Journal: Scientific Reports

Article Title: Functional NMDA receptors are expressed by human pulmonary artery smooth muscle cells

doi: 10.1038/s41598-021-87667-0

Figure Lengend Snippet: Functional NMDA receptors are localized on the surface of HPASMCs. Immunofluorescence staining was performed on non-permeabilized HPASMCs. Both GluN1 and GluN2 (2B and 2D) subunits were detected on the surface of HPASMCs ( A , B ). GluN1 co-localized with GluN2B ( A ) and GluN2D ( B ) on the surface of HPASMCs, respectively. Results are representative images taken from at least three separate experiments. Scale bar = 100 µM. ( C , D ) are representative scatter plots of each fluorescent pixel from confocal images with GluN1 green fluorescent intensity along the x -axis and GluN2 red fluorescent intensity along the y -axis. The shaded area in the upper right quadrant represents colocalized pixels above background with the associated Pearson correlation coefficient indicated for all colocalized pixels. ( E ) Representative traces for cultured HPASMCs in response to 100 μM NMDA and 10 μM glycine treatment. Cells were clamped at − 50 mV and whole cell recordings were performed. ( F ) NMDA and glycine treatment evoked an inward whole-cell current of 10.9 ± 1.7 pA (Mean ± SE, **p < 0.01, n = 16). ( G ) NMDA and glycine treatment evoked increased AUC compared with baseline (*p < 0.05, n = 16).

Article Snippet: After 3 washes in TBS, sections were permeabilized and pre-blocked followed by primary antibody incubation at the following concentrations: platelet endothelial cell adhesion molecule (PECAM-1) (ab28364, Abcam, Cambridge, MA) (1/150), GluN1 (556308, BD Pharmingen, San Jose, CA) (1/1000) (this reference contains characterization of the antibody against GluN1), GluN2A (AGC-002, Alomone, Jerusalem, Israel) (1/20,000), GluN2B (AGC-003, Alomone, Jerusalem, Israel) (1/3000), GluN2C (ab105146, Abcam, Cambridge, MA) (1/30,000) or GluN2D (ab186816, Abcam, Cambridge, MA) (1/10,000).

Techniques: Functional Assay, Immunofluorescence, Staining, Cell Culture

Journal: Health

Article Title: Long-Term Impact of Maternal Protein Malnutrition on Learning and Memory Abilities and DNA Methylating Profiles of the Nervous System in Offspring Rats

doi: 10.4236/health.2014.615239

Figure Lengend Snippet: Figure 6. Protein expressions in 8-week-old offspring rats’ brain tissue. (a)-(e) show Western blotting and immunohisto- chemical results for NMDAR1, NMDAR2B, DR1, DR2 and GR; (f) The reference factor was the data gathered on the five types of target gene mRNAs of the control group. demethylation within mature neurons and suppresses synaptic function [23]. Thus, several changes in methyla- tion in the hippocampus of 8-week-old offspring rats was found, and had an effect on the methylation status of the Grin2b promoter; it also produced a decrease in the expression of NMDAR2B in offspring, which played an important role in the LTP effect. The LTP effect is the basis of learning and memory in the hippocampus, the key structure in controlling spatial orientation [24] [25], which is susceptible to damage under acute and chronic stress [26]. The embryonic and fetal period is the most vulnerable stage during pregnancy. It was speculated that thein utero effect on the hippocampus might be the early stage of cognitive and learning dysfunction of offspring, as previous studies have shown that this anatomical abnormality may lead to deficits in hippocampal-related behaviors [27]-[29]. The LTP effect involved histone modification and DNA methylation [30]-[33]. Additional- ly, the key receptors, NMDA receptors, are involved in a wide range of learning and memory activities, synaptic plasticity, neural development, ischemic brain injury, neurodegeneration, epilepsy, cancer and many other im- portant physiological and pathological processes [34]-[37]. Also, there was an adverse impact on the learning pattern in both 8-week-old and 1-year-old offspring, ac- companied by a lower birth weight associated with catch-up growth, increased serum glucose level, reduced se- rum corticosterone and ACTH levels of offspring, and morphological changes in cerebral histiocytes, including the fetal hippocampus, similar to the findings of human epidemiology studies [38]-[40].

Article Snippet: Dropped antibody NMDAR1, NMDAR2B, DR1 (Wuhan Boster Company BA0612, BA0614, BA08031:100), DR2 (Wuhan Boster Company BA0804 1:20), GR (Wuhan Boster Company BA0895 1:40) were diluted by Triton-x-100 TBS (PH = 7) and applied onto the sections, with TBS as a negative control.

Techniques: Western Blot, Control, Methylation, Produced, Expressing, Modification, DNA Methylation Assay