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Identification of differentially expressed genes in dorsal root ganglion in early diabetic rats.

ABSTRACT: To screen and identify differentially expressed genes in the dorsal root ganglion (DRG) in early experimental diabetic rats.Diabetic model rats were induced by single intraperitoneal injection of streptozotocin (STZ). At the second week after STZ injection, the sensory nerve conduction velocities (SNCV) of sciatic nerve were measured as an indicator of neuropathy. The technique of silver-staining mRNA differential display polymerase chain reaction (DD-PCR) was used to detect the levels of differentially expressed genes in rat DRG. The cDNA fragments that displayed differentially were identified by reverse-hybridization, cloned and sequenced subsequently, and then confirmed by Northern blot.The SNCV in the diabetic model group [n = 9, (45.25+/-10.38) m/s] reduced obviously compared with the control group [n = 8, (60.10+/-11.92) m/s] (P < 0.05). Seven distinct cDNA clones, one was up-regulated gene and the others were down-regulated ones, were isolated by silver-staining mRNA differential display method and confirmed by Northern blot. According to the results of sequence alignment with GenBank data, majority of the clones had no significant sequence similarity to previously reported genes except only one that showed high homology to 6-pyruvoyl-tetrahydropterin synthase mRNA (accession No. BC059140), which had not been reported to relate to diabetic neuropathy.These differentially expressed genes in the diabetic DRG may contribute to the pathogenesis of diabetic peripheral neuropathy.

SUBMITTER: Zhu Q

PROVIDER: S-EPMC5552583 | biostudies-literature | 2008 Aug

REPOSITORIES: biostudies-literature

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Identification of differentially expressed genes in dorsal root ganglion in early diabetic rats.

Zhu Qing Q Gu Jin-Hua JH Zhu Hong-Yan HY Xu Ji-Liang JL

Neuroscience bulletin 20080801 4

<h4>Objective</h4>To screen and identify differentially expressed genes in the dorsal root ganglion (DRG) in early experimental diabetic rats.<h4>Methods</h4>Diabetic model rats were induced by single intraperitoneal injection of streptozotocin (STZ). At the second week after STZ injection, the sensory nerve conduction velocities (SNCV) of sciatic nerve were measured as an indicator of neuropathy. The technique of silver-staining mRNA differential display polymerase chain reaction (DD-PCR) was u ...[more]

PMID: 18668150

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Project description:BackgroundDuring the development of the central nervous system (CNS), patterning processes along the dorsoventral (DV) axis of the neural tube generate different neuronal subtypes. As development progresses these neurons are arranged into functional units with varying cytoarchitecture, such as laminae or nuclei for efficient relaying of information. Early in development ventral and dorsal regions are similar in size and structure. Different proliferation rates and cell migration patterns are likely to result in the formation of laminae or nuclei, eventually. However, the underlying molecular mechanisms that establish these different structural arrangements are not well understood.We undertook a differential display polymerase chain reaction (DD-PCR) screen to identify genes with distinct expression patterns between dorsal and ventral regions of the chick midbrain in order to identify genes which regulate the sculpturing of such divergent neuronal organisation. We focused on the DV axis of the early chick midbrain since mesencephalic alar plate and basal plate develop into laminae and nuclei, respectively.ResultsWe identified 53 differentially expressed bands in our initial screen. Twenty-six of these could be assigned to specific genes and we could unambiguously show the differential expression of five of the isolated cDNAs in vivo by in situ mRNA expression analysis. Additionally, we verified differential levels of expression of a selected number of genes by using reverse transcriptase (RT) PCR method with gene-specific primers.One of these genes, QR1, has been previously cloned and we present here a detailed study of its early developmental time course and pattern of expression providing some insights into its possible function. Our phylogenetic analysis of QR1 shows that it is the chick orthologue of Sparc-like 1/Hevin/Mast9 gene in mice, rats, dogs and humans, a protein involved in cell adhesion.ConclusionThis study reveals some possible networks, which might be involved in directing the difference in neuronal specification and cytoarchitecture observed in the brain.

Project description:BACKGROUND: The different sensory modalities temperature, pain, touch and muscle proprioception are carried by somatosensory neurons of the dorsal root ganglia. Study of this system is hampered by the lack of molecular markers for many of these neuronal sub-types. In order to detect genes expressed in sub-populations of somatosensory neurons, gene profiling was carried out on wild-type and TrkA mutant neonatal dorsal root ganglia (DRG) using SAGE (serial analysis of gene expression) methodology. Thermo-nociceptors constitute up to 80 % of the neurons in the DRG. In TrkA mutant DRGs, the nociceptor sub-class of sensory neurons is lost due to absence of nerve growth factor survival signaling through its receptor TrkA. Thus, comparison of wild-type and TrkA mutants allows the identification of transcripts preferentially expressed in the nociceptor or mechano-proprioceptor subclasses, respectively. RESULTS: Our comparison revealed 240 genes differentially expressed between the two tissues (P < 0.01). Some of these genes, CGRP, Scn10a are known markers of sensory neuron sub-types. Several potential markers of sub-populations, Dok4, Crip2 and Grik1/GluR5 were further analyzed by quantitative RT-PCR and double labeling with TrkA,-B,-C, c-ret, parvalbumin and isolectin B4, known markers of DRG neuron sub-types. Expression of Grik1/GluR5 was restricted to the isolectin B4+ nociceptive population, while Dok4 and Crip2 had broader expression profiles. Crip2 expression was however excluded from the proprioceptor sub-population. CONCLUSION: We have identified and characterized the detailed expression patterns of three genes in the developing DRG, placing them in the context of the known major neuronal sub-types defined by molecular markers. Further analysis of differentially expressed genes in this tissue promises to extend our knowledge of the molecular diversity of different cell types and forms the basis for understanding their particular functional specificities.

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Identification of differentially expressed genes in dorsal root ganglion in early diabetic rats.

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Identification of differentially expressed genes in dorsal root ganglion in early diabetic rats.

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