Biomedical subjects

J M Hutson

Publications and source records attributed to J M Hutson.

At least 37 records · Page 2Linked to original sources

PubMed2002

The role of insulin 3, testosterone, Müllerian inhibiting substance and relaxin in rat gubernacular growth.

Transabdominal testicular descent is influenced by various anatomical and hormonal factors and is mediated by gubernacular enlargement and regression of the cranial suspensory ligament, but its mechanism remains controversial. The aim of this study was to determine which hormones have a direct effect on the proliferation of cells in the day 17 fetal rat gubernaculum in vitro, using an organ culture system. The effects of synthetic rat insulin 3 (INSL3), inactive INSL3, dihydrotestosterone (DHT), DHT+INSL3, human Müllerian inhibiting substance (hMIS), hMIS+INSL3 and human gene 2 relaxin were tested, together with co-culture with fetal rat testis. Cell proliferation was assessed using a bromodeoxyuridine labelling index. The results showed that MIS and relaxin have a mild effect on gubernacular growth, whilst INSL3 and DHT have a more marked effect. The combination of MIS+INSL3 showed an effect close to that of co-culture with testis. However, the most pronounced effect was caused by DHT+INSL3. RT-PCR analysis indicated that the fetal rat gubernaculum strongly expresses putative INSL3 receptors, weakly expresses MIS type II receptors and does not express relaxin receptors. In conclusion, a number of different hormones directly influence growth of the gubernaculum in vitro, including the recently reported hormone INSL3. INSL3 shows a direct stimulatory effect on the swelling reaction, while DHT and MIS may have roles in augmenting this growth.

Animals↗

PubMed2002

Effect of an anti-androgen on testicular descent and inguinal closure in a marsupial, the tammar wallaby (Macropus eugenii).

Androgens are essential for testicular descent in eutherian mammals, but little is known about its hormonal control in marsupials. This study reports the effects of daily treatment with the anti-androgen flutamide (10 mg kg(-1)) from day 9 to day 75 after birth on the descent of the testis and inguinal closure in tammar wallabies. By day 75 after birth, the testes of control males had descended and the prostate gland was well developed. The testes of all flutamide-treated males had passed through the inguinal canal and were situated in the base of the scrotum. Three of the nine flutamide-treated males had unilateral inguinal hernias. The size of the inguinal canal, regardless of whether a hernia was present, was significantly wider than that of control males. Development of the prostate gland was significantly inhibited. By day 75 after birth, the phallus was significantly longer in control males than in females, whereas the phallus of flutamide-treated males was similar to that of control females. In flutamide-treated males, the lumbar 1 dorsal root ganglia was feminized and significantly fewer cell bodies expressed calcitonin gene- related peptide. As the anti-androgen treatment resulted in a reduction in the number of calcitonin gene-related peptide-positive cell bodies in the dorsal root ganglion supplying the genitofemoral nerve, the process of inguinal closure in tammar wallabies may be mediated by calcitonin gene-related peptide via the genitofemoral nerve, as indicated in humans. Flutamide treatment inhibited development of the prostate gland and phallus, which are both androgen-dependent structures, but it did not affect the normal descent of the testis, indicating that testicular descent can proceed when the action of androgens is blocked.

Androgen Antagonists↗

PubMed2001

Chemical synthesis and biological activity of rat INSL3.

The recently identified protein, insulin 3 (INSL3), has structural features that make it a bona fide member of the insulin superfamily. Its predicted amino acid sequence contains the classic two-peptide chain (A- and B-) structure with conserved cysteine residues that results in a disulphide bond disposition identical to that of insulin. Recently, the generation of insl3 knockout mice has demonstrated that testicular descent is blocked due to the failure of a specific ligament, the gubernaculum, to develop. The mechanism by which INSL3 exerts its action on the gubernaculum is currently unknown. The purpose of this study was to, for the first time, synthesize rat INSL3 and test its action on organ cultures of foetal rat gubernaculum. INSL3 also contains a cassette of residues Arg-X-X-X-Arg within the B-chain, a motif that is essential for characteristic activity of another related member of the superfamily, relaxin. Hence, the relaxin activity of rat INSL3 was also tested in two different relaxin bioassays. The primary structure of rat INSL3 was determined by deduction from its cDNA sequence and successfully prepared by solid phase peptide synthesis of the two constituent chains followed by their combination in solution. Following confirmation of its chemical integrity by a variety of analytical techniques, circular dichroism spectroscopy confirmed the presence of high beta-turn and alpha-helical content, with a remarkable spectral similarity to the synthetic ovine INSL3 peptide and to synthetic rat relaxin. The synthetic rat INSL3 bound with very low affinity to rat relaxin receptors and had no activity in a relaxin bioassay. Furthermore, it did not augment or antagonize relaxin activity. The rat INSL3 did however induce growth of foetal rat gubernaculum in whole organ cultures demonstrating that INSL3 has a direct action on this structure.

Amino Acid Sequence↗

PubMed2001

Early development of the gubernaculum and cremaster sac in estrogen receptor knockout mice.

AIM exogenous estrogen causes gubernacular atrophy and cryptorchidism in fetal rodents. Mice with an estrogen receptor-alpha (ERalpha) disrupted gene mutation (alphaERKO) were studied to determine whether ablation of endogenous estrogen action, through ERalpha, had an effect on gubernacular development. Serial sagittal sections were made of the pelvis in fetal and day 7 postnatal wild-type and alphaERKO mice with the estrogen receptor-alpha "knockout" gene mutation. Wild-type (n = 24), heterozygote (n = 13) and alphaERKO mice (n = 12) were sacrificed at 16, 17 and 18 days fetal life and at 7 days postnatally. The size of the gubernaculum, cremaster muscle, cremaster sac, and the width of the sac at both ends in day 7 mice were quantitated by computer analysis. Visually and statistically the ERKO mice could not be separated from the wild-type mice during fetal life. At day 7 postnatally, a thicker cremaster sac was noted morphologically, and also a statistically significant difference was seen in the width of the cremaster sac at the sac's tip. Sac area, cremaster muscle area and the width of the sac at the sac's end did not differ significantly. Overall there is minimal phenotypic change observed in the alphaERKO mouse compared to wild-type at the early developmental stages investigated. However, at postnatal day 7, there is a difference in the width of the cremasteric sac tip. This suggests that the effect of ERalpha, and thus signaling on the developing gubernaculum, occurs late in development. Alternatively, an action from the recently discovered ERbeta may be involved. Exploration of a betaERKO and the double knock-out alphaERKO/betaERKO mouse should be informative in evaluating the effect of endogenous estrogens in gubernacular development.

Animals↗

PubMed2001

Investigation of the intra-abdominal oesophagus and hiatus in fetal rats with oesophageal atresia and tracheo-oesophageal fistula.

After surgical management of their oesophageal atresia (OA) and tracheo-oesophageal fistula (TOF), most patients exhibit evidence of gastro-oesophageal reflux (GOR) and many have oesophagitis. However, the aetiology of the GOR is still controversial. This study was undertaken to document whether there are congenital abnormalities in the intra-abdominal oesophagus and the hiatus in the fetal rat with OA and TOF following exposure to adriamycin (ADR). Time-pregnant rats were injected daily with either saline or 2 mg/kg ADR intraperitoneally on gestational days (GD) 6-9. The fetuses (n = 56) from 8 litters were harvested on GD 21 for examination. The length of the oesophagus between the diaphragmatic crura and the gastro-oesophageal junction (GOJ) and the sizes of the stomach and the oesophageal hiatus were measured under a dissecting microscope. The length of the oesophagus between the diaphragmatic crura and the GOJ in the ADR-treated fetuses (0.85 +/- 0.37 mm) was significantly shorter than in control fetuses (2.41 +/- 0.32 mm) (P < 0.0001). The size of the stomach in ADR-treated fetuses (5.30 +/- 1.01 mm) was significantly smaller than in the controls (8.07 +/- 0.49 mm) (P < 0.001). Moreover, the size of the oesophageal hiatus in ADR-treated fetuses (1.16 +/- 0.43 mm) was markedly larger than in the controls (0.32 +/- 0.1 mm) (P < 0.0001). These results showed that the congenital abnormalities in ADR-treated rat fetuses may account for the oesophageal functional disorders seen after surgical correction in patients who have OA and TOF.

Abnormalities, Drug-Induced↗

PubMed2001

Enlargement of the processus vaginalis during testicular descent in rats.

The role of the processus vaginalis (PV) during inguinoscrotal testicular descent remains controversial. Some authors propose passive dragging of the PV by the migrating testis, while others suggest active elongation. In addition, the exact site of growth is unknown. Our aim was to determine whether the PV actively proliferates at its tip or stretches passively during the inguinoscrotal phase of descent in the rat. Gubernacula were removed from Sprague-Dawley (SD) rats and congenitally-cryptorchid TS mutants. Animals (at days 3, 7, 10, and 11) were treated with bromo-uridine deoxyribose (BUdR) 2 h before death. BUdR incorporation into newly-synthesised DNA served as a marker for cell division. The gubernacula were processed for haematoxylin and eosin and immunoperoxidase staining. Three sites were examined: (1) the tip of the PV on either side of the gubernacular bulb; (2) the proximal gubernacular cord; and (3) the proximal parietal PV. At each site, 50 adjacent cells were counted and the number of positive cells recorded. The highest BUdR labelling in SD rats was at the tip (site 1) on day 3 (17/50) compared with sites 2 (11/50) and 3 (9/50) (P < 0.05). Labelling decreased by 7 and 11 days to similar levels in all three sites. In TS rats, labelling rates were lower at day 3 and were highest at the tip at day 11. These results suggest active growth of the caudal tip of the PV during testicular descent. In normal rats, the growth rate slows as the testis approaches the scrotum. By contrast, in TS rats growth continued longer. We propose that the PV elongates actively from the tip to allow the intraperitoneal testis to leave the abdomen in a special peritoneal diverticulum.

Animals↗

PubMed2001

Clusters containing open-shell molecules: minimum-energy structures and low-lying isomers of ArnCH (X 2 pi), n = 1 to 15.

The size evolution of the equilibrium structures of open-shell ArnCH (X 2 pi) Van der Waals clusters is investigated for n = 1 to 15. We describe a method for combining pair potentials for Ar-CH and Ar-Ar interactions to obtain potential energy surfaces for ArnCH clusters. For each cluster size considered, the global and a few energetically close local minima are calculated using simulated annealing followed by a direct minimization scheme. Ar2CH is found to have an unusually stable planar structure, which persists as a motif in larger ArnCH clusters and has a strong effect on their optimal geometries. The lowest-energy isomers of ArnCH with n = 3 to 11 have all Ar atoms in a shell around CH. The only exception is Ar4CH, where the fully solvated isomer is 3 cm-1 higher in energy than the optimal isomer with CH bound to the surface of the Ar4 tetrahedron. For n = 7 to 11, the minimum-energy structure of ArnCH derives from the global minimum of the Arn + 1 cluster, by replacing the Ar atom at the bottom of the pentagonal bipyramid with CH. The lowest-energy structure of Ar12CH is that of the optimal icosahedral Ar13 cluster, with CH replacing one of the Ar atoms on the cluster surface. This structure supports the proposition based on the spectroscopic data, that for ArnCH clusters with about 10 to 50 Ar atoms CH resides on the surface of Arn.

Journal Article↗

PubMed2001

Gastrointestinal transit and anorectal manometry in children with colonic substance P deficiency.

BACKGROUND AND AIMS: Severe intractable constipation in children may be associated with a reduction of substance P (SP)- containing fibers in colonic circular muscle. The aim of this study was to characterize gastrointestinal transit (GIT), anorectal manometry (ARM) and electromyographic (EMG) changes in these children. METHODS: Seromuscular laparoscopic biopsies of the colon were obtained from 35 children with severe constipation. Immunofluorescent staining for SP and vasoactive intestinal peptide (VIP) were then performed on these specimens. The cohort of patients studied included a SP-deficient group (SPD, n = 25) who had reduced numbers of SP-immunoreactive nerve fibers. The other group consisted of patients with normal staining for both SP and VIP (SPN, n = 10). Gastrointestinal transit studies (gastric emptying, orocecal and colonic transit) suitable for analysis were available for 17 patients (SPD, n = 9 and SPN, n = 8). The colon was divided into segments and radioactivity counts in each segment were expressed as a percentage of the total colonic count at each time point (6, 24, 32 and 48 h). The geometric center (GC), ARM, EMG, clinical and demographic data characteristics of both groups of patients were compared. RESULTS: There were no differences in demographic data, gastric emptying, orocecal transit or geometric center of transit in the colon between the two patient groups. The ARM and EMG studies suggested that the SPN group have a higher mean threshold volume of balloon distension required to initiate a rectoanal inhibitory reflex, and a higher incidence of anismus; however, this did not reach statistical significance. CONCLUSIONS: These data suggest a trend that the SPN patients have a greater problem with obstructive defecation and abnormal rectal sensation than those with SPD. We were unable to confirm any defect in colonic transit in the SPD patients compared with the SPN group.

Anal Canal↗

PubMed2001

Slow transit constipation in children.

Patients with chronic constipation that fails to respond to treatment remain a challenge for paediatricians and surgeons. Ongoing work in our institution suggests that a number of children with intractable symptoms have slow transit constipation, which has only been described recently in paediatrics. Common features of slow transit are: delayed passage of the first meconium stool beyond 24 h of age, symptoms of severe constipation within a year, or treatment-resistant 'encopresis' at 2-3 years, soft stools despite infrequent bowel actions, and delay in colonic transit on a transit study. A proportion of children with slow transit constipation have an abnormality of intestinal innervation associated with the dysfunctional colonic motility, recognized as intestinal neuronal dysplasia (IND). Intestinal neuronal dysplasia type B, the most common variant of IND, is defined on rectal biopsy by hyperplasia of the submucosal plexus. On laparoscopic colon muscle biopsy, many specimens show reduced numbers of excitatory substance P-immunoreactive nerve fibres in the circular muscle. Functional markers of the nerves allow new diagnostic criteria to be developed which may also allow a more rational approach to treatment. The aetiology remains obscure and the optimal management poorly defined, although subtotal colectomy, proximal colostomy or appendicostomy (for antegrade enemas) have been tried. Once the anatomy and physiology of the colon in children with slow colonic transit is better understood, we will have defined not only a new form of constipation, but also will be able to consider new therapies.

Child↗

PubMed2001

Ontogeny of bladder agenesis in rats induced by adriamycin.

OBJECTIVES: To determine the anatomical steps leading to bladder agenesis in rats prenatally exposed as fetuses on gestational days (GD) 6-9 to adriamycin. MATERIALS AND METHODS: Timed-pregnant Sprague-Dawley rats were injected intraperitoneally with adriamycin at 2 mg/kg (n = 28) on GD 6-9 (vaginal plug = day 0). The control group (n = 21) received saline. Fetuses were harvested on GD 10, 11, 12, 13, 14, 15 and 16. Serial paraffin sections were prepared from a minimum of 10 experimental and five control fetuses at each gestational age, and stained with either trichrome or haematoxylin and eosin, and examined by light microscopy. RESULTS: In the control group the urorectal septum first became visible and the mesonephric ducts apparently abutting the anterior cloaca on GD 12. The presumptive urinary bladder was clearly defined on GD 14. On GD 15, the common excretory ducts became incorporated into the newly formed urogenital sinus and the ureters opened into the bladder. In the treated animals, beginning on GD 11, the undivided cloaca was noticeably smaller and by GD 13-14, the vesical extension of the urogenital sinus was conspicuously absent. Instead, opposite ureters joined to drain directly into the proximal blind-ending urethra or the persistent distal urogenital sinus. CONCLUSIONS: Prenatal exposure of rat fetuses to adriamycin resulted in primary agenesis rather than secondary resorption of the bladder. The ontogeny showed that the mechanism underpinning bladder development is unique and is under the influence of factors that can be targeted by adriamycin. Further work will elucidate the unique nature of bladder organogenesis and should have important applications in future research into artificial bladders.

Abnormalities, Drug-Induced↗

PubMed2001

Antegrade continence enemas in the treatment of slow-transit constipation.

BACKGROUND/PURPOSE: Children with anorectal dysfunction can now be treated by antegrade continence enema (ACE), as described Malone et al. Those with idiopathic constipation, however, are not thought to be suitable for this treatment. Over 150 children attend the authors' department with proven slow transit constipation (mostly proven on nuclear transit/X-ray study), and the authors reviewed the outcome in the 40 of these who have had the ACE procedure. Families completed a questionnaire and attended interview with an independent assessor. METHODS: Of the 40 patients, 32 patients were assessed. Follow-up ranged from 3 to 54 months (median, 18 months) and age ranged from 5 to 17 years (median age, 10 years). Three of 32 stomas were no longer in use. Frequency of soiling was reduced significantly in 20 patients, and a further 6 patients were clean (P <.01). Abdominal pains were relieved significantly (P <.05), and appetite and mood improved. RESULTS: Stomal complications were frequent, (stenosis in 16 of 29, mucus leak in 20 of 29, fecal leak 3 of 29, catheter-related pain in 20 of 29). Slow evacuation (12 of 29) and pain with enema (17 of 20) also were common. CONCLUSION: Malone appendicostomy does improve the well being of patients with slow transit constipation, but the advantages are less dramatic than in children with normal motility.

Adolescent↗

PubMed2001

Undescended testis is accompanied by calcitonin gene related peptide accumulation within the sensory nucleus of the genitofemoral nerve in trans-scrotal rats.

PURPOSE: Recent data suggest that calcitonin gene related peptide (CGRP) released from the sensory branch of the genitofemoral nerve may regulate testicular descent. We studied the number of CGRP immunoreactive cells in the sensory nucleus of the genitofemoral nerve (L1 to L2 dorsal root ganglia) in cryptorchid trans-scrotal rats. Four-week-old trans-scrotal rats with unilateral undescended testis underwent bilateral genitofemoral nerve dissection and retrograde nerve labeling with the fluorescent dye 4,6-diamidino-2-phenylindole (DAPI). Animals were sacrificed 48 hours later and the L1 to L2 dorsal root ganglia were removed. Serial sections were obtained and double fluorescent labeled with antibody to CGRP. Retrograde labeled and CGRP immunoreactive cells were counted using an epi-fluorescent microscope. In the 6 male trans-scrotal rats evaluated we noted a mean plus or minus standard deviation of 1,272 +/- 98 retrograde labeled dorsal root ganglion cells ipsilateral to a fully descended testicle, including 98 +/- 34 that were also CGRP immunoreactive. On the side of the undescended testis there was a mean of 1,600 +/- 304 DAPI positive cells and 160 +/- 51 CGRP immunoreactive, DAPI labeled cells. The difference was significant (p <0.02). This study shows that in trans-scrotal rats the sensory nucleus of the genitofemoral nerve contains more CGRP immunoreactive cells ipsilateral to an undescended testis than on the contralateral side, highlighting the significance of CGRP supply through the sensory branch of the genitofemoral nerve for testicular descent.

Animals↗

PubMed2001

Leydig insulin-like hormone, gubernacular development and testicular descent.

PURPOSE: Testicular descent is controlled by 2 morphological and hormonal steps. Transabdominal testicular descent is mediated by gubernacular swelling and regression of the cranial suspensory ligament. Müllerian inhibiting substance (MIS) has been proposed to stimulate the swelling but this remains controversial. Recently, a mouse mutant for Leydig insulin-like hormone (Insl3) was found to have undescended testis and deficient gubernaculum. We examine the testicular position of Insl3 mutant mice and the development of gubernacula. MATERIALS AND METHODS: Mice with Insl3 homozygotes (-/-), heterozygotes (+/-) and wild-types (+/+) were examined at embryonic day 16.5 and birth. Macroscopic dissections and measurements of the testicular position, as well as microscopic analysis (hematoxylin and eosin, and Masson's trichrome) were performed. RESULTS: Of the mice 11 Insl3 homozygote males had significantly impaired testicular descent at embryonic day 16.5 and birth (p <0.01), and the cord was thin and elongated, while 14 heterozygotes and 7 wild-types had normal testicular descent. Microscopically, the gubernaculum of Insl3 homozygotes was small with some muscle development but no central core of mesenchyme at embryonic day 16.5. On the other hand, heterozygotes and wild-types had normal gubernacular development with a swelling reaction. CONCLUSIONS: Insl3 mutants show feminized gubernaculum with deficient mesenchymal core. Insl3 appears to have some role in the gubernacular swelling reaction in mice.

Animals↗

PubMed2000

The suspensory ligament of the clitoris: connective tissue supports of the erectile tissues of the female urogenital region.

We aimed to define the gross anatomy of the supporting structures of the clitoris. We performed a dissection of the perineum of a series of 22 female and four male cadavers. Specific dissection of the clitoral and penile suspensory ligament complex was performed in four female and two male cadavers. Serial written observations and photography were used to document the findings. Our findings were then compared with the anatomical description of these structures in the historical and current anatomical literature. The suspensory ligament of clitoris consistently displayed two components: a superficial fibro-fatty structure extending from a broad base within the mons pubis to converge on the body of the clitoris and extending into the labia majora: in addition there is a deep component with a narrow origin on the symphysis pubis extending to the body and the bulbs of the clitoris. The supporting structures of the clitoris are more substantial and complex than previously described. Their shape, extent, and orientation are different from analogous structures of the penis, the suspensory ligament of which was found as described in the literature.

Adolescent↗

PubMed2000

Apoptotic cell death and fertility in three unilateral cryptorchid rat models.

Three rat strains have been studied, using a sensitive apoptotic detection method for germ-cell degeneration, to resolve the controversy regarding the effect of cryptorchidism on the contralateral descended testis (CDT). Sprague Dawley and Buffalo rats were made cryptorchid by operation at 20-22 days of age, while trans-scrotal (T-S) rats were a congenitally unilateral cryptorchid strain. Sham operated rats or normal T-S littermates were used as controls. Experiments were performed over a period ranging from 2 weeks to 18 months. Testis weight was assayed, as was the detection of apoptosis by agarose gel laddering and immunohistochemistry by using the TUNEL method. Labeled cells in 150 cross-sectioned testis tubules were counted on the TUNEL stained slides and the mean number of labeled cells per tubule was calculated. Paternity studies on Sprague Dawley and T-S rats were carried out at 12 and 24 weeks of age to assess fertility by the resultant number of pregnancies and litter sizes. Both Sprague Dawley and T-S rat models showed a biphasic distribution of apoptosis levels. This biphasic distribution was not observed in Buffalo rats as they were only studied at later time points (12-20 weeks). A significant effect on either testis weight or apoptosis in the CDT compared with the control descended testis (P > or = 0.1) has not been found in these three cryptorchid models, and the present results are discussed with reference to observations of other researchers in rodents and humans. While the cryptorchid testis showed a high level of labeled apoptotic cells per tubule in all rat strains, fertility was not affected and remained the same as controls at 12 and 24 weeks. There was, however, a marked strain difference in fertility in T-S as compared with Sprague Dawley rats. After 24 weeks of cryptorchidism, both control and cryptorchid T-S rats had a 44% pregnancy incidence compared with a 90% pregnancy incidence in Sprague Dawley rats. In addition, litter size in T-S control and cryptorchid rats were small compared with those of Sprague Dawley rats at 12 and 24 weeks.

Animals↗

PubMed2000

Balanitis xerotica obliterans with urethral stricture after hypospadias repair.

Three cases of urethral stricture due to balanitis xerotica obliterans (BXO) after hypospadias repair are reported. The first patient showed white, dense scarring on the prepuce before the hypospadias repair and developed a stricture of the urethra after the operation. The second and the third were uneventful for 6 and 2 years, respectively, after the hypospadias repair, and then developed urethral strictures. Pathologic diagnosis of the stenotic lesion is essential. Complete excision of the affected urethra with topical steroid ointment or sublesional triamcinolone injection is recommended for this condition. Although the complication of BXO after hypospadias repair is rare (3 out of 796 cases with hypospadias in our series), surgeons need to be aware of this condition as a cause for late onset of urethral problems.

Balanitis↗

PubMed2000

Fusion of childhood inguinal hernia induced by HGF and CGRP via an epithelial transition.

BACKGROUND/PURPOSE: Recent evidence has suggested that calcitonin gene-related peptide (CGRP), which is released from the genitofemoral nerve, may trigger fusion of the patent processus vaginalis in children with inguinal hernia. The purpose of this study was to determine whether CGRP triggers the release of mesenchymal factors leading to subsequent fusion of the processus vaginalis. METHODS: The response of cultured epithelial cells derived from the patent processus vaginalis was analysed by a novel in vitro culture system. Epithelial cells lining fresh hernial sacs (removed at inguinal herniotomy) were detached enzymatically and cultured for 72 hours on Micropore filters, in the presence of either 100 ng/mL hepatocyte growth factor (HGF), 7.4 x 10(-6) mol/L CGRP (amino acids 1 to 37), 7.4 x 10(-6) mol/L CGRP (8 to 37) antagonist, 10% fetal calf serum (FCS), or serum-free medium (SFM) alone. Transformation from an epithelial cell morphology to motile mesenchymal fibroblast-like cells was assessed by an average migration score (AMS), ranging from 0 with no sign of migration, to 3 with greater than 75% of cells migrating. Confocal microscopy was used to record changes in expression of epithelial (cytokeratin) and mesenchymal (vimentin) markers, as well as actin. Beta-catenin also was examined because it is part of the molecular complex that links cadherins to actin resulting in cell-cell adhesion. RESULTS: Epithelial and mesenchymal markers underwent either down-regulation or up-regulation as epithelial cell sheets broke apart and individual cells commenced migration. The AMS after 72 hours of culture was 0.22 with SFM (control); with FCS the score was 1.4 (P < .01). The AMS score with CGRP (1 to 37) was 0.55 (P = .165) and with its analogue, CGRP (8 to 37), which is a competitive inhibitor, 0.67 (P = .309). Neither was significant. HGF caused a significant increase in the AMS to 1.56 (P = .01). CONCLUSION: Both HGF and FCS (which contains various undefined peptides and growth factors) produced transformation of hernial sac epithelial cells, whereas CGRP and its inactive analogue did not. CGRP receptors are localised to mesenchymal fibroblasts within the processus vaginalis connective tissue, suggesting that CGRP could act indirectly via HGF, which, in turn, promotes fusion of the processus vaginalis. In the future, a nonsurgical treatment of inguinal herniae in children might be possible by the local administration of agents which promote fusion.

Actins↗

PubMed2000

Does the sensory nucleus of the genitofemoral nerve have a role in testicular descent?

BACKGROUND/PURPOSE: A role for the genitofemoral nerve (GFN) and its neurotransmitter, CGRP, in testicular descent has been well established. The exact mechanism, however, by which circulating androgens act on the GFN is not yet known. The authors studied the sensory nucleus of the GFN (L1-L2 dorsal root ganglia [DRG]) to determine whether it is sexually dimorphic and able to be influenced by intrauterine antiandrogen treatment. METHODS: Sprague-Dawley rats were injected daily with 100 mg/kg/d of the antiandrogen flutamide on day 16 to 19 of pregnancy. Control animals were treated with vehicle only. At the age of 2 to 3 days the newborn rats underwent unilateral dissection of the GFN. The proximal end was labelled with fluorescent dye, diamidinophenyl indole. The rats were killed 48 hours later, and the relevant ganglia (L1,L2) were removed. Cryostat frozen serial sections were cut, and retrogradely labelled fluorescent cells were counted under an epifluorescence microscope. In 32 animals, the cells were double fluorescent labelled with antibody to CGRP and FITC. RESULTS: Of 75 rats evaluated, the mean number of the DAPI-positive, retrogradely labelled cells in the control groups was 266 +/- 55 in the male, and 230 +/- 67 in the female as opposed to 186 +/- 45 and 161 +/- 35 in the flutamide-treated male and female groups, respectively. In 32 animals the DRG sections were double labelled for CGRP. The number of CGRP plus DAPI-positive cells were as follows: control males, 60 +/-12; control females, 50 +/- 9; flutamide males, 36 +/- 8; flutamide females, 40 +/- 10. CONCLUSIONS: These findings show a sexual dimorphism in the number of GFN cell bodies in the DRG. Flutamide decreases the number of GFN cell bodies in the DRG of both males and females. Our results are consistent with a role for circulating androgens acting on the sensory nucleus of the GFN (DRG) instead of the motor nucleus as previously thought. The release of CGRP from the nerve endings may occur via the sensory branch of the GFN.

Androgen Antagonists↗