To eliminate blood contamination before surgeries, I usually use ice-cold 1 X PBS to perfuse a mouse. To fix mouse tissues before immunohistochemistry, I use 4 or 10% formalin to perfuse mouse.
1. Anesthetize the mouse with isoflurane in the hood.
2. Open the mouse from the abdominal.
3. Cut open the diaphragm along both side and expose the heart. Make sure not cutting major vessels.
4. Remove a drop of heparin in a 1 ml syringe with 26 G needle. Draw blood from the left ventricle in the apex of the heart.
5. Cut open the right atrium using a scissor at right angle. Make sure not cutting other parts of the heart. You will notice blood flowing from the right atrium,
6. Remove a drop of heparin in a 10 ml syringe and draw 10 ml ice-cold PBS. Use 23G needle to stick into the left ventricle. Note the best angle is paralleling the edge between left and right ventricle.
7. Drain the PBS into the heart. You will see blood flowing out from right atrium.
8. Try one more time with another 10 ml PBS if necessary. Use 4 or 10% formalin instead of PBS when fixing mouse tissues.
9. The liver turns yellow slowly. The sign for good perfusion is the yellowish color of kidney.
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SIRT6 links glucose and lipid metabolism
SIRT1, a histone deacetylase, has drawn considerable attention due to its important roles in metabolic regulation and longevity. SIRT1 is induced by fasting and suppressed by feeding. SIRT1 increases glucose production and beta oxidation in liver to meet energy needs during food deprivation. Another member of SIRT family, SIRT6 follows similar expression patterns as SIRT1. The function of SIRT6 in metabolic regulation remains unknown.
Kim and colleagues found SIRT1 can regulate SIRT6 expression in liver. Specifically, SIRT1 functions together with FOXO3a and stimulates SIRT6 transcription during fasting. The activation of SIRT6 directly suppresses gene expression of metabolic enzymes involved in triglyceride synthesis and glycolysis in liver. Deficiency of SIRT6 causes increases of glucose utilization, reduced beta oxidation and as a result, fatty liver. More importantly, Kim et al found the expression of SIRT6 is decreased in human fatty liver samples which indicates SIRT6 may play critical roles in liver steatosis in clinical settings. This study landed in Cell Metabolism of this month.
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