scholarly journals Differentiated PDGFRα-Positive Cells: A Novel In-Vitro Model for Functional Studies of Neuronal Nitric Oxide Synthase

2021 ◽  
Vol 22 (7) ◽  
pp. 3514
Author(s):  
Bashair M. Mussa ◽  
Amir Ali Khan ◽  
Ankita Srivastava ◽  
Sallam Hasan Abdallah
Keyword(s):  
Nitric Oxide ◽  
Survival Rate ◽  
Growth Factor ◽  
Nitric Oxide Synthase ◽  
Connective Tissue ◽  
Neuronal Nitric Oxide Synthase ◽  
Tissue Growth ◽  
No Donor ◽  
Functional Studies ◽  
Oxide Synthase

It is evident that depletion of interstitial cells and dysfunction of nitric oxide (NO) pathways are key players in development of several gastrointestinal (GI) motility disorders such as diabetic gastroparesis (DGP). One of the main limitations of DGP research is the lack of isolation methods that are specific to interstitial cells, and therefore conducting functional studies is not feasible. The present study aims (i) to differentiate telomerase transformed mesenchymal stromal cells (iMSCs) into platelet-derived growth factor receptor-α-positive cells (PDGFRα-positive cells) using connective tissue growth factor (CTGF) and L-ascorbic acids; (ii) to investigate the effects of NO donor and inhibitor on the survival rate of differentiated PDGFRα-positive cells; and (iii) to evaluate the impact of increased glucose concentrations, mimicking diabetic hyperglycemia, on the gene expression of neuronal nitric oxide synthase (nNOS). A fibroblastic differentiation-induction medium supplemented with connective tissue growth factor was used to differentiate iMSCs into PDGFRα-positive cells. The medium was changed every day for 21 days to maintain the biological activity of the growth factors. Gene and protein expression, scanning electron and confocal microscopy, and flow cytometry analysis of several markers were conducted to confirm the differentiation process. Methyl tetrazolium cell viability, nitrite measurement assays, and immunostaining were used to investigate the effects of NO on PDGFRα-positive cells. The present study, for the first time, demonstrated the differentiation of iMSCs into PDGFRα-positive cells. The outcomes of the functional studies showed that SNAP (NO donor) increased the survival rate of differentiated PDGFRα-positive cells whereas LNNA (NO inhibitor) attenuated these effects. Further experimentations revealed that hyperglycemia produced a significant increase in expression of nNOS in PDGFRα-positive cells. Differentiation of iMSCs into PDGFRα-positive cells is a novel model to conduct functional studies and to investigate the involvement of NO pathways. This will help in identifying new therapeutic targets for treatment of DGP.

Neuronal nitric oxide synthase (NOS-I) knockout increases the survival rate of neural cells in the hippocampus independentlyof BDNF

2007 ◽  
Vol 35 (2) ◽  
pp. 261-271 ◽  
Author(s):  
Sabrina Fritzen ◽  
Angelika Schmitt ◽  
Katharina Köth ◽  
Claudia Sommer ◽  
Klaus-Peter Lesch ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Survival Rate ◽  
Nitric Oxide Synthase ◽  
Neuronal Nitric Oxide Synthase ◽  
Neural Cells ◽  
Oxide Synthase

scholarly journals Urethral dysfunction in a rat model of chemically induced prostatic inflammation: potential involvement of the MRP5 pump

2020 ◽  
Vol 318 (3) ◽  
pp. F754-F762
Author(s):  
Eduardo C. Alexandre ◽  
Nailong Cao ◽  
Shinsuke Mizoguchi ◽  
Tetsuichi Saito ◽  
Masahiro Kurobe ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Growth Factor ◽  
Nitric Oxide Synthase ◽  
Transforming Growth Factor ◽  
Hypoxia Inducible Factor ◽  
Neuronal Nitric Oxide Synthase ◽  
Transforming Growth Factor Β ◽  
Interleukin 1Β ◽  
Functional Assays ◽  
Oxide Synthase

Prostate inflammation (PI) is a clinical condition associated with infection and/or inflammation of the prostate. It is a common disease frequently associated to lower urinary tract (LUT) symptoms. The urethra is an understudied structure in the LUT and plays a fundamental role in the urinary cycle. Here, we proposed to evaluate the effect of PI on the urethra tissue. Male Sprague-Dawley rats were used, and PI was induced by formalin injection into the ventral lobes of the prostate. The pelvic urethra at the prostatic level was harvested for histological analysis, contraction (electrical field stimulation and phenylephrine), and relaxation (sodium nitroprusside/MK-571) experiments. Various gene targets [cytochrome c oxidase subunit 2, transforming growth factor-β1, interleukin-1β, hypoxia-inducible factor-1α, α1A-adrenoceptor, inositol 1,4,5-trisphosphate receptor type 1, voltage-gated Ca2+ channel subunit-α1D, neuronal nitric oxide synthase, soluble guanylyl cyclase, phosphodiesterase 5A, protein kinase CGMP-dependent 1, and multidrug resistance-associated protein 5 (MRP5; ATP-binding cassette subfamily C member 5)] were quantified, and cGMP levels were measured. No histological changes were detected, and functional assays revealed decreased contraction and increased relaxation of urethras from the PI group. The addition of MK-571 to functional assays increased urethral relaxation. Genes associated with inflammation were upregulated in urethras from the PI group, such as cytochrome oxidase c subunit 2, transforming growth factor-β1, interleukin-1β, and hypoxia-inducible factor-1α. We also found increased expression of L-type Ca2+ channels and the neuronal nitric oxide synthase enzyme and decreased expression of the MRP5 pump. Finally, cGMP production was enhanced in urethral tissue of PI animals. The results indicate that PI is associated with proinflammatory gene expression in the urethra without histologically evident inflammation and that PI produces a dysfunctional urethra and MRP5 pump downregulation, which results in cGMP accumulation inside the cell. These findings would help to better understand LUT dysfunctions associated with PI and the role of MRP pumps in the control of LUT function.

scholarly journals Neuronal nitric oxide synthase regulation of calcium cycling in ventricular cardiomyocytes is independent of Cav1.2 channel modulation under basal conditions

2019 ◽  
Vol 472 (1) ◽  
pp. 61-74 ◽  
Author(s):  
Janine Ebner ◽  
Michal Cagalinec ◽  
Helmut Kubista ◽  
Hannes Todt ◽  
Petra L. Szabo ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Single Channel ◽  
Neuronal Nitric Oxide Synthase ◽  
Inhibitory Effect ◽  
Open Probability ◽  
No Donor ◽  
Ventricular Cardiomyocytes ◽  
Nos Inhibitors ◽  
Oxide Synthase

AbstractNeuronal nitric oxide synthase (nNOS) is considered a regulator of Cav1.2 L-type Ca2+ channels and downstream Ca2+ cycling in the heart. The commonest view is that nitric oxide (NO), generated by nNOS activity in cardiomyocytes, reduces the currents through Cav1.2 channels. This gives rise to a diminished Ca2+ release from the sarcoplasmic reticulum, and finally reduced contractility. Here, we report that nNOS inhibitor substances significantly increase intracellular Ca2+ transients in ventricular cardiomyocytes derived from adult mouse and rat hearts. This is consistent with an inhibitory effect of nNOS/NO activity on Ca2+ cycling and contractility. Whole cell currents through L-type Ca2+ channels in rodent myocytes, on the other hand, were not substantially affected by the application of various NOS inhibitors, or application of a NO donor substance. Moreover, the presence of NO donors had no effect on the single-channel open probability of purified human Cav1.2 channel protein reconstituted in artificial liposomes. These results indicate that nNOS/NO activity does not directly modify Cav1.2 channel function. We conclude that—against the currently prevailing view—basal Cav1.2 channel activity in ventricular cardiomyocytes is not substantially regulated by nNOS activity and NO. Hence, nNOS/NO inhibition of Ca2+ cycling and contractility occurs independently of direct regulation of Cav1.2 channels by NO.

Sign in / Sign up

Export Citation Format

Share Document