Differentiation of brown adipose tissue and biogenesis of thermogenic mitochondria in situ and in cell culture

doi:10.1016/0005-2728(90)90258-6

Biochimica et Biophysica Acta (BBA) - Bioenergetics

Volume 1018, Issues 2–3, 25 July 1990, Pages 243-247

https://doi.org/10.1016/0005-2728(90)90258-6 Get rights and content

Abstract

Differentiation and biogenesis of mitochondria in brown adipose tissue (BAT) was studied in situ and in cell culture by Western blotting, enzyme activity measurements, [³⁵S]methionine incorporation and immunofluorescence microscopy. In different rodent species the perinatal development of BAT thermogenic function resulted from the formation of thermogenic mitochondria which replaced the preexisting nonthermogenic mitochondria. Their biogenesis was characterized by the sudden appearance and rapid increase of the uncoupling protein (UCP), increase of cytochrome oxidase (COX) and decrease of H⁺-ATPase. In primary cell culture, differentation of precursor cells from mouse BAT to typical multilocular adipocytes was accompanied by increasing content of COX and H⁺-ATPase. A selective synthesis of UCP was induced by activation of β-adrenergic receptors or by elevated levels of cellular cAMP. UCP was quantitatively incorporated into mitochondria and within 24 h after stimulation reached near physiological concentration. Both in situ and in cell culture, the conditions enabling the expression of UCP gene were accompanied by activation of intracellular thyroxine 5′-deiodinase.

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Cited by (21)

Mitochondrial diseases and ATPase defects of nuclear origin
2004, Biochimica et Biophysica Acta - Bioenergetics
Citation Excerpt :
The cellular content of ATPase differs in mammalian tissues and it may vary also during ontogenetic development. Physiologically ATPase is down-regulated about 10-fold, relative to other OXPHOS complexes, in thermogenic brown fat where the electrochemical potential of proton gradient is dissipated in the form of heat by the uncoupling protein UCP1 [34]. Interestingly, the two-dimensional electrophoretic pattern of OXPHOS proteins in brown fat is almost identical with that found in patients with ATPase deficiency.
Dysfunctions of the F₁F_o-ATPase complex cause severe mitochondrial diseases affecting primarily the paediatric population. While in the maternally inherited ATPase defects due to mtDNA mutations in the ATP6 gene the enzyme is structurally and functionally modified, in ATPase defects of nuclear origin mitochondria contain a decreased amount of otherwise normal enzyme. In this case biosynthesis of ATPase is down-regulated due to a block at the early stage of enzyme assembly—formation of the F₁ catalytic part. The pathogenetic mechanism implicates dysfunction of Atp12 or other F₁-specific assembly factors. For cellular energetics, however, the negative consequences may be quite similar irrespective of whether the ATPase dysfunction is of mitochondrial or nuclear origin.
Inverse distribution of uncoupling proteins expression and oxidative capacity in mature adipocytes and stromal-vascular fractions of rat white and brown adipose tissues
1999, FEBS Letters
To investigate relationships between the uncoupling protein (UCP) family and oxidative metabolism in fat pads, we measured the cytochrome oxidase activity, used as an index of oxidative capacity, and the mRNA content encoding UCP1, UCP2 and UCP3. Most oxidative potential was found in the stromal-vascular fraction (SVF) of brown fat and in mature adipocytes of white fat (inguinal and periovarian). Considering the whole fat pads, the oxidative potential observed in mature white adipocytes fraction was not negligible compared with that of brown adipocytes fraction. UCP1 and UCP3 were expressed exclusively in mature brown adipocytes. Whatever the deposit, UCP2 mRNA was mainly localized in the SVF. These results indicate that, in fat, high oxidative potential is not necessarily linked to high UCPs transcripts content and point out the oxidative capacity of SVF from brown fat.
Physiological approach to maturation of brown adipocytes in primary cell culture
1997, Biochimica et Biophysica Acta - Molecular Cell Research
Molecular and metabolic aspects of differentiation of brown adipocytes of the Djungarian hamster (Phodopus sungorus) were studied in primary culture. Expression of uncoupling protein and lipoprotein lipase were investigated by Western and Northern blotting and indirect immuno-fluorescence microscopy. The activity of 5′-deiodinase type II was determined by a radioactive enzyme assay. Activity of cytochrome-c-oxidase and cell respiration rates were measured with a Clark electrode. We evaluated functional differences of developmental stages by measuring the reaction to β-adrenergic stimulation throughout the differentiation process. The results show that differentiation of hamster brown adipocytes is an at least two-step development with physiologically discriminable cell types. Generation of triiodothyronine (T3) from thyroxine by activation of the 5′deiodinase occurs in immature brown adipocytes and is mediated primarily by β1- rather than β3-adrenergic receptors. The thermogenic capacity is subsequently increased in mature brown adipocytes. β-Adrenergic receptor stimulation increases UCP expression of mature adipocytes but is not able to recruit new brown adipocytes.
Expression of interleukin-1α and interleukin-1 receptor type I genes in murine brown adipose tissue
1993, FEBS Letters
At thermoneutral conditions, high steady-state levels of transcripts for both IL-1α and its receptor IL-1RtI were found in specialized thermogenic organ, brown adipose tissue (BAT) of adult mice, as compared with the levels in lymph nodes, brain and spleen. A pronounced decrease of IL-1α mRNA level in BAT was observed after lipopolysaccharide (LPS) administration and after exposure to cold. Likewise, LPS decreased the IL-1 RtI mRNA level and depressed also the expression of cold-inducible genes for the BAT-specific heat-producing uncoupling protein and for lipoprotein lipase. It is concluded that, besides the centrally-mediated effects, there exists a direct peripheral interaction of IL-1 cytokines with BAT cells.
Differentiation of Brown Adipose Cells in Three-Dimensional Collagen Gel Culture
1993, Pathology Research and Practice
Brown adipose cells are heat-producing cells through non-shivering thermogenesis by in tram itochondrial “thermogenin”. This specific protein is a marker for their cellular differentiation. It has long been known that cultured brown adipose cells in monolayer rapidly lose the thermogenin bioactivity. In this study, we cultured brown adipose cells in three-dimensional type I collagen gel matrix. Under this culture condition, they were able to survive, and differentiated morphologically and functionally for a long period of time, especially exhibited the characteristic immunohistochemical activity of thermogenin. These findings suggest that brown adipose cells differentiate in type I collagen gel. In this condition, cholera toxin or BRL 37344, one of beta₃-adrenoceptor agonists, specifically stimulated the brown adipose cells.
Regulation of UCP gene expression in brown adipocytes differentiated in primary culture. Effects of a new β-adrenoceptor agonist
1992, Molecular and Cellular Endocrinology
Primary cultures of precursor cells from mouse and rat brown adipose tissue (BAT) were used to study the effect of a new β-agonist (ICI D7114) on the uncoupling protein (UCP) gene expression. ICI 215001 (the active metabolite of D7114) increased the expression of UCP and its mRNA in brown adipocytes differentiating in vitro in a dose-dependent manner. This stimulating effect was not inhibited by propranolol, a non-specific β-antagonist, but was partially reduced by bupranolol, a β₃-antagonist. No expression of UCP mRNA was ever induced by ICI 215001 in white adipocytes differentiated in vitro. It was concluded that the drug could affect the brown adipose cells through a β₃-pathway. It could clearly modulate the expression of UCP in brown adipocytes differentiated in vitro, but was not able by itself to turn on the gene.

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ColloquiaDifferentiation of brown adipose tissue and biogenesis of thermogenic mitochondria in situ and in cell culture

Abstract

Comp. Biochem. Physiol.

Biochim. Biophys. Acta

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Biochim. Biophys. Acta

Physiol. Rev.

Biochemistry

Biol. Neonate

Biochim. Biophys. Acta

J. Biol. Chem.

Colloquia
Differentiation of brown adipose tissue and biogenesis of thermogenic mitochondria in situ and in cell culture