Cryptorchidism in homozygous <i>Adamts16^mutant</i> rats.

<p>Representative images exposing rat testes from age-matched 50 day old rats. Black arrows point to smaller, undescended testes in homozygous <i>Adamts16^mutant</i> rats.</p

Immuno-histochemical staining for cilia and proliferating cells within the seminiferous tubules.

<p>Fifty-day old age-matched rat testis sections were immunostained for cilia using α-tubulin (green), proliferating cells with PCNA (red) and nuclei (blue).</p

Detection of <i>Adamts16</i> mRNA within the rat testes.

<p>Presence of <i>Adamts16</i> mRNA in the testes of age-matched 30 day old animals was determined by <i>in-situ</i> hybridization as described under the methods section. <i>β-actin</i> was used as a control.</p

TUNEL staining for apoptosis and images of sperms within the seminiferous tubules.

<p>Fifty-day old age-matched rat testis sections were assessed for apoptosis as described under the methods section. Apoptotic cells are stained green.</p

Temporal histological changes in rat testes.

<p>Testes from n = 3 animals each from the various groups of animals were observed under (a) 10X and (b) 40X magnification post PSH staining as described under the methods section. Representative images are shown.</p

Comparisons of normalized testes weights.

<p>Bars represent weights of testes from age-matched rats (n = 3/group at each time point) normalized to their body weights. *p<0.05.</p

Sildenafil inhibits endothelin-induced vasoconstriction but not cellular proliferation in smooth muscle cells. a.

<p>To identify distribution of phosphodiesterase (PDE) subtypes, semi-quantitative RT-PCR was performed. Mouse brain homogenate was used as an internal control. <b>b.</b> To examine the roles of PDE5 in smooth muscle cells, we applied sildenafil on basilar arteries. Sildenafil significantly decreases endothelin-induced cytosolic calcium increase and vascular constriction. <b>c.</b> Unlike cilostazol, sildenafil does not have any effect on endothelin-induced cellular proliferation of smooth muscle cells. N = 3.</p

Cilostazol inhibits endothelin-induced calcium increase and vasoconstriction in mouse intracranial arteries. a.

<p>Cytosolic calcium of basilar arteries are studied <i>ex vivo</i>. Segment of artery is imaged and recorded before and after endothelin (ET) treatment in the presence or absence of cilostazol. Artery segment is pseudocolored to denote changes in cytosolic free calcium level from low to high (hi) as indicated in the color bar. <b>b.</b> Micrographs show the pons region of the whole brain before and after endothelin treatment in the presence or absence of cilostazol and/or H-89. The vascular reflection line denoting vascular tone (constriction) of intracranial arteries is adequately visible. Arrowheads point at the anterior inferior cerebellar arteries or basilar arteries to compare vascular tone before and after treatment. Sagittal sections of mouse brain are obtained at the pons, near the interpeduncular fossa boundary. A standard H&E staining is shown in the far right. Red box indicates the area of interest, where basilar artery is located. Arrows denote basilar artery constriction. Bar = 50 µm <b>c.</b> Cilostazol significantly decreases endothelin-induced cytosolic calcium increase. Administration of H-89 blocks the effect of cilostazol <b>d.</b> Changes in diameter denoting vascular constriction are expressed in percentage relative to baseline diameter prior to endothelin treatment. N = 5.</p

Cilostazol inhibits endothelin-induced cell proliferation in mouse femoral arteries.

<p>Effects of endothelin (ET) in the presence or absence of cilostazol and PKA inhibitor (H-89) on cell proliferation are studied with immunolabeling in femoral arteries <i>ex vivo</i>. Segment of artery is imaged and recorded for phase contrast to indicate the morphology of the artery, dapi to visualize cell nucleus, α-actin to outline contractile smooth muscle cells and BrdU to determine cellular proliferation property of the vascular cells. Endothelin increases cell proliferation as indicated by BrdU staining and cell growth into the lumen of the artery. Cilostazol blocks cell proliferation induced by endothelin, and H-89 inhibits efficacy of cilostazol. White box denotes a larger magnification. Asterisk indicates significant difference in fluorescence intensity per area within the smooth muscle cells, compared to control groups. N = 4.</p

Primer Sequences.

<p>Primer Sequences.</p