulin plus dexamethasone for 72 h resulted in an approximately sevenfold increase in ACC mRNA, a fivefold increase in total enzyme activity and a marked increase in the proportion of the enzyme in the active state when compared with explants cultured with no added hormones for the same period. However, there was a lag of between 32 and 48 h before marked induction of any of these parameters by insulin plus dexamethasone was observed. Induction of the alpha-tubulin gene paralleled that of the ACC gene, suggesting that cytoskeletal rearrangements are associated with the aquisition of sensitivity to insulin plus dexamethasone. These results demonstrate that the reduction in lipogenic capacity in ovine adipose tissue during lactation is related to repression of the ACC gene, both at the level of steady-state mRNA abundance and possibly at translation, as well as to suppression of the mechanisms that regulate the proportion of ACC in the active state, and these are further related to the marked insensitivity of these parameters to insulin plus dexamethasone in vitro.

Acetyl-CoA Carboxylase↗
PubMed1996

Relationships between hypothalamic neuropeptide Y and food intake in the lactating rat.

NPYergic neurons in the hypothalamic arcuate nucleus (ARC) that project to the paraventricular nucleus (PVN) are postulated to regulate food intake and energy balance. This projection is overactive in lactation and is thought to drive hyperphagia in this condition. We have explored further the relationship between hypothalamic NPY and food intake in lactation and tested the hypothesis that hypoinsulinemia is the stimulus to NPY neuronal activity. Compared with nonlactating controls (n = 10), freely fed lactating rats (n = 9) showed significantly increased (p < 0.05) NPY levels in the ARC and medial preoptic area (MPO), but there was no significant increase in whole hypothalamic NPY mRNA levels. Lactating rats (n = 8) that were restricted to control rats' food intake for 3 days showed generally higher hypothalamic NPY levels, with significantly higher concentrations than controls (p < 0.05) in the ARC, MPO, PVN, and lateral hypothalamic area (LHA); NPY mRNA levels were also significantly increased (p < 0.05). Across all three experimental groups, there was a significant inverse correlation between plasma insulin concentration and hypothalamic NPY mRNA levels (r = -0.39, p < 0.01). We conclude that the ARC-PVN projection is overactive in lactation and that this is not a consequence of hyperphagia. Hypoinsulinemia may stimulate these neurons, as it is thought to do in other conditions of energy deficit.

Animals↗
PubMed1996

Insulin-like growth factors-I and -II, somatotropin, prolactin, and placental lactogen are not acute effectors of lipolysis in ruminants.

The acute regulation of lipolysis in the adipose tissue of ruminants was evaluated with lactating cows (n = 4) and growing ewe lambs (n = 11). Subcutaneous adipose tissue was obtained by biopsy or at slaughter and was incubated with varying concentrations of biologically active insulin-like growth factors-I and -II (IGF-I, IGF-II), somatotropin (ST), prolactin (PRL), or placental lactogen (PL) to determine the effect of these hormones on lipolysis. Complimentary studies were conducted to examine the effects of IGF-I and IGF-II on the acute regulation of lipolysis in adipose tissue from lactating ewes and wethers under a variety of incubation conditions. Isoproterenol (ISO), a beta-adrenergic agonist known to rapidly stimulate lipolysis, was used as a positive control. Incubation with ISO for 3 hr resulted in a significant increase in the rates of lipolysis. However, there was no stimulation of lipolysis over the 3-hr incubation at any concentration or under any conditions for IGF-I or IGF-II. Furthermore, ST, PRL, or PL had no acute effects on the rates of lipolysis in adipose tissue. These data demonstrate that IGF-I, IGF-II, ST, PRL, and PL are not acute effectors of the lipolytic rate in the adipose tissue of ruminants.

Adipose Tissue↗
PubMed1996

GH inhibition of lipogenesis and stimulation of lipolysis in sheep adipose tissue: involvement of protein serine phosphorylation and dephosphorylation and phospholipase C.

The intracellular signalling systems involved in the chronic insulin-antagonistic, anti-lipogenic effects and also the lipolytic effect of GH have been investigated in sheep adipose tissue in an in vitro tissue culture system. During culture, chronic exposure to GH decreased the rate of lipogenesis and prevented the increase in lipogenesis induced by insulin. GH also increased glycerol release into the culture medium. GH had no acute, insulin-like effect on lipogenesis in sheep adipose tissue. Pretreatment with phorbol ester to down-regulate isoforms of protein kinase C or addition of the protein serine kinase inhibitor staurosporine decreased the anti-lipogenic effect of GH while the protein serine kinase inhibitor H7 eliminated it completely. Pretreatment with phorbol ester or addition of H7 also decreased the insulin-antagonistic effect of GH on lipogenesis. Addition of the protein serine phosphatase inhibitor okadaic acid or the phosphatidyl choline phospholipase C inhibitor D609 both diminished the anti-lipogenic and insulin-antagonistic effects of GH. Chronic exposure of adipose tissue to GH had no effect on the total activity of acetyl CoA carboxylase or its activation status but it did diminish the increase in activation status induced by insulin. H7 and okadaic acid also diminished the increase in activation status of acetyl CoA carboxylase induced by insulin but did not alter the effect of GH on this variable. Okadaic acid decreased total acetyl CoA carboxylase activity. Pretreatment with phorbol ester or the addition of H7, staurosporine or okadaic acid increased glycerol release into the culture medium to the same extent as GH itself; the effects of GH and these various agents were not additive. These studies suggest that the anti-lipogenic, insulin-antagonistic effects of GH involve both protein serine kinases and phosphatases, possibly including one or more isoforms of protein kinase C, and a phosphatidyl choline-specific phospholipase C. Comparison with studies by others on the GH enhancement of preadipocyte differentiation and prolactin stimulation of lipogenesis in mammary tissue suggests involvement of protein kinase C at an early stage in all three systems. In contrast, effects of okadaic acid vary with the system, suggesting the involvement of protein serine phosphatase activity in a late stage of the action of GH. The effects of GH on lipogenesis and lipolysis do not occur via identical mechanisms.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗