Fetal Brown Adipose Tissue Research Articles

The effect of maternal arginine supplementation on the development of the thermogenic program in the ovine fetus

Brown adipose tissue (BAT) is a specialised fat store that is metabolised by the newborn lamb to ensure effective adaptation to the cold challenge of the extra-uterine environment. Increasing BAT reserves therefore has the potential to increase neonatal thermogenesis and survival. It is established that arginine supplementation can increase fetal BAT stores but the biological mechanisms involved are unclear. The objective of this study was to test the hypothesis that increased fetal BAT stores resulting from maternal arginine supplementation is mediated by activation of the thermogenic program. Brown adipose tissue was collected from fetuses of ewes supplemented with arginine from 100 to 140 days of gestation. Increased peri-renal fat stores in fetuses from arginine-supplemented ewes was associated with an increase in uncoupling protein 1 (UCP-1) and PRD1-BF-1-RIZ1 homologous domain containing protein-16 expression, but not proliferator-activated receptor gamma or proliferator-activated receptor gamma-co-activator-1α in BAT. The activity of UCP-1 is regulated by hormones including cortisol and thyroid hormones. Cortisol level in fetuses from supplemented sheep was 68% greater than those from control ewes, indicating that cortisol may control upregulation of UCP-1 expression in the ovine neonate. The DNA and RNA concentration in BAT of both groups suggest that increased peri-renal fat stores is not associated with an increase in cell number or number of ribosomes, but rather an increase in the size of individual fat cells. Collectively, these results indicate that maternal arginine supplementation during mid to late gestation improved the thermoregulatory ability of lambs and this could potentially increase their survival in early life.

Expression and Subcellular Localization of Estrogen Receptors α and β in Human Fetal Brown Adipose Tissue

Brown adipose tissue (BAT) has the unique ability of generating heat due to the expression of mitochondrial uncoupling protein 1 (UCP1). A recent discovery regarding functional BAT in adult humans has increased interest in the molecular pathways of BAT development and functionality. An important role for estrogen in white adipose tissue was shown, but the possible role of estrogen in human fetal BAT (fBAT) is unclear. The objective of this study was to determine whether human fBAT expresses estrogen receptor α (ERα) and ERβ. In addition, we examined their localization as well as their correlation with crucial proteins involved in BAT differentiation, proliferation, mitochondriogenesis and thermogenesis including peroxisome proliferator-activated receptor γ (PPARγ), proliferating cell nuclear antigen (PCNA), PPARγ-coactivator-1α (PGC-1α), and UCP1. The fBAT was obtained from 4 human male fetuses aged 15, 17, 20, and 23 weeks gestation. ERα and ERβ expression was assessed using Western blotting, immunohistochemistry, and immunocytochemistry. Possible correlations with PPARγ, PCNA, PGC-1α, and UCP1 were examined by double immunofluorescence. Both ERα and ERβ were expressed in human fBAT, with ERα being dominant. Unlike ERβ, which was present only in mature brown adipocytes, we detected ERα in mature adipocytes, preadipocytes, mesenchymal and endothelial cells. In addition, double immunofluorescence supported the notion that differentiation in fBAT probably involves ERα. Immunocytochemical analysis revealed mitochondrial localization of both receptors. The expression of both ERα and ERβ in human fBAT suggests a role for estrogen in its development, primarily via ERα. In addition, our results indicate that fBAT mitochondria could be targeted by estrogens and pointed out the possible role of both ERs in mitochondriogenesis.

Impacts of arginine nutrition on embryonic and fetal development in mammals

Embryonic loss and intrauterine growth restriction (IUGR) are significant problems in humans and other animals. Results from studies involving pigs and sheep have indicated that limited uterine capacity and placental insufficiency are major factors contributing to suboptimal reproduction in mammals. Our discovery of the unusual abundance of the arginine family of amino acids in porcine and ovine allantoic fluids during early gestation led to the novel hypothesis that arginine plays an important role in conceptus (embryo and extra-embryonic membranes) development. Arginine is metabolized to ornithine, proline, and nitric oxide, with each having important physiological functions. Nitric oxide is a vasodilator and angiogenic factor, whereas ornithine and proline are substrates for uterine and placental synthesis of polyamines that are key regulators of gene expression, protein synthesis, and angiogenesis. Additionally, arginine activates the mechanistic (mammalian) target of rapamycin cell signaling pathway to stimulate protein synthesis in the placenta, uterus, and fetus. Thus, dietary supplementation with 0.83 % L-arginine to gilts consuming 2 kg of a typical gestation diet between either days 14 and 28 or between days 30 and 114 of pregnancy increases the number of live-born piglets and litter birth weight. Similar results have been reported for gestating rats and ewes. In sheep, arginine also stimulates development of fetal brown adipose tissue. Furthermore, oral administration of arginine to women with IUGR has been reported to enhance fetal growth. Collectively, enhancement of uterine as well as placental growth and function through dietary arginine supplementation provides an effective solution to improving embryonic and fetal survival and growth.

Pref-1 in brown adipose tissue: specific involvement in brown adipocyte differentiation and regulatory role of C/EBPδ

Pref-1 (pre-adipocyte factor-1) is known to play a central role in regulating white adipocyte differentiation, but the role of Pref-1 in BAT (brown adipose tissue) has not been analysed. In the present study we found that Pref-1 expression is high in fetal BAT and declines progressively after birth. However, Pref-1-null mice showed unaltered fetal development of BAT, but exhibited signs of over-activation of BAT thermogenesis in the post-natal period. In C/EBP (CCAAT/enhancer-binding protein) α-null mice, a rodent model of impaired fetal BAT differentiation, Pref-1 was dramatically overexpressed, in association with reduced expression of the Ucp1 (uncoupling protein 1) gene, a BAT-specific marker of thermogenic differentiation. In brown adipocyte cell culture models, Pref-1 was mostly expressed in pre-adipocytes and declined with brown adipocyte differentiation. The transcription factor C/EBPδ activated the Pref-1 gene transcription in brown adipocytes, through binding to the proximal promoter region. Accordingly, siRNA (small interfering RNA)-induced C/EBPδ knockdown led to reduced Pref-1 gene expression. This effect is consistent with the observed overexpression of C/EBPδ in C/EBPα-null BAT and high expression of C/EBPδ in brown pre-adipocytes. Dexamethasone treatment of brown pre-adipocytes suppressed Pref-1 down-regulation occurring throughout the brown adipocyte differentiation process, increased the expression of C/EBPδ and strongly impaired expression of the thermogenic markers UCP1 and PGC-1α [PPARγ (peroxisome-proliferator-activated receptor γ) co-activator-α]. However, it did not alter normal fat accumulation or expression of non-BAT-specific genes. Collectively, these results specifically implicate Pref-1 in controlling the thermogenic gene expression program in BAT, and identify C/EBPδ as a novel transcriptional regulator of Pref-1 gene expression that may be related to the specific role of glucocorticoids in BAT differentiation.

Arginine nutrition and fetal brown adipose tissue development in diet-induced obese sheep

The global incidence of human obesity has more than doubled over the past three decades. An ovine model of obesity was developed to determine effects of maternal obesity and arginine supplementation on maternal, placental, and fetal parameters of growth, health, and well being. One-hundred-twenty days prior to embryo transfer, ewes were fed either ad libitum (n = 10) to induce obesity or 100% National Research Council-recommended nutrient requirements (n = 10) as controls. Embryos from superovulated ewes with normal body condition were transferred to the uterus of control-fed and obese ewes on day 5.5 post-estrus to generate genetically similar singleton pregnancies. Beginning on day 100 of gestation, obese ewes received intravenous administration of saline or L-arginine-HCl three times daily (81 mg arginine/kg body weight/day) to day 125, whereas control-fed ewes received saline. Fetal growth was assessed at necropsy on day 125. Maternal obesity increased (1) percentages of maternal and fetal carcass lipids and (2) concentrations of leptin, insulin, glucose, glutamate, leucine, lysine and threonine in maternal plasma while reducing (1) concentrations of progesterone, glycine and serine in maternal plasma and (2) amniotic and allantoic fluid volumes. Administration of L-arginine to obese ewes increased arginine and ornithine concentrations in maternal and fetal plasma, amniotic fluid volume, protein content in maternal carcass, and fetal brown adipose tissue (+60%), while reducing maternal lipid content and circulating leptin levels. Fetal or placental weight did not differ among treatments. Results indicate that arginine treatment beneficially reduces maternal adiposity and enhances fetal brown adipose tissue development in obese ewes.

Bir Çocukta Spermatik Kord Hibernoması

Hibernomas are rare benign neoplasms derived from the remnants of fetal brown adipose tissue, seen mainly in adults. Most of them are located in areas that normally include brown-fat remnants. Although these tumors are considered benign and malignant transformation has not been reported, they tend to grow into large sizes. If a hibernoma is detected during inguinoscrotal approaches, it should be excised totally because of the risk of growth. To date, recurrence has not been described following complete excision, which is therefore recommended as the treatment of choice. We would like to present an 11-month-old child with hibernoma of the spermatic cord which was surgically treated. One child presenting such a lesion has previously been reported in the literature.

Arginine nutrition and fetal brown adipose tissue development in nutrient-restricted sheep

Intrauterine growth restriction is a significant problem worldwide, resulting in increased rates of neonatal morbidity and mortality, as well as increased risks for metabolic and cardiovascular disease. The present study investigated the role of maternal undernutrition and L-arginine administration on fetal growth and development. Embryo transfer was utilized to generate genetically similar singleton pregnancies. On Day 35 of gestation, ewes were assigned to receive either 50 or 100% of their nutritional requirements. Ewes received i.v. injections of either saline or L-arginine three times daily from Day 100 to Day 125. Fetal growth was assessed at necropsy on Day 125. Maternal dietary manipulation altered circulating concentrations of leptin, progesterone, and amino acids in maternal plasma. Fetal weight was reduced in nutrient-restricted ewes on Day 125 compared with 100% fed ewes. Compared with saline-treated underfed ewes, maternal L-arginine administration did not affect fetal weight but increased weight of the fetal pancreas by 32% and fetal peri-renal brown adipose tissue mass by 48%. These results indicate that L-arginine administration enhanced fetal pancreatic and brown adipose tissue development. The postnatal effects of increased pancreatic and brown adipose tissue growth warrant further study.

Hibernoma of the axillary region: a rare benign adipocytic tumor

Hibernoma is a rare benign tumor considered to arise from remnants of fetal brown adipose tissue. It tends to occur in sites where brown fat persists beyond fetal life, such as the interscapular region, but can occur in sites where brown fat is usually absent in adults. Clinicallywell, hibernomas are slow-growing, asymptomatic tumors. However, unlike lipomas, MRI findings sometimes mislead clinicians to diagnose a malignant neoplasm. We describe a 63-year-old male with an axillary hibernoma involving the brachial neurovascular bundles and mimicking a well-differentiated liposarcoma, from which it should be distinguished.

Giant hibernoma of the neck: A case report

Hibernoma is a rare benign tumour arising from remnants of fetal brown adipose tissue. We reported a rare case of hibernoma located in the neck. We also discuss the radiographic findings, pathologic features, and management of hibernomas.

Resection of intrapericardial hibernoma associated with constrictive pericarditis

Hibernoma is a benign soft-tissue tumor, which arises from the remnants of fetal brown adipose tissue. Out of less than 300 cases of hibernoma described to date, pericardial sac is an unusual localization to develop. When the tumor increases in size, this benign pathology can lead to compression of cardiac chambers and cause life-threatening complications in an asymptomatic patient. Here, the authors present the case of a 20-year-old male who underwent an operation for the treatment of constrictive pericarditis, in which an intrapericardial sessile lesion over diaphragmatic surface of pericardial sac was incidentally discovered. The tumor was excised and diagnosed as hibernoma. No recurrence was evident two years after the procedure.

Hibernoma subescapular. Presentación de un caso y revisión de la bibliografía

Hibernomas are uncommon neoplasms composed of brown adipose tissue. The largest database found contains 170 cases. In humans, this type of tumor arises from the remnants of fetal brown adipose tissue. Incomplete excision may result in continued growth and recurrence, even in tumors appearing to be histologically benign. The interest of the case reported herein lies in detection of the tumor during axillary lymphadenectomy for a right breast carcinoma.

Adenohibernoma of the breast

Dear Sir, Brown fat is mainly found in infants and children and gradually disappears from most sites of the body with increasing age. During adulthood, deposits of brown fat persist around kidneys, adrenal glands, and aorta and within the mediastinum and neck. Tumors composed of brown fat (hibernomas) are rare in breast [1, 2, 4, 5]. A rare case of hamartoma containing brown fat has been reported by Garijo et al. [5]. Adenohibernoma, hibernoma-containing glands, is also a very rare lesion as one case only has been described until now [2]. Here, we are reporting a second case. The patient, a 57-year-old lady, underwent excisional biopsy 2 cm across due to right breast lesion detected mammographically. The family history was unremarkable. Histologically, in addition to tubular carcinoma, an incidental lesion was detected nearby which had not been detected on mammograms. This latter lesion was well circumscribed and measuring 3 mm. It was formed partly by a fibroepithelial proliferation reminiscent of fibroadenoma with myxoid stroma (Fig. 1). Nuclear atypia and epithelial proliferation were not present. The remaining part of the nodular lesion showed dilated glands immersed within fat tissue, which showed uniform, normochromatic, and centrally located nuclei with vacuolated granular eosinophilic cytoplasms consistent with brown fat (Fig. 2). Accordingly, the vacuolated granular brown fat cells were positive with avidin–biotin complex peroxidase for S-100 protein (Dako, polyclonal, 1:1500; Fig. 3) and negative for wide spectrum keratin (Dako, MNF116, 1:200) and cluster of differentiation (CD)68 (Neomarkers, monoclonal, MAC387, 1:200). There was no residual lesion within the re-excision specimen which consisted of fibrofatty tissue measuring 4 cm. Seven lymph nodes from lower axillary level were reactive. There is no evidence of disease after a 6-year follow-up. Adenolipomas are lesions composed of glands immersed in adult classical adipous tissue and are closely associated with hamartomas. Both lesions are fibroglandular and are well circumscribed [3] and the amount of fat is by definition conspicuous in adenolipomas compared with hamartomas. Hamartomas are lesions that result from faulty development in an organ, composed of an abnormal mixture of normally occurring tissue elements or an abnormal proportion of a single element normally present at that site. Brown fat is not a normal constituent of adult breast tissue and in this respect adenohibernomas cannot be regarded as hamartomas. Damiani and Panarelli [2] suggested a hamartomatous origin from remnants of fetal brown adipose tissue or, alternatively, the possibility of de novo mixed proliferation showing brown fat differentiation in its mesenchymal component for adenohibernomas. In addition, the focus that the granular cells are associated to a fibroadenoma is a further proof that the adenohibernoma is not a hamartoma. This situation was favored by Fisher et al. [3] who believed that fibroadenomas and hamartomas were Virchows Arch (2008) 452:351–352 DOI 10.1007/s00428-007-0553-x

Chronic umbilical cord compression results in accelerated maturation of lung and brown adipose tissue in the sheep fetus during late gestation

Umbilical cord compression (UCC) sufficient to reduce umbilical blood flow by 30% for 3 days, results in increased fetal plasma cortisol and catecholamines that are likely to promote maturation of the fetal lung and brown adipose tissue (BAT). We determined the effect of UCC on the abundance of uncoupling protein (UCP)1 (BAT only) and -2, glucocorticoid receptor (GR), and 11beta-hydroxysteroid dehydrogenase (11beta-HSD)1 and -2 mRNA, and mitochondrial protein voltage-dependent anion channel (VDAC) and cytochrome c in these tissues. At 118 +/- 2 days of gestation (dGA; term approximately 145 days), 14 fetuses were chronically instrumented. Eight fetuses were then subjected to 3 days of UCC from 125 dGA, and the remaining fetuses were sham operated. All fetuses were then exposed to two 1-h episodes of hypoxemia at 130 +/- 1 and 134 +/- 1 dGA before tissue sampling at 137 +/- 2 dGA. In both tissues, UCC upregulated UCP2 and GR mRNA, plus VDAC and cytochrome c mitochondrial proteins. In lung, UCC increased 11beta-HSD1 mRNA but decreased 11beta-HSD2 mRNA abundance, a pattern reversed for BAT. UCC increased UCP1 mRNA and its translated protein in BAT. UCP2, GR, 11beta-HSD1 and -2 mRNA, plus VDAC and cytochrome c protein abundance were all significantly correlated with fetal plasma cortisol and catecholamine levels, but not thyroid hormone concentrations, in the lung and BAT of UCC fetuses. In conclusion, chronic UCC results in precocious maturation of the fetal lung and BAT mitochondria, an adaptation largely mediated by the surge in fetal plasma cortisol and catecholamines that accompanies UCC.

Pleural hibernoma. Treatment by videothoracoscopic surgery

Hibernoma is a rare, benign tumor originating from fetal brown adipose tissue. Although it is usually found in the interscapular region, it has also been reported in sites where brown fat is less common. Diagnosis of hibernoma is based on histological examination of an excised specimen. Since it is difficult to differentiate preoperatively between hibernoma and other soft-tissue tumors, such as lipoma or, more importantly, liposarcoma, total surgical excision is the treatment of choice. This paper offers what would appear to be the first reported case of pleural hibernoma treated by videothoracoscopic surgery.

Effect of maternal nutrition on brown adipose tissue and its prolactin receptor status in the fetal lamb.

We investigated the influence of maternal nutritional enhancement during the second half of gestation on prolactin receptor (PRLR) abundance in fetal brown adipose tissue (BAT) and liver close to term (i.e. 141-144 d gestation). Ewes were provided with 100% (i.e. control; n = 8) or 150% (i.e. well-fed; n = 7) of their metabolic requirements from 80 to 144 d gestation. Crude plasma membranes were prepared from fetal BAT and hepatic tissue, and individual molecular weight isoforms for the long and short forms of the PRLR were detected by immunoblotting. Mitochondrial preparations were prepared from BAT to measure the amount of the BAT-specific mitochondrial uncoupling protein-1 and its thermogenic activity (i.e. guanosine 5'-diphosphate binding). Fetuses sampled from well-fed ewes were heavier (controls, 3927 +/- 196 g; well-fed, 4783 +/- 219 g; p = 0.01) but possessed less BAT per kilogram body weight (controls, 5.92 +/- 0.43 g/kg; well-fed, 3.85 +/- 0.19 g/kg; p = 0.001), which had a greater uncoupling protein-1 abundance (controls, 56 +/- 5% of reference; well-fed, 78 +/- 9% of reference; p < 0.01) and higher thermogenic activity (controls, 157 +/- 41 pmol guanosine 5'-diphosphate per milligram mitochondrial protein; well-fed, 352 +/- 36 pmol guanosine 5'-diphosphate per milligram mitochondrial protein; p < 0.01) than controls. Multiple isoforms of the long and short forms of the P1LR were detected in all tissues. BAT from well-fed fetuses had a higher abundance of the 15-kD isoform of the long form of the PRLR (controls, 1.6 +/- 0.4 densitometric units; well-fed, 16.3 +/- 2.0 densitometric units; p < 0.001). This isoform was not detected in hepatic tissue. Maternal nutrient intake had no effect on any other isoforms of the PRLR in BAT or liver. In conclusion, increasing the quantity of feed provided in late gestation acts to promote fetal weight and BAT maturation, the combination of which will enhance neonatal viability.

Insulin receptor substrate (IRS) proteins IRS-1 and IRS-2 differential signaling in the insulin/insulin-like growth factor-I pathways in fetal brown adipocytes.

In the present study we have investigated the contribution of the insulin receptor substrate proteins (IRS-1 and IRS-2) to the insulin/insulin like growth factor I (IGF-I)-signaling pathways in fetal rat brown adipocytes, a model that expresses both insulin and IGF-I receptors. Insulin/IGF-I rapidly stimulated IRS-1 and IRS-2 tyrosine phosphorylation, their association with p85alpha, and IRS-1- and IRS-2-associated phosphatidylinositol (PI) 3-kinase activation to the same extent, the effect of insulin being stronger than the effect of IGF-I at the same physiological dose (10 nM). Furthermore, insulin/IGF-I stimulated IRS-1-associated Grb-2 phosphorylation. However, IRS-2-associated Grb-2 phosphorylation was barely detected. Pull-down experiments with glutathione-S-transferase-fusion proteins containing SH2-domains of p85alpha revealed a strong association between IRS-1 and IRS-2 with p85alpha in response to insulin/IGF-I, the insulin effect being stronger than IGF-I. However, the Grb-2-SH2 domain showed functional differences. While a strong association between IRS-1/Grb-2 was found, IRS-2/Grb-2 association was virtually absent in response to insulin/IGF-I, as also demonstrated in competition studies with a phosphopeptide containing the phosphotyrosine 895 residue within the putative Grb-2-binding domain. Finally, insulin/IGF-I stimulated tyrosine phosphorylation of the three SHC proteins (46, 52, and 66 kDa). Moreover, insulin/IGF-I markedly increased the amount of Grb-2-associated SHC proteins by the same extent. Our results suggest that both IRS-1 and IRS-2 are required for phosphatidylinositol 3-kinase activation that leads to adipogenic and thermogenic differentiation of fetal brown adipose tissue; meanwhile, IRS-1 and SHC, but not IRS-2, associate with Grb-2 leading to the ras-mitogen-activated protein kinase-signaling pathway required for fetal brown adipocyte proliferation.

Suppressive action of endogenous adenosine on ovine fetal nonshivering thermogenesis.

Nonshivering thermogenesis is not initiated when the fetal sheep is cooled in utero but appears to require the removal of an inhibitor of placental origin at birth. To test whether adenosine is such an inhibitor, we examined the effect of the adenosine antagonist theophylline on the initiation of nonshivering thermogenesis during sequential cooling, ventilation, and umbilical cord occlusion in utero. Theophylline (18 mg/kg bolus and 0.6 mg.kg-1.min-1 thereafter) was infused for 90 min before and 90 min after cord occlusion. Theophylline enhanced the nonshivering thermogenic free fatty acid (FFA) and glycerol responses before cord occlusion, raising FFA concentrations 99% to 415 +/- 60 mueq/l (P < 0.01) and glycerol levels 87% to 526 +/- 135 mumol/l (P < 0.05). These FFA (P < 0.001) and glycerol (P < 0.05) concentrations were significantly greater than the corresponding period during the birth-simulation control. Umbilical cord occlusion did not alter FFA levels but induced a 41% rise in glycerol concentrations to 774 +/- 203 mumol/l (P < 0.05). The increases in nonshivering thermogenic indexes after the administration of the adenosine-receptor antagonist suggest that the quiescent state of ovine fetal brown adipose tissue may result, in part, from the tonic inhibitory actions of adenosine and that a decrease in adenosine concentrations enhances nonshivering thermogenesis. However, the further rise after umbilical cord occlusion suggests that at least one other inhibitor of placental origin inhibits nonshivering thermogenesis before birth.

Regulation of uncoupling protein messenger ribonucleic acid and 5'-deiodinase activity by thyroid hormones in fetal brown adipose tissue.

We studied the regulation of type II 5'-deiodinase (5'D-II) activity and uncoupling protein (UCP) messenger RNA (mRNA) by thyroid hormones in fetal brown adipose tissue (BAT). Fetuses were obtained from hypothyroid pregnant rats infused with increasing doses of T4 or T3. Infusion of T4 into hypothyroid pregnant rats increased T4 and T3 concentrations and inhibited 5'D-II activity in fetal BAT. In contrast, infusion of T3 increased BAT 5'D-II activities at low, normal, or high BAT T3 concentrations. The relationship between thyroid hormone concentrations in fetal BAT and plasma showed that BAT T3 concentrations are relatively stable, increasing less than 2-fold over a wide range of circulating T4 (3-fold) or T3 (8-fold) concentrations. Most T3 in fetal BAT are locally derived from T4 and not from plasma T3. UCP mRNA expression decreased to 30% of control values in hypothyroid fetuses. UCP mRNA levels were restored to normal in parallel with BAT T3 concentrations after the infusion of either T4 or T3. UCP mRNA levels correlate well with BAT T3 concentrations. Supraphysiological doses of T4 did not further increase either BAT T3 or UCP mRNA levels. T3 might regulate basal UCP mRNA expression during fetal life.

Regulation of uncoupling protein messenger ribonucleic acid and 5'- deiodinase activity by thyroid hormones in fetal brown adipose tissue

Regulation of uncoupling protein messenger ribonucleic acid and 5'- deiodinase activity by thyroid hormones in fetal brown adipose tissue

Scalp hibernoma: Case report and literature review

Hibernomas are uncommon, benign neoplasms derived from the remnants of fetal brown adipose tissue. A review of the world literature revealed 105 cases, with the interscapular location the most common. Typically, hibernomas are asymptomatic and slow-growing. Adequate treatment usually consists of simple excision. We describe the second documented case of a scalp hibernoma.