Tunicamycin-Induced Renal Injury Model
Objective: To examine the role of CHOP transcription factor in programmed cell death and cellular regeneration in response to endoplasmic reticulum (ER) stress-induced renal injury using a tunicamycin-induced nephrotoxicity model
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Materials1
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Protocol Steps
Animal preparation and genotyping
Obtain mice with defined chop genotypes including wild-type (chop +/+), heterozygous (chop +/-), and knockout (chop -/-) animals
Note: Three genotype groups were compared in this study
View evidence from paper
“mice with defined chop genotypes to a single sublethal intraperitoneal injection of tunicamycin”
Tunicamycin administration
Administer a single sublethal intraperitoneal injection of tunicamycin to induce ER stress
Note: Injection is intraperitoneal and sublethal dose
View evidence from paper
“exposing mice with defined chop genotypes to a single sublethal intraperitoneal injection of tunicamycin”
Clinical observation
Monitor mice for development of severe illness characterized by transient renal insufficiency
Note: Severe illness with transient renal insufficiency was observed as the primary phenotype
View evidence from paper
“resulted in a severe illness characterized by transient renal insufficiency”
Tissue collection and histological analysis
Collect kidney tissue and perform histological examination to assess for acute tubular necrosis and cellular changes
Note: In chop +/+ and chop +/- mice, histological picture similar to human acute tubular necrosis was observed
View evidence from paper
“development of a histological picture similar to the human condition known as acute tubular necrosis”
CHOP expression analysis
Assess early expression of CHOP in the proximal tubules of kidney tissue
Note: Early CHOP expression in proximal tubules was observed in chop +/+ and chop +/- mice
View evidence from paper
“associated with the early expression of CHOP in the proximal tubules”
TUNEL assay for programmed cell death detection
Perform TUNEL staining to identify and quantify programmed cell death (apoptotic) cells in kidney tissue
Note: TUNEL-positive cells serve as a marker for programmed cell death; chop -/- animals exhibited fourfold lower levels compared to wild-type
View evidence from paper
“The proximal tubule epithelium of chop -/- animals exhibited fourfold lower levels of TUNEL-positive cells (a marker for programmed cell death)”
Assessment of cellular regeneration
Evaluate evidence of cellular regeneration in kidney tissue following ER stress-induced injury
Note: chop -/- animals showed significantly less evidence for subsequent regeneration despite severe renal dysfunction
View evidence from paper
“significantly less evidence for subsequent regeneration”