Regulation of MAP2, GFAP, and calcium in the CA3 Region Following Kainic Acid Exposure to organotypic hippocampal slice culture [version 3; peer review: 2 approved]
Background Neurodegeneration due to neurotoxicity is one of the phenomena in temporal lobe epilepsy. Experimentally, hippocampal excitotoxicity process can occur due to kainic acid exposure, especially in the CA3 area. Neuronal death, astrocyte reactivity and increased calcium also occur in hippocam...
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2025-01-01
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| author | Aulanni'am Aulanni'am Kusworini Handono Ettie Rukmigarsari Hidayat Sujuti Machlusil Husna |
| author_facet | Aulanni'am Aulanni'am Kusworini Handono Ettie Rukmigarsari Hidayat Sujuti Machlusil Husna |
| author_sort | Aulanni'am Aulanni'am |
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| description | Background Neurodegeneration due to neurotoxicity is one of the phenomena in temporal lobe epilepsy. Experimentally, hippocampal excitotoxicity process can occur due to kainic acid exposure, especially in the CA3 area. Neuronal death, astrocyte reactivity and increased calcium also occur in hippocampal excitotoxicity, but few studies have investigated immediate effect after kainic acid exposure. The organotypic hippocampal slice culture (OHSC) is a useful model for studying the neurodegeneration process, but there are still many protocol differences. In this study, minor modifications were made in the OHSC protocol. Methods OHSC was obtained from three healthy wild type Wistar rats aged P10. Healthy culture slices were obtained and lasted up to 10 days in vitro (DIV 10). Bath application of kainic acid for 48 hours in DIV 10 followed by observation of its initial effects on neurons, astrocytes, and calcium via the expression of MAP2, GFAP, and intracellular calcium imaging, subsequently. Results After 48 h of kainic acid administration, there was a significant increase in intracellular calcium intensity (p = 0.006 < α), accompanied by a significant decrease in MAP2 (p = 0.003 < α) and GFAP (p = 0.010 < α) expression. Conclusion These findings suggest early neuronal and astrocyte damage at the initial onset of hippocampal injury. This implies that astrocyte damage occurs early before an increase in GFAP that characterizes reactive astrogliosis found in other studies. Damage to neurons and astrocytes may be associated with increased intracellular calcium. It is necessary to develop further research regarding regulation of calcium, MAP2, and GFAP at a spatial time after exposure to kainic acid and strategies to reduce damage caused by excitotoxicity. |
| format | Article |
| id | doaj-art-7a80dc847ff44a11aa85185732f90d3f |
| institution | Kabale University |
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| language | English |
| publishDate | 2025-01-01 |
| publisher | F1000 Research Ltd |
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| spelling | doaj-art-7a80dc847ff44a11aa85185732f90d3f2025-01-15T01:00:01ZengF1000 Research LtdF1000Research2046-14022025-01-0112176262Regulation of MAP2, GFAP, and calcium in the CA3 Region Following Kainic Acid Exposure to organotypic hippocampal slice culture [version 3; peer review: 2 approved]Aulanni'am Aulanni'am0https://orcid.org/0000-0002-8715-4592Kusworini Handono1Ettie Rukmigarsari2Hidayat Sujuti3Machlusil Husna4https://orcid.org/0000-0001-5373-505XDepartment of Chemistry, Faculty of Sciences Universitas Brawijaya, Malang, East Java, IndonesiaDepartment of Clinical Pathology, Faculty of Medicine Universitas Brawijaya / dr. Saiful Anwar General Hospital, Malang, East Java, IndonesiaFaculty of Teaching and Education Sciences, Islamic University of Malang, Malang, East Java, IndonesiaDepartment of Ophthalmology, Faculty of Medicine Universitas Brawijaya / dr. Saiful Anwar General Hospital, Malang, East Java, IndonesiaDoctoral Program of Medical Science, Faculty of Medicine, Universitas Brawijaya, Malang, East Java, IndonesiaBackground Neurodegeneration due to neurotoxicity is one of the phenomena in temporal lobe epilepsy. Experimentally, hippocampal excitotoxicity process can occur due to kainic acid exposure, especially in the CA3 area. Neuronal death, astrocyte reactivity and increased calcium also occur in hippocampal excitotoxicity, but few studies have investigated immediate effect after kainic acid exposure. The organotypic hippocampal slice culture (OHSC) is a useful model for studying the neurodegeneration process, but there are still many protocol differences. In this study, minor modifications were made in the OHSC protocol. Methods OHSC was obtained from three healthy wild type Wistar rats aged P10. Healthy culture slices were obtained and lasted up to 10 days in vitro (DIV 10). Bath application of kainic acid for 48 hours in DIV 10 followed by observation of its initial effects on neurons, astrocytes, and calcium via the expression of MAP2, GFAP, and intracellular calcium imaging, subsequently. Results After 48 h of kainic acid administration, there was a significant increase in intracellular calcium intensity (p = 0.006 < α), accompanied by a significant decrease in MAP2 (p = 0.003 < α) and GFAP (p = 0.010 < α) expression. Conclusion These findings suggest early neuronal and astrocyte damage at the initial onset of hippocampal injury. This implies that astrocyte damage occurs early before an increase in GFAP that characterizes reactive astrogliosis found in other studies. Damage to neurons and astrocytes may be associated with increased intracellular calcium. It is necessary to develop further research regarding regulation of calcium, MAP2, and GFAP at a spatial time after exposure to kainic acid and strategies to reduce damage caused by excitotoxicity.https://f1000research.com/articles/12-47/v3organotypic hippocampal slice culture MAP2 GFAP intracellular calcium excitotoxicity kainic acideng |
| spellingShingle | Aulanni'am Aulanni'am Kusworini Handono Ettie Rukmigarsari Hidayat Sujuti Machlusil Husna Regulation of MAP2, GFAP, and calcium in the CA3 Region Following Kainic Acid Exposure to organotypic hippocampal slice culture [version 3; peer review: 2 approved] F1000Research organotypic hippocampal slice culture MAP2 GFAP intracellular calcium excitotoxicity kainic acid eng |
| title | Regulation of MAP2, GFAP, and calcium in the CA3 Region Following Kainic Acid Exposure to organotypic hippocampal slice culture [version 3; peer review: 2 approved] |
| title_full | Regulation of MAP2, GFAP, and calcium in the CA3 Region Following Kainic Acid Exposure to organotypic hippocampal slice culture [version 3; peer review: 2 approved] |
| title_fullStr | Regulation of MAP2, GFAP, and calcium in the CA3 Region Following Kainic Acid Exposure to organotypic hippocampal slice culture [version 3; peer review: 2 approved] |
| title_full_unstemmed | Regulation of MAP2, GFAP, and calcium in the CA3 Region Following Kainic Acid Exposure to organotypic hippocampal slice culture [version 3; peer review: 2 approved] |
| title_short | Regulation of MAP2, GFAP, and calcium in the CA3 Region Following Kainic Acid Exposure to organotypic hippocampal slice culture [version 3; peer review: 2 approved] |
| title_sort | regulation of map2 gfap and calcium in the ca3 region following kainic acid exposure to organotypic hippocampal slice culture version 3 peer review 2 approved |
| topic | organotypic hippocampal slice culture MAP2 GFAP intracellular calcium excitotoxicity kainic acid eng |
| url | https://f1000research.com/articles/12-47/v3 |
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