Morphological,histochemical and stereological analysis of the female canine M. urethralis

The M. urethralis was morphologically investigated in ten medium-sized female dogs of different breeds and age, as well as histochemically and stereologically analysed in a homogeneous group of five female beagles. Macroscopically, the muscle was essentially confined to the distal third and most strongly developed in the fourth quarter of the urethra. Here, it surrounded the urethra transversely at the ventral and lateral aspects, passing with its caudal fibres dorsally onto the vagina. The muscle fibres were assembled in groups of different sizes and usually separated by thick connective tissue septae. Based on the myofibrillar actomyosin ATPase (mATPase) reaction, type I and two main subtype II fibres could be differentiated. Type II fibres were, however, indistinguishable by their metabolic enzyme activities since both subclasses displayed oxidative-glycolytic properties. The subtype II fibres containing the more acidlabile mATPase activity were classified as IIA, whereas the other main subtype was designated IIS (subtype) and considered as peculiar to the dog. In addition, the investigation revealed a rare fibre type exhibiting the histochemical profile of IIC fibres. There was no evidence of classical glycolytic IIB fibres. The M. urethralis was composed of 24% type I and 76% type II fibres with an average diameter of 34.9 and 28.5 m, respectively. Connective tissue constituted 52.8% of the total muscle volume. Due to the predominance of fast twitch II fibres, the urethral muscle is not designed to maintain a high tone over long periods of time. Its primary role is to function rapidly and intermittently guaranteeing urinary continence during stress situations when other continence factors are overburdened.     

AChE-positive innervation of the ureters, urinary bladder, and urethra in pigs

Histochemical method of KARNOWSKY and ROOTS (1964) was used to discover the AChE-positive nerves. These nerve fibres were found in all layers of all organs under study. The ureter was weakly innervated, while the urinary bladder and the urethra possessed strong AChE-positive innervation. AChE-positive fibres were most abundant in the bladder trigone. Muscular membrane was the best supplied layer, both in the urinary bladder and in the urethra. Part of AChE-positive nerves was connected with the blood vessels in all organs under discussion.     

Adrenergic innervation of the lower urinary tract in cattle

Studies were carried out on 8 sexually immature male calves. Sections of the ureters, urinary bladder, and urethra were cut with a freezing microtome and the method of Ky?s?la et al. (1980) was used to visualize the adrenergic nerve fibres. It was found that bovine ureters possessed weak innervation; most of the nerves was located in the muscular membrane, and only in the paravesical part, sparse nerve fibres were found in the submucosa of this one. Apex of the urinary bladder was more weakly supplied with the adrenergic nerves than the corpus, whereas bladder's trigonum and cervix possessed numerous nerve fibres in both muscular and submucosal membranes. The distribution pattern of adrenergic nerves in the urethra was similar to that of urinary bladder's cervix. The presence of adregeneric nerve fibres was found in submucosal layer of both the urinary bladder and the urethra. Part of the nerves was connected with blood vessels of the organs under study.     

Adrenergic innervation of the ureters, urinary bladder, and urethra in pigs

Studies were conducted on 4 sexually mature and 4 immature pigs. Scraps of the ureters, urinary bladder, and urethra were cut with a freezing microtome. Fluorescence method of Torre and Surgeon (1976) was used to reveal the adrenergic innervation. It was found that the ureters were weakly supplied with the adrenergic nerves; most of the nerves were located in the muscular and submucosal membranes. Apex of the urinary bladder possessed the weakest innervation. More nerves were found in particular layers of the bladder corpus whereas bladder trigonum and cervix possessed numerous nerves. Adrenergic innervation of the urethra was similar to that of the urinary bladder's cervix. Adrenergic nerves were present in the serous and muscular membranes of both the urinary bladder and the urethra. Part of the nerve fibres was connected with blood vessels of the organs under study.     

Adrenergic, nitrergic and peptidergic innervation of the urethral muscle in the boar

In this study, the innervation of the urethral muscle in adult male pigs was investigated using combined NADPH-diaphorase (NADPH-d) histochemistry and immunocytochemistry. Nerve fibres supplying the urethral muscle were found to show NADPH-d activity and they also expressed immunoreactivity to catecholamine synthesising enzymes including tyrosine hydoxylase (TH) and dopamine-beta-hydroxylase (DbetaH) as well as to: vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY). Different subpopulations of the nerve fibres (NADPH-d positive, TH-, DbetaH-, VIP- and NPY-immunoreactive (IR), but also NADPH-d/VIP- and NADPH-d/NPY-IR) were disclosed. These nerve fibres were observed not only to run among muscle fibres of the urethral muscle, but also within extrinsic nerve trunks. Moreover, in the organ studied, numerous ganglia were found. The intramural ganglia, composed of a few to 30 neurons were located in the proximal, middle and distal regions of the pelvic urethra. In the vicinity of the urethral muscle, there were mainly small ganglia containing two to several neurons, but also larger ganglia consisting of up to tens neurons were encountered in the connective tissue surrounding the pelvic urethra. In the ganglia observed in the neighbourhood of the urethral muscle, different subpopulations of nerve cells were found, namely: catecholaminergic, nitrergic, VIP-IR, NPY-IR and also NADPH-d/DbetaH-, NADPH-d/VIP- and NADPH-d/NPY-positive. Possible sources of the innervation for this muscle were also discussed.     

Relationships between spindle density, muscle architecture and fibre type composition in different parts of the rabbit masseter.

The occurrence of muscle spindles was studied in the masseter muscle of rabbits by light-microscopy of whole muscle sections. The distribution of the spindles appeared to be heterogeneous. Most spindles lay in the anterior/deep and mid-parts of the masseter. The lateral/superficial part contained only a few muscle spindles. The distribution of the spindles is correlated to the distribution of slow twitch (type I) extrafusal fibres. This means that spindles, like type I fibres might be involved in the control of fine movements and posture. Spindle density and type I fibre density increase with distance from the temporomandibular joint. This could mean that spindles are involved in controlling bite force.     

Degeneration of adrenergic nerves in the urinary bladder during pregnancy

The adrenergic innervation of the urinary bladder of normal female and pregnant rats has been studied using a fluorescence histochemical method. The bladder is richly innervated by adrenergic nerve fibres as is evidenced by the presence of numerous adrenergic nerves in the adventitia, musculosa and submucosa. However, adrenergic nerve cells could not be observed. During pregnancy, adrenergic nerve fibres showed signs of degeneration, as most of the nerve fibres disappeared and the surviving fibres were much swollen. 10 days after parturition the pattern and density of adrenergic innervation became almost similar to those of the control animals.     

Automated cytometry of fibre size and spatial distribution in the superficial masseter muscle of the rat at three ages

Summary An image cytometry program was applied to sections of the superficial masseter muscle of female and male 21-, 42- and 105-day-old rats. Lesser diameter and spatial distribution of more than 6000 muscle fibres were automatically measured in digital images from muscle sections stained for myofibrillar ATPase activity.In this muscle, only type 2A, 2B and 2C fibres were observed. At the three ages and in both sexes, 2A fibres were found to have the largest diameter and were the most frequent (> 54%). In the 21-day-old animals, females presented larger diameters than did males; in contrast, in the 105-day-old animals, the three fibre types were larger in males than in females. At all ages and in both sexes, type 2A occupied 32 to 80% more cross-sectional area than type 2B. Most images (98%) showed a random spatial distribution of their fibre types.All fibre types grew in diameter with age. The coefficient of variation of the diameter was age-independent and remained under 23%. The finding of an age-independent variable could have a practical application: an increase of the coefficient of variation (> 25%) can be considered as pathological, even without a perfect age-matched control.     

The development of fibre types in grafts of a slow tonic avian muscle, the dorsocutaneous latissimus dorsi

The dorsocutaneous (DLD) and anterior (ALD) latissimus dorsii are both homogeneous slow tonic muscles. Autografts of mature DLD were attached onto the ALD of chickens to study regeneration of slow tonic muscle fibres innervated exclusively by slow tonic nerves. Fifty-three grafts were examined from 3 to 231 days after implantation for myosin ATPase, and for heavy chains of fast myosin. New muscle fibres in grafts were initially type 1 (slow) or type 2 (fast twitch). Tonic type 3 fibres were slow to differentiate and were not seen within 59 days. From 105 days many fibres were type 3A and type 1 were no longer apparent. However, type 2 fibres persisted and appeared to be present instead of type 3B fibres even after 8 months.     

The fine structure of the guinea-pig muscle spindle

Summary Nuclear bag and nuclear chain intrafusal fibres are present in guinea-pig muscle spindles. Unlike muscle spindles in other species two types of nuclear chain fibre seem to be present. The electron microscopical appearance of one type of nuclear chain fibre is similar to that of nuclear bag fibres.It is suggested that under tension the nuclei of small nuclear bag fibres become sufficiently displaced to form nuclear chain-like fibres. The frequent occurrence of fibres which combine some of the properties of both nuclear bag and nuclear chain fibres indicates the possible occurrence of a third type of intrafusal fibre.The sensory innervation of guinea-pig muscle spindles is similar to that of the cat and the rat. Three types of motor nerve ending which could be classified according to the complexity of their subneural apparatus were seen.     

No classical type IIB fibres in dog skeletal muscle

Summary To analyse the fibre type composition of adult dog skeletal muscle, enzyme histochemistry, immunohistochemistry for type I, IIA and IIB myosins, and peptide mapping of myosin heavy chains isolated from typed single fibres were combined. Subdivision of type II fibres into two main classes according to the activity of the m-ATPase after acidic and alkaline preincubation proved to be rather difficult and was only consistently achieved after a very careful adjustment of the systems used. One of these sub-classes of type II fibres stained more strongly for m-ATPase activity after acidic and alkaline preincubation, was oxidative-glycolytic and showed a strong reaction with an anti-type IIA myosin. The other one, however, although unreactive with anti-IIA myosin, was also oxidative-glycolytic, and only showed a faint reaction with an anti-type IIB myosin. Peptide mapping of the myosin heavy chains of typed single fibres revealed two populations of heavy chains among the type II fibre group. Thus, in dog muscle, we are confronted with the presence of two main classes of type II fibres, both oxidative-glycolytic, but differing in the structure of their myosin heavy chains. In contrast to some reports in the literature, no classical type IIB fibres could be detected.     

Relationship between myosin heavy chain IId isoform and fibre types in soleus muscle of the rat after hindlimb suspension

The relationship between the myosin heavy chain (HC) IId isoform and histochemically defined fibre types was investigated in the rat soleus muscle after hindlimb suspension. After 4 weeks of suspension, right and left muscles were removed and fibre type composition and total fibre number were examined by histochemical myosin adenosine triphosphatase staining sections. Myosin HC isoforms were analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. After the suspension, there was a significant decrease in the percentage of type I fibres and a concomitant increase in that of type IIa fibres. However, the total number of fibres was not affected by suspension. The synthesis of HC IId isoform, which was not found in the control, and the decrease in the ratio of slow type myosin heavy chain isoform (HC I) were observed after suspension. These results would may suggest that the change of fibre type composition was caused by a shift from type I to IIa fibres after suspension. Furthermore, it could be suggested that the synthesis of HC IId isoform occurred during the stage of type shift from type I to IIa fibres.     

Engineering of pre‐vascularized urethral patch with muscle flaps and hypoxia‐activated hUCMSCs improves its therapeutic outcome

Tissue engineering has brought new hopes for urethral reconstruction. However, the absence of pre‐vascularization and the subsequent degradation of materials often lead to the failure of in vivo application. In this study, with the assistance of hypoxia‐activated human umbilical cord mesenchymal stem cells (hUCMSCs), pedicled muscle flaps were used as materials and pre‐incubated in ventral penile subcutaneous cavity of rabbit for 3 weeks to prepare a pre‐vascularized urethral construct. We found that small vessels and muscle fibres were scattered in the construct after 3 weeks' pre‐incubation. The construct presented a fibrous reticular structure, which was similar to that of the corpus spongiosum under microscope examination. The produced constructs were then used as a patch graft for reconstruction of the defective rabbit urethra (experimental group), natural muscular patch was used as control (control group). Twelve weeks after the reconstructive surgery, urethrography and urethroscope inspections showed wide calibres of the reconstructed urethra in the experimental group. Histopathological studies revealed that fibrous connective tissues and abundant muscle fibres constituted the main body of the patch‐grafted urethra. In contrast, in the control group, only adipose tissue was found in the stenosis‐reconstructed urethra, replacing the originally grafted muscular tissue. To our knowledge, this is the first report that successfully constructed a pre‐vascularized urethral construct by using hypoxia‐activated hUCMSC and pedicled muscle flaps. More importantly, the pre‐vascularized construct showed a good performance in urethral reconstruction when applied in vivo. The study provided a novel strategy for tissue engineering of pre‐vascularized urethral construct for the defective urethra, representing a further advancement in urethral reconstruction.     

Sex-Related Differences in Urethra Development in Human Embryos

Virtually no information on the chronology of prenatal development of the human urinary tract and the sex-related differences in the emergence of urinary tract topography during embryonic development is presently available. The aim of our work was to study sex-related differences in urethra development in human embryos and early fetuses. Forty-nine preparations of human embryos and early fetuses without external signs of anatomical abnormalities were studied in order to achieve the aim and fulfill the objectives of the study. Embryos and early fetuses were divided into six groups according to gestational age and parietococcygeal length. The complex of adequate methods of morphological research used in the study included preparation and microscopy of serial histological and anatomical sections of human embryos, including female and male urinary tracts, preparation of 3D-reconstruction models, and morphometry. The formation of prostatic urethra, a derivative of the urogenital sinus, was shown to occur at the beginning of the ninth week of embryogenesis, and the primordium of the internal sphincter of the urinary tract was formed at the end of the tenth week. Formation of the terminal part of spongy urethra took place during weeks 10–11 and involved funnel-like protrusion of the ectoderm from the top of the balanus towards the urethra lumen. The secondary ventral displacement of the urethral opening does not occur in female fetuses, and, therefore, only the prostatic urethra is a homolog of the female urinary tract. The pelvic part of the urogenital sinus was transformed into the prostatic urethra and the membranous urethra of the male at the end of the first stage of fetal development. Elongation of the genital tubercle (a penis primordium) and formation of the urethral ridge walls that involved the urogenital folds occurred at the same time.     

Ureaplasmas and mycoplasmas in chimpanzees of various breeding capacities

Adult chimpanzees (24 male, 76 female) with low and high rates of conception were examined for ureaplasmas, arginine-metabolizing mycoplasmas and chlamydiae. Ureaplasmas were isolated from the throat of only 1 male and 1 female animal, but from the urethra of 29% of the males and from the vagina of 95% of the females. Mycoplasmas were isolated from the throat more often than were ureaplasmas, but from the genital tract with about the same frequency as ureaplasmas. The numbers of organisms, of either type, isolated from the vagina were larger than the numbers isolated from the male urethra. Chlamydiae were not isolated from any animal. The occurrence of ureaplasmas and mycoplasmas and the numbers of these organisms isolated were similar in animals with low or high rates of conception. Furthermore, no association was noted between the organisms in the lower genital tract and the occurrence of abortion and/or stillbirth.     

A new method to correlate histology with biomechanical properties in urethral tissue. An in-vitro study using light microscopy and scanning acoustic microscopy

When surveying the classical biomechanical theory of flow and resistance, the passive elastic properties of the urethra seems to be important for the transport of urine though the urethra. The aim of this study was to show that scanning acoustic microscopy (SAM) is a suitable methodology for investigating elastic properties of the urethra, and that it can be used to correlate elastic properties to histological areas. One 40 kg female pig and one 2 kg male rabbit comprised the material. A SAM2000 was used at a working frequency of 1000 MHz. Sections of nominal 3 micrometer thickness fixed urethral tissue were prepared for SAM and stained for light microscopy. The histological layers of the urethra were evident in the SAM image, and showed highly variable values of elastic properties. The layers seen with SAM correlated well with those seen with light microscopy. In conclusion, we have provided the first images of the microelastic properties of the urethra and correlated them to histology.     

Polymorphism of myofibrillar proteins of rabbit skeletal-muscle fibres. An electrophoretic study of single fibres.

Rabbit predominantly fast-twitch-fibre and predominantly slow-twitch-fibre skeletal muscles of the hind limbs, the psoas, the diaphragm and the masseter muscles were fibre-typed by one-dimensional polyacrylamide-gel electrophoresis of the myofibrillar proteins of chemically skinned single fibres. Investigation of the distribution of fast-twitch-fibre and slow-twitch-fibre isoforms of myosin light chains and the type of myosin heavy chains, based on peptide ''maps'' published in Cleveland. Fischer, Kirschner & Laemmli [(1977) J. Biol. Chem. 252, 1102-1106], allowed a classification of muscle fibres into four classes, corresponding to histochemical types I, IIA, IIB and IIC. Type I fibres with a pure slow-twitch-type of myosin were found to be characterized by a unique set of isoforms of troponins I, C and T, in agreement with the immunological data of Dhoot & Perry [(1979) Nature (London) 278, 714-718], by predominance of the beta-tropomyosin subunit and by the presence of a small amount of an additional tropomyosin subunit, apparently dissimilar from fast-twitch-fibre alpha-tropomyosin subunit. The myofibrillar composition of type IIB fast-twitch white fibres was the mirror image of that found for slow-twitch fibres in that the fast-twitch-fibre isoforms only of the troponin subunits were present and the alpha-tropomyosin subunit predominated. Type IIA fast-twitch red fibres showed a troponin subunit composition identical with that of type IIB fast-twitch white fibres. On the other hand, a unique type of myosin heavy chains was found to be associated with type IIA fibres. Furthermore, the myosin light-chain composition of these fibres was invariably characterized by a small amount of LC3F light chain and by a pattern that was either a pure fast-twitch-fibre light-chain pattern or a hybrid LC1F/LC2F/LC3F/LC1Sb light-chain pattern. By these criteria type IIA fibres could be distinguished from type IIC intermediate fibres, which showed coexistence of fast-twitch-fibre and slow-twitch-fibre forms of myosin light chains and of troponin subunits.     

Structural variations in pili expressed during gonococcal infection

Gonococci were cultured from the urethra of male patients and from the cervix and urethra of their female partners. SDS-PAGE of cell lysates from within each group of consorts showed that outer membrane protein I remained constant but considerable variations were seen in the apparent molecular weight of protein II. Pili were purified from the isolates of some groups of consorts. In each case the pili expressed by the isolates from the female cervix and urethra differed in subunit molecular weight and were usually also distinct from the pili expressed by the isolate from the male partner.     

Congenital muscle fibre type disproportion: clinical, morphological and biochemical findings in children

Muscle biopsies from quadriceps femoris muscle of normal subjects and subjects with symptoms of congenital myopathy, aged 1-3 years, were examined for morphological and biochemical differences. Four patients showed clinical signs of Congenital Fibre Type Disproportion (CFTD) as described originally by Brooke (1973), while the other cases showed only signs of hypotonia and diffuse weakness as described by Clancy et al. (1980). No morphological differences between patients with different clinical signs were found in any biopsy sample from the quadriceps femoris muscle, as regards fibre size; type I fibres were always smaller than type II fibres. The difference in fibre size was more than 15% in all cases, and the variability coefficient of the larger type II fibres was less than 250. Nevertheless, as regards fibre occurrence, two patients showed more type I fibres than type II fibres, four patients showed more type II fibres than type I fibres and one patient had a marked type II fibre predominance. Enzyme activities assayed in the crude mitochondrial fraction showed no abnormalities between normal subjects and patients. An increase in the activity of lactate dehydrogenase in the crude extract was found in all cases.     

鳗鲡繁殖生物学研究 Ⅱ.下海雌鳗脑垂体超显微构造的研究

神经垂体主要由神经分泌纤维、脑垂体细胞和微血管组成。神经分泌纤维主要是无髓鞘神经纤维,也有一些是有髓鞘神经纤维。神经垂体中还有一些多层体构造。神经分泌纤维有两个基本类型:A型纤维含有直径为1250—1750Å的神经分泌颗粒;B型纤维含有直径为450—1000Å的颗粒状囊泡。腺垂体的分泌细胞按其超显微构造的特点和所含的分泌颗粒大小不同可以区分为六个类型:催乳激素分泌细胞、促甲状腺激素分泌细胞,促肾上腺皮质激素分泌细胞、促生长激素分泌细胞、促性腺激素分泌细胞和后腺垂体的分泌细胞。