Epigenetic change in IGF2R is associated with fetal overgrowth after sheep embryo culture.

Manipulation or non-physiological embryo culture environments can lead to defective fetal programming in livestock. Our demonstration of reduced fetal methylation and expression of ovine IGF2R suggests pre-implantation embryo procedures may be vulnerable to epigenetic alterations in imprinted genes. This highlights the potential benefits of epigenetic diagnostic screening in developing embryo procedures.

Regulators of adipocyte precursor cells.

Lean and adipose tissue growth are two of the most important targets for manipulation in commercial livestock. Adipose tissue growth occurs by both hyperplasia and hypertrophy. The processes involved in adipocyte hypertrophy are relatively well understood but much less is known about adipocyte hyperplasia.

Decreased muscle cell proliferation in chicks with a deletion in the GH receptor gene.

The increase in muscle weight in neonatal animals is a consequence of increased protein accretion and DNA content. GH increases protein accretion but direct effects of GH on myogenic cell proliferation have not been demonstrated. Sex-linked dwarfism in the chick is caused by mutation or deletion in the GH receptor gene and has provided a useful model to study the physiological consequences of GH insensitivity.

Regulation of chick muscle satellite cells by fibroblast growth factors: interaction with insulin-like growth factor-I and heparin.

This study describes the effect of acidic and basic fibroblast growth factor (FGF) on DNA synthesis in chick satellite cells in vitro and interactions with insulin-like growth factor-I (IGF-I) and exogenous heparin. Basic bFGF stimulated incorporation of [3H]thymidine into DNA with a half-maximum concentration (ED50) of 3.23 +/- 0.

Molecular events in adipocyte development.

Adipocyte hyperplasia occurs by the proliferation and differentiation of adipocyte precursor cells or preadipocytes. Although the process of commitment to the adipocyte lineage is poorly understood, a great deal of information has accumulated about the processes and regulatory mechanisms involved in preadipocyte differentiation. The differentiation of preadipocytes is known to be characterized by increased transcription of a number of specific genes.

Decreased fat content and increased lean in pigs treated with antibodies to adipocyte plasma membranes.

Antibodies were prepared in sheep against purified plasma membranes from pig adipocytes. Western (immuno) blotting revealed reactions of the antisera with a large number of proteins in adipocyte plasma membranes but remarkably few in plasma membranes from muscle, kidney, liver, lung, brain, spleen, and erythrocytes. This illustrated the high degree of specificity the serum had for adipose tissue.

Regulation of adipocyte precursor DNA synthesis by acidic and basic fibroblast growth factors: interaction with heparin and other growth factors.

The development of adipose tissue is dependent on the growth and differentiation of fibroblast-like adipocyte precursor cells. Culture of adipocyte precursor cells in vitro has provided an ideal system for identifying potential regulators of proliferation and differentiation. We have demonstrated that both acidic fibroblast growth factor (aFGF) and basic fibroblast growth factor (bFGF) stimulate chicken adipocyte precursor DNA synthesis in a dose-dependent manner up to a concentration of 100 micrograms aFGF/l and 1 microgram bFGF/l.

Comparison of the rates of proliferation of adipocyte precursor cells derived from two lines of chicken which differ in their rates of adipose tissue development.

1. Putative adipocyte precursor cells were isolated from the white adipose tissue of young broiler and layer chickens and cultured in vitro. 2.

Multiple growth factor mRNAs are expressed in chicken adipocyte precursor cells.

We have examined the expression of growth factor genes in primary cultures of chicken adipocyte precursors. RNA was extracted from proliferating and differentiated cells, reversed transcribed and amplified by PCR using gene specific primers. The identity of the PCR products was confirmed by restriction mapping.

Effects of transforming growth factor-alpha on chicken adipocyte precursor cells in vitro.

The hyperplastic capacity of adipose tissue resides in a group of fibroblast-like adipocyte precursor cells. There is evidence to suggest that their proliferation and differentiation is regulated by insulin-like growth factor-I (IGF-I) and transforming growth factor-beta (TGF-beta) but there is less information about other growth factors which may also participate in adipocyte precursor cell hyperplasia. Transforming growth factor-alpha (TGF-alpha) is a 50 amino acid polypeptide which has been shown to stimulate proliferation in both neoplastic and normal cell types acting through the epidermal growth factor (EGF) receptor.

Adipose tissue lipogenesis and fat deposition in leaner broiler chickens.

Rates of hepatic lipogenesis and secretion of plasma triglyceride-rich lipoproteins in 6- to 7-wk-old broiler chickens were similar to the overall rate of fat deposition in these birds, although approximately 20% of [14C]-labeled VLDL was oxidized to CO2 within 8 h. Only 6-7% of VLDL and portomicron triglyceride was taken up by the abdominal fat pad, but this proportion of total triglyceride flux could account for about 80-85% of the total fatty acids accumulating in that depot. The rate of lipogenesis in adipose tissue was much lower than that in the liver, but it could account for much of the remaining fatty acids.

Identification of chicken (Gallus domesticus) adipocyte plasma membrane and differentiation specific proteins using SDS-PAGE and western blotting.

1. Affinity-purified adipocyte membrane proteins were used to raise antisera in two sheep. 2.

Regulation of DNA synthesis in chicken adipocyte precursor cells by insulin-like growth factors, platelet-derived growth factor and transforming growth factor-beta.

Adipose tissue growth can occur by both hypertrophy and hyperplasia. The capacity for adipocyte hyperplasia in vivo resides in a population of fibroblast-like adipocyte precursor cells but the regulation of the proliferation of these cells by growth factors has not been well characterized. This study was designed to determine the effects of the insulin-like growth factors (IGF-I and IGF-II), platelet-derived growth factor (PDGF) and transforming growth factor-beta 1 (TGF-beta 1) added alone or together on the proliferation of primary adipocyte precursor cells in vitro.

Effects of transforming growth factor beta 1 and basic fibroblast growth factor on lipoprotein lipase activity in primary cultures of chicken (Gallus domesticus) adipocyte precursors.

1. The effect of TGF-beta and bFGF on lipoprotein lipase activity in chicken adipocyte precursors was investigated. 2.

Cytotoxic antibodies to chicken adipocytes and their precursors: lack of tissue specificity.

1. Antisera raised in sheep against chicken adipocyte plasma membranes recognised adipocyte, liver and red blood cell membranes in enzyme immunoassays (EIA). 2.

The effects of macrophage-derived cytokines on lipid metabolism in chicken (Gallus domesticus) hepatocytes and adipocytes.

1. The effects of avian and mammalian cytokines on avian lipid metabolism were compared using cultured chicken hepatocytes and adipocytes. 2.

Biochemical indicators of fatness in meat-type chickens: lack of correlation between lipoprotein lipase activity in post-heparin plasma and body fat.

1. Possible relationships between fatness and lipoprotein lipase activity in adipose tissue and plasma from heparinised birds were examined in 7-week-old male and female broilers. 2.

Effect of Escherichia coli endotoxin on tissue lipoprotein lipase activities in chickens.

1. Four-week-old broiler chickens were injected intravenously with from 0.01 to 1 mg of E.

Contribution of lipoprotein lipase to differences in fatness between broiler and layer-strain chickens.

The growth of abdominal fat in chickens from broiler and layer-strains up to 10 weeks of age was measured and compared with changes in plasma very low density lipoprotein (VLDL) concentration and tissue lipoprotein lipase activities. The growth of abdominal fat in broilers was much more rapid than in layer-strain chickens. Plasma VLDL concentrations in the two strains were similar up to 5 weeks of age but thereafter concentrations tended to be higher in broilers.

Can phosphorylation of phosphatidate phosphohydrolase by a cyclic AMP-dependent mechanism regulate its activity and subcellular distribution and control hepatic glycerolipid synthesis?

Incubating the particle-free supernatant of rat liver with alkaline phosphatase decreased the activity of phosphatidate phosphohydrolase by 21-29%. When the particle-free supernatant was incubated with various combinations of Mg2+, ATP, cyclic AMP and cyclic AMP-dependent protein kinase this failed to alter significantly phosphatidate phosphohydrolase activity under the conditions employed. The incubation of hepatocytes in monolayer culture with 0.

Partial purification and characterization of the soluble phosphatidate phosphohydrolase of rat liver.

A method is described by which the Mg2+-stimulated phosphatidate phosphohydrolase can be purified from the soluble fraction of liver from ethanol-treated rats. The increase in specific activity was about 416-fold. This involved purification by adsorption on calcium phosphate, chromatography on DE-52 DEAE-cellulose, separation on Ultrogel AcA-34 and chromatography on CM-Sepharose 6B.