Reduced Levels of mRNA Encoding Endoskeletal and Growth-Associated Proteins in Sensory Ganglia in Experimental Diabetes

Abstract

This study investigated changes in levels of mRNAs encoding the three neurofilament (NF) proteins NF-L (low), NF-M (medium), and NF-H (high) and two growth-associated proteins, GAP-43 and T alpha 1 alpha-tubulin, in lumbar dorsal root ganglia of control and streptozocin-induced diabetic rats. After 8 weeks of diabetes the animals were killed, and total RNA was isolated from the L4 and L5 dorsal root ganglia and subjected to Northern blotting, with constant amounts of total RNA loaded onto each lane. A truncated sense RNA for GAP-43 was included as an internal standard during RNA isolation to enable accurate quantification of mRNA levels. The filters were probed sequentially with 32P-labeled cDNAs encoding NF-L, NF-M, NF-H, GAP-43, T alpha 1 alpha-tubulin, and citrate synthase. Hybridizing RNAs were detected by autoradiography and quantified by image analysis. Hybridization signals were normalized to those of the internal standard. In diabetes, NF-L mRNA levels (2.5- and 4-kilobase [kb] transcripts) were decreased by 35 (P = 0.002) and 34% (P < 0.001), respectively, the NF-H mRNA level was decreased by 65% (P < 0.001), but the NF-M mRNA remained unchanged. T alpha 1 alpha-tubulin and GAP-43 mRNA levels were reduced by 56 (P < 0.001) and 30% (P < 0.05), respectively. Levels of citrate synthase mRNA were unchanged. These data indicate a selective defect of expression of growth-associated and endoskeletal proteins in experimentally induced diabetes.