Lora Nunes

Interstitial cystitis : increased sympathetic innervation and related neuropeptide synthesis

To investigate the possibility of a neural deterioration of the bladder wall in interstitial cystitis, bladder tissue from 10 patients with interstitial cystitis was compared with that from 10 control subjects by means of immunohistochemistry. An enhanced innervation of the bladder in the submucosa and detrusor muscle was found to represent an increase of sympathetic but not cholinergic neurons. In interstitial cystitis the number of neurons positive for vasoactive intestinal polypeptide and neuropeptide Y was higher and carried a larger number of axonal varicosities, whereas the number of neurons positive for substance P and calcitonin-gene-related peptide was not significantly different in both groups. We conclude that interstitial cystitis is associated with increased sympathetic outflow into the bladder and altered metabolism of vasoactive intestinal polypeptide and neuropeptide Y. Since similar changes have been observed in other inflammatory diseases of a presumably autoimmune nature, such as rheumatoid arthritis, Crohns disease and colitis ulcerosa, the pathophysiology of interstitial cystitis may share common pathways with the latter. Experience in these diseases may facilitate a better understanding of the pathophysiology of interstitial cystitis and suggest new therapeutic concepts.

REGENERATION OF NITRIC OXIDE SYNTHASE-CONTAINING NERVES AFTER CAVERNOUS NERVE NEUROTOMY IN THE RAT

In patients who recover erectile function after radical prostatectomy (with preservation of at least 1 neurovascular bundle), a recovery time of 6 to 18 months is not uncommon. As this is also the usual time required for regeneration of spinal nerves, we believe that regeneration of cavernous nerves, partially damaged inadvertently, may be responsible. In a rat model, we examined the long-term effect of unilateral and bilateral cavernous nerve transection on the nonadrenergic/noncholinergic (NANC) nervous system and erectile function. In 31 rats, nitric oxide synthase (NOS), the enzyme that catalyzes nitric oxide production, was identified in penile nerve fibers from a mid-shaft segment with nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase staining and antibody to neuronal NOS. Animals were divided into three groups: 5 rats underwent pelvic exploration without transection of cavernous nerves (sham group); 13 rats underwent unilateral neurotomy of a 5-mm. segment of the cavernous nerve; and 13 rats underwent bilateral neurotomy. After bilateral ablation, the NOS-positive nerve fibers were significantly decreased at 3 weeks and remained so at 6 months; no erectile response could be elicited by pelvic nerve stimulation. After unilateral ablation, the NOS-positive nerve fibers were similarly decreased on the side of the neurotomy at 3 weeks, but by 6 months the number had increased significantly and approximated the level on the contralateral side. Furthermore, electrostimulation of the intact side induced a greater intracavernous pressure response at 6 months than at 3 weeks (N.B. the rat has an incomplete septum). Fibers positive for NOS were also identified in the dorsal nerve. The staining pattern diminished as rapidly and significantly on the side of neurotomy as in tissue from the corpus cavernosum. However, regeneration was not seen. To our knowledge, this is the first demonstration of regeneration of NOS-containing nerves after cavernous nerve neurotomy. Our findings support the reports by others that unilateral nerve-sparing is sufficient to preserve erectile function.

AGE DECREASES NITRIC OXIDE SYNTHASE-CONTAINING NERVE FIBERS IN THE RAT PENIS

PURPOSE To study the effect of aging on erectile function in a rat model. MATERIALS AND METHODS We investigated: 1) the number and distribution of nerve fibers within the corpus cavernosum and dorsal nerve containing vasoactive intestinal polypeptide (VIP) and nitric oxide synthase (NOS); and 2) the erectile response to apomorphine (a central dopamine receptor agonist), electrostimulation of the cavernous nerve, and intracorporeal papaverine injection. RESULTS The number of NOS-containing nerve fibers was significantly less in the old rats (24 months) than in the young (2.5 months) and intermediate (8.5 months)-aged (63.3 +/- 3.35 vs. 135.1 +/- 10.88 [p < or = 0.0002] and 127.8 +/- 11.65 [p < or = 0.0002]). The number of erections induced by apomorphine was significantly less in the old rats than in the young (1.0 +/- 3.1 vs. 3.6 +/- 0.26; p < 0.002). With electrostimulation, the latency period before the onset of the intracavernous pressure rise was noted to increase with age (2.3 +/- 0.24 sec. for the young vs. 6.77 +/- 0.98 sec. for the old, p < or = 0.0001). The maximal intracavernous pressure after intracavernous papaverine injection decreased with age. CONCLUSION The erectile mechanism appears to remain intact as rats age, but the response to central and peripheral stimulation decreases. The reduction in NOS-containing nerve fibers might account for these observations.

Semen manipulation: improved sperm recovery and function with a two-layer Percoll gradient

The authors compared a simple, two-layer Percoll density gradient technique with the swim-up technique for semen preparation in 128 men. In samples from normospermic (n = 55), oligospermic (n = 26), and asthenospermic (n = 29) men, the Percoll technique significantly improved yield, percent motility, and absolute number of motile sperm recovered, but in samples from oligoasthenospermic men (n = 18), only percent motility was improved. The Percoll density gradient also selected sperm with markedly improved function as assessed by both the sperm penetration assay and the fertility index. In 37 samples negative on the sperm penetration assay when processed with the swim-up technique, 19 (51%) became positive when processed with the Percoll technique. The Percoll density gradient is an improved method for semen manipulation as it allows greater recovery of sperm with higher motility and improved sperm function.

The effect of neural embryonic stem cell therapy in a rat model of cavernosal nerve injury

To isolate embryonic stem cells that have differentiated along the neuronal cell line, and to assess whether injecting these neural stem cells into the corpus cavernosum influences cavernosal nerve regeneration and functional status.

Homologous acellular matrix graft for urethral reconstruction in the rabbit : Histological and functional evaluation

OBJECTIVE To evaluate urethral replacement by a free homologous graft of acellular urethral matrix in a rabbit model. MATERIALS AND METHODS In 30 male New Zealand rabbits, a 0.8 to 1.1 cm. segment of the urethra was resected, replaced with an acellular matrix graft of 1.0 to 1.5 cm. (mean 1.3 cm.), and placed on an 8F feeding tube. Additionally 4 animals underwent sham operation. At varying intervals before sacrifice (from 10 days to 8 months), the animals underwent urodynamic evaluation and retrograde urethrography (for which 4 untreated rabbits served as control). The grafted specimens were prepared for evaluation histologically and by reverse-transcription polymerase chain reaction (RT-PCR). RESULTS In all animals, the acellular matrix graft remained in its original position. Histological examination showed complete epithelialization and progressive vessel infiltration. At 3 months, smooth muscle bundles were first observed infiltrating the matrix at the end-to-end anastomosis; after 6 months, the smooth muscle bundles had grown into one-third of the matrix. Urodynamics did not detect any difference between the control and matrix-grafted animals in bladder volume, leak-point pressure and residual volume. RT-PCR detected an increase in IGF mRNA in the graft between week 3 and month 6 and in HB-EGF mRNA after day 10 through month 3. TGF-alpha mRNA was not detected; TGF-beta mRNA was unchanged from normal urethral tissue. By 8 months, the host and implant could not be differentiated by urethrography. CONCLUSION The acellular urethral matrix allows single-stage urethral reconstruction. All tissue components were seen in the grafted matrix after 3 months, with further improvement over time; however, the smooth muscle in the matrix was less than in normal rabbit urethra and was not well oriented. RT-PCR revealed the importance of time-dependent growth factor influences during regeneration.

An animal model to study lower urinary tract symptoms and erectile dysfunction: the hyperlipidaemic rat

To present evidence that rats fed a high‐fat diet could serve as a useful animal model to study both lower urinary tract symptoms (LUTS) and erectile dysfunction (ED), as recent epidemiological studies have shown a strong association between LUTS and ED but the physiological basis behind this relationship is unknown.

Free ureteral replacement in rats: regeneration of ureteral wall components in the acellular matrix graft

Abstract Objectives To evaluate ureteral replacement by a free homologous graft of acellular matrix in a rat model. Methods In 30 male Sprague-Dawley rats, a 0.3 to 0.8-cm midsegment of the left ureter was resected and replaced with an acellular matrix graft of equal length placed on a polyethylene stent. The animals were killed at varying intervals, and the grafted specimens were prepared for light and electron microscopy. Results In all animals, the acellular matrix graft remained in its original position without evidence of incrustation or infection, and histologic examination showed complete epithelialization and progressive infiltration by vessels. At 10 weeks, smooth muscle fibers were observed; at 12 weeks, nerve fibers were first detected; at 4 months, smooth muscle cells had assumed regular configuration. Conclusions The ureteral acellular matrix graft appears to promote the regeneration of all ureteral wall components.

The effect of vascular endothelial growth factor and brain-derived neurotrophic factor on cavernosal nerve regeneration in a nerve-crush rat model

To test the hypothesis that an intracavernosal injection with brain‐derived neurotrophin factor (BDNF) and vascular endothelial growth factor (VEGF) can facilitate nerve regeneration and recovery of erectile function after cavernosal nerve injury.

DECREASED TRABECULAR SMOOTH MUSCLE AND CAVEOLIN-1 EXPRESSION IN THE PENILE TISSUE OF AGED RATS

PURPOSE Because decreased trabecular smooth muscle content is reportedly associated with vasculogenic impotence in men, we performed a rodent study to investigate the effect of aging on trabecular smooth muscle content and caveolin-1 protein expression in penile smooth muscle cells. MATERIALS AND METHODS In 6 young (age 3 months) and 6 old (age 24 months) rats erectile function was evaluated by cavernous nerve stimulation. At sacrifice penile tissue samples were collected for Western blot analysis, Massons trichrome staining, caveolin-1 immunostaining and electron microscopy. The percent of smooth muscle in the trabecular tissue was assessed by computer assisted image analysis. RESULTS In the aged rats mean intracavernous pressure plus or minus standard deviation was decreased (70 +/- 8.8 versus 107 +/- 12.3 cm. water) and the latency period was increased (7.8 +/- 1.2 versus 4.5 +/- 0.5 seconds) significantly compared to values in the young rats (p <0.001). The mean ratio of trabecular smooth muscle-to-connective tissue was also significantly altered in old versus young rats (27% +/- 2.9% versus 42.1% +/- 5.1%, p <0.001). Immunostaining for caveolin-1 was noted in each group in the sarcolemma of smooth muscle cells and endothelium of trabecular sinusoids but the staining pattern was less intense and the percent of smooth muscle positive for caveolin-1 was decreased in aged versus young rats (17.9% +/- 2.5% versus 27.5% +/- 3.6%, p <0.001). Moreover, young trabecular smooth muscle cells had more caveolae in the sarcolemma on electron microscopy and a higher expression of caveolin-1 protein on Western blot analysis. In contrast, higher endothelial nitric oxide synthase protein expression was noted in the penile tissue of old rats. CONCLUSIONS In these aged rats the decreased ratio of trabecular smooth muscle-to-collagen and the reduced expression of caveolin-1 may contribute to erectile dysfunction.