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J A Gosling

Publications and source records attributed to J A Gosling.

At least 19 recordsLinked to original sources

Structure and autonomic innervation of the human vas deferens: a review.

The motor innervation of the smooth muscle coat of the human vas deferens is predominantly noradrenergic in type while a less dense and differently distributed presumptive cholinergic innervation is also in evidence, although the precise role of the latter is undetermined. Immunohistochemical studies have confirmed the presence of catecholamine-synthesizing enzymes tyrosine hydroxylase (TH) and dopamine beta hydroxylase (DbetaH) in the majority of fine, varicose intramuscular nerves, about two-thirds of which also contain neuropeptide Y (NPY). Minor populations of noradrenergic nerves contain enkephalin (ENK), galanin (GAL), somatostatin (SOM), or nitric oxide synthase (NOS). The presumptive cholinergic intramuscular nerves contain vasoactive intestinal polypeptide (VIP) and NPY. The subepithelial nerves of the vas deferens are assumed to have a secretomotor function and are rich in acetylcholinesterase and NPY, many also containing either VIP or NOS. The muscle coat of the human vas deferens is poorly differentiated until after birth, the intramuscular nerves in the fetus being relatively thick and non-varicose. Development of a subepithelial nerve plexus lags behind that in the muscle coat but its density in the neonatal vas deferens resembles that seen in the adult. Observations on specimens of human vas deferens obtained at vasovasostomy carried out 1 to 15 years after vasectomy have shown a marked reduction in the density of noradrenergic nerves in the muscle coat of the testicular portion while that in the urethral portion remains unaltered. Furthermore, the subepithelial secretomotor nerves degenerate in the testicular portion. These long-term changes in the pattern of innervation of the vas deferens consequent upon vasectomy may have profound effects upon the outcome of vasovasostomy with respect to subsequent sperm maturation, transport, and viability.

Autonomic Nervous System

The structure and autonomic innervation of the vesico-ureteric junction in cases of primary ureteric reflux.

OBJECTIVE: To determine the histological structure and autonomic innervation of the vesico-ureteric junction (VUJ) in cases of primary ureteric reflux, to compare the results with those reported previously for non-refluxing VUJs and thus determine possible structural anomalies which could be responsible for vesico-ureteric reflux (VUR). MATERIALS AND METHODS: Nineteen specimens of VUJs with reflux were obtained from four males and 10 females undergoing ureteric reimplantation. Serial frozen sections were cut and stained either with Masson's trichrome or immunostained for protein gene product (PGP 9.5), a general nerve marker, dopamine beta hydroxylase (D beta H), a marker for noradrenergic nerves, or neuropeptide Y (NPY). RESULTS: Longitudinally orientated muscle bundles of narrow diameter accompanied the ureter to its termination at the ureteric orifice. Where they merged with those of the superficial trigone. In some specimens, an additional muscle component was identified, situated on the outer aspect of the ureteric muscle coat, consisting of an incomplete layer of distinctive muscle whose constituent cells formed an interlacing mesh-work. Occasionally, specimens included detrusor muscle which consisted of relatively large diameter compact bundles separated from the ureter by a connective tissue sheath. D beta H- and NPY-immunoreactive (-IR) nerves were numerous among the ureteric muscle bundles. Many NPY-IR nerves occurred within the detrusor muscle while D beta H-IR nerves were mainly perivascular. When present, the outer muscle component was richly innervated by both D beta H- and NPY-IR nerves. PGP immunoreactivity varied among regions in the same section, PGP-IR nerves frequently being less numerous than those containing D beta H or NPY. CONCLUSION: These results indicate that the morphology of the VUJ in cases of VUR is indistinguishable from the normal VUJ described previously. Furthermore, the density and distribution of D beta H- and NPY-IR nerves is identical to that in controls. However, the antigenicity of PGP in the reflux specimens appears to be masked in some regions by an unknown factor.

Autonomic Nervous System

The vesico-ureteric junction in three cases of primary obstructive megaureter associated with ectopic ureteric insertion.

OBJECTIVE: To examine the histological structure and autonomic innervation of the vesico-ureteric junction (VUJ) in three cases of primary obstructive megaureter occurring in association with ectopic ureteric insertion. PATIENTS AND METHODS: Specimens of VUJ were obtained from one male and two females with primary obstructive megaureter and ectopic ureteric insertions. Serial frozen sections were cut and stained with either Masson's trichrome or immunostained for protein gene product (PGP 9.5; a general nerve marker) or dopamine beta-hydroxylase (DbetaH), a marker for noradrenergic nerves. RESULTS: In each case examined the terminal ureter was encircled by a thick collar of smooth muscle on the inside of which lay normal narrow-diameter longitudinally orientated ureteric muscle bundles. Loose connective tissue separated the surrounding detrusor muscle from this abnormal muscle collar, which was formed of distinctive smooth muscle cells arranged in a meshwork. The autonomic innervation of the ureteric and detrusor muscle bundles was similar to that previously reported for the normal VUJ while the thick muscle collar was richly innervated by DbetaH-immunoreactive noradrenergic nerves. CONCLUSION: The presence of an additional smooth muscle collar surrounding the terminal ureter may impede the normal flow of urine from ureter to bladder and be the cause of megaureter in these patients. We propose that the presence of this muscular collar arises from the developmental anomaly and suggest that these cases represent a subgroup that requires early surgical treatment.

Child

Postmortem changes in the immunohistochemical demonstration of nerves in human ventricular myocardium.

In order to delineate the effects of death on the immunofluorescence of autonomic nerves supplying the human ventricular myocardium, we studied percutaneous myocardial samples obtained postmortem from 5 individuals within 3 h of death. Subsequent samples were obtained daily from the same individuals up to a total of 5-11 d. The antibodies employed included those against protein gene product 9.5 to demonstrate nervous tissue, dopamine beta-hydroxylase and tyrosine hydroxylase to reveal catecholaminergic neural tissue and neuropeptide Y. An indirect immunofluorescence technique using the avidin-biotin method was employed. The density of myocardial protein gene product 9.5 immunoreactive nerves declined on the 7th day, and became markedly diminished by the 11th day. Immunoreactive dopamine beta-hydroxylase nerves decreased on the 5th day, and were difficult to identify by the 9th day. The density of tyrosine hydroxylase and neuropeptide Y containing nerves rapidly diminished on the 3rd and 4th days, and became undetectable by the 7th and 8th days, respectively. The present results indicate that, depending on the type of antibodies used, immunohistochemical techniques can be used on human hearts obtained up to within 6 d of death to study cardiac innervation.

Aged

Immunohistochemical characteristics of human paraganglion cells and sensory corpuscles associated with the urinary bladder. A developmental study in the male fetus, neonate and infant.

Triple label immunohistochemistry was used to study the coexistence of the catecholamine-synthesising enzymes dopamine beta-hydroxylase (DBH) and tyrosine hydroxylase (TH) and several neuropeptides including neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), substance P (SP), calcitonin gene-related peptide (CGRP), somatostatin (SOM) and galanin (GAL) as well as nitric oxide synthase (NOS) in developing pelvic paraganglion cells in a series of human male fetal, neonatal and infant specimens ranging in age from 13 wk of gestation to 3 y postnatal. 13-20 wk old fetal specimens possessed large clusters of paraganglion cells lying lateral to the urinary bladder and prostate gland which were intensely DBH-immunoreactive (-IR) but lacked TH, NOS and the neuropeptides investigated. With increasing fetal age small clusters of paraganglion cells were observed in the muscle coat of the urinary bladder. At 23 wk of gestation occasional paraganglion cells were NOS or NPY-IR while at 26 wk of gestation the majority of paraganglion cells were TH-IR and a few were SOM or GAL-IR. Some postnatal paraganglia within the bladder musculature contained cells which were all VIP, SP or CGRP-IR while others displayed coexistence of NOS and NPY, SP and CGRP, or NPY and VIP. The presence of NOS in certain paraganglion cells indicates their capacity to generate nitric oxide (NO). These results show that human paraganglion cells develop different phenotypes possibly dependent upon their location within the bladder wall. A delicate plexus of branching varicose nerves was observed in the fetal paraganglia which increased in density with increasing gestational age. The majority of these nerves were VIP-IR while others were CGRP, SP, NPY, NOS or GAL-IR. The presence of nerve terminals adjacent to the paraganglion cells implies a neural influence on the functional activity of the paraganglia. Some paraganglia in the late fetal and early postnatal specimens contained Timofeew's sensory corpuscles, resembling pacinian corpuscles in their morphology. The central nerve fibre of these corpuscles displayed immunoreactivity for SP, CGRP and NOS, the latter indicating a possible role for NO in afferent transmission from the urinary bladder. In addition, a few corpuscles were penetrated by a noradrenergic nerve fibre immunoreactive for NPY and TH, which may have a modulatory role on the sensory receptor.

Calcitonin Gene-Related Peptide

Co-localization of nitric oxide synthase, neuropeptides and tyrosine hydroxylase in nerves supplying the human post-natal vas deferens and seminal vesicle.

OBJECTIVE: To determine the distribution and patterns of co-localization of nitric oxide synthase (NOS), neuropeptides and tyrosine hydroxylase (TH) in intrinsic nerves of the human post-natal vas deferens and seminal vesicle. MATERIALS AND METHODS: Double and triple immunolabelling methods were used in tissue from 10 male infants and children (age range 2 months to 3 years) obtained at post-mortem examinations carried out within 12 h of death. RESULTS: Most nerves supplying the muscle coat of either organ were TH-immunoreactive (-IR), most of which also contained neuropeptide Y (NPY) while a smaller proportion contained both NPY and NOS. Minor populations of the TH/NPY-IR intramuscular nerves contained calcitonin gene-related peptide (CGRP), galanin (GAL), met-enkephalin (m-ENK) or vasoactive intestinal polypeptide (VIP). Non-TH-IR intramuscular nerves were relatively infrequent and most contained NPY and either VIP or NOS. Presumptive secretomotor nerves formed subepithelial plexuses in both organs, most of which contained NPY co-localized with either VIP or NOS, with minor populations containing CGRP and/or GAL. TH- and substance P (SP) -IR nerves were not observed subepithelially. Perivascular nerve plexuses were mainly formed by TH-IR varicose nerves, most of which contained co-localized NPY and CGRP, with a smaller proportion containing NPY and NOS and minor populations containing VIP, m-ENK, SP or GAL. CONCLUSION: These results indicate that the autonomic control of the human vas deferens and seminal vesicle is provided by several immunohistochemically distinct nerve populations. Furthermore, NOS is present in a proportion of both the noradrenergic and non-noradrenergic nerves. Pharmacological studies are now required to elucidate the precise roles of nitric oxide and neuropeptides in the functional control of these organs.

Biomarkers

A double-label immunohistochemical study of intramural ganglia from the human male urinary bladder neck.

Double-label immunocytochemistry was used to investigate the colocalisation of various neuropeptides and the enzymes nitric oxide synthase (NOS) and tyrosine hydroxylase (TH) in intramural ganglia of the human male urinary bladder neck and trigone. Postmortem specimens were obtained from 7 male infants and children ranging in age from 2 mo to 3 y who had died as a result of cot death or accidental trauma. On average 60% of the intramural neurons were non-TH-immunoreactive (-IR) (i.e. presumptive cholinergic) and 40% were TH- and D beta H-IR (i.e. noradrenergic). Within the non-TH-IR population, calcitonin gene-related peptide (CGRP) was found in 65% of cells, neuropeptide Y (NPY) in 90%, nitric oxide synthase (NOS) in 45%, somatostatin (SOM) in 90%, and vasoactive intestinal polypeptide (VIP) in 40%. The corresponding values for the TH-IR neurons were CGRP (54%), NPY (70%), NOS (58%), SOM (73%) and VIP (40%). All the observed bombesin (BOM)-immunoreactivity was colocalised with TH while 90% of VIP and almost all the CGRP was colocalised with NPY. Less than 5% of neurons were immunoreactive for substance P (SP) or met-enkephalin (m-ENK) and some of these also contained TH. Varicose nerve fibres were seen in close proximity to some of the intramural neurons, the majority of such varicosities showing immunoreactivity to CGRP, VIP or TH. Less common were pericellular varicosities immunoreactive to NPY, SOM or SP. These results demonstrate the neurochemical heterogeneity of intramural neurons in the human bladder neck and provide indirect evidence for the complexity of the peripheral innervation of the human urinary bladder.

Calcitonin Gene-Related Peptide

Modification of bladder structure in response to outflow obstruction and ageing.

The morphology of the normal bladder has been compared with that of the obstructed and the ageing bladder, using both light and electron microscopy. In the normal bladder the smooth muscle cells are closely packed together with relatively little intervening connective tissue. The smooth muscle bundles are innervated predominantly by autonomic presumptive cholinergic nerve fibres, while adrenergic nerves are usually observed in association with blood vessels. In the obstructed bladder, many smooth muscle cells are surrounded by large amounts of connective tissue and some of the muscle cells change their function from contracting to seemingly being involved in collagen synthesis. A significant reduction in the innervation of the smooth muscle cells is observed. In the ageing bladder, the smooth muscle cells have a normal morphology which is in contrast to the findings in the obstructed bladder. A reduction in the innervation of the ageing bladder is observed, although to a considerably lesser extent than that observed in the obstructed bladder. As the morphology of the obstructed bladder differs from that of the ageing bladder, it is unlikely that the changes seen in the obstructed bladder simply reflect a function of age.

Adult

Co-localisation of tyrosine hydroxylase, nitric oxide synthase and neuropeptides in neurons of the human postnatal male pelvic ganglia.

Double-label immunocytochemistry was used to investigate the co-localisation of neuropeptides and the enzyme nitric oxide synthase (NOS) with tyrosine hydroxylase (TH) in autonomic ganglia of the human postnatal male pelvic plexus. Postmortem specimens were obtained from six male infants and children ranging in age from 2 to 12 months who had died as a result of cot death or accidental trauma. On average, ganglia lying adjacent to the neck of the urinary bladder contained 45% of neurons which were TH-immunoreactive (-IR) while ganglia situated adjacent to the posterior and lateral aspects of the prostate gland contained 67% of neurons which were TH-IR. All the TH-IR neurons also contained dopamine beta-hydroxylase and were considered to be noradrenergic in type. On average, 61% of TH-IR neurons in bladder ganglia contained NOS, compared with 77% of non-TH-IR neurons (based on counts of over 1,000 cells in each case), while the percentages of TH- and non-TH-IR neurons containing neuropeptides were: calcitonin gene-related peptide (CGRP) (30%; 11%), neuropeptide Y (NPY) (66%; 92%), somatostatin (SOM) (70%; 29%), substance P (SP) (64%; 46%), vasoactive intestinal polypeptide (VIP) (64%; 83%). The equivalent values for TH- and non-TH-IR neurons in prostatic ganglia were NOS (38%; 59%), CGRP (55%; 18%), NPY (62%, 65%), SOM (14%, 20%), SP (13%, 8%) and VIP (42%; 82%). Varicose nerve fibers within the ganglia were seen forming pericellular arborizations around many of the ganglion cells, the most numerous containing TH-, CGRP-, NPY-, SOM- or VIP-immunoreactivity. Less common were pericellular varicosities containing SP-immunoreactivity while terminals containing NOS were not observed. No correlation could be detected between the peptide contents of the ganglion cells and of the associated pericellular terminals. However, the peptide content of the ganglion cells found in association with the urinary bladder and prostate gland correlates well with the previously documented coexistence of enzymes and neuropeptides in the intrinsic nerve fibers supplying these two regions of the human postnatal male genitourinary system.

Dopamine beta-Hydroxylase

Nitric oxide synthase and tyrosine hydroxylase are colocalized in nerves supplying the postnatal human male genitourinary organs.

PURPOSE: The objective of this study was to examine the distribution of nitric oxide synthase (NOS) and the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH) in nerve fibers supplying the human neonatal male genitourinary organs. MATERIALS AND METHODS: An indirect double label immunofluorescence technique was employed on specimens obtained from infants and children at postmortem examination. RESULTS: Many nerve fibers immunoreactive for both NOS and TH were observed in the muscle coat of the vas deferens and the seminal vesicle, within the fibromuscular stroma of the prostate gland and at the bladder neck, and also formed perivascular plexuses in each of these organs. Double-labeled nerves occurred less frequently in the intramural ureters and superficial trigone while similar nerves in the bladder body were relatively sparse. Numerous nerves immunoreactive for NOS but not TH were observed at the base of the epithelium of each organ examined. Four types of autonomic ganglion cell were observed in nearby pelvic ganglia: those which contained NOS and TH, those which contained NOS alone, those which contained TH alone and those which contained neither NOS nor TH. CONCLUSION: The results indicate that many of the noradrenergic nerves as well as non-noradrenergic nerves supplying the male genitourinary organs have the capacity to synthesize nitric oxide (NO) and that NO may play a significant role in the autonomic control of both the urinary and genital organs in the postnatal human male.

Cadaver

A quantitative histological evaluation of the dilated ureter of childhood. II: Ectopia, posterior urethral valves and the prune belly syndrome.

A quantitative histological study was performed on specimens of 33 ureters obtained from 14 male and 19 female patients 5 days to 14 years old (mean age 1.2 years). A high resolution color image video analysis system was used to quantify and compare collagen and smooth muscle components of the muscularis layers to normal control ureters from patients of similar ages. In comparing ureters with ectopia (7), ureters with ectopic ureteroceles (20) and control ureters (4) there was not a statistically different collagen-to-smooth muscle ratio among the groups. In the patients with posterior urethral valves the amount of collagen and smooth muscle was not statistically different from controls (p > 0.01), although the collagen-to-smooth muscle ratio was increased. The 4 patients with the prune belly syndrome had a collagen-to-smooth muscle ratio that was markedly elevated (1.21 versus 0.39) compared with controls. When this group was analyzed as 2 separate groups (obstructed versus refluxing ureters) the difference was more apparent (p < 0.004). Ureters with ectopia or ectopic ureteroceles and ureters associated with posterior urethral valves had similar quantitative amounts of smooth muscle (60%, 56% and 52%, respectively). In patients with the prune belly syndrome obstructed ureters had 65% muscle and refluxing ureters had 38% muscle on evaluation. The percentage of collagen was 33% in ureters with ectopia, 37% in those with ureteroceles and 48% in those associated with posterior urethral valves compared with 23% in controls. In the group with the prune belly syndrome there was 30% collagen in obstructed ureters and 62% collagen in refluxing ureters. Our findings demonstrate that while these dilated ureters had different etiologies the overall quantitative composition of collagen-to-smooth muscle ratios was similar except in refluxing ureters associated with the prune belly syndrome. Our study provides further insight into the pathological nature of such ureters and considerations for surgical repair.

Adolescent

Immunohistochemical localization of neuromarkers and neuropeptides in human fetal and neonatal urinary bladder.

OBJECTIVE: To use immunohistochemical techniques to determine the spatial and temporal distribution of a variety of neuropeptides in the human fetal and neonatal urinary bladder. MATERIALS AND METHODS: Thirteen pre-natal specimens ranging in gestational age from 17 to 35 weeks were acquired following abortion or miscarriage. In addition two post-natal specimens aged 8 and 12 weeks were obtained at post-mortem and were included in this study. The overall innervation of each specimen was visualized using the general nerve marker protein gene product 9.5 (PGP). Localization of dopamine-beta-hydroxylase (DBH) and tyrosine hydroxylase (TH) revealed putative noradrenergic nerves. The neuropeptides studied included neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), substance P (SP), and calcitonin gene-related peptide (CGRP). RESULTS: At 17 weeks a rich plexus of PGP and NPY-containing nerves was present throughout the detrusor muscle coat. As gestational age increased, VIP, SP and CGRP-containing nerves were observed with increasing frequency although SP and CGRP were mainly confined to perivascular nerve plexuses. TH- and DBH-containing nerves were first observed in the intramural ureters at 30 weeks and the detrusor muscle at 35 weeks and were relatively numerous in the intramural ureters and muscle of the superficial trigone in the two post-natal specimens. PGP-containing nerves were first observed beneath the bladder epithelium at 23 weeks and gradually became more numerous with increasing age. Occasional NPY, VIP, SP and CGRP-containing nerves were observed in the submucosa but TH- and DBH-immunostained nerves were especially numerous in the mucosa of the trigone in the two post-natal specimens, many such nerves being unrelated to the vascular supply. CONCLUSIONS: The bladder detrusor possesses a rich autonomic innervation by 17 weeks of gestation and this presumptive cholinergic innervation is associated with NPY immunoreactivity. Presumptive noradrenergic nerves appear relatively late in pre-natal development and mainly supply the intramural ureters and superficial trigone. A submucosal plexus of nerves has been demonstrated, the functional significance of which remains uncertain.

Calcitonin Gene-Related Peptide

Development of peptide-containing nerves in the human fetal vas deferens and seminal vesicle.

OBJECTIVE: To use immunohistochemical methods to study the developing autonomic innervation of the human fetal vas deferens and seminal vesicle. MATERIAL AND METHODS: Thirteen pre-natal specimens ranging in gestational age from 13 to 30 weeks were acquired following abortion or miscarriage. The overall innervation of each specimen was visualized using protein gene product 9.5 (PGP), a general nerve marker, while the onset and development of specific neuropeptide-containing sub-populations were investigated using antisera to neuropeptide Y (NPY), vasoactive intestinal peptide (VIP), substance P (SP), calcitonin gene related peptide (CGRP), bombesin (BOM), somatostatin (SOM), and met-enkephalin (ENK). In addition the occurrence and distribution of presumptive noradrenergic nerves was studied using antisera to dopamine-beta-hydroxylase (D beta H) and tyrosine hydroxylase (TH). RESULTS: At 13 weeks numerous PGP, D beta H, TH, NPY and ENK immunoreactive (-IR) nerve trunks were present in the adventitia of the vas deferens and seminal vesicle but at this stage nerve fibres were not present in the smooth muscle coat of either organ. By 17 weeks, fine PGP-, D beta H, and TH-IR nerve fibres had penetrated the outer aspect of the muscle coat of the seminal vesicle but not the vas deferens. At 20 weeks a branching network of PGP-, D beta H- and TH-IR nerve fibres occurred throughout the full thickness of the muscle coat of the seminal vesicle while similar nerves were present only in the outer half of the muscle coat of the vas deferens. At 23 weeks the full thickness of the muscle coat of the vas deferens was richly innervated by a branching plexus of PGP-IR nerves. Many of these adventitial and intramuscular nerves were immunoreactive for D beta H or TH while some were immunoreactive for either NPY or ENK. Occasional adventitial nerves were immunoreactive for SP or CGRP, these being first observed at 20 weeks. VIP-IR nerves were extremely rare in the muscle coat of either organ, being first observed at 17 weeks in the seminal vesicle and at 20 weeks in the vas deferens where they mainly formed perivascular plexuses. PGP-IR nerves were first observed in the submucosa of the seminal vesicle at 20 weeks and in the vas deferens at 21 weeks. Some of these nerves were perivascular in location while other formed a subepithelial plexus which increased in density with increasing gestational age. At 22 weeks of gestation some of the submucosal nerves were immunoreactive for SP or NPY, while at 30 weeks NPY-IR nerves formed the majority of subepithelial nerves. Occasional VIP-IR subepithelial nerves were first observed at 26 weeks but were extremely rare even at 30 weeks. Submucosal nerves immunoreactive for CGRP, D beta H, TH or ENK did not occur in any of the specimens examined. CONCLUSION: (i) From 13 weeks gestation autonomic nerves develop in the muscle coat of the fetal seminal vesicle and vas deferens, being denser in the seminal vesicle than the vas deferens up to 23 weeks gestation. (ii) The majority of the intramuscular nerves in either organ contain D beta H, TH, NPY and ENK and are presumably noradrenergic in type. (iii) A subepithelial nerve plexus develops around 20 weeks gestation and contains NPY but not VIP, unlike the adult organs. (iv) Scattered neuroendocrine cells immunoreactive for SOM are present in the mucosa of the seminal vesicle from 23 weeks of gestation.

Autonomic Nervous System

The innervation of the human myocardium at birth.

In order to delineate the type and distribution of autonomic nerves within the atrial and ventricular myocardium of the neonatal human heart, numerous samples of atrial and ventricular myocardium from 4 neonatal human hearts with no cardiac anomaly, freshly obtained at necropsy, were processed and studied using immunohistochemical and enzyme histochemical techniques. The antisera included those used to demonstrate protein gene product (PGP) 9.5 as a general neural marker, dopamine beta-hydroxylase (DBH) and tyrosine hydroxylase (TH) as indicators for presumptive sympathetic neural tissue, and neuropeptide Y (NPY). A histochemical technique was used to reveal tissue cholinesterase activity. Numerous PGP-immunoreactive (PGP-IR) nerves were seen in the atrial myocardium, forming perivascular plexuses and lying in close apposition to myocardial cells. Fewer PGP-IR nerves were found amongst the myocardium of the ventricles. Both DBH-IR and TH-IR nerves demonstrated a similar pattern of distribution as that of PGP-IR nerves; in the atria, however, they were less numerous, while in the ventricles, their density approximated to that of PGP-IR nerves. Relatively few NPY-IR nerves were observed either in the atrial or the ventricular myocardium. The density of acetylcholinesterase (AChE) positive nerves in the walls of the atria was less than that of PGP-IR nerves although their distribution patterns were similar. In the ventricles, AChE positive nerves were rarely observed. It is concluded that the neonatal human heart possesses a rich supply of autonomic nerves. The atria possess at least two populations of nerves, presumably sympathetic and vagal, whereas the walls of the ventricles are innervated principally by presumptive sympathetic nerves.

Acetylcholinesterase

An immunohistochemical study of the innervation of the ureterovesical junction in infancy and childhood.

OBJECTIVE: To use histological and immunohistochemical methods to study the structure and innervation of the human ureterovesical junction (UVJ). MATERIALS AND METHODS: A series of 24 post-natal specimens taken from patients ranging in age from 1 month to 6 years were examined. Routine histological slides were stained with Masson's trichrome. In addition, an indirect immunohistochemical method was used to study the occurrence and distribution of nerves immunoreactive for the neuropeptides vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), substance P (SP) and calcitonin gene-related peptide (CGRP). Immunoreactivity to tyrosine hydroxylase (TH), dopamine-B-hydroxylase (DBH) and to protein gene product (PGP) 9.5, a general nerve marker, were also studied. RESULTS: The UVJ comprised a ureteric muscle component (the intramural ureter) and a detrusor component (the immediately adjacent region of the urinary bladder). In the majority of specimens a third or intermediate layer was also present. This additional component consisted of tightly-packed smooth muscle cells which formed an incomplete layer that partially surrounded the juxta-vesical and intramural parts of the ureter. Numerous PGP-, VIP-, NPY, DBH- and TH- like immunoreactive (-LIR) nerves were associated with the smooth muscle bundles which comprised the intramural ureter. Such nerves ran in the connective tissue separating ureteric smooth muscle bundles and rarely coursed amongst individual smooth muscle cell comprising each bundle. SP- and CGRP- containing nerves were rarely observed in association with the intramural ureter and none were detected in the ureteric submucosa. The intermediate muscle layer was richly innervated by PGP-, TH-, DBH- and NPY- containing nerves which ran amongst the smooth muscle cells comprising this layer. VIP-, SP- and CGRP-LIR nerves were not observed within the intermediate layer. The detrusor component of the UVJ was innervated by PGP-, NPY- and VIP-LIR nerves which frequently extended between the smooth muscle cells forming the detrusor muscle bundles. TH-, DBH-, SP- and CGRP-LIR nerve fibres were rarely encountered. CONCLUSION: These findings indicate that noradrenergic nerves play a major role in the control of the ureteric component of the UVJ. In addition, the present results form baseline morphological data with which to compare the results of future studies on the structure of the UVJ in cases of vesicoureteric reflux.

Child

The intramural innervation of the human vas deferens and seminal vesicle in infants and children.

Immunohistochemical methods were used to study the autonomic innervation of the vas deferens and seminal vesicle in a series of human postnatal specimens ranging in age from 1 month to 3 years. The occurrence and distribution of nerves immunoreactive for the neuropeptides vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) substance P (SP) and calcitonin gene-related peptide (CGRP) were investigated. In addition immunoreactivity to tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and to protein gene product (PGP 9.5), a general nerve marker were also studied. A neurohistochemical method was used to localise acetylcholinesterase. The results obtained from either organ were similar. Regardless of age, a rich plexus of nerve fibres immunoreactive for PGP 9.5 was present both within the muscle coat and also beneath the epithelium of the vas deferens and seminal vesicle. Some acetylcholinesterase containing nerves occurred in the muscle coat but the majority were found under the epithelium in the connective tissue of the mucosa. TH and DBH-containing nerves (presumably noradrenergic in type) formed dense intramuscular plexuses but none occurred subepithelially. In contrast NPY-containing nerves formed a less dense intramuscular plexus and were also observed beneath the epithelium. Thus while NPY may occur in some of the intramuscular noradrenergic nerve fibres it is clearly not confined to this type of nerve in either the vas deferens or the seminal vesicle. SP- and CGRP-containing nerves were extremely infrequent and, when observed, were confined to the muscle coat.(ABSTRACT TRUNCATED AT 250 WORDS)

Autonomic Nervous System

Ultrastructural observations on cystitis cystica in human bladder urothelium.

Cystitis cystica, a common urothelial pathology whose aetiology, morphology and clinical significance are poorly understood, affects the human urinary bladder and trigone in both sexes. We have studied the fine structure of urothelial cysts in 11 patients diagnosed cystoscopically as suffering from cystitis cystica. Several abnormal features were observed in the adjacent urothelium, including large intracellular vacuoles (4 patients), Brunn's nest (5), lymphocyte infiltration (10) and generally disorganised urothelial architecture (10). Squamous metaplasia was observed in one case. The wall of each cyst consisted of a 2-3 layered epithelium with either tall columnar or flattened cells lining the fluid-filled lumen. Both types of lining cell possessed short microvilli, while the columnar type also contained numerous membrane-bound, electron dense secretory granules in the apical cytoplasm. Rough endoplasmic reticulum, mitochondria and Golgi membranes were plentiful in the surface cells. Junctional complexes joined adjacent lining cells. The deeper cells contained relatively fewer organelles, while a basal lamina separated the cyst wall from the underlying connective tissue.

Aged

Histological and histochemical study of the vesicoureteric junction in infancy and childhood.

The morphology of the vesicoureteric junction was compared using 19 post mortem specimens obtained from male and female Afro-American and Caucasian children with an age range of 1 to 72 months (mean 4). All specimens were serially sectioned and the tissues processed using standard histological and histochemical techniques (acetylcholinesterase ¿AChE¿ and pseudocholinesterase ¿PChE¿). The results failed to reveal any differences in the structure of the vesicoureteric junction with respect to age, sex and ethnic origins. The vesicoureteric junction comprised 3 histologically and histochemically distinct smooth muscle components. Ureteric muscle formed a complete inner layer rich in PChE which continued beyond the ureteric orifices to merge distally with the superficial trigone. An intermediate layer of muscle was also demonstrated whose constituent muscle cells possessed specific histological features and which was rich in both AChE and PChE, which is distinct from that derived from the ureter and detrusor. The presence of detrusor muscle on the outer aspect of the juxtavesical segment of ureter rich in AChE was also confirmed. While this study, using histochemical studies in infants and children, did not reveal any differences in the structure of the vesicoureteric junction with respect to age, sex or ethnic origin, an intermediate layer of muscle was identified with histochemical characteristics more like that of the male genital tract than that derived from the ureter or detrusor muscle.

Acetylcholinesterase