Journal: Brain : a journal of neurology

Article Title: Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia.

doi: 10.1093/brain/awr338

Figure Lengend Snippet: Figure 3 Three hours of hypoxia and 6 h of reperfusion lead to significant endothelial cell death in vitro in rat brain microvascular endothelial cells (RBMVEC; A). Non-specific gap junction blocker carbenoxolone, non-specific hemichannel blocker LaCl3, and connexin43 (Cx43) mimetic peptide protected endothelial cells against hypoxic injury, with the number of viable cells significantly higher than no treatment, while scrambled peptide did not have any protective effects. The number of viable cells was expressed as percentage of the control without hypoxia. Hypoxia and reperfusion also lead to significant propidium iodide dye uptake into primary rat brain microvascular endothelial cells indicating open hemichannels (B). Carbenoxolone, LaCl3 and connexin43 mimetic peptide significantly prevented dye uptake compared to no treatment, indicating hemichannel closure, while scrambled peptide did not have any effect. Stars denote statistical significance when compared to the control group or compared between groups in brackets; P 5 0.05.

Article Snippet: Rat brain microvascular endothelial cells (R840K-05a, Cell Applications) were plated into 24-well plates (1 105 cells/well) in rat brain microvascular endothelial cell growth media (R819K-500, Cell Applications) and allowed to settle for 16 h. Medium was then removed and replaced with Dulbecco’s Modified Eagle’s Medium/F12 containing 0.5% foetal bovine serum and 1% glutamine.

Techniques: In Vitro, Control

Journal: Oncotarget

Article Title: Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells

doi: 10.18632/oncotarget.12720

Figure Lengend Snippet: A. Light field of SSCs cultured with melatonin and GDNF, bar=50 μm. B. Cell density after being cultured with different cell mediums at 48 h. The initial number was 5*10 4 . C. QRT-PCR and western blot analysis of proliferation, self-renewal and Sertoli cell markers. D. Western blot analysis of proliferation and Sertoli cell markers. *, P<0.05,**, P<0.01.

Article Snippet: GDNF levels were determined by using a Human glial cell line-derived neurotrophic factor (GDNF) ELISA Kit (BOSTER).

Techniques: Cell Culture, Quantitative RT-PCR, Western Blot

Journal: Oncotarget

Article Title: Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells

doi: 10.18632/oncotarget.12720

Figure Lengend Snippet: A. ELISA analysis of GDNF levels in the SSCs medium. B. Western Blot analysis of phosphorylation levels of AKT and ERK. *, P<0.05,**, P<0.01.

Article Snippet: GDNF levels were determined by using a Human glial cell line-derived neurotrophic factor (GDNF) ELISA Kit (BOSTER).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Phospho-proteomics

Journal: Croatian Medical Journal

Article Title: Uroguanylin increases Ca 2+ concentration in astrocytes via guanylate cyclase C-independent signaling pathway

doi: 10.3325/cmj.2021.62.250

Figure Lengend Snippet: Uroguanylin (UGN, 100 nM) changes the H + and HCO 3 - transport in primary astrocyte cultures. UGN increases the Na + /H + exchanger activity measured by ammonia pulse ( A ) and HCO 3 - transport ( B ); summarized effects ( C ). The mechanism of cytoplasm alkalinization due to the activation of Na + /H + exchanger and HCO 3 - transport is presented. The mean of the slopes of the control experiments was set to 100%. The results are shown as mean ± standard deviation (SD), n = 3-5, primary culture isolated from three newborn animals. Bar represents 100 s. Asterisk indicates significant difference compared with controls, P < 0.05. Continuous line – control; dashed line – UGN.

Article Snippet: Supernatant-containing cells were collected and centrifuged for 6 min at 300 g. The supernatant solution was removed, and cell pellet was subjected to magnetic activated cell separation, as previously described (Astrocytes Isolation Starter Kit, Cat. No. 130-096-054, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) ( ).

Techniques: Activity Assay, Activation Assay, Control, Standard Deviation, Isolation

Journal: Croatian Medical Journal

Article Title: Uroguanylin increases Ca 2+ concentration in astrocytes via guanylate cyclase C-independent signaling pathway

doi: 10.3325/cmj.2021.62.250

Figure Lengend Snippet: Guanylate cyclase C (GC-C) is expressed in neurons of different brain regions. GC-C (green) co-localized with a neuronal nuclei marker (NeuN = red, left panel) but not with the astrocytes marker, glial fibrillary acidic protein (GFAP = red, right panel) in ( A ) the hypothalamus, ( B ) cerebellar cortex, ( C ) cerebellar deep nuclei, and ( D ) cerebral cortex of wild-type mice. GC-C knockout (KO) mice were used as a negative control. Bar represents 50 μm.

Article Snippet: Supernatant-containing cells were collected and centrifuged for 6 min at 300 g. The supernatant solution was removed, and cell pellet was subjected to magnetic activated cell separation, as previously described (Astrocytes Isolation Starter Kit, Cat. No. 130-096-054, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) ( ).

Techniques: Marker, Knock-Out, Negative Control

Journal: Croatian Medical Journal

Article Title: Uroguanylin increases Ca 2+ concentration in astrocytes via guanylate cyclase C-independent signaling pathway

doi: 10.3325/cmj.2021.62.250

Figure Lengend Snippet: Uroguanylin (UGN) Ca 2+ signaling pathway in the cortical astrocytes of brain slices. mRNA of guanylate cyclase C (GC-C) (341 bp) is expressed in the hypothalamus (H), midbrain (MB), cerebral cortex (Cx), and cerebellum (Cb) but not in astrocytes (A) ( A ). M – marker, In – intestine as positive control, Neg – negative control, glyceraldehyde 3-phosphate dehydrogenase (GAPDH, 235 bp) was used as cDNA control. Ca 2+ measurements were performed only in SR101-positive cells ( B ) of GC-C knockout (KO) (left, the number of animals is 3, the number of brain slices is 6, the number of cells is 32) and UGN KO (right, the number of animals is 3, the number of brain slices is 5, the number of cells is 14) cerebral cortex, and the results are compared with those of their wild-type littermates (GC-C WT: the number of animals is 4, the number of brain slices is 6, the number of cells is 28; UGN WT: the number of animals is 3, the number of brain slices is 5, the number of cells is 17). In neurons of the cerebral cortex of WT animals, UGN did not affect the intracellular Ca 2+ concentration (the number of animals is 4, the number of brain slices is 8, the number of cells is 43). Hyperkalemia of 30 mM was used as a positive control ( C ). The results are shown as mean ± standard deviation (SD) ΔF/F 0 is change of light output in time (ΔF) over initial brightness (F 0 ). Bar represents 10 s.

Article Snippet: Supernatant-containing cells were collected and centrifuged for 6 min at 300 g. The supernatant solution was removed, and cell pellet was subjected to magnetic activated cell separation, as previously described (Astrocytes Isolation Starter Kit, Cat. No. 130-096-054, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) ( ).

Techniques: Marker, Positive Control, Negative Control, Control, Knock-Out, Concentration Assay, Standard Deviation

Journal: Croatian Medical Journal

Article Title: Uroguanylin increases Ca 2+ concentration in astrocytes via guanylate cyclase C-independent signaling pathway

doi: 10.3325/cmj.2021.62.250

Figure Lengend Snippet: Uroguanylin (UGN) Ca 2+ signaling pathway in primary astrocyte cultures. An electrophysiological recording showing an effect of UGN (10 nM) opposite to that of membrane permeable cyclic guanosine monophosphate (cGMP, 100 μM) on astrocyte membrane potential – original trace. Dashed line represents starting membrane potential ( A ). Guanylin (GN, 10 nM) and UGN hyperpolarized, while membrane permeable cGMP depolarized astrocytes ( B ). Hyperpolarization caused by guanylin peptides was negatively correlated with the starting membrane potential ( C ). UGN (100 nM) and bradykinin (BK, 1 μM – positive control) increased intracellular Ca 2+ concentration ( D ). The results are shown as mean ± standard deviation (SD). The number of experiments is shown in parentheses. Octothorpe indicates a significant difference between cGMP effects and effects of guanylin peptides, at P < 0.05. Asterisk indicates a significant difference between UGN and BK effects, at P < 0.05. ΔF/F 0 is change of light output in time (ΔF) over initial brightness (F 0 ). Bar represents 10 s.

Article Snippet: Supernatant-containing cells were collected and centrifuged for 6 min at 300 g. The supernatant solution was removed, and cell pellet was subjected to magnetic activated cell separation, as previously described (Astrocytes Isolation Starter Kit, Cat. No. 130-096-054, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) ( ).

Techniques: Membrane, Positive Control, Concentration Assay, Standard Deviation