GAD Gene Expression Research Articles

Glutamic acid decarboxylase gene expression in the dopamine-denervated striatum: effects of intrastriatal fetal nigral transplants or chronic apomorphine treatment.

Glutamic acid decarboxylase mRNA expression was studied in the striatum of rats subjected to a unilateral 6-hydroxydopamine lesion of the nigrostriatal dopamine projection, followed by either intrastriatal transplants of fetal nigral neurons or chronic apomorphine treatment. For in situ hybridization histochemistry, a 35S-labelled cRNA probe selective for the mRNA encoding the 67-kDa isoform of the enzyme (GAD67) was used. The results show that fetal dopaminergic grafts in the dopamine-denervated striatum restore an inhibitory control on GAD67 gene expression in the host neurons and also counteract the up-regulation of GAD67 mRNA levels induced by chronic apomorphine treatment.

Developmental expression of GABAA receptor subunit and GAD genes in mouse somatosensory barrel cortex

In situ hybridization histochemistry with radioactive cRNA probes was used to study patterns of gene expression for alpha1, alpha2, alpha4, alpha5, beta1, beta2, and gamma2 subunit mRNAs of typeAgamma aminobutyric acid (GABA(A)) receptors and for 67-kDa glutamic acid decarboxylase (GAD67) mRNA in mouse barrel cortex during the period (postnatal days 1-12; P1-P12) when thalamocortical innervation of layer IV barrels is occurring. The alpha1, beta2, and gamma2 subunit mRNAs increased substantially with age, especially in layers V and VI, and throughout the period studied, invariably had the same laminar-specific patterns of expression. All three mRNAs were highly expressed in the dense cortical plate at P1. In layer IV after differentiation of barrels, they were expressed in cells of both barrel walls and hollows but especially in the walls. The alpha2, alpha4, alpha5, and beta1 subunit mRNAs were expressed at lower levels and had different laminar patterns of distribution; alpha2 and alpha4 showed switches between layers over time; alpha5 was invariably associated with the subplate or its derivative, beta1 with layer IV. Levels of alpha2 mRNA did not change over time; alpha4 and beta1 mRNAs increased and alpha5 decreased. GAD67 mRNA was highest in layer I at P1 and progressively increased in other layers. These results suggest that postnatal development of GABA(A) receptors is mainly directed at the production of receptors assembled from alpha1, beta2, and gamma2 subunits, with beta1 contributing in layer IV. Other subunits may be associated with receptors involved in trophic actions of GABA during development and may give GABA(A) receptor-mediated responses in the developing cortex their particular physiological profile.

Changes in brain GAD mRNA in neuroleptic-treated rats

The present study examined the effect of a subchronic systemic administration of the glutamate metabotropic mGluR5 receptor antagonist MPEP on l-DOPA-induced dyskinesias and striatal gene expression in adult rats with a unilateral 6-OHDA lesion of dopamine neurons. The daily systemic administration of l-DOPA for 2 weeks induced a gradual increase in limb dyskinesia and axial dystonia. The subchronic systemic co-administration of MPEP reduced the severity of limb dyskinesia and axial dystonia over the whole duration of l-DOPA treatment. Subchronic l-DOPA administration was paralleled by a significant increase in mRNA levels of the two isoforms of the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD67 and GAD65) and preprodynorphin (PPD). Single cell analysis on emulsion radioautographs indicated that l-DOPA-induced increases in GAD67 occurred predominantly in preproenkephalin-unlabeled striatonigral and, to a lesser extent, in preproenkephalin-labeled striatopallidal neurons. MPEP completely reversed the effects of l-DOPA on GAD67 and reduced the increases in GAD65 and PPD mRNA levels in striatonigral neurons. MPEP also reversed the small l-DOPA-induced increase in GAD67 mRNA levels in striatopallidal neurons. Altogether, the findings support the idea that the relative efficacy of mGluR5 receptor antagonists to oppose l-DOPA-induced abnormal involuntary movements involves an ability to oppose increases in GAD gene expression and GABA-mediated signaling in striatonigral and striatopallidal neurons. The results also confirm the potential usefulness of antagonists of mGluR5 receptors as adjuncts in the treatment of l-DOPA-induced dyskinesia in patients with Parkinson's disease.

GABAergic modulation of striatal peptide expression in rats and the alterations induced by dopamine antagonist treatment

GABAergic modulation of enkephalin, substance P and glutamic acid decarboxylase (GAD67) gene expression and the alterations induced by dopamine receptor blockade were studied in the rat striatum. Following subchronic treatment with the GABA-A agonist muscimol, the GABA-B agonist baclofen or the GABA transaminase inhibitor γ-vinyl GABA there were no significant changes in striatal peptide and GAD67 gene expression. Following repeated administration of the D-2 antagonists, eticlopride and haloperidol, there was an increase in enkephalin and GAD67 mRNA levels and parallel decrease in that of substance P. These were unaffected by co-administration of γ-vinyl GABA. The D-1 antagonist, SCH 23390 administered alone or together with γ-vinyl GABA did not alter peptide or GAD67 mRNA levels. It seems that pharmacological stimulation of GABA receptors has little effect on enkephalin, substance P or GAD67 mRNA expression in striatal output neurons.

Glutamic acid decarboxylase gene expression in thalamic reticular neurons transplanted as a cell suspension in the adult thalamus

The goal of the present study was to determine whether alterations in neuronal morphology and connections in thalamic grafts were accompanied by changes in the expression of mRNA encoding glutamic acid decarboxylase (GAD), the key enzyme in the synthesis of GABA, the normal neurotransmitter of neurons of the thalamic reticular nucleus. Cell suspensions of rat fetal tissue containing both thalamic reticular nucleus and ventrobasal primordia were transplanted into the excitotoxically lesioned somatosensory thalamus of adult rats. Levels of messenger RNA (mRNA) encoding GAD (Mr 67,000; GAD67) were measured 7 days to 4 months following transplantation via quantitative in situ hybridization with 35S-radiolabeled antisense RNAs. Expression of GAD67 mRNA in the thalamic reticular nucleus was analyzed in parallel in rat pups between 0 and 30 days postnatally, and in adult animals. As already observed with immunohistochemistry, transplanted neurons of the thalamic reticular nucleus did not group in specific clusters but rather mingled with unlabeled (putatively ventrobasal) neurons. Levels of labelling for GAD67 mRNA per neuron increased over time and reached adult levels during the third week post-grafting, i.e. 2 weeks after the theoretical birthdate of the neurons (grafted at embryonic days 15–16). Similar values were observed and a plateau was reached at similar time points during normal ontogeny. The results suggest that, in contrast to morphology and size of the neuronal cell bodies, gene expression of GAD67 develops normally despite the ectopic location of neurons of the thalamic reticular nucleus in the somatosensory thalamus, the abnormal connectivity and the lack of segregation from non-GABAergic neurons. Therefore, dissociation and transplantation of the tissue, resulting in disruption of the extracellular matrix and alteration of cellular interactions, differentially affects various aspects of neuronal maturation. Because neuronal activity in the graft is similar to that in the adult thalamic reticular nucleus, the results further suggest that, as shown in other experimental systems, GAD67 mRNA levels in the graft are regulated in relation to the functional activity of GABAergic neurons.

Quantitative in situ hybridization analysis of glutamic acid decarboxylase messenger RNA in developing rat cerebellum

The appearance and relative amounts of GAD mRNA in rat cerebellar neurons during postnatal development was studied by in situ hybridization. GAD mRNA content within all GABAergic neurons increased during the first month of postnatal development, but the degree and time course of the increase varied among different neuronal types. In newborn rats, GAD mRNA was present only in the prenatally-formed Purkinje and Golgi cells. GAD mRNA in Golgi cells had reached adult levels by postnatal day 14, while GAD mRNA levels in Purkinje cells reached adult levels one week later. Most basket cells expressed GAD mRNA by postnatal day 14, and final levels were attained one week later. Stellate cells in the bottom two-thirds of the molecular layer attained their final GAD mRNA content by postnatal day 21 whereas stellate cells in close proximity to the pial surface were not yet mature at this age. No GAD mRNA was detected within the external granular layer at any time during development. In adult rat, approximately 40% of cerebellar GAD mRNA was contained within the Purkinje cell population, 38% within the stellate cells, 17% within the basket cells, and only 5% within the Golgi cells. Increases in GAD mRNA within GABAergic neurons during cerebellar development correlated with the timing of neuronal maturation and synaptogenesis in these cell populations, suggesting that synaptic activity affects GAD gene expression in developing cerebellum.