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Partial deafferentation of the developing rat spinal cord delays the spontaneous repression of GAD67 mRNAs in spinal cells.

Early and ubiquitous detection of GABA in the rat spinal cord before the occurrence of synaptogenesis has led to the concept of a neurotrophic role of GABA, in addition to a promoting effect on neurite extension and neurodevelopment. The aim of this study was to further establish, in vivo, evidence for a link between the maturation of spinal cord innervation and the regulation of several isoforms of the synthetic enzymes of GABA, the glutamic acid decarboxylases GAD65, GAD67, and EP10, the embryonic truncated form of GAD67. Neonatal capsaicin treatment was used to induce a specific loss of afferent fibers (unmyelinated C fibers, thin myelinated fibers A delta) to the dorsal horn. The regulation of various GAD mRNAs was investigated using sensitive techniques such as RT-PCR and in situ hybridization. The sensitivity of the methods was further enhanced by the use of a gaseous detector (beta-imager) to quantitate the mRNAs species. After neonatal capsaicin treatment, higher levels of GAD67 mRNA were detected transiently during the postnatal development of the rat spinal cord. A maximum two-fold increase of GAD67 mRNA was found on the day following the capsaicin injection and reached control values within 3 weeks. In contrast, GAD65 mRNA levels remained low and were unaffected by the treatment, and EP10 was not detected. In addition, we have found a similar upregulation, with the same time course, of the cytoskeletal protein beta-actin. The capsaicin-induction of mRNA synthesis was, however, two-fold greater for beta-actin than for GAD67. Moreover, since this upregulation of GAD67 mRNA coincides with the sprouting of unaffected afferent fibers and of 5HT axons, one can hypothesize that GAD67 participates in the structural plasticity occurring in reaction to the capsaicin-induced partial deafferentation.