The effect of sodium thiopental as a GABA mimetic drug in neonatal period on expression of GAD65 and GAD67 genes in hippocampus of newborn and adult male rats

Document Type : Original Article


1 Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran

2 Embryonic and Stem Cell Biology and Biotechnology Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

3 Rayan Center for Neuroscience and Behavior, Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran


Objective(s): Development of the nervous system in human and most animals is continued after the birth. Critical role of this period in generation and specialization of the neuronal circuits is confirmed in numerous studies. Any pharmacological intervention in this period may result in structural, functional or behavioral abnormalities. In this study, sodium thiopental a GABA mimetic drug was administrated to newborn rats and their GAD65 and GAD67 expression in hippocampus was evaluated before and after puberty.
Materials and Methods: Newborn male Wistar rats were received sodium thiopental (35 mg/kg) daily for 11 days (from 4 to 14 days after birth). Expression of GAD65 and GAD67 in their hippocampus was compared with control groups in 15 and 45 days after birth with RT-qPCR method.
Results: Significant down regulation of GAD65 and GAD67 gene expression was observed in treated rats compared with control group in 45 days after birth animals. But no significant difference was shown between experimental and control groups 15 days after birth animals.
Conclusion: The effect of sodium thiopental on GAD65 and GAD67 expression only at adult rats showed a latent period of influence which can be attributed to dosage or intension of sodium thiopental neurotoxicity. Significant down regulation of GAD65 and GAD67 showed unwanted effect of sodium thiopental as GABA mimetic drug in critical period of development.


1.Zachmann M TP, Nyhan WL. The occurrence of gammaaminobutyric acid in human tissues other than brain. J Biol Chem 1966; 241:1355–1358.
2.Otsuka M, Iversen LL, Hall ZW, Kravitz EA. Release of gamma-aminobutyric acid from inhibitory nerves of lobster. Proc Natl Acad Sci U S A 1966; 56:1110-1115.
3.Meldrum B. Pharmacology of GABA. Clin Neuro-pharmacol 1982; 5:293-316.
4.Guidotti A, Corda MG, Wise BC, Vaccarino F, Costa E. GABAergic synapses. Supramolecular organization and biochemical regulation. Neuropharmacol 1983; 22:1471-1479.
5.Ouellet L, de Villers-Sidani E. Trajectory of the main GABAergic interneuron populations from early develop-ment to old age in the rat primary auditory cortex. Front Neuroanat. 2014; 8:40.
6.Bormann J. Electrophysiology of GABAA and GABAB receptor subtypes. Trends  Neurosci1988; 11:112-116.
7.Macdonald RL, Twyman RE, Ryan-Jastrow T, Angelotti TP. Regulation of GABAA receptor channels by anticonvulsant and convulsant drugs and by phosphorylation. Epilepsy Res 1992; 9:265-277.
8.Ruiz A, Campanac E, Scott RS, Rusakov DA, Kullmann DM. Presynaptic GABAA receptors enhance transmission and LTP induction at hippocampal mossy fiber synapses. Nat Neurosci 2010; 13:431-438.
9.Nicoll RA, Eccles JC, Oshima T, Rubia F. Prolongation of hippocampal inhibitory postsynaptic potentials by barbiturates. Nature 1975; 258:625-627.
10.Narahashi T, Arakawa O, Brunner EA, Nakahiro M, Nishio M, Ogata N, et al. Modulation of GABA receptor-channel complex by alcohols and general anesthetics. Adv Biochem Psychopharmacol 1992; 47:325-334.
11.Mhatre M, Ticku MK. Chronic ethanol treatment upregulates the GABA receptor beta subunit expression. Brain Res Mol Brain Res 1994; 23:246-252.
12.Curtis DR, Duggan AW, Felix D, Johnston GAR. Bicuculline and central GABA receptors. Nature 1970; 228:676-677.
13.Engel D, Pahner I, Schulze K, Frahm C, Jarry H, Ahnert-Hilger G, et al. Plasticity of rat central inhibitory synapses through GABA metabolism. J  Physiol 2001; 535:473-482.
14.Peng L, Hertz L, Huang R, Sonnewald U, Petersen SB, Westergaard N, et al. Utilization of Glutamine and of TCA Cycle Constituents as Precursors for Transmitter Glutamate and GABA. Dev Neurosci 1993; 15:367-377.
15.Schousboe A, Westergaard N, Sonnewald U, Petersen SB, Huang R, Peng L, et al. Glutamate and glutamine metabolism and compartmentation in astrocytes. Dev Neurosci 1993; 15:359-366.
16.Erlander MG, Tillakaratne NJ, Feldblum S, Patel N, Tobin AJ. Two genes encode distinct glutamate decarboxylases. Neuron 1991; 7:91-100.
17.Jensen K, Chiu CS, Sokolova I, Lester HA, Mody I. GABA transporter-1 (GAT1)-deficient mice: differential tonic activation of GABAA versus GABAB receptors in the hippocampus. J  Neurophysiol 2003; 90:2690-701.
18.Asada H, Kawamura Y, Maruyama K, Kume H, Ding R, Ji FY, et al. Mice lacking the 65 kDa isoform of glutamic acid decarboxylase (GAD65) maintain normal levels of GAD67 and GABA in their brains but are susceptible to seizures. Biochem Biophys Res Commun 1996; 229:891-895.
19.Kash SF, Johnson RS, Tecott LH, Noebels JL, Mayfield RD, Hanahan D, et al. Epilepsy in mice deficient in the 65-kDa isoform of glutamic acid decarboxylase. Proc Natl Acad Sci U S A 1997; 94:14060-14065.
20.Kash SF, Tecott LH, Hodge C, Baekkeskov S. Increased anxiety and altered responses to anxiolytics in mice deficient in the 65-kDa isoform of glutamic acid decarboxylase. Proc Natl Acad Sci U S A 1999; 96:1698-1703.
21.Heldt SA, Green A, Ressler KJ. Prepulse inhibition deficits in GAD65 knockout mice and the effect of antipsychotic treatment. Neuropsychopharmacology 2004; 29:1610-1619.
22.Wu H, Jin Y, Buddhala C, Osterhaus G, Cohen E, Jin H, et al. Role of glutamate decarboxylase (GAD) isoform, GAD65, in GABA synthesis and transport into synaptic vesicles-Evidence from GAD65-knockout mice studies. Brain Res  2007; 1154:80-83.
23.Asada H, Kawamura Y, Maruyama K, Kume H, Ding RG, Kanbara N, et al. Cleft palate and decreased brain gamma-aminobutyric acid in mice lacking the 67-kDa isoform of glutamic acid decarboxylase. Proc Natl Acad Sci U S A 1997; 94:6496-6499.
24.Hyde TM, Lipska BK, Ali T, Mathew SV, Law AJ, Metitiri OE, et al. Expression of GABA signaling molecules KCC2, NKCC1, and GAD1 in cortical development and schizophrenia. J  Neurosci 2011; 31:11088-11095.
25.Charych EI, Liu F, Moss SJ, Brandon NJ. GABA(A) receptors and their associated proteins: implications in the etiology and treatment of schizophrenia and related disorders. Neuropharmacology 2009; 57:481-495.
26.Akbarian S, Huntsman MM, Kim JJ, Tafazzoli A, Potkin SG, Bunney WE Jr, et al. GABAA receptor subunit gene expression in human prefrontal cortex: comparison of schizophrenics and controls. Cereb Cortex. 1995; 5:550-560.
27.Heckers S, Konradi C. GABAergic mechanisms of hippocampal hyperactivity in schizophrenia. Schizophr Res 2015; 167:4-11.
28.Marenco S, Weinberger DR. The neurodevelopmental hypothesis of schizophrenia: following a trail of evidence from cradle to grave. Dev Psychopathol 2000; 12:501-527.
29.Leinekugel X, Tseeb V, Ben-Ari Y, Bregestovski P. Synaptic GABAA activation induces Ca2+ rise in pyramidal cells and interneurons from rat neonatal hippocampal slices. J Physiol 1995; 487:319-329.
30.LoTurco JJ, Owens DF, Heath MJ, Davis MB, Kriegstein AR. GABA and glutamate depolarize cortical progenitor cells and inhibit DNA synthesis. Neuron 1995; 15:1287-1298.
31.Obrietan K, van den Pol AN. GABA neurotransmission in the hypothalamus: developmental reversal from Ca2+ elevating to depressing. J Neurosci 1995; 15:5065-5077.
32.Ganguly K, Schinder AF, Wong ST, Poo M. GABA itself promotes the developmental switch of neuronal GABAergic responses from excitation to inhibition. Cell 2001; 105:521-532.
33.Ben-Ari Y. Excitatory actions of gaba during development: the nature of the nurture. Nat Rev Neurosci 2002; 3:728-739.
34.Perrot-Sinal TS, Auger AP, McCarthy MM. Excitatory actions of GABA in developing brain are mediated by l-type Ca2+ channels and dependent on age, sex, and brain region. Neuroscience 2003; 116:995-1003.
35.Nuñez JL, McCarthy MM. Evidence for an extended duration of GABA-mediated excitation in the developing male versus female hippocampus. Dev neurobiol 2007; 67:1879-1890.
36.Ikonomidou C, Bosch F, Miksa M, Bittigau P, Vockler J, Dikranian K, et al. Blockade of NMDA receptors and apoptotic neurodegeneration in the developing brain. Science 1999; 283:70-74.
37.Gargiulo S, Greco A, Gramanzini M, Esposito S, Affuso A, Brunetti A, et al. Mice anesthesia, analgesia, and care, Part I: anesthetic considerations in preclinical research. ILAR J 2012; 53:E55-E69.
38.Kaila K. Ionic basis of GABAA receptor channel function in the nervous system. Prog Neurobiol 1994; 42:489-537.
39.Yamada J, Okabe A, Toyoda H, Kilb W, Luhmann HJ, Fukuda A. Cl- uptake promoting depolarizing GABA actions in immature rat neocortical neurones is mediated by NKCC1. J Physiol 2004; 557:829-841.
40.Blaesse P, Airaksinen MS, Rivera C, Kaila K. Cation-chloride cotransporters and neuronal function. Neuron 2009; 61:820-838.
41.Owens DF, Kriegstein AR. Is there more to GABA than synaptic inhibition? Nat Rev Neurosci 2002; 3:715-727.
42.Zhang Z, Sun QQ. The balance between excitation and inhibition and functional sensory processing in the somatosensory cortex. Int Rev Neurobiol 2011; 97:305-333.