Suppression of c-Fos Protein and mRNA Expression in Pentylenetetrazole-Induced Kindled Mouse Brain by Isoxylitones Shabana Usman Simjee & Farzana Shaheen & M. Iqbal Choudhary & Atta-ur Rahman & Siddiqua Jamall & Syed Uzair Ali Shah & Noureen Khan & Nurul Kabir & Nadeem Ashraf Received: 4 August 2011 /Accepted: 7 November 2011 /Published online: 15 December 2011 # Springer Science+Business Media, LLC 2011 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 combina- tion 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 isoxyli- tone (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. Keywords Epilepsy . Kindling . Isoxylitones . Delphinium denudatum . c-fos mRNA . c-Fos protein 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 S. U. Simjee : F. Shaheen : M. I. Choudhary : A.-u. Rahman : S. U. A. Shah : N. Khan : N. Ashraf H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences (ICCBS), University of Karachi, Karachi 75270, Pakistan S. U. Simjee (*) : M. I. Choudhary : A.-u. Rahman : N. Kabir Dr. Panjwani Center for Molecular Medicine and Drug Research, ICCBS, University of Karachi, Karachi 75270, Pakistan e-mail: shabana.simjee@iccs.edu S. U. Simjee e-mail: sh01us@hotmail.com S. Jamall Department of Biochemistry, University of Karachi, Karachi 75270, Pakistan J Mol Neurosci (2012) 47:559–570 DOI 10.1007/s12031-011-9674-4