Yazar "Bolat, Busenur" seçeneğine göre listele

Listeleniyor 1 - 2 / 2
  • Küçük Resim
    Öğe
    A proteomic analysis for profiling NeuroD2 related changes in N2A neuroblastoma cell line
    (Inönü University Faculty of Medicine, 2023) Beker, Merve; Bolat, Busenur; Aslan, Feyza Şule; Özbay, Elif; Kaya, Sare Burcu
    Aim: NeuroD2 transcription factor is a key regulator of neurogenin-NeuroD signaling network and induces neuronal development, differentiation, neurogenesis and calcium de- pendent signaling. NeuroD2 regulates expression of survival and plasticity related proteins in neurons. Surprisingly, inhibition of NeuroD2 causes an increase in apoptotic cell death. Even though previous studies found out important data about NeuroD2 function, molec- ular interactions of NeuroD2 behind all of these impacts remains elusive. For this reason, it was aimed to shed light on the proteome profile of NeuroD2 based changes in the N2A cell line. Materials and Methods: NeuroD2 over-expression and NeuroD2 inhibition groups were constructed via lentiviral vectors. Mouse N2A cell line was transfected with the given vectors and incubated for 6 hours. After incubation samples were prepared for proteomic analyses with Filter Aided Sample Preparation (FASP) protocol and LC-MS/MS analysis was carried out. Results: Under conditions of overexpression and inhibition, detected proteins were fil- tered according to significant cut off values. The filtered proteins were further investigated to exhibit a coherent expression in each situation. Eventually, increased NeuroD2 activity was accompanied by an increase in N-alpha-acetyltransferase 25 (NAA25), and Synapto- brevin homolog (YKT6). On the other hand, when NeuroD2 was suppressed, expression of Cytoplasmic Dynein 1 Light Intermediate Chain 1, Kinesin-Like Protein (KIF-11), Leucine-tRNA Ligase (LARS1), and Ubiquitin-Associated Protein 2 (UBA2) were found to be upregulated with a reverse action. Conclusion: Up-regulations of the proteins Cytoplasmic Dynein 1, KIF11, LARS1, and UBA2 suggested that these proteins might be controlled by inhibition of NeuroD2. In this contex it can be said that, axonal transport, neuronal signaling, and activity of PI3K/AKT pathway can be indirectly regulated by NeuroD2.
  • Küçük Resim
    Öğe
    Striatal dopaminergic neurons as a potential target for GDNF based ischemic stroke therapy
    (Turkey Clinics, 2022) Beker, Mustafa Çağlar; Beker, Merve; Çağlayan, Ahmet Burak; Bolat, Busenur; Kılıç, Ülkan; Köse, Gamze; Kılıç, Ertuğrul
    Background/aim: Glial cell-line-derived neurotrophic factor (GDNF) is a well-known regulatory neurotrophic factor on dopaminergic neurons. Several pathologies have been documented so far in case of any impairment in the dopaminergic system. This study aimed to investigate the potential protective role of lentiviral GNDF delivery on the small population of tyrosine hydroxylase (TH) positive dopamine producing striatal neurons after ischemic stroke. Materials and methods: Fourteen C57BL/6J male mice (8–10 weeks) were intracerebrally treated with lentiviral GDNF (Lv-GDNF) or vehicle. Ten days after injections, cerebral ischemia was induced by blockage of the middle cerebral artery. Animals were terminated 72 h after ischemia, and their brains were taken for histological and molecular investigations. Following confirmation of GDNF overexpression, TH immunostaining and immunoblotting were used to evaluate the role of GDNF on dopaminergic neurons. Next, Fluro Jade C staining was implemented to examine the degree of neuronal degeneration at the damaged parenchyma. Results: Neither the amount of TH positive dopaminergic neurons nor the expression of TH changed in the Lv-GDNF treated animals comparing to the vehicle group. On the other hand, GDNF exposure caused a significant increase in the expression of Nurr1, an essential transcription factor for dopaminergic neurons and Gap43, growth and plasticity promoting protein, in the ischemic striatum. Treatment with Lv-GDNF gave rise to a significant reduction in the number of degenerated neurons. Finally, enhanced GDNF expression also induced expression of an important stress-related transcription factor NF-?B as well as the nitric oxide synthase enzymes iNOS and nNOS in the contralesional hemisphere. Conclusion: Considering these results together, GDNF’s impact on the survival of striatal dopaminergic neurons is not outstanding for its neuroprotective role. However, it seems that GDNF conducts several signaling pathways by acting on key transcription factors and shows its protective feature by fine-tuning the degeneration-related processes.