Mukaddes Pala1, Ismail Meral2, Nilgun Pala Acikgoz3, Senay Gorucu Yilmaz4, Semra Karaca Okur5, Seyma Acar6, Yalcin Polat7, Fahri Akbas8. 1. Department of Physiology, Faculty of Medicine, Malatya Turgut Ozal University, Malatya, Turkey. mukaddes.pala@ozal.edu.tr. 2. Department of Physiology, Faculty of Medicine, Bezmialem Vakif University, Istanbul, Turkey. 3. Department of Neurology, Faculty of Medicine, Bezmialem Vakif University, Istanbul, Turkey. 4. Department of Nutrition and Dietetics, Gaziantep University, Gaziantep, Turkey. 5. Mehmet Akif Ersoy Thoracic and Cardiovascular Surgery Training and Research Hospital, Istanbul, Turkey. 6. Family Health Center, Sancaktepe No. 1, Istanbul, Turkey. 7. Department of Pathology, Faculty of Medicine, Biruni University, Istanbul, Turkey. 8. Department of Medical Biology, Faculty of Medicine, Bezmialem Vakif University, Istanbul, Turkey.
Abstract
BACKGROUND: Epileptogenesis is a process that results in neurons firing abnormally, causing seizures. Increasing evidence has shown that miRNAs expressed in the epileptic hippocampus are involved in epileptogenesis. We demonstrated the expression changes of miRNAs that may be effective in epileptogenesis in silico analysis in the kindling model created with Pentylenetetrazole (PTZ). Thus, we aimed to identify the target genes responsible for epileptogenesis. METHODS AND RESULTS: Fifteen male Wistar-albino rats (200-230 g) were randomly divided into two groups control (n = 6) and PTZ (n = 9). The control group received 0.5 ml saline, and the PTZ group (35 mg/kg i.p.) intraperitoneally (i.p.) (11 times, every other day) to induce tonic-clonic seizures. Seizures were observed and scored 30 min after PTZ injection. After the last dose of PTZ (75 mg/kg) administration, the hippocampus tissues of the rats were removed by anesthesia. Analysis of miRNAs was performed with the Affymetrix gene chip miRNA sequence (728 miRNA) and confirmed by the Real-Time Polymerase Chain Reaction (Real-Time PCR) method (29 miRNAs). We evaluated the expression change of the target gene of miRNA, whose expression change was detected using in silico analysis, by q-RT PCR. Eight miRNAs with changes in expression were detected. Of these miRNAs, miR-342-p was downregulated in the PTZ group and was statistically significant (p < 0.005). Ultimately, we determined that the target gene of miR-342-p is a metabotropic glutamate receptor 2 (GRM2) and that GRM2 expression is upregulated. CONCLUSIONS: Downregulation of miR-342-3p in the PTZ kindling model may result in the upregulation of GRM2.
BACKGROUND: Epileptogenesis is a process that results in neurons firing abnormally, causing seizures. Increasing evidence has shown that miRNAs expressed in the epileptic hippocampus are involved in epileptogenesis. We demonstrated the expression changes of miRNAs that may be effective in epileptogenesis in silico analysis in the kindling model created with Pentylenetetrazole (PTZ). Thus, we aimed to identify the target genes responsible for epileptogenesis. METHODS AND RESULTS: Fifteen male Wistar-albino rats (200-230 g) were randomly divided into two groups control (n = 6) and PTZ (n = 9). The control group received 0.5 ml saline, and the PTZ group (35 mg/kg i.p.) intraperitoneally (i.p.) (11 times, every other day) to induce tonic-clonic seizures. Seizures were observed and scored 30 min after PTZ injection. After the last dose of PTZ (75 mg/kg) administration, the hippocampus tissues of the rats were removed by anesthesia. Analysis of miRNAs was performed with the Affymetrix gene chip miRNA sequence (728 miRNA) and confirmed by the Real-Time Polymerase Chain Reaction (Real-Time PCR) method (29 miRNAs). We evaluated the expression change of the target gene of miRNA, whose expression change was detected using in silico analysis, by q-RT PCR. Eight miRNAs with changes in expression were detected. Of these miRNAs, miR-342-p was downregulated in the PTZ group and was statistically significant (p < 0.005). Ultimately, we determined that the target gene of miR-342-p is a metabotropic glutamate receptor 2 (GRM2) and that GRM2 expression is upregulated. CONCLUSIONS: Downregulation of miR-342-3p in the PTZ kindling model may result in the upregulation of GRM2.
Authors: Jyun-You Liou; Elliot H Smith; Lisa M Bateman; Samuel L Bruce; Guy M McKhann; Robert R Goodman; Ronald G Emerson; Catherine A Schevon; L F Abbott Journal: Elife Date: 2020-03-23 Impact factor: 8.140
Authors: Adam Armada-Moreira; Joana I Gomes; Carolina Campos Pina; Oksana K Savchak; Joana Gonçalves-Ribeiro; Nádia Rei; Sara Pinto; Tatiana P Morais; Robertta Silva Martins; Filipa F Ribeiro; Ana M Sebastião; Vincenzo Crunelli; Sandra H Vaz Journal: Front Cell Neurosci Date: 2020-04-24 Impact factor: 5.505