Intranuclear paracrystals observed in striated muscle specific LIM protein-deficient mouse cardiomyocytes.

A paracrystalline structure was observed within left ventricular cardiomyocyte nuclei of MLP(-/-) mice. The paracrystal possessed cross lines, approximately 8.0 micro m long and 0.3 micro m wide, with a slender spindle shape and a periodicity of 13 nm. Paracrystals were best observed along the longitudinal orientation of myofibrils and were detected in less than 10% of the nuclei observed. One dimension of the protein unit forming the paracrystal was 8.5 nm long. The electron density of the paracrystal appeared to be slightly higher than that of heterochromatin, suggesting that RNA-associated proteins are constituents of the paracrystal. This is the first report of intranuclear paracrystals in cardiomyocytes, which appear to be unique to MLP(-/-) mice.

Effect of beta-aminopropionitrile and hyaluronic acid on repair of collagenase-induced injury of the rabbit Achilles tendon.

Collagenase was injected into the Achilles tendon of both hind legs of 10 clinically normal adult male New Zealand white rabbits. One month after induction of the injury, beta-aminoproprionitrile (BAPN) or hyaluronic acid (HA) was injected into the tendon core of the right hind leg of each rabbit, the left hind leg being left untreated. The treatment effects were evaluated by electron microscopy and analysis of the glycosaminoglycan (GAG) content of samples at 2 and 6 months post-treatment. At 2 months, collagen fibrils in tendons from both hind legs were relatively small in diameter, irregularly arranged, and interspersed with abundant active tenocytes as compared with those in normal tendon uninjured by collagenase. In the matrix, the amount of HA increased, but chondroitin-6-sulphate was eliminated. At 6 months, BAPN-treated tendons had small-diameter, regularly arranged collagen fibrils. HA-treated tendons, on the other hand, had large diameters, as well as regularly arranged collagen fibrils by comparison with non-treated tendon. The results suggest that HA, unlike BAPN, promoted healing.

Characteristics of dorsal lingual papillae of the Bactrian camel (Camelus bactrianus).

The morphology of dorsal lingual papillae of the Bactrian camel (Camelus bactrianus) was studied by using light and scanning electron microscopy. Filiform and lenticular papillae were considered as mechanical papillae but fungiform and vallate papillae were considered as gustatory papillae. Filiform papillae were distributed mostly in the anterior two-thirds region of the tongue. Each filiform papilla consisted of one primary papilla and a few smaller secondary papillae. Lenticular papillae were distributed on the torus linguae. The larger papillae were arranged in two parallel lines medially whereas the smaller papillae were laterally located. Most of the fungiform papillae were found on the lateral margins of the anterior two-thirds of the tongue. These papillae were small and round. Intra-epithelial taste buds were located on the dorsal surface of each papilla. The vallate papillae were arranged in two rows on each rim of the torus linguae. Each round- and flat-shaped vallate papilla was surrounded by a prominent gustatory groove and an annular pad. A few taste buds were observed in the lateral epithelium of the papillae. The keratinization of the covering stratified squamous epithelium of the mechanical lingual papillae was relatively thicker than those of the gustatory papillae. The lingual papillae of the Bactrian camel exhibited some different characteristics from other domestic ruminants. These morphological characteristics of the tongue of the Bactrian camel might have evolved to assist the camel in prehension and manipulating of the inorganic stiff plants that grow in its environment and therefore might relate to the feed and feeding habits of the animal.

Prevalence, vasculature, and innervation of myocardial bridges in dogs.

OBJECTIVE: To report gross anatomic examination of the canine myocardial bridge (MB), a muscular band found above the coronary artery (CA), with respect to its occurrence, location, vascularization, and innervation. SAMPLE POPULATION: 629 canine hearts obtained within 1 to 3 hours after euthanasia. PROCEDURE: After an incision was made at the left fifth intercostal space, the pericardial sac was cut open, and if an MB was present, the heart, lungs, and annexed structures were removed together and subsequently subjected to macroscopic examination of MB musculature and innervation after formalin fixation. Vascular casting was performed by use of methyl methacrylate perfusion. RESULTS: Of the 629 canine hearts examined, 189 (30%) had MB, occurrence of which was independent of sex, age, and breed. Among 13 MB-containing specimens examined in detail, there was great variation in thickness (0.11 to 2.24 mm; mean, 0.45 mm) of MB and distance (24 to 236 microns; mean, 103 microns) between the MB and the paraconal interventricular branch of the left CA (PIBL). One pair or 2 pairs of blood vessels from the PIBL supplied the MB muscle. Venous blood returned to the coronary circulation via the branches of the great coronary vein coursing on both sides of the PIBL, in close contact with the PIBL and the groove wall. The 2 veins rejoined at the upper portion of the PIBL and passed obliquely to the coronary groove under the left auricle, and finally drained the blood through the coronary sinus into the right ventricle. Innervation to the MB muscle was derived from nerve branches of the middle cervical ganglion and left vagus nerve. CONCLUSION: Prevalence and localization of MB in dogs and human beings are similar. Vascularization of the MB muscle originates from the PIBL. The cervical ganglion and vagus nerve control the MB muscle.

Equine synovial villi: distinctive structural organization of vasculature and novel nerve endings.

The structural arrangement and cellular distribution of endothelial and lining cells of the synovial villi were studied in the equine palmar/plantar recess of the metacarpo- and metatarsophalangeal joints by light microscopy and electron microscopy. The extent and distribution of blood vessels varied with villous shape and length. The majority of vessels formed concentric circles in cross and longitudinal sections and probably are arranged in a convoluted, spiral or helical pattern. The villi do not contain smooth muscle cells or typical capillaries as observed in other organs. Under the electron microscope, the endothelium is surrounded by connective tissue and discontinuous circular cells, presumably fibroblasts. The outermost layer was sometimes surrounded by type A and/or B synovial cells. The lumen of the blood vessels at the top of villus appeared to be constricted in most cases, with a diameter of about 12 +/- 3 microns. Blood vessels formed by more than six endothelial cells in the middle portion of villus generally were not constricted. Well-developed cytoplasmic processes extended into the lumen of blood vessels. The constriction of blood vessels with no apparent smooth muscle presence and the observation of numerous intermediate filaments in the cytoplasm of the endothelial cells suggests that these villous blood vessels constrict through contraction of their own endothelial cells. Lining cells were distributed unevenly even within a single villus; the villous lining cells seemed to have directional preferences with domination of synovial type A cells. Surprisingly, structures resembling myelinated nerve ends (approximately 0.2 microns) were observed between juxtaposed endothelial cells as well as directly on an endothelial cell, suggesting that these nerve endings may be a sensor detector of either pressure or temperature or have a proprioceptive-like function. Synovial villi have a distinctive structural arrangement of vessels, lining cells, and nerve endings.

A novel phasic contraction induced by dithiothreitol in frog skeletal muscle.

1. Dithiothreitol (DTT), at 50-100 mM, induced a phasic reversible contraction of frog skeletal muscle. 2. Exposure of single fibers to nifedipine (20 microM), an L-type Ca2+ antagonist, blocked the twitch and tetanus tensions but never affected the DTT-induced contraction. 3. DTT also produced a phasic contraction in fibers where voltage sensors were inactivated in the presence of high K+ concentration (190 mM). 4. A fiber was mechanically skinned after observation of DTT-induced contraction. The skinned fiber contracted in response to a DTT concentration similar to that required to produce contraction in intact fibers before skinning. 5. In skinned fibers, DTT, at 100 or 200 mM, inhibited the accumulation of Ca2+ by SR, but not Ca2+ ATPase activity. 6. These results suggest that a high concentration of DTT triggers Ca2+ efflux from the SR through action on the Ca2+ release channel and/or closely associated proteins, such as triadin and FK-506 binding protein.

Cardiac rod body: hypertrophic Z-line in an aged pony.

Numerous rod bodies were found in a heart sample from a 33-year-old pony by a conventional electron-microscopic technique. The rod bodies were concentrated in localized areas of both atria and ventricles, without a specific pattern of distribution. The rods appear to have a typical crystalline-like structure which presumably contains actin backbone filaments and alpha-actinin as major protein constituents. Diminution, fragmentation, and disorganization of the myofibrils, random expansion of electron-dense materials, especially at the fasciae adherens of the intercalated disc, an increase of intercellular space, frequently associated with connective tissue elements, and accumulation of glycogen granules, lipofuscin, mitochondria, and nonviable organelles in the rod body-containing cells suggested that rod-containing myocytes are in a degenerating stage and that rod body formation occurs in atrophic cardiomyocytes rather than normal or hypertrophic cardiomyocytes in the aged pony heart.

Myocardial bridging as a factor in heart disorders: critical review and hypothesis.

The purpose of this report is to review the previous and current methods for the detection of a MB and to summarize results of work by other investigators as well as our own recent morphological studies with emphasis on work since 1983. We will discuss the presumable association of MBs with heart disorders, on the basis of significant publications. We describe the importance of both basic and clinical research for the assessment of MB influence in various heart disorders. Aspects covered are: (1) historical background of MBs and the relationship between the MB and CA; (2) general characteristics of MBs; (3) methods for studying MBs; (4) compressive effect of the MB on the CA; (5) clinical symptoms including myocardial ischemia, angina pectoris, myocardial infarction, development of atherosclerosis and thrombosis, ventricular fibrillation and sudden death; (6) treatment of MB-associated heart disorders by resection of MBs and (7) future research trends.

Myocardial bridge muscle on left anterior descending coronary artery differs from subepicardial myocardium of the left ventricle in dogs.

The myocardial bridge (MB) is a muscle band found sporadically above the coronary artery (CA) in humans and certain animals such as the dog, cat and sheep. The purpose of our study was to compare the structure of the MB muscle with that of tissue from the subepicardial myocardium. The histological studies included toluidine blue staining of 1-micron-thick sections and Gomori's trichrome staining of canine cardiac samples. The MB muscle of the dog heart is characterized by a distinctive spatial arrangement, with individual fibers separated by substantial elements of intercellular connective tissue in cross-section. Longitudinally, the long, slender fibers are aligned continuously with intermediation of intercalated disks lying perpendicular to the long axis of the fibers. In other regions of the left ventricular subepicardial myocardium, each myocyte is tightly packed in transverse view. There is great variation in the thickness (0.11-2.24 mm, average 0.45 mm) of MBs and the distance (24-236 microns, average 103 microns) between MBs and the left anterior descending coronary artery (LAD) among the 13 affected dogs examined, with no apparent relationship between the occurrence of MBs and either age or sex. These results on MB alignment suggest that the MB muscle generates force along the long axis of the fiber orientation as skeletal muscle does, and with minimal constriction of the CA; if so, the function of MB myocytes may differ from that of common cardiac myocytes, as does the structure. Then, the long-supposed downward compressive force of MBs on the LAD would be minimal in most cases; however, when the MB produces a systolic narrowing of the LAD known as the milking effect, the degree of lateral compression and its influence should depend not only on the substantial size of the MB muscle but also on the distance between the MB muscle and LAD. The environment surrounding the LAD may be a crucial factor in determining whether the MB influences the induction of heart disorders or not.

Is there a protease that preferentially cleaves the M-line in partially dehydrated muscle?

When a partially dehydrated muscle fibre bundle (PDM, 65% H(2)O, pH 5.5, at 4 °C) was treated with a supernatant fraction (M-line-cleaving fraction: MCF) of muscle homogenate for 5 hr, the M-lines disappeared. MCF was extracted from rabbit skeletal muscles by homogenization with 15 mM HCl containing 0.5 M NaCl (pH 3.7), fractionated with 25-65% (NH(4))(2)SO(4) and clarified by Sephadex G-75. Rabbit psoas PDM was obtained with an osmotic dehydration sheet and glycerinated. One end of the bisected fibre bundle was incubated with 10 mM Na-acetate (pH 5.5), 1 mM EDTA, 5 mM ß-mercaptoethanol (ß-MCE), 150 mM KCl, 10 mM NaN(3) with MCF at 25 °C for 5 hr, the muscle being stretched and relaxed several times. The other end was incubated in the same solution, except that MCF was omitted (control). Electron microscopy showed the myofibrils broken down at the M-line in the presence of MCF. The myofilaments were closely packed near the Z-line and flared out at both ends near the centre of the sarcomere (bow-tie shape). Thus, the Z-line is not the only target of proteases and structural decomposition can also occur at the M-line under specific conditions. An M-line cleaving protease may exist in the MCF muscle extract.

DNase I interaction on muscle Z-line.

The effect of deoxyribonuclease I on muscle Z-line structures was re-examined. Under conditions of deoxyribonuclease I activation (presence of the divalent cation Ca2+ and Mg2+), a deoxyribonuclease I preparation did not affect Z-line structure if phenylmethylsulfonylfluoride, an inhibitor of serine proteases, was also present. In the absence of protease inhibitor, both Z-lines and M-lines were digested, even in the presence of EDTA and EGTA as inhibitors of deoxyribonuclease I. These electron microscopic observations were consistent with the following results from sodium dodecyl sulphate gel electrophoresis: when the protease was inhibited but deoxyribonuclease I was activated, myofibrillar proteins remained essentially intact. However, degradation of proteins in both rabbit psoas and chicken pectoralis myofibrils was observed in the presence of deoxyribonuclease I inhibitors when the protease inhibitor was absent. Our data strongly suggest that the interaction of deoxyribonuclease I with Z-line proteins previously reported is most likely due to contamination of the deoxyribonuclease I fraction by the serine-type proteases.

Hypertrophic Z-line observed in aged one-humped camel (Camelus dromedarius).

The ultrastructure of cardiac muscle from 3 aged one-humped camels (Camelus dromedarius) was examined to see if aged camels also develop hypertrophic Z-lines as observed in the heart of aged humans and other animals. The results proved that the aged camel heart develops hypertrophic Z-lines together with other age-associated alterations, including loss of myofibrils and myofilaments, increase of intermyofibrillar spaces and intercellular spaces, and disruption of mitochondrial cristae and intercalated disc structure. Hypertrophic Z-lines are somewhat smaller than those of other species, but occur both in the atria and ventricles. Although the majority of cardiac muscle samples revealed a normal structure, the finding of hypertrophic Z-lines in camel heart is the first such observation in a ruminant animal. The anomalous camel muscle resembles that of other species in terms of cell alterations commonly associated with aged hearts.

Proximal paraconal interventricular myocardial bridge in dog: ultrastructural characterization.

The ultrastructure of the dog myocardial bridges (MB), named here the proximal paraconal interventricular-myocardial bridge (PPI-MB), was examined to determine if its structure is identical to that of ordinary cardiac muscle. This study is preparatory to the establishment of PPI-MB as a physical and physiological model system to examine the influence of MB on coronary flow in cardiac function. Five of 46 dog hearts examined possessed PPI-MB, the occurrence of which was independent of sex, age and breed. The dog PPI-MB was localized in a limited area in the proximal third of the paraconal interventricular groove. PPI-MB myocytes and myofibrils were arranged quite straight and smooth, with minimum branching. PPI-MB myocytes formed side-by-side cell junctions through the mediation of mitochondria. Connective tissue elements were involved in terminal cell adhesion to form the myomyous junction beside the intercalated disc. The excitation conductive system of sarcoplasmic reticulum-T tubule often showed a triad. Numerous nerve axons or nerve ending-like structures appeared. Many structural views deviated somewhat from common cardiac muscle structure. This paper represents the first detailed description of PPI-MB muscle.