Journal: International Journal of Molecular Sciences

Article Title: Effects of Varying Glucose Concentrations on ACE2′s Hypothalamic Expression and Its Potential Relation to COVID-19-Associated Neurological Dysfunction

doi: 10.3390/ijms23179645

Figure Lengend Snippet: Cell viability of hypothalamic neurons under various glucose concentrations at different time points. ( A ) Increased glucose concentrations (mg/L) of 5400, 10,800, 16,200, and 21,600 (please check) at 24, 48 and 72 h time points enhanced the viability of cells significantly, with maximum percentage (~200%) observed at the highest glucose concentration of 21,600 mg/L compared to the control condition (4500 mg/L). ( B ) Low glucose concentrations (mg/L) of 2000, 900, 500, and 200 at 24, 48 and 72 h time points affected the viability of cells adversely, with a maximum reduction observed at the lowest glucose concentration of 200 mg/L at longest exposure of 72 h as compared to the control condition (4500 mg/L). The effects of decreasing glucose concentrations on the viability of hypothalamic neurons were published previously by our group. Data is represented as mean ± SEM ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Embryonic mouse hypothalamic cells (mHypoE-N39; Cedarlane, Burlington, ON, Canada) were grown in Dulbecco’s Modified Eagles Medium (DMEM 6429; Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma Aldrich, St. Louis, MO, USA) and 1% penicillin-streptomycin (Sigma Aldrich, St. Louis, MO, USA) and maintained in a humidified atmosphere at 37 °C with 5% CO 2 .

Techniques: Concentration Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Effects of Varying Glucose Concentrations on ACE2′s Hypothalamic Expression and Its Potential Relation to COVID-19-Associated Neurological Dysfunction

doi: 10.3390/ijms23179645

Figure Lengend Snippet: Gene expression of ACE2 in hypothalamic neurons under various glucose concentrations at different time points. ( A ) Increase in glucose concentrations (mg/L) of 5400, 10,800, 16,200, and 21600 at 24, 48 and 72 h time points showed an increase in ACE2 ′s gene expression, with significant fold changes observed at the higher concentrations of 10,800, 16,200, and 21,600 mg/L after longest exposure of 72 h as compared to the control condition (4500 mg/L). ( B ) Decreasing glucose concentrations (mg/L) of 2000, 900, 500, and 200 at 24, 48 and 72 h time points also showed an increase in gene expression of ACE2 , with significant fold changes observed at the lower concentrations of 500 and 200 mg/L at 72 h time point, as compared to the control condition (4500 mg/L). Data is represented as mean ± SEM ( n = 4, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Embryonic mouse hypothalamic cells (mHypoE-N39; Cedarlane, Burlington, ON, Canada) were grown in Dulbecco’s Modified Eagles Medium (DMEM 6429; Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma Aldrich, St. Louis, MO, USA) and 1% penicillin-streptomycin (Sigma Aldrich, St. Louis, MO, USA) and maintained in a humidified atmosphere at 37 °C with 5% CO 2 .

Techniques: Gene Expression, Control

Journal: International Journal of Molecular Sciences

Article Title: Effects of Varying Glucose Concentrations on ACE2′s Hypothalamic Expression and Its Potential Relation to COVID-19-Associated Neurological Dysfunction

doi: 10.3390/ijms23179645

Figure Lengend Snippet: Protein expression of ACE2 in hypothalamic neurons under various glucose concentrations at different time points. ( A ) With increasing glucose concentrations (mg/L) of 5400, 10,800, 16,200, and 21,600, ACE2 (120 kDa) showed an upward trend in protein expression, with significant upregulation ( p < 0.05) observed at the highest concentration of 21,600 mg/L after 72 h exposure as compared to the control condition (4500 mg/L). ( B ) Histogram with relative fold change for ACE2′s protein expression ( n = 3) with increasing glucose concentrations compared to loading control (β-actin; 40 kDa). ( C ) With decreasing glucose concentrations (mg/L) of 2000, 900, 500, and 200, ACE2′s protein expression increased significantly ( p < 0.05) at longest exposure of 72 h and at the lowest concentrations of 500 and 200 mg/L glucose, as compared to the control condition (4500 mg/L). ( D ) Histogram with relative fold change for ACE2′s protein expression ( n = 3) with decreasing glucose concentrations compared to loading control (β-actin). Data is represented as mean ± SEM (* p < 0.05). Full blot images are provided in .

Article Snippet: Embryonic mouse hypothalamic cells (mHypoE-N39; Cedarlane, Burlington, ON, Canada) were grown in Dulbecco’s Modified Eagles Medium (DMEM 6429; Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma Aldrich, St. Louis, MO, USA) and 1% penicillin-streptomycin (Sigma Aldrich, St. Louis, MO, USA) and maintained in a humidified atmosphere at 37 °C with 5% CO 2 .

Techniques: Expressing, Concentration Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Effects of Varying Glucose Concentrations on ACE2′s Hypothalamic Expression and Its Potential Relation to COVID-19-Associated Neurological Dysfunction

doi: 10.3390/ijms23179645

Figure Lengend Snippet: Gene expression of TMPRSS2 in hypothalamic neurons under various glucose concentrations at different time points. ( A ) Increase in glucose concentrations (mg/L) of 5400, 10,800, 16,200, and 21,600 at 24, 48 and 72 h time points showed that TMPRSS2 expression increased slightly at 24 h and significantly after 48 h but decreased after 72 h, as compared to the control condition (4500 mg/L). ( B ) With decreasing glucose concentrations (mg/L) of 2000, 900, 500, and 200, at the lowest concentrations of 900, 500, and 200 mg/L, TMPRSS2 expression was observed to decrease significantly compared to the control condition (4500 mg/L). Data is represented as mean ± SEM ( n = 4, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Embryonic mouse hypothalamic cells (mHypoE-N39; Cedarlane, Burlington, ON, Canada) were grown in Dulbecco’s Modified Eagles Medium (DMEM 6429; Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma Aldrich, St. Louis, MO, USA) and 1% penicillin-streptomycin (Sigma Aldrich, St. Louis, MO, USA) and maintained in a humidified atmosphere at 37 °C with 5% CO 2 .

Techniques: Gene Expression, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Effects of Varying Glucose Concentrations on ACE2′s Hypothalamic Expression and Its Potential Relation to COVID-19-Associated Neurological Dysfunction

doi: 10.3390/ijms23179645

Figure Lengend Snippet: Protein expression of TMPRSS2 in hypothalamic neurons under various concentrations of glucose at different time points. ( A ) With increasing glucose concentrations (mg/L) of 5400, 10,800, 16,200, and 21,600, protein expression of TMPRSS2 (54 kDa) showed an increase after 24h, which with longer exposures of 48 and 72 h, decreased as compared to the control condition (4500 mg/L) which was not significant. ( B ) Histogram with relative fold change for TMPRSS2′s protein expression ( n = 3) with increasing glucose concentrations compared to loading control (β-actin; 40 kDa). ( C ) With decreasing glucose concentrations (mg/L) of 2000, 900, 500, and 200, there was an increase in TMPRSS2′s protein expression after 24 h, whereas the opposite trend was observed after 48 h. However, after more prolonged exposure to 72 h, its expression first increased (at 2000 and 900 mg/L) and then decreased (at 500 and 200 mg/L concentrations), as compared to the control condition (4500 mg/L) but was not significant. ( D ) Histogram with relative fold change for TMPRSS2′s protein expression ( n = 3) with decreasing glucose concentrations compared to loading control (β-actin). Data are represented as mean ± SEM. Full blot images are provided in .

Article Snippet: Embryonic mouse hypothalamic cells (mHypoE-N39; Cedarlane, Burlington, ON, Canada) were grown in Dulbecco’s Modified Eagles Medium (DMEM 6429; Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma Aldrich, St. Louis, MO, USA) and 1% penicillin-streptomycin (Sigma Aldrich, St. Louis, MO, USA) and maintained in a humidified atmosphere at 37 °C with 5% CO 2 .

Techniques: Expressing, Control