A method for the isolation of lipid droplet fractions from decapsulated rat adrenals

Mrotek, James J.; Mathewm, J.K.; Curtis, Joseph C.; Johansson, K. R.

doi:10.1016/0039-128x(81)90037-4

Cited by 26 publications

(6 citation statements)

References 30 publications

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“…Preparation of lipid droplet fractions for Western blot analysis 3T3-L1 preadipocytes were incubated with SE at the indicated times (0, 2, 6, and 18 h), and total cell homogenates, fatcake, and cytosolic fractions were obtained according to the methods of Clifford et al (2000) and Mrotek et al (1981). The cells were lysed in 50 mM TRIS-HCl buffer, pH 7.4, containing 225 mM sucrose, 1 mM EDTA, 1 mM benzamidine, 1 mg/ml pepstatin, 1 mg/ml leupeptin, 1 mg/ml antipain, and 50 mM NaF.…”

Section: Induction Of Lipid Droplet Formationmentioning

confidence: 99%

Temporal and spatial assembly of lipid droplet-associated proteins in 3T3-L1 preadipocytes

Wang

Hwang

Greenberg

et al. 2003

Histochemistry and Cell Biology

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This study aimed to investigate the relationship between newly formed lipid droplets and lipid droplet surface proteins, including perilipin, adipose differentiation related protein (ADRP), and p200 kDa protein (p200) in 3T3-L1 preadipocytes, during lipogenesis. Sterol ester was used to induce nascent lipid droplets in 3T3-L1 preadipocytes and the sequence of lipids and lipid droplet surface proteins was studied using a combination of immunohistochemistry and Nile red staining/Oil red O. We demonstrated that, although most growing lipid droplets appeared to have a lipid core surrounded by a fluorescent rim of ADRP, perilipin, and p200, tiny protein aggregates of ADRP, perilipin, or p200 could also be found to occur in the absence of lipid accumulation. In addition, ADRP associated with nascent lipid droplets prior to that of perilipin or p200. We provide evidence that lipid droplet surface proteins, especially ADRP and perilipin, are important in serving as a nucleation center for the assembly of lipid to form nascent lipid droplets.

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Section: Induction Of Lipid Droplet Formationmentioning

confidence: 99%

Temporal and spatial assembly of lipid droplet-associated proteins in 3T3-L1 preadipocytes

Wang

Hwang

Greenberg

et al. 2003

Histochemistry and Cell Biology

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“…The intracellular lipid droplets of adrenocortical cells were purified as described in a previous report (19). Each adrenal gland was trimmed to remove fat and homogenized in 1 ml of a cold 0.25 M sucrose solution with a Teflon/glass homogenizer on ice.…”

Section: Isolation Of Intracellular Lipid Dropletsmentioning

confidence: 99%

Association of Globular β-Actin with Intracellular Lipid Droplets in Rat Adrenocortical Cells and Adipocytes

Fong

Liao

et al. 2001

Biochemical and Biophysical Research Communications

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“…Since cholesterol and cholesterol ester are major components of lipid droplets in Leydig cells (Mrotek et al 1981), the cholesterol contents were quantified using Leydig cells isolated from the mice. The levels of total cholesterol were significantly decreased in Leydig cells isolated from the 2,4-D-treated wild-type mice compared with the control wild-type mice (Fig.…”

Section: Resultsmentioning

confidence: 99%

PPARα-dependent cholesterol/testosterone disruption in Leydig cells mediates 2,4-dichlorophenoxyacetic acid-induced testicular toxicity in mice

et al. 2016

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It was reported that 2,4-dichlorophenoxyacetic acid (2,4-D), a commonly used herbicide and a possible endocrine disruptor, can disturb spermatogenesis, but the precise mechanism is not understood. Since 2,4-D is a weak peroxisome proliferator in hepatocytes and peroxisome proliferator-activated receptor α (PPARα) is also expressed in Leydig cells, this study aimed to investigate the link between PPARα and 2,4-D-mediated testicular dysfunction. 2,4-D (130 mg/kg/day) was administered to wild-type and Ppara-null mice for 2 weeks, and the alterations in testis and testosterone/cholesterol metabolism in Leydig cells were examined. Treatment with 2,4-D markedly decreased testicular testosterone in wild-type mice, leading to degeneration of spermatocytes and Sertoli cells. The 2,4-D decreased cholesterol levels in Leydig cells of wild-type mice through down-regulating the expression of 3-hydroxy-3-methylglutaryl coenzyme A synthase 1 and reductase, involved in de novo cholesterogenesis. However, the mRNAs encoding the important proteins involved in testosterone synthesis were unchanged by 2,4-D except for CYP17A1, indicating that exhausted cholesterol levels in the cells is a main reason for reduced testicular testosterone. Additionally, pregnancy rate and the number of pups between 2,4-D-treated wild-type male mice and untreated female mice were significantly lower compared with those between untreated couples. These phenomena were not observed in 2,4-D-treated Ppara-null males. Collectively, these results suggest a critical role for PPARα in 2,4-D-induced testicular toxicity due to disruption of cholesterol/testosterone homeostasis in Leydig cells. This study yields novel insights into the possible mechanism of testicular dysfunction and male infertility caused by 2,4-D.

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A method for the isolation of lipid droplet fractions from decapsulated rat adrenals

Cited by 26 publications

References 30 publications

Temporal and spatial assembly of lipid droplet-associated proteins in 3T3-L1 preadipocytes

Temporal and spatial assembly of lipid droplet-associated proteins in 3T3-L1 preadipocytes

Association of Globular β-Actin with Intracellular Lipid Droplets in Rat Adrenocortical Cells and Adipocytes

PPARα-dependent cholesterol/testosterone disruption in Leydig cells mediates 2,4-dichlorophenoxyacetic acid-induced testicular toxicity in mice

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