Melatonin-Based Therapeutics for Atherosclerotic Lesions and Beyond: Focusing on Macrophage Mitophagy

The Cellular Origin in Atheromatous Lesion

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010006800x ◽

1970 ◽

Vol 28 ◽

pp. 202-203

Author(s):

T. M. Murad ◽

H. A. I. Newman ◽

K. F. Kern

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Rabbit Aorta ◽

Mixed Population ◽

The Other ◽

Cellular Origin ◽

Ultrastructural Studies ◽

Atherosclerotic Lesions ◽

Show Evidence ◽

Early Lesion

The origin of lipid containing cells in atheromatous lesion has been disputed. Geer in his study on atheromatous lesions of rabbit aorta, suggested that the early lesion is composed mainly of lipid-laden macrophages and the later lesion has a mixed population of macrophages and smooth muscle cells. Parker on the other hand, was able to show evidence that the rabbit lesion is primarily composed of lipid-laden cells of smooth muscle origin. The above studies and many others were done on an intact lesion without any attempt of cellular isolation previous to their ultrastructural studies. Cell isolation procedures have been established for atherosclerotic lesions through collagenase and elastase digestion Therefore this procedure can be utilized to identify the cells involved in rabbit atheroma.

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En face dual fluorescence staining of fatty streaks allows observation of initiation and progression of atherosclerotic lesions

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100140695 ◽

1995 ◽

Vol 53 ◽

pp. 864-865

Author(s):

Anne M. Klinkner ◽

Crystal R. Waites ◽

Peter J. Bugelski ◽

William D. Kerns

Keyword(s):

Fluorescent Dyes ◽

Nile Red ◽

Dimethyl Formamide ◽

High Cholesterol Diet ◽

Methods Development ◽

Atherosclerotic Lesions ◽

En Face ◽

Biochemical Evaluation ◽

Stock Solutions ◽

Dual Staining

A primary effort in the understanding of the progression of atherosclerotic disease has been methods development for visualization of the atherosclerotic plaque. We introduce a new method for the qualitative analysis of lipids in atherosclerotic fatty streaks which also retains those lipids for biochemical evaluation. An original aspect of the process is the ability to view an entire fatty streak en face, selectively stained for specific lipid classes within the lesion.New Zealand white rabbits were fed a high cholesterol diet(0.15%-0.3% for 14 wks). The aorta was removed and fixed in Carson's phosphate buffered formaldehyde followed by dual staining in the fluorescent dyes Nile red and filipin. Stock solutions of nile red(0.5mg/ml acetone) and filipin(2.5mg/ml dimethyl formamide) were prepared and kept at -20°C; all subsequent steps were at RT. 0.5cm × 1.0cm pieces of aorta were trimmed and adventitia removed. The pieces were then washed 3×15 min in PBS w/o CaMg, soaked in Nile red(NR)/filipin(Fl) stain(100(il NR stock + 200μl Fl stock in 10 ml PBS for 30 min, washed in PBS 3×30 min, rinsed with distilled water, mounted(Crystal Mount, Biomedia) and coverslipped and viewed by fluorescence microscopy.

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