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Suppression of c-Fos Protein and mRNA Expression in Pentylenetetrazole-Induced Kindled Mouse Brain by Isoxylitones

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Abstract

An early immediate gene c-fos has been proposed as the gene responsible for turning on molecular events that might underlie the long-term neural changes occurring during kindling. We have evaluated the effects of novel anticonvulsant isomeric compounds isoxylitones [(E/Z)-2-propanone-1,3,5,5-trimethyl-2-cyclohexen-1-ylidine] on the c-Fos protein and mRNA expression in the brain samples of kindled mice and compared it with the normal and untreated kindled groups. Kindling was induced in male NMRI mice by repeated administration of sub-convulsive dose (50 mg/kg) of pentylenetetrazole (PTZ) until a seizure score of 4–5 was achieved. The c-Fos expression was quantified by combination of immunohistochemistry and RT-PCR protocols. Both the immunohistochemical and RT-PCR analysis revealed a marked increase in the expression of c-fos mRNA and protein in the brain regions tested in case of PTZ-kindled control group compared to normal control. In contrast, the isoxylitone (30 mg/kg)-treated group demonstrated significant reduction of c-Fos expression compared to PTZ-kindled control animals. However, low expression of c-fos mRNA was only detected in the thalamus of the isoxylitone-treated brain samples. Based on these observations, we suggest that isoxylitones may have the capacity to control the seizure pattern by mechanism such as the suppression of c-Fos protein and mRNA levels in different regions of the brain. Further investigations to explore the mechanism of action of these compounds are under process.

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Abbreviations

BLA:

Basolateral amygdala

CeL:

Lateral division of the central nucleus of amygdala

CeM:

Medial division of the central nucleus of amygdala

DLG:

Dorsal lateral geniculate nucleus of thalamus

CMT:

Centromedial nucleus of thalamus

PVT:

Paraventricular nucleus of thalamus

IEGs:

Immediate early genes

IDV:

Integrated density value

PBS:

Phosphate buffer saline

PTZ:

Pentylenetetrazole

RT-PCR:

Reverse transcriptase polymerase chain reaction

ROI:

Region of interest

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GABA:

Gamma aminobutyric acid

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Acknowledgment

This work was supported by grant 20-855/RND/07/540 from the Higher Education Commission, Pakistan, and a grant from the Organization for the Prohibition of Chemical Weapons (OPCW; S/228/2000).

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Authors and Affiliations

  1. H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences (ICCBS), University of Karachi, Karachi, 75270, Pakistan

    Shabana Usman Simjee, Farzana Shaheen, M. Iqbal Choudhary, Atta-ur Rahman, Syed Uzair Ali Shah, Noureen Khan & Nadeem Ashraf

  2. Dr. Panjwani Center for Molecular Medicine and Drug Research, ICCBS, University of Karachi, Karachi, 75270, Pakistan

    Shabana Usman Simjee, M. Iqbal Choudhary, Atta-ur Rahman & Nurul Kabir

  3. Department of Biochemistry, University of Karachi, Karachi, 75270, Pakistan

    Siddiqua Jamall

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  1. Shabana Usman Simjee
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Simjee, S.U., Shaheen, F., Choudhary, M.I. et al. Suppression of c-Fos Protein and mRNA Expression in Pentylenetetrazole-Induced Kindled Mouse Brain by Isoxylitones. J Mol Neurosci 47, 559–570 (2012). https://doi.org/10.1007/s12031-011-9674-4

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