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M Manabe

Publications and source records attributed to M Manabe.

At least 37 records · Page 2Linked to original sources
PubMed1990

Tissue-specific distribution of a novel component of epithelial basement membranes.

Monoclonal antibodies against basement membrane (BM) were generated using the matrix deposited by cultured rabbit corneal epithelial cells as immunogen. BM antibodies were identified by immunofluorescent staining of frozen tissue sections and of extracellular matrix of living cultured cells. BM localization was confirmed by immunoelectron microscopy. Antibody AE26 immunoprecipitates a 140,000 Mr component from radiolabeled corneal epithelial cells and recognizes this component plus a 95,000 Mr band on Western blots. The antigen resists extraction by high and low salt and by nonionic detergents, but is solubilized in 4 M urea/1% mercaptoethanol. On isoelectric focusing and nonequilibrium pH gradient gels, AE26 antigen migrates to the acidic region (pI less than 3). The molecule is destroyed by trypsin, but is insensitive to bacterial collagenase. In frozen tissue sections, AE26 stains only BM of stratified epithelia plus trachea, ureter, lung, and intestine, but no other epithelial or nonepithelial BM. AE26 antigen is detected on Western blots of cornea, skin, and lung extracts, but not liver, kidney, or muscle, indicating that this is not due to masking of the epitope. This tissue distribution is different from any previously described BM molecule. Although we have not ruled out the possibility that AE26 recognizes a modification or fragment of a known BM component (particularly entactin), the acidic pI, collagenase resistance, and unusual tissue specificity suggest that AE26 recognizes a new BM protein. The BM heterogeneity demonstrated by AE26 may play a structural role or provide positional signals to the overlying epithelium.

Animals↗
PubMed1990

Uroplakin I: a 27-kD protein associated with the asymmetric unit membrane of mammalian urothelium.

The luminal surface of mammalian urothelium is covered with numerous plaques (also known as the asymmetric unit membrane or AUM) composed of semi-crystalline, hexagonal arrays of 12-nm protein particles. Despite the presumed importance of these plaques in stabilizing the urothelial surface during bladder distention, relatively little is known about their protein composition. Using a mouse mAb, AE31, we have identified a 27-kD protein that is urothelium-specific and is differentially expressed in superficial umbrella cells. This protein (pI approximately 5.8) partitions into the detergent phase during Triton X-114 phase separation. Pulse-chase experiments using cultured bovine urothelial cells showed that this protein is synthesized as a 32-kD precursor that is processed through a 30-kD intermediate, to the mature 27-kD form. In cytoplasmic vesicles containing immature AUM, the AE31 epitope is detected in patches on the cytoplasmic side, but in mature, apical AUM it is detected exclusively on the luminal side. This suggests an unusual translocation of the AE31 epitope during AUM maturation; more data are required, however, to substantiate this interpretation. Immunoaffinity purification of the 27-kD protein results in the copurification in approximately molar ratio of a 15-kD protein, as well as a small and variable amount of a 47-kD protein. Immunoblotting data indicate that these three proteins are immunologically distinguishable. This copurified 15-kD protein is relative basic (pI approximately 8.0). Like the 27-kD protein, it is urothelium-specific and is present mainly in the umbrella cells. Together, our data indicate that a 27-kD protein is urothelial plaque-associated (uroplakin I). Based on complex formation data, we provisionally name the 15-kD protein uroplakin II; additional data will be required to determine whether this and the 47-kD protein are integral parts of AUM. The identification of these AUM-associated and -related proteins, plus the availability of a culture system capable of synthesizing and processing some of these molecules, offer new opportunities for studying the detailed structure, assembly, and function of asymmetrical unit membrane.

Animals↗
PubMed1990

Assessing the differentiation state of cultured bovine urothelial cells: elevated synthesis of stratification-related K5 and K6 keratins and persistent expression of uroplakin I.

Although significant progress has recently been made in culturing mammalian urothelial cells, relatively little is known about their biochemical differentiation. In this paper, we assessed the differentiation state of cultured bovine urothelial cells by analyzing their keratins and a cell surface marker, uroplakin I. Urothelial cells were serially cultured either in a serum-free medium, or in a serum-containing medium in the presence of 3T3 feeder cells, with similar results. Despite their stratified appearance, both normal urothelium and cultured urothelial cells synthesize mainly K8, K18 and K19, keratins that are typically seen in simple epithelia. However, cultured urothelial cells synthesize a greatly increased amount of K5 and K6 keratins, which are usually expressed by stratified epithelia but present only in trace amounts in normal urothelium. These data indicate that, as far as keratin synthesis is concerned, cultured urothelial cells undergo an altered pattern of differentiation towards a more 'stratified phenotype'; this unusual finding has interesting implications for urothelial evolution. In the meantime, many superficial cells in cultured urothelial colonies make uroplakin I, a 27 x 10(3) Mr protein subunit of the asymmetrical unit membrane (AUM) characteristic of urothelial (superficial) umbrella cells. These results indicate that cultured urothelial cells undergo, at least in part, AUM biogenesis. Cultured urothelial cells thus provide a useful experimental model system for studying certain early steps of AUM formation.

Animals↗
PubMed1990

[Nutritional condition of epidermolysis bullosa patients assessed from body height and weight].

Exact body height and weight data of patient with epidermolysis bullosa simplex (EBS) and recessive dystrophic epidermolysis bullosa (RDEB) were collected through a nation-wide epidemiologic investigation concerning epidermolysis bullosa (EB) patients. This data was then compared with the averaged body height and weight of age and sex matched normal controls. The nutritional condition of the EB patients was evaluated statistically. The data showed that height of female EBS patients was less than the controls (p less than 0.01), and furthermore, that height and weight of male and female RDEB patients was also less than the controls (p less than 0.01). These results indicated that EB, especially RDEB patients, were in the poor nutritional condition, and that the adequate nutritional management might significantly influence the prognosis of these patients.

Adolescent↗
PubMed1990

[Epidural anesthesia with high dose fentanyl for abdominal surgery].

An epidural catheter was inserted at T9-L2 interspace and 10 micrograms.kg-1 fentanyl with (E+) or without (E-) epinephrine 1:100,000 was given for 82 elective abdominal surgeries. N2O 66%, enflurane and muscle relaxant were used as needed. The onset and the duration of the action were estimated to be approximately 15 minutes and 4 hours, respectively. Anesthesia was maintained with enflurane below 0.4% (0.22 +/- 0.09%) in 70 patients (85.4%). E+ group needed significantly lower concentration of enflurane than E- group. There was no severe hemodynamic change during the operation. Systolic pressure, diastolic pressure and heart rate during the operation were 115.2 +/- 16.0 mmHg, 69.4 +/- 10.8 mmHg and 74.2 +/- 11.4 min-1, respectively, each of which was about 18% less than the values on arrival in the operating room. Sixty-one patients (82.5%) woke rapidly. Almost all patients felt well and had no pain during the recovery period. Naloxone 0.05-4 mg was administered intravenously in 21 patients (31.7%) whose respiratory rate was below 10 min-1. The patients with shorter operation time (shorter than 2.5 hours) needed more naloxone. Troubles of respiratory depression did not occur in the recovery room and in the ward in both naloxone and non-naloxone groups. This anesthesia method which induces mild depression of blood pressure and heart rate may be indicated for patients with ischemic heart disease or with poor cardiac function, but has no advantages in patients with poor respiratory function who need early extubation after a short operation.

Abdomen↗
PubMed1989

Excitation and inhibition of medullary inspiratory neurons by two types of burst inspiratory neurons in the cat.

In Nembutal-anesthetized and artificially ventilated cats, we studied the connectivity of burst inspiratory (I) neurons in the Bötzinger complex and the ventral respiratory group (VRG) with spike-triggered averaging methods. Burst I neurons exhibited tonic (I-TON) or decrementing (I-DEC) firing patterns. Spikes of I-TON neurons induced monosynaptic EPSPs in intracellularly recorded I neurons of both the VRG and the dorsal respiratory group (DRG). Spikes of I-DEC neurons induced monosynaptic inhibitory postsynaptic potentials (IPSPs) in both VRG and DRG I neurons.

Action Potentials↗
PubMed1989

Axonal trajectory and terminal distribution of inspiratory neurons of the dorsal respiratory group in the cat's medulla.

In Nembutal-anesthetized and artificially ventilated cats, we studied the morphological properties of the inspiratory neurons of the dorsal respiratory group (DRG) with HRP intracellular staining. A total of 37 neurons were stained and their axonal trajectories and terminal distribution in the medulla were analyzed. The somata were located predominantly in the ventrolateral region of the nucleus of the solitary tract and were distributed between 2,300 mum rostral and 700 mum caudal to the obex. Most (26/33) of the neurons tested were antidromically activated by the stimulation of the contralateral (n = 24) or ipsilateral (n = 2) cervical cord. To examine the existence of collateral branches in the brainstem, we traced axonal trajectories in 28 neurons. In most cases, the stem axons issuing from the cells of origin coursed ventrally and then turned medially to cross the midline without giving off any axon collaterals. However, six neurons had axonal collaterals in the brain stem ipsilateral to the somata. At least four types of collateralization were observed. The stem axon of the first type bifurcated at the area ipsilateral and ventral to the cell body. One branch crossed the midline to project to the spinal cord, and the other, thinner branch descended caudally in the ipsilateral medullary reticular formation without distributing any terminals. The axon of the second type projected to the contralateral spinal cord and distributed collateral branches with terminal boutons in the ipsilateral ventral respiratory group (VRG). The third type projected to the contralateral spinal cord and distributed terminal boutons in the medial part of the nucleus of the solitary tract (NTS) and its vicinity. The fourth type distributed numerous branches with terminal boutons in and around the ventral part of the NTS and the VRG area. This study indicates that some inspiratory neurons of the DRG influence not only spinal respiratory neurons but also medullary respiratory neurons in the vicinity of the DRG and the VRG.

Animals↗
PubMed1989

Morphology of augmenting inspiratory neurons of the ventral respiratory group in the cat.

The present study examined, in Nembutal-anesthetized and artificially ventilated cats, the morphologic properties of the inspiratory neurons of the ventral respiratory group (VRG). Horseradish peroxidase (HRP) was injected into 21 augmenting inspiratory or late inspiratory neurons with peak firing rates in the late inspiratory phase. The majority of the stained neurons were antidromically activated by stimulation of the cervical cord. Thirteen somata, located within or around the nucleus ambiguus (AMB), between 100 microns caudally and 2,000 microns rostrally to the obex, were stained. In ten cases, the stem axons issuing from the cells of origin coursed medially to cross the midline without giving off any axonal collaterals. Three neurons gave rise to axonal collaterals on the ipsilateral side, distributing boutons in the medullary reticular formation, in the vicinity of the AMB, hypoglossal nucleus, solitary tract, and dorsal motor nucleus of the vagus. In eight neurons, only the axons were labeled; in four of these, which were antidromically activated from the spinal cord, the stem axons crossed the midline 2,000-3,000 microns rostral to the obex and descended in the reticular formation around the AMB down to the cervical cord. They issued several axonal collaterals, distributing terminal boutons at the level of the caudal end of the retrofacial nucleus and about 1,000 microns rostral and caudal from the obex. Terminals were found mainly in and around the AMB, and a few were found in the vicinity of the dorsal motor nucleus of the vagus. The remaining four nonactivated axons distributed their terminal boutons widely in the reticular formation around the AMB. Thus, the augmenting inspiratory neurons of the VRG were shown to project not only to the spinal cord, but also to the VRG, hypoglossal nucleus, and dorsal motor nucleus of the vagus.

Action Potentials↗
PubMed1989

Monosynaptic excitation of medullary inspiratory neurons by bulbospinal inspiratory neurons of the ventral respiratory group in the cat.

In Nembutal-anesthetized, immobilized, and artificially ventilated cats, we studied the connectivity of medullary collaterals of bulbospinal inspiratory (BS-I) neurons in the ventral respiratory group (VRG). BS-I neurons which projected to the contralateral spinal cord were isolated extracellularly and intracellular recordings were made from the respiratory neurons in the contralateral VRG. The intracellular membrane potentials were averaged using extracellular spikes of the BS-I neurons as triggers (spike-triggered averaging method). Fast-rising and short-lasting depolarizing potentials locked to the triggering spikes were obtained and shown to be unitary EPSPs induced monosynaptically by the medullary collaterals of BS-I neurons. A total of 137 pairs were analyzed and unitary EPSPs were found in 11 pairs. Four BS-I neurons and 7 inspiratory vagal motoneurons received EPSPs from the medullary collaterals of BS-I neurons. These findings suggest that 1) BS-I neurons in the VRG drive medullary motoneurons of accessory respiratory muscles and phrenic or intercostal motoneurons simultaneously, 2) BS-I neurons on both sides synchronize via the excitatory connections, and 3) the augmenting firing pattern of BS-I neurons might partly be produced by this reexcitatory connection within the population of BS-I neurons.

Action Potentials↗
PubMed1989

Expression of hair-related keratins in a soft epithelium: subpopulations of human and mouse dorsal tongue keratinocytes express keratin markers for hair-, skin- and esophageal-types of differentiation.

The dorsal surfaces of mammalian tongues are covered with numerous projections known as filiform papillae whose morphology varies in different species. Using a panel of monoclonal antibodies to keratins as probes, we have established that, in both human and mouse, the interpapillary epithelia express mainly the "esophageal-type" keratins, while the papillary epithelia express "skin-type" keratins as well as some keratins reacting with a monoclonal antibody (AE13) to hair keratins. The AE13-reactive proteins of the mouse were found to be very similar to those of authentic mouse hair keratins. However, the corresponding protein of human tongue appears to be different from all known human keratins. This protein has a MW of 51K; it is relatively acidic; it is sulfhydryl-rich, as revealed by iodoacetic acid-induced charge and apparent size shift; it shares an epitope with all the known acidic human hair keratins; and it is associated with keratin fibrils in vivo. This protein may therefore be regarded as a novel type I "hard" keratin. These data establish that mammalian dorsal tongue epithelia can be divided into at least three compartments that undergo mainly "esophageal-", "skin-" and "hair"-types of differentiation. Different keratin filaments, e.g., those of the esophageal- and hair-types, exhibit strikingly different degrees of lateral aggregation, which can potentially account for the different physical strength and rigidity of various cellular compartments. Our data also suggest the possibility that variations in papillary structure in human and mouse may arise from different spatial arrangements of specific keratinocytes, and/or from the expression of specialized hair-related keratins.

Animals↗
PubMed1989

Association of a basic 25K protein with membrane coating granules of human epidermis.

Keratinocytes of the upper granular layers contain unique round-to-oval granules, 100-500 nm in diameter, in their peripheral cytoplasm. These granules (known as membrane coating granules [MCG], or lamellar granules) fuse with the apical cell surface of uppermost granular cells and discharge their contents into the intercellular space, where they are believed to play a role in establishing the permeability barrier of the epidermis and possibly in regulating the orderly desquamation of terminally differentiated keratinocytes. Using two monoclonal antibodies originally prepared against hair follicle antigens, we have identified a 25K epidermal protein in association with both MCG-like granules in the peripheral cytoplasm of granular cells as well as MCG-derived intercellular material. This protein is relatively basic (pI greater than 8), largely aqueous soluble, methionine deficient, and is relatively abundant in epidermis (comprising up to approximately 0.1% of soluble proteins). Its distribution is restricted to the granular layer of keratinized (cornified) stratified squamous epithelia. The identification of this protein component opens new avenues for studying the molecular mechanisms underlying the establishment of permeability barrier and/or regulation of desquamation.

Animals↗
PubMed1989

Basement membrane heterogeneity and variation in corneal epithelial differentiation.

We have previously shown that the expression of a major 64-Kda keratin (K3) in corneal epithelium is site-related. It is found suprabasally in limbal epithelium, but uniformly (basal cells included) in central corneal epithelium. In the present study, we used a panel of antibodies against various components of corneal epithelial basement membrane to investigate a possible correlation between basement membrane heterogeneity and differential (basal vs. suprabasal) K3 keratin expression. One of these antibodies, AE27, stains human conjunctival basement membrane weakly, limbal basement membrane heterogeneously, and central corneal basement membrane strongly. Basal cells resting on basement membrane that stains strongly with AE27 tend to stain with monoclonal antibody AE5, which recognizes keratin K3. Basal cells on basement membrane staining weakly with AE27 tend not to stain with AE5. No such correlation exists between AE5 staining and type IV collagen, which is detectable immunohistochemically in conjunctival and limbal basement membrane, but not in corneal basement membrane overlying Bowman's layer. These results suggest that basement membrane of human corneal/conjunctival epithelium can be divided into at least three domains: the conjunctival basement membrane (type IV collagen-positive, AE27-weak), the limbal basement membrane (type IV collagen-positive, AE27-strong), and corneal basement membrane (type IV collagen-negative, AE27-strong). The results also raise the possibility that basement membrane heterogeneity may play a functional role in regulating keratin expression and other aspects of differentiation of corneal epithelium; more experiments are needed to test this hypothesis.

Animals↗
PubMed1989

[A study on the estimation of prevalence of epidermolysis bullosa in Japan].

An estimation of the prevalence of epidermolysis bullosa was carried out in Japan. A questionnaire was sent by mail to the departments of dermatology and pediatrics in 644 main hospitals throughout Japan requesting confirmation of the number of epidermolysis bullosa cases experienced. The response rate was 63.0% or 406 hospitals. Based on the secondary questionnaire survey, we confirmed 393 cases of epidermolysis bullosa, and then divided them into the five following groups: "simplex" type 182 (48.2%), "junctional" type 23 (6.0%), "dominant dystrophic" type 72 (18.7%), "recessive dystrophic" type 109 (28.2%), unclassified 7 (1.8%). Using the respondent rate by geographic region and clinical department, the estimated prevalence of each type of epidermolysis bullosa was calculated as follows: "simplex" type 340-470 (0.29-0.40 per 100,000 population), "junctional" type 18-24 (0.015-0.020), "dominant dystrophic" type 130-180 (0.11-0.15) and "recessive dystrophic" type 180-250 (0.15-0.21). The total number of epidermolysis bullosa patients in Japan was estimated as being more than 670 in 1983.

Epidermolysis Bullosa↗
PubMed1989

[Surgical treatment of esophageal stenosis with recessive dystrophic epidermolysis bullosa--esophageal dilatation using balloon catheter method].

We report the details of our treatment of 5 patients with recessive dystrophic epidermolysis bullosa (RDEB). Good results were obtained by esophageal dilatation using the balloon catheter method. Prior to the operation, we corrected the patients' nutritional status with oral supplementation of Hinex-R as well as intravenous drip infusion of Intra-lipid, based on the nutrition index obtained by body weight, serum albumin, rapid turnover proteins, and blood minimum essence. After sufficient anesthesia of the nasopharyngeal mucosa with xylocain spray, we inserted a Microvasive Rigiflex Balloon Dilator into the stenotic region while scanning the upper GI. Then we dilated gradually by barium injection. The patients' condition improved after this treatment and follow up X-ray examination revealed no recurrence of stenosis. There were several advantages to this method: 1) It was safer and could be done repeatedly. 2) It caused less stress for patients. 3) Its effectiveness appeared rapidly. We analyzed the frequency and severity of esophageal stenosis of 12 cases with EB in our department. Ten out of 12 cases complained of subjective esophageal symptoms. Esophageal stenosis was observed in all 8 cases examined by esophagogram. We concluded that this surgical procedure for esophageal stenosis was as important as digit reconstruction in the management of RDEB.

Adolescent↗
PubMed1988

Distribution of medullary respiratory neurons in the rat.

In Nembutal-anesthetized and spontaneously breathing rats, a total of 226 respiratory neurons were recorded in the medulla extending from the caudal end of the facial nucleus to 1 mm caudal to the obex. They were classified into inspiratory (I) and expiratory (E) neurons by their temporal relationships to diaphragm EMGs. One hundred and seventeen I and 108 E neurons were identified. I and E neurons were further classified into augmenting, decrementing, and other types based on their firing patterns. Almost all the respiratory neurons recorded were located around the nucleus ambiguus and the nucleus retroambigualis, corresponding to the ventral respiratory group (VRG) of the cat. On the other hand, only a few respiratory neurons were identified around the ventrolateral nucleus of the solitary tract, corresponding to the dorsal respiratory group of the cat. In the VRG, 3 subgroups were distinguished rostrocaudally. One group of E neurons was located ventrally to the rostral part of the nucleus ambiguus, presumably corresponding to the Bötzinger complex defined in the cat. Another group of E neurons extended caudally beyond the obex, from the caudal portion of the nucleus ambiguus through the nucleus retroambigualis. Between these two groups of E neurons, an assembly of predominantly I neurons existed in the vicinity of the nucleus ambiguus. These characteristics of distributions were basically similar to those of the VRG of the cat.

Action Potentials↗
PubMed1988

Efferent projections of inspiratory neurons of the ventral respiratory group. A dual labeling study in the rat.

The efferent projections of the medullary respiratory neurons of the rat were studied using an anterograde tracer, Phaseolus vulgaris leucoagglutinin (PHA-L). In Nembutal-anesthetized rats, PHA-L was iontophoretically applied to (1) the area of inspiratory neurons of the ventral respiratory group (VRG) around the nucleus ambiguus, or (2) the area ventrolateral to the solitary tract. In addition, a fluorescence retrograde tracer, Fast blue (FB), was injected into the cervical phrenic nerve several days after the PHA-L injection. When PHA-L was injected into the area of predominantly inspiratory neurons of VRG, dense PHA-L-labeled axons were observed bilaterally in the spinal cord: the ipsilateral projections were noticeably denser than the contralateral ones. Fine axonal branches were distributed around a column of the phrenic motoneurons and boutons were observed on the somata of the FB-labeled motoneurons, suggesting monosynaptic connections between VRG inspiratory neurons and phrenic motoneurons. On the other hand, when PHA-L was injected into the area ventrolateral to the solitary tract, only a few descending axons to the spinal cord were seen bilaterally. No contacts between the PHA-L-labeled axons and the FB-labeled phrenic motoneurons were observed. The brainstem projections of the VRG were found bilaterally in the nuclei ambigui, Cajal's interstitial nuclei of the solitary nucleus, the solitary nuclei, the hypoglossal nuclei, the Kölliker-Fuse's nuclei, and the subcoeruleus areas.

Action Potentials↗