A quantitative assessment of the density of the protein gene product 9.5 (PGP9.5), the neural cell adhesion molecule (NCAM), and the low-affinity nerve growth factor receptor (NGFR) expressing nerve fibers in the circular muscle layer in the colon was carried out by morphometric analyses from 13 patients with Hirschsprung's disease (HD). The difference in the nerve fiber density between the ganglionic and aganglionic segments was compared by calculating the ratio of the sum of the areas occupied by positively stained nerve fibers per unit area of the muscle after immunohistochemical staining on paraffin embedded tissue sections using computer software. There was an obvious difference in the density of the PGP9.5 stained nerve fibers between the ganglionic (0.0380 +/- 0.0171) and aganglionic segments (0.0143 +/- 0.01661). The NCAM-positive nerve fibers were fewer in number than those of both the PGP9.5-positive fibers and NCAM-positive fibers, which were also markedly lower in number in the aganglionic segment (0.0066 +/- 0.0076) than in the ganglionic segment (0.0230 +/- 0.0195). Immunostaining for low-affinity NGFR revealed much fainter staining in the ganglionic and aganglionic segment without a statistically significant difference in their density. Considering the fact that PGP9.5 is a very sensitive marker for nerve fibers, the results of this study reaffirm the innervation failure of the proper muscle in HD. The decreased NCAM expression level in the aganglionic segment appears to be caused not by the selective down-regulation of NCAM expression among the nerve fibers but by a markedly reduced number of nerve fibers.
Colon/*innervation ; Hirschsprung Disease/*pathology ; Human ; Muscle, Smooth/*innervation ; Nerve Fibers/*chemistry/pathology ; Neural Cell Adhesion Molecules/*analysis ; Receptor, Nerve Growth Factor/analysis ; Thiolester Hydrolases/*analysis

OBJECTIVETo investigate the role of ICC-like cells in bladder neuromodulation in rat urinary bladder.

METHODS14 SD rats and 1 guinea pig were sacrificed in this study. The ultra structural relationships among interstitial cells, nerves and detrusor smooth muscle cells (DSMCs) of urinary bladder were investigated by transmission electron microscopy (TEM). c-kit immunofluorescence was used to identify ICC-like cells in SD rat urinary bladder and the structural relationship between ICC-like cells and nerve terminals was studied by immunofluorescence (double-label).

RESULTSGap junction between ICC-like cells and DSMCs was confirmed by TEM. ICC-like cells were very close apposition with nerve terminals under TEM. ICC-like cells were identified to exist in sub-urothelium layer, along the longitude of smooth muscle bundles and among detrusor smooth muscle in SD rat urinary bladder by c-kit immunofluorescence. Double-labeled tissue with c-kit and PGP9.5 antibodies also showed that ICC-like cells were very close apposition with nerve terminals in SD rat bladder.

CONCLUSIONSMorphological study indicated that ICC-like cells in rat urinary bladder may play an important role in detrusor neuromodulation. Further study on function will be helpful for elucidating the mechanism of bladder neuromodulation clearly.

Animals ; Cells, Cultured ; Female ; Gap Junctions ; Guinea Pigs ; Male ; Muscle, Smooth ; innervation ; Myocytes, Smooth Muscle ; cytology ; ultrastructure ; Nerve Endings ; ultrastructure ; Rats ; Rats, Sprague-Dawley ; Urinary Bladder ; cytology ; innervation ; ultrastructure

OBJECTIVETo assess the clinical significance of electrophysiological tests of bulbocavernosus reflex (BCR), ischiocavernosus reflex (ICR) and pudenda somatosensory evoked potential (PSEP) for patients with erectile dysfunction (ED).

METHODSElectrophysiological tests of BCR, ICR and PSEP were performed for 232 ED patients with IIEF-5 scores of 2-20 (10.39 +/- 5.52), another 21 normal volunteer controls underwent the same tests, and the results were compared.

RESULTSAbnormal results, such as prolonged and advanced latencies, were found in 94 (40.5%) of the patients, which suggested neurotic ED with different degrees of cauda equine nerve injuries.

CONCLUSIONElectrophysiological tests of BCR, ICR and PSEP can objectively reveal the functional state of the cauda equine nerve and offer an important support to the diagnosis of nerve-mediated ED.

Adult ; Aged ; Case-Control Studies ; Electrophysiology ; Erectile Dysfunction ; physiopathology ; Evoked Potentials, Somatosensory ; Humans ; Male ; Middle Aged ; Muscle, Smooth ; physiopathology ; Penis ; innervation ; physiopathology ; Reflex ; physiology

Acetylcholine ; metabolism ; physiology ; Animals ; Cholinergic Agonists ; pharmacology ; Male ; Muscle Contraction ; drug effects ; physiology ; Muscle Relaxation ; drug effects ; physiology ; Muscle, Smooth ; drug effects ; pathology ; physiopathology ; Rabbits ; Random Allocation ; Receptors, Cholinergic ; physiology ; Synaptic Transmission ; drug effects ; Urinary Bladder ; drug effects ; innervation ; physiopathology

The concept of muscle rehabilitation after nerve injury is not a novel idea and is practiced in many branches of medicine, including urology. Bladder rehabilitation after spinal cord injury is universally practiced. The erectile dysfunction (ED) experienced after radical prostatectomy (RP) is increasingly recognized as being primarily neurogenic followed by secondary penile smooth muscle (SM) changes. There is unfortunately no standard approach to penile rehabilitation after RP because controlled prospective human studies are not available. This article reviews the epidemiology, experimental pathophysiological models, rationale for penile rehabilitation, and currently published rehabilitation strategies.
Alprostadil ; administration & dosage ; Animals ; Erectile Dysfunction ; etiology ; rehabilitation ; Humans ; Male ; Muscle, Smooth ; physiopathology ; Penile Erection ; physiology ; Penis ; innervation ; Peripheral Nerve Injuries ; Phosphodiesterase 5 Inhibitors ; Phosphodiesterase Inhibitors ; administration & dosage ; Prostatectomy ; adverse effects

Animals ; Enteric Nervous System ; cytology ; Gastrointestinal Diseases ; etiology ; Gastrointestinal Motility ; physiology ; Humans ; Intestinal Mucosa ; cytology ; Intestines ; cytology ; innervation ; Muscle, Smooth ; cytology ; physiology ; Myenteric Plexus ; cytology ; Proto-Oncogene Proteins c-kit ; analysis

Objective: To explore the effects of immediate and delayed intracavernous injection of bone marrow mesenchymal stem cells (BM-MSCs) on neurogenic erectile dysfunction (NED) induced by bilateral cavernous nerve injury in Sprague-Dawley (SD) rats. METHODS: BM-MSCs isolated from male SD rats were cultured and identified. Twenty-eight 8-week-old male SD rats were randomly divided into four groups, sham operation, NED model control, BM-MSCs immediate, and BM-MSCs delayed, and NED models were established in the latter three groups by crushing the bilateral cavernous nerves. The rats in the sham operation and model control groups were injected intracavernously with placebo while those in the latter two with BM-MSCs immediately or 2 weeks after modeling. At 12 weeks after operation, the penile function of the rats was assessed according to the penile intracavernous pressure (ICP), mean arterial pressure (MAP), and ICP/MAP ratio obtained from different groups of rats. Then, all the animals were sacrificed and the penile cavernosal tissue collected for histological analysis. RESULTS: At 12 weeks after modeling, both ICP and ICP/MAP were significantly increased in the BM-MSCs immediate and delayed groups as compared with those in the model control (P <0.05), and so were the ratio of smooth muscle to collagen (P <0.05) and the smooth muscle content in the corpus cavernosum (P <0.05), and the number of neurofilament (NF)-positive nerve fibers (P <0.05) and the expression of neuronal nitric oxide synthase (nNOS) in the dorsal nerves of the midshaft penis (P <0.05). CONCLUSIONS Intracavernous injection of BM-MSCs can improve erectile function in rats with bilateral cavernous nerve injury by elevating the smooth muscle-collagen ratio and smooth muscle content in the corpus cavernosum and thus preventing its fibrosis as well as by increasing the number of NF-positive nerve fibers and expression of nNOS in the penile dorsal nerves.
Animals ; Disease Models, Animal ; Erectile Dysfunction ; enzymology ; etiology ; therapy ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Muscle, Smooth ; Nitric Oxide Synthase Type I ; metabolism ; Penile Erection ; physiology ; Penis ; enzymology ; innervation ; Pudendal Nerve ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo identify and separate the ventral root from dorsal root, which is the key for success of the artificial somatic-autonomic reflex pathway procedure for neurogenic bladder after spinal cord injury (SCI). Here we report the results of intra-operating room monitoring with 10 paralyzed patients.

METHODSTen male volunteers with complete suprasacral SCI underwent the artificial somatic-autonomic procedure under general anesthesia. Vastus medialis, tibialis anticus and gastrocnemius medialis of the left lower limb were monitored for electromyogram (EMG) activities resulted from L4, L5, and S1 stimulation respectively to differentiate the ventral root from dorsal root. A Laborie Urodynamics system was connected with a three channel urodynamic catheter inserted into the bladder. The L2 and L3 roots were stimulated separately while the intravesical pressure was monitored to evaluate the function of each root.

RESULTSThe thresholds of stimulation on ventral root were 0.02 ms duration, 0.2-0.4 mA, (mean 0.3 mA+/-0.07 mA), compared with 0.2-0.4 ms duration, 1.5-3 mA (mean 2.3 mA+/-0.5 mA) for dorsal root (P<0.01) to cause revoked potentials and EMG. Electrical stimulation on L4 roots resulted in the EMG being recorded mainly on vastus medialis, while stimulation on L5 or S1 roots caused electrical activities of tibialis anticus or gastrocnemius medialis respectively. The continuous stimulation for about 3-5 seconds on S2 or S3 ventral root (0.02 ms, 20 Hz, and 0.4 mA) could resulted in bladder detrusor contraction, but the strongest bladder contraction over 50 cm H2O was usually caused by stimulation on S3 ventral root in 7 of the 10 patients.

CONCLUSIONSIntra-operating room electrophysiological monitoring is of great help to identify and separate ventral root from dorsal root, and to select the appropriate sacral ventral root for best bladder reinnervation. Different parameters and thresholds on different roots are the most important factors to keep in mind to avoid damaging the roots and to assure the best results.

Adult ; Autonomic Pathways ; physiopathology ; Electric Stimulation Therapy ; methods ; Electromyography ; Electrophysiology ; methods ; Humans ; Male ; Muscle Contraction ; Muscle, Skeletal ; physiopathology ; Muscle, Smooth ; physiopathology ; Reflex ; Spinal Cord Injuries ; complications ; physiopathology ; Spinal Nerve Roots ; physiopathology ; surgery ; Thigh ; Urinary Bladder ; innervation ; physiopathology ; Urinary Bladder, Neurogenic ; etiology ; physiopathology ; surgery

Nitric oxide (NO) is a non-adrenergic, non-cholinergic neurotransmitter found in the enteric nervous system that plays a role in a variety of enteropathies, including inflammatory bowel disease. Alteration of nitrergic neurons has been reported to be dependent on the manner by which inflammation is caused. However, this observed alteration has not been reported with acetic acid-induced colitis. Therefore, the purpose of the current study was to investigate changes in nitrergic neuromuscular transmission in experimental colitis in a rat model. Distal colitis was induced by intracolonic administration of 4% acetic acid in the rat. Animals were sacrificed at 4 h and 48 h postacetic acid treatment. Myeloperoxidase activity was significantly increased in the acetic acid-treated groups. However, the response to 60 mM KCl was not significantly different in the three groups studied. The amplitude of phasic contractions was increased by Nomega-nitro-L-arginine methyl ester (L-NAME) in the normal control group, but not in the acetic acid-treated groups. Spontaneous contractions disappeared during electrical field stimulation (EFS) in normal group. However, for the colitis groups, these contractions initially disappeared, and then reappeared during EFS. Moreover, the observed disappearance was diminished by L-NAME; this suggests that these responses were NO-mediated. In addition, the number of NADPH-diaphorase positive nerve cell bodies, in the myenteric plexus, was not altered in the distal colon; whereas the area of NADPH-diaphorase positive fibers, in the circular muscle layer, was decreased in the acetic acidtreated groups. These results suggest that NO-mediated inhibitory neural input, to the circular muscle, was decreased in the acetic acid-treated groups.
Acetic Acid/toxicity ; Animals ; Colitis/chemically induced/*pathology/*physiopathology ; Colon/drug effects/enzymology/*innervation/pathology ; Indicators and Reagents/toxicity ; Male ; Muscle Contraction/drug effects ; Muscle, Smooth/drug effects/metabolism ; Myenteric Plexus/pathology ; NADPH Dehydrogenase/metabolism ; NG-Nitroarginine Methyl Ester/pharmacology ; Neuromuscular Junction/drug effects/*metabolism ; Nitrergic Neurons/drug effects/*metabolism ; Nitric Oxide/*metabolism ; Peroxidase/metabolism ; Potassium Chloride/pharmacology ; Rats ; Rats, Sprague-Dawley

AIMTo investigate alterations of smooth muscle cells and collagen fibers in corpus cavernosum following cavernous neurectomy and its relation to the expression of transforming growth factor-beta1 (TGF-beta1).

METHODSTen adult male SD rats (neurectomy group) were subject to a bilateral cavernous nerve (CN) resection aseptically under an operating microscope, with 6 sham-operated rats as the control. Fifteen weeks after the operation, the penile specimens were collected and prepared for quantitative-analyzing of ratio of smooth muscle to collagen fibers in corpus cavernosum with confocal microscopy, and for detecting the expression of TGF-beta1 by RT-PCR and western-blot.

RESULTSSmooth muscle cells that show red color after fluorescent-labeling with tetramethylrhodamine isothiocyanate-phalloidin and collagen fibers that produce green autofluorescence after paraformaldehyde fixation were clearly identified under the confocal microscope. Quantification of fluorescent intensity showed that the ratio of smooth muscle to collagen fibers in corpus cavernosum in neurectomy group was 0.265 +/- 0.125, which was significantly lower than that in sham-operated group (0.760 +/- 0.196, P<0.01). RT-PCR and western-blot analyses revealed a significantly higher expression of TGF-beta1 in the penile tissues of the neurectomy animals than that in sham-operated group.

CONCLUSIONBilateral ablation of CN can lead to fibrosis of corpus cavernosum, which may be related to an increased expression of TGF-beta1 induced by hypoxia in cavernous tissue after denervation.

Actins ; metabolism ; Animals ; Apomorphine ; Blotting, Western ; Collagen ; metabolism ; DNA Primers ; Dopamine Agonists ; Fibrosis ; Fluorescent Dyes ; Male ; Muscle Denervation ; Muscle, Smooth ; pathology ; Penile Erection ; drug effects ; Penis ; innervation ; pathology ; Prostatectomy ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1