HT22 hücre farklılaşması insülin reseptör seviyelerini azaltır

Abstract

Amaç: Beyin, yaygın insülin reseptörü (IR) ekspresyonu olan insüline duyarlı bir organ olarak kabul edilmektedir. Beyindeki IR sinyali; nöronal gelişim, beslenme davranışı, vücut ağırlığı, dikkat, öğrenme ve hafıza gibi bilişsel süreçler için gereklidir. Primer fare hipokampal nöronal hücrelerinden ölümsüzleştirilen ana HT4 hücrelerinden türetilen HT22 hücreleri, insülin sinyali ile ilgili araştırmalarda kullanılmaktadır. Bununla birlikte, bu hücrelerin insülin sinyallemesindeki rolü bilinmemektedir. Bu çalışmada, nörobazal besiyeri kullanılarak farklılaştırılmış hücrelerde IR düzeylerini incelemeyi amaçladık. Gereç ve yöntem: Çalışma için öncelikle, hücreler 24 saat boyunca 2x105 hücre/kuyu olacak şekilde 6 oyuklu plakalara ekildi. Hücreler %80 konfluansa ulaştıktan sonra normal büyüme ortamı farklılaşma ortamı ile değiştirildi ve hücreler 72 saat 370C'de %5 CO2'de inkübe edildi. İnsülin reseptörünün (IR) ekspresyonunu belirlemek için western blot prosedürü kullanıldı. Bulgular: Sonuçlarımız, HT22 hücrelerinin farklılaşmasının nörit büyümesini desteklediğini göstermektedir. Ayrıca, IR protein seviyeleri farklılaşmış HT22 hücrelerinde önemli ölçüde azalmıştır. Sonuç: Bu bulgu, IR sinyalinin önemli olduğu durumlarda nörobazal ortamın kullanımının dikkatli bir şekilde değerlendirilmesini gerektirebilir.

Keywords

• HT22 hücre
• farklılaşma
• insülin reseptörü

HT22 cell differentiation reduces insulin receptor levels

Abstract

Purpose: The brain is an insulin-sensitive organ and has widespread insulin receptor (IR) expression. IR signaling in the brain is essential for neuronal development, feeding behavior, body weight, and cognitive processes such as attention, learning, and memory. HT22 cells, which are derived from parent HT4 cells that are immortalized from primary mouse hippocampal neuronal cells are used in research related to insulin signaling. However, the role of these cells in insulin signaling is not known. In this study, we aimed to examine IR levels in cells differentiated using neurobasal medium. Material and methods: For the study, briefly, the cells were seeded in 6-well plates at 2x105 cells/well for 24 h. After the cells reached 80% confluence, the normal growth medium was replaced with a differentiation medium and the cells were incubated for 72 hours at 370C in 5% CO2. Western blot procedure was used to determine the expression of the IR. Result: Our results show that differentiation of HT22 cells stimulates neurite outgrowth. Furthermore, IR protein levels were significantly downregulated in differentiated HT22 cells. Conclusion: This finding may require careful consideration of the use of neurobasal medium in conditions where IR signaling is important.

Keywords

• HT22 cell
• differentiation
• insulin receptor

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