1. Regulation of Neuronal Nitric Oxide Synthase in Rat Adrenal Medulla
by Naoharu Iwai, Kazumitsu Hanai, Ikuo Tooyama, Yoshihisa Kitamura, and Masahiko Kinoshita
Hypertension
Volume 25(3):
March 1, 1995
Copyright © American Heart Association, Inc. All rights reserved.

2. Northern blots show neuronal nitric oxide synthase (nNOS) mRNA expression levels in 24-week-old spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY).
Northern blots show neuronal nitric oxide synthase (nNOS) mRNA expression levels in 24-week-old spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Expression of nNOS mRNA was determined in the cerebellum, adrenal gland, and brain stem (pons plus medulla oblongata) in 4-, 16-, and 24-week-old SHR and WKY. No significant differences in expression levels were observed between 4- and 16-week-old SHR and WKY (data not shown). A 12- and 5-kb (lower faint band) nNOS mRNA are visible. Lanes 1 through 6 correspond to RNAs obtained from WKY (1) and SHR (2) cerebellum, WKY (3) and SHR (4) decapsular portion of the adrenal gland, and WKY (5) and SHR (6) brain stem.
Naoharu Iwai et al. Hypertension. 1995;25:
Copyright © American Heart Association, Inc. All rights reserved.

3. Top, Northern blots show neuronal nitric oxide synthase (nNOS) mRNA expression levels in the decapsular portion of the adrenal gland of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY).
Top, Northern blots show neuronal nitric oxide synthase (nNOS) mRNA expression levels in the decapsular portion of the adrenal gland of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Lanes 1 through 12 correspond to RNAs obtained from 4-week-old SHR (1 and 2) and WKY (3 and 4), 16-week-old SHR (5 and 6) and WKY (7 and 8), and 24-week-old SHR (9 and 10) and WKY (11 and 12). Bottom, Bar graph shows summary of the analyses (n=4 in each group). Expression levels were determined by densitometer.
Naoharu Iwai et al. Hypertension. 1995;25:
Copyright © American Heart Association, Inc. All rights reserved.

4. Top, Blots show effects of the angiotensin II antagonist TCV-116 on neuronal nitric oxide synthase (nNOS) mRNA expression in the decapsular portion of the adrenal gland of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY).
Top, Blots show effects of the angiotensin II antagonist TCV-116 on neuronal nitric oxide synthase (nNOS) mRNA expression in the decapsular portion of the adrenal gland of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Lanes 1 through 8 correspond to RNAs obtained from 24-week-old control SHR (1 and 2) and WKY (3 and 4) and 24-week-old TCV-treated SHR (5 and 6) and WKY (7 and 8). Because of double hybridization with nNOS and GAPDH cDNAs, background signals from ribosomal RNAs were visible. Bottom, Bar graph shows summary of the analyses (n=4 in each group).
Naoharu Iwai et al. Hypertension. 1995;25:
Copyright © American Heart Association, Inc. All rights reserved.

5. Top, Blots show effects of hydralazine on neuronal nitric oxide synthase (nNOS) mRNA expression in the decapsular portion of the adrenal gland of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY).
Top, Blots show effects of hydralazine on neuronal nitric oxide synthase (nNOS) mRNA expression in the decapsular portion of the adrenal gland of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Lanes 1 through 8 correspond to RNAs obtained from 24-week-old control (1 and 2) and hydralazine-treated WKY (3 and 4) and 24-week-old control (5 and 6) and hydralazine-treated SHR (7 and 8). Bottom, Bar graph shows summary of the analyses (n=4 in each group).
Naoharu Iwai et al. Hypertension. 1995;25:
Copyright © American Heart Association, Inc. All rights reserved.

6. Left, Northern blots show effects of reserpine on neuronal nitric oxide synthase (nNOS) mRNA expression in the decapsular portion of the adrenal gland of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY).
Left, Northern blots show effects of reserpine on neuronal nitric oxide synthase (nNOS) mRNA expression in the decapsular portion of the adrenal gland of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Lanes 1 through 4 correspond to RNAs obtained from 24-week-old reserpine-treated (1) and vehicle-treated SHR (2), and 24-week-old reserpine-treated (3) and vehicle-treated WKY (4). Right, Bar graph shows summary of the analysis (n=5 in each group). Reserpine treatment was associated with a dramatic increase in nNOS mRNA expression.
Naoharu Iwai et al. Hypertension. 1995;25:
Copyright © American Heart Association, Inc. All rights reserved.

7. Blots show effects of reserpine treatment on neuronal nitric oxide synthase (nNOS) mRNA expression in the decapsular portion of the adrenal gland of 24-week-old Wistar-Kyoto rats (WKY).
Blots show effects of reserpine treatment on neuronal nitric oxide synthase (nNOS) mRNA expression in the decapsular portion of the adrenal gland of 24-week-old Wistar-Kyoto rats (WKY). The dose response and time course of changes in nNOS mRNA expression were investigated. Lanes 1 through 10 correspond to RNAs obtained from control WKY (1 and 2), WKY treated with 1 mg/kg reserpine for 24 hours (3 and 4), WKY treated with 5 mg/kg reserpine for 24 hours (5 and 6), WKY treated with 1 mg/kg reserpine for 48 hours (7 and 8), and WKY treated with 5 mg/kg reserpine for 48 hours (9 and 10).
Naoharu Iwai et al. Hypertension. 1995;25:
Copyright © American Heart Association, Inc. All rights reserved.

8. Top, Western blots show analysis of neuronal nitric oxide synthase (nNOS) expression in the decapsular portion of the adrenal gland (D-AD) of 24-week-old Wistar-Kyoto rats (WKY).
Top, Western blots show analysis of neuronal nitric oxide synthase (nNOS) expression in the decapsular portion of the adrenal gland (D-AD) of 24-week-old Wistar-Kyoto rats (WKY). WKY were treated with reserpine (5 mg/kg) and killed 24 hours later. Cytosols of the D-AD (30 μg) were analyzed with sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Lanes 1 through 8 correspond to molecular weight markers (myosin, 200 kD; β-galactosidase, 116 kD) (1), 5 mmol/L NADPH–eluted fraction from a 2′,5′-ADP–agarose column (0.5 μg) (2), cytosols of the D-AD from control WKY (3 through 5), and cytosols of the D-AD from reserpine-treated WKY (6 through 8). A dramatic increase in the intensity of immunoreactive nNOS protein was observed in reserpine-treated WKY. Bottom, Blots show validity of the antibody against nNOS. Left, Steps in the purification of nNOS from rat cerebella. The gel was stained with Coomassie blue. Lane 1, molecular weight markers; lane 2, cytosol of rat cerebella (20 μg); lane 3, 5 mmol/L NADPH–eluted fraction from a 2′,5′-ADP–agarose column (2 μg); and lane 4, 5 mmol/L EGTA–eluted fraction from a calmodulin-agarose column (1 μg). Arrow indicates nNOS protein. Right, Western blot analysis of purified nNOS by the antibody used in the present study. Lane 1, cytosol of rat cerebella (20 μg); lane 2, 5 mmol/L NADPH–eluted fraction from a 2′,5′-ADP–agarose column (0.5 μg); and lane 3, 5 mmol/L EGTA–eluted fraction from a calmodulin-agarose column (0.13 μg).
Naoharu Iwai et al. Hypertension. 1995;25:
Copyright © American Heart Association, Inc. All rights reserved.

9. Photomicrographs show NADPH-diaphorase–positive ganglionic cells in the adrenal medulla of Wistar-Kyoto rats treated with vehicle (A) and reserpine (B).
Photomicrographs show NADPH-diaphorase–positive ganglionic cells in the adrenal medulla of Wistar-Kyoto rats treated with vehicle (A) and reserpine (B). NADPH-diaphorase–positive cells and fibers can be seen in both cases. No significant difference in the distribution pattern was detected between control and reserpine-treated rats. Original magnification ×400.
Naoharu Iwai et al. Hypertension. 1995;25:
Copyright © American Heart Association, Inc. All rights reserved.