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Adult and embryonic GAD transcripts are spatiotemporally regulated during postnatal development in the rat brain.

Popp A, Urbach A, Witte OW, Frahm C - PLoS ONE (2009)

Bottom Line: Radioactive in situ hybridizations confirmed the occurrence of embryonic GAD67 transcripts in the olfactory bulb and furthermore detected their localization mainly in the subventricular zone and the rostral migratory stream.Embryonic GAD67 transcripts can hardly be detected in the adult brain, except for specific regions associated with neurogenesis and high synaptic plasticity.Therefore a functional role in processes like proliferation, migration or synaptogenesis is suggested.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Friedrich-Schiller-University, Jena, Germany.

ABSTRACT

Background: GABA (gamma-aminobutyric acid), the main inhibitory neurotransmitter in the brain, is synthesized by glutamic acid decarboxylase (GAD). GAD exists in two adult isoforms, GAD65 and GAD67. During embryonic brain development at least two additional transcripts exist, I-80 and I-86, which are distinguished by insertions of 80 or 86 bp into GAD67 mRNA, respectively. Though it was described that embryonic GAD67 transcripts are not detectable during adulthood there are evidences suggesting re-expression under certain pathological conditions in the adult brain. In the present study we systematically analyzed for the first time the spatiotemporal distribution of different GADs with emphasis on embryonic GAD67 mRNAs in the postnatal brain using highly sensitive methods.

Methodology/principal findings: QPCR was used to precisely investigate the postnatal expression level of GAD related mRNAs in cortex, hippocampus, cerebellum, and olfactory bulb of rats from P1 throughout adulthood. Within the first three postnatal weeks the expression of both GAD65 and GAD67 mRNAs reached adult levels in hippocampus, cortex, and cerebellum. The olfactory bulb showed by far the highest expression of GAD65 as well as GAD67 transcripts. Embryonic GAD67 splice variants were still detectable at birth. They continuously declined to barely detectable levels during postnatal development in all investigated regions with exception of a comparatively high expression in the olfactory bulb. Radioactive in situ hybridizations confirmed the occurrence of embryonic GAD67 transcripts in the olfactory bulb and furthermore detected their localization mainly in the subventricular zone and the rostral migratory stream.

Conclusions/significance: Embryonic GAD67 transcripts can hardly be detected in the adult brain, except for specific regions associated with neurogenesis and high synaptic plasticity. Therefore a functional role in processes like proliferation, migration or synaptogenesis is suggested.

Show MeSH
Expression of embryonic splice variants in rat cortex (Ctx), hippocampus (HC), cerebellum (Cbl), and olfactory bulb (OB) at different postnatal ages.Probes were examined for embryonic GAD67 transcript expression by qPCR. Embryonic transcripts were jointly (EGAD) and separately (I-80 and I-86) amplified. Numbers of transcripts were calculated per 1000 transcripts of GAPDH. Data represent mean±SEM of five individual animals. *P≤0.05, the comparison of regions at each analyzed postnatal day (left table) and the comparison of ages within each region (right table) as analysed by one-way ANOVA followed by Tukey's post hoc test.
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pone-0004371-g003: Expression of embryonic splice variants in rat cortex (Ctx), hippocampus (HC), cerebellum (Cbl), and olfactory bulb (OB) at different postnatal ages.Probes were examined for embryonic GAD67 transcript expression by qPCR. Embryonic transcripts were jointly (EGAD) and separately (I-80 and I-86) amplified. Numbers of transcripts were calculated per 1000 transcripts of GAPDH. Data represent mean±SEM of five individual animals. *P≤0.05, the comparison of regions at each analyzed postnatal day (left table) and the comparison of ages within each region (right table) as analysed by one-way ANOVA followed by Tukey's post hoc test.

Mentions: Embryonic GAD67 splice variants were detectable from P1 on, but the transcript level continuously declined until adulthood (Fig. 3). Transcripts were barely detectable in cortical and hippocampal regions from P7 on whereas the reduction was decelerated in cerebellum and OB (Fig. 3). On P7 the expression of EGAD in the cerebellum was significantly higher compared to cortex and hippocampus, whereas the three regions displayed the same expression level on P21. The OB showed by far the highest transcript level at all investigated ages (Fig. 3). Discrimination of EGAD revealed about eight times more transcripts of I-86 than I-80 at all investigated ages and in all regions indicating that embryonic transcripts are dominated by I-86 (Fig. 3). The course of expression and the relative distribution of both transcripts across the evaluated regions were similar and resembled that of EGAD.


Adult and embryonic GAD transcripts are spatiotemporally regulated during postnatal development in the rat brain.

Popp A, Urbach A, Witte OW, Frahm C - PLoS ONE (2009)

Expression of embryonic splice variants in rat cortex (Ctx), hippocampus (HC), cerebellum (Cbl), and olfactory bulb (OB) at different postnatal ages.Probes were examined for embryonic GAD67 transcript expression by qPCR. Embryonic transcripts were jointly (EGAD) and separately (I-80 and I-86) amplified. Numbers of transcripts were calculated per 1000 transcripts of GAPDH. Data represent mean±SEM of five individual animals. *P≤0.05, the comparison of regions at each analyzed postnatal day (left table) and the comparison of ages within each region (right table) as analysed by one-way ANOVA followed by Tukey's post hoc test.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2629816&req=5

pone-0004371-g003: Expression of embryonic splice variants in rat cortex (Ctx), hippocampus (HC), cerebellum (Cbl), and olfactory bulb (OB) at different postnatal ages.Probes were examined for embryonic GAD67 transcript expression by qPCR. Embryonic transcripts were jointly (EGAD) and separately (I-80 and I-86) amplified. Numbers of transcripts were calculated per 1000 transcripts of GAPDH. Data represent mean±SEM of five individual animals. *P≤0.05, the comparison of regions at each analyzed postnatal day (left table) and the comparison of ages within each region (right table) as analysed by one-way ANOVA followed by Tukey's post hoc test.
Mentions: Embryonic GAD67 splice variants were detectable from P1 on, but the transcript level continuously declined until adulthood (Fig. 3). Transcripts were barely detectable in cortical and hippocampal regions from P7 on whereas the reduction was decelerated in cerebellum and OB (Fig. 3). On P7 the expression of EGAD in the cerebellum was significantly higher compared to cortex and hippocampus, whereas the three regions displayed the same expression level on P21. The OB showed by far the highest transcript level at all investigated ages (Fig. 3). Discrimination of EGAD revealed about eight times more transcripts of I-86 than I-80 at all investigated ages and in all regions indicating that embryonic transcripts are dominated by I-86 (Fig. 3). The course of expression and the relative distribution of both transcripts across the evaluated regions were similar and resembled that of EGAD.

Bottom Line: Radioactive in situ hybridizations confirmed the occurrence of embryonic GAD67 transcripts in the olfactory bulb and furthermore detected their localization mainly in the subventricular zone and the rostral migratory stream.Embryonic GAD67 transcripts can hardly be detected in the adult brain, except for specific regions associated with neurogenesis and high synaptic plasticity.Therefore a functional role in processes like proliferation, migration or synaptogenesis is suggested.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Friedrich-Schiller-University, Jena, Germany.

ABSTRACT

Background: GABA (gamma-aminobutyric acid), the main inhibitory neurotransmitter in the brain, is synthesized by glutamic acid decarboxylase (GAD). GAD exists in two adult isoforms, GAD65 and GAD67. During embryonic brain development at least two additional transcripts exist, I-80 and I-86, which are distinguished by insertions of 80 or 86 bp into GAD67 mRNA, respectively. Though it was described that embryonic GAD67 transcripts are not detectable during adulthood there are evidences suggesting re-expression under certain pathological conditions in the adult brain. In the present study we systematically analyzed for the first time the spatiotemporal distribution of different GADs with emphasis on embryonic GAD67 mRNAs in the postnatal brain using highly sensitive methods.

Methodology/principal findings: QPCR was used to precisely investigate the postnatal expression level of GAD related mRNAs in cortex, hippocampus, cerebellum, and olfactory bulb of rats from P1 throughout adulthood. Within the first three postnatal weeks the expression of both GAD65 and GAD67 mRNAs reached adult levels in hippocampus, cortex, and cerebellum. The olfactory bulb showed by far the highest expression of GAD65 as well as GAD67 transcripts. Embryonic GAD67 splice variants were still detectable at birth. They continuously declined to barely detectable levels during postnatal development in all investigated regions with exception of a comparatively high expression in the olfactory bulb. Radioactive in situ hybridizations confirmed the occurrence of embryonic GAD67 transcripts in the olfactory bulb and furthermore detected their localization mainly in the subventricular zone and the rostral migratory stream.

Conclusions/significance: Embryonic GAD67 transcripts can hardly be detected in the adult brain, except for specific regions associated with neurogenesis and high synaptic plasticity. Therefore a functional role in processes like proliferation, migration or synaptogenesis is suggested.

Show MeSH