Nei, Chong Pei
(2018)
Changes of GABA receptor and immunoreactivity modification of IP3 & BDNF-TrkB in KA-induced rat epileptogenic hippocampal neuronal culture.
Masters thesis, Universiti Sains Malaysia.
Abstract
INTRODUCTION: Epilepsy seizure is a common condition occurring in
approximately 50 million people worldwide. Epilepsy seizure is a transient occurrence
of sign and or symptoms due to abnormal excessive or hypersynchronous activity in the
brain. Studies showed GABAAR supporting the role of BDNF-TrkB as a neuroprotective
factor in the hippocampus and BDNF-TrkB triggers one of the downstream signalling
IP3R which is involved in epilepsy seizure. However, past studies focus on in vivo and
in vitro organotypic hippocampal slices culture. It is unclear whether the findings of the
current study would extrapolate in vitro dissociated primary rat hippocampal neuron
culture.
OBJECTIVE: To study the changes of γ-Amino butyric acid A receptor
(GABAAR), Brain derived neurotrophic factor (BDNF), Tyrosine kinase receptor B
(TrkB) and Inositol 1, 4, 5 triphosphate receptor (IP3R) in normal and epileptogenic
hippocampal neuron.METHOD: In this study, chemo-convulsant method, kainic acid (KA) induced
seizure applied to (E-18) rat hippocampus neuron. The E-18 rat hippocampus neuronwas cultured based on the optimised method by using poly-L-lysine coated glass
coverslips (Todd et al., 2013). The day in vitro (DIV), 14 culture neurons induced with
0.5μMKA for 30 (KA30), 60 (KA60) and 90 (KA)minutes respectively. The normal and
KA treated neuron underwent cell viability, neurite outgrowth density assessment and
immunocytochemistry assessment respectively.
RESULTS: Finding showed no significant effects of GABAA α1-containing
receptors (GABAA α1 containing R )and BDNF of the KA treatment to the normal
neuron culture. The TrkB receptor was significantly decreased by the treatment (F (3, 8)
= 8.761, p < 0.01). The KA30, KA60 and KA 90 were significant decreased compared to
the control group with no KA treated culture. The IP3R was elevated significantly y the
treatment of KA (F (3, 8) = 6.954, p < 0.05) .
CONCLUSIONS: The KA administration on hippocampal neuron culture
showed an increase expression in the TrkB (BDNF receptor) and IP3R significantly
whereas, the immunoreactivity of BDNF and GABAA α1 containing R showed a
decreased expression that is not statistically significant. Altogether, our study showed
that KA administration on in vitro rat hippocampal neuron induces epileptogenesis
which, results in changes in the GABAA α1 containing R, BDNF, TrkB and IP3R.
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