Figure 1725

Schematic representation of differential display. In a complex organ like the kidney, ischemic renal injury triggers altered expression of various cell factors and vascular components. Depending on the severity of the insult, expression of these genes can vary in individual cells, leading to their death, survival, or proliferation. A better understanding of the various factors and the signal transduction pathways transduced by them that contribute to cell death can lead to development of therapeutic strategies to interfere with the process of cell death. Similarly, identification of factors that are involved in initiating cell migration, dedifferentiation, and proliferation may lead to therapy aimed at accelerating the regeneration program. To identify the various factors involved in cell injury and regeneration, powerful methods for identification and cloning of differentially expressed genes are critical. One such method that has been used extensively by several laboratories is the differential display polymerase chain reaction (DD-PCR).

In this schematic, mRNA is derived from kidneys of sham-operated (controls) and ischemia-injured rats, some pretreated with insulin-like growth factor (IGF-I). The mRNAs are reverse transcribed using an anchored deoxy thymidine-oligonucleotide (oligo-dT) primer (Example: dT[12]-MX, where M represent G, A, or C, and X represents one of the four nucleotides). An anchored primer limits the reverse transcription to a subset of mRNAs. The first strand cDNA is then PCR amplified using an arbitrary 10 nucleotide-oligomer primer and the anchored primer. The PCR reaction is performed in the presence of radioactive or fluorescence-labeled nucleotides, so that the amplified fragments can be displayed on a sequencing gel. Bands of interest can be excised from the gel and used for further characterization. ARF—acute renal failure.


Sham +IGF-1

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