Tissue- and development-specific expression of multiple alternatively spliced transcripts of rat neuronal nitric oxide synthase.

Lee, Min Ae; Cai, Le; Hübner, Norbert; Lee, Young Ae; Lindpaintner, Klaus

doi:10.1172/jci119673

Cited by 79 publications

(72 citation statements)

References 37 publications

(37 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…All genes were analyzed in triplicate, and a mean value was calculated; values were subsequently normalized to ␤-actin levels for each cDNA sample. Importantly, PC12 cell ␤-actin expression was not effected by NGF treatment, as had similarly been demonstrated previously (Lee et al, 1997). Detection of the PCR product was monitored by measuring the increase in fluorescence caused by the binding of SYBR green dye to the double-stranded DNA product using the ABI PRISM 7700 sequence detection system (PerkinElmer, Waltham, MA).…”

Section: Methodsmentioning

confidence: 58%

Coregulation of Natively Expressed Pertussis Toxin-Sensitive Muscarinic Receptors with G-Protein-Activated Potassium Channels

Clancy¹,

Boyer²,

Slesinger³

2007

Journal of Neuroscience

View full text Add to dashboard Cite

show abstract

Section: Methodsmentioning

confidence: 58%

Coregulation of Natively Expressed Pertussis Toxin-Sensitive Muscarinic Receptors with G-Protein-Activated Potassium Channels

Clancy¹,

Boyer²,

Slesinger³

2007

Journal of Neuroscience

View full text Add to dashboard Cite

show abstract

“…nNOS is alternatively spliced (22)(23)(24). The alternatively spliced forms are designated nNOS-β and nNOS-γ, whereas the predominant wild-type nNOS is designated nNOSα (25)(26)(27).…”

Section: Resultsmentioning

confidence: 99%

Cyclic AMP-dependent phosphorylation of neuronal nitric oxide synthase mediates penile erection

Hurt

Sezen²,

Lagoda³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Nitric oxide (NO) generated by neuronal NO synthase (nNOS) initiates penile erection, but has not been thought to participate in the sustained erection required for normal sexual performance. We now show that cAMP-dependent phosphorylation of nNOS mediates erectile physiology, including sustained erection. nNOS is phosphorylated by cAMP-dependent protein kinase (PKA) at serine(S)1412. Electrical stimulation of the penile innervation increases S1412 phosphorylation that is blocked by PKA inhibitors but not by PI3-kinase/Akt inhibitors. Stimulation of cAMP formation by forskolin also activates nNOS phosphorylation. Sustained penile erection elicited by either intracavernous forskolin injection, or augmented by forskolin during cavernous nerve electrical stimulation, is prevented by the NOS inhibitor L-NAME or in nNOS-deleted mice. Thus, nNOS mediates both initiation and maintenance of penile erection, implying unique approaches for treating erectile dysfunction.gasotransmitter | smooth muscle relaxation | endothelial NOS | cyclic GMP | phosphoantibody N itric oxide (NO) is well established as a mediator of penile erection (1, 2). Neuronal NO synthase (nNOS) is highly localized to the penile innervation (3, 4). Electrical stimulation of the cavernous nerve (CN) to the penis elicits penile erection, which is abolished with NOS inhibitors and markedly reduced in nNOS-α-deleted mice (nNOSα −/− ) (5-7). Neuronal depolarizationinduced production of NO reflects calcium entry activating calmodulin associated with nNOS to stimulate NO formation (8,9). This activation is short-lived and thus is believed to account only for the initiation of penile erection (10, 11). Sessa and colleagues (12) established that increased blood flow and associated shear stress, acting via PI3-kinase, augments the activity of the serine protein kinase Akt, which phosphorylates vascular endothelial NOS (eNOS) at serine(S)1179, causing prolonged NO formation at resting intracellular calcium levels (13). The increased penile blood flow through cavernosal vessels initiated by nNOS activation similarly stimulates penile Akt to phosphorylate eNOS, thereby promoting sustained maximal penile erection (14).nNOS possesses a phosphorylation consensus sequence at S1412 that closely resembles the sequence surrounding eNOS-S1179, and is thought to be a target for Akt in some systems (15-21). We wondered whether nNOS phosphorylation at S1412 might regulate penile erection. Using a highly selective antibody for phospho-S1412-nNOS (P-nNOS) we report electrically stimulated nNOS phosphorylation via cAMP-dependent protein kinase (PKA) and not through Akt. P-nNOS activity contributes to sustained erection in concert with phospho-eNOS stimulation, and the neuronal and endothelial NOS activities are independently regulated by separate signaling cascades. We propose a model integrating the posttranslational phospho-stimulation of normal erectile physiology. ResultsElectrical Stimulation of Penile Innervation Augments Phosphorylation of nNOS. We developed a C-terminal ...

show abstract

“…Several different isoforms of NOS have been identified that are tissue and organ specific. NOS-I has been localized primarily to neuronal cells 15 but is also expressed in smooth muscle of the corpora and in the epithelium of the urethra; 16 it is Ca 2þ dependent 17 and can be subdivided into three distinct transcripts, NOS-Ia, NOS-Ib and NOS-Ic 18 that have distinct 5 0 untranslated first exons that arise from alternative splicing to a common second exon NOS-Ia and NOS-Ib have been identified in the penis while NOS-Ic has not. 16 NOS-II is often expressed in cells following immunological stimulation (expressed in cytokine induced macrophages) as a host defense mechanism, is Ca 2þ independent and plays a role in the inflammatory process.…”

Section: Introductionmentioning

confidence: 99%

Analysis of NOS isoform changes in a post radical prostatectomy model of erectile dysfunction

et al. 2001

View full text Add to dashboard Cite

Optimal treatment of erectile dysfunction (ED) following radical prostatectomy remains a subject of much controversy and is a significant concern for prostate cancer patients requiring surgical intervention. Neural stimulation involving nitric oxide synthase (NOS) is a crucial aspect of the normal erection process. In this study NOS isoform interaction was evaluated to improve our understanding of molecular changes pertaining to erection post radical prostatectomy. Bilateral cavernous nerve (CN) resected and control adult male Sprague-Dawley rats were killed 7, 14 and 21 days after injury. RT-PCR, in situ hybridization, Western blot and immunohistochemical analysis were used to evaluate changes in NOS isoform expression and distribution. NOS-I protein was dramatically decreased after CN injury while NOS-III and NOS-II remained unchanged. A profound decrease in smooth muscle and endothelium was observed in the corpora. To our knowledge this is the first report of differential altered NOS isoform protein abundance under conditions which mimic radical prostatectomy. These results show the importance of maintaining at least partial innervation of the penis after surgical intervention and that endothelial and smooth muscle changes resulting from loss of innervation may account for the ED observed in prostatectomy patients.

show abstract

Tissue- and development-specific expression of multiple alternatively spliced transcripts of rat neuronal nitric oxide synthase.

Cited by 79 publications

References 37 publications

Coregulation of Natively Expressed Pertussis Toxin-Sensitive Muscarinic Receptors with G-Protein-Activated Potassium Channels

Coregulation of Natively Expressed Pertussis Toxin-Sensitive Muscarinic Receptors with G-Protein-Activated Potassium Channels

Cyclic AMP-dependent phosphorylation of neuronal nitric oxide synthase mediates penile erection

Analysis of NOS isoform changes in a post radical prostatectomy model of erectile dysfunction

Contact Info

Product

Resources

About