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F Hata

Publications and source records attributed to F Hata.

At least 55 records · Page 3Linked to original sources

High-resolution immunogold cytochemistry indicates that AQP4 is concentrated along the basal membrane of parietal cell in rat stomach.

Gastric parietal cells secrete hydrochloric acid in stomach. Because the secreted HCl solution is isotonic with the plasma fluid, it should accompany the water transport across the membranes of parietal cells. Aquaporins (AQPs) are water channel proteins that play the central role in the cellular handling of water in various mammalian tissues. Using immunocytochemistry, we found that AQP4 was expressed only in parietal cells of rat gastric mucosa. Immunogold electron microscopy study further demonstrated that AQP4 was mostly localized at the basal membrane of parietal cells. In the basal membrane, AQP4 was prominently enriched on the portion contacting with the basement membrane surrounding gastric glands. These results suggest that the contact between basement membrane and basal membrane may generate the signal involved in the targeting of AQP4 in gastric parietal cells.

Animals↗

Critical role of sulfenic acid formation of thiols in the inactivation of glyceraldehyde-3-phosphate dehydrogenase by nitric oxide.

The relationship between possible modifications of the thiol groups of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) by nitric oxide (NO) and modified enzyme activity was examined. There are 16 free thiols, including 4 active site thiols, in a tetramer of GAPDH molecule. NO donors, sodium nitroprusside (SNP), and S-nitroso-N-acetyl-DL-penicillamine (SNAP) decreased the number of free thiols with a concomitant inhibition of GAPDH activity in a concentration- and time-dependent manner. After treatment for 30 min, free thiols were maximally decreased to 8-10 per GAPDH tetramer and enzyme activity was also inhibited to 5-10% of control activity. In the presence of 30 mM dithiothreitol (DTT), these effects were completely blocked. Since similar results were obtained in the case of hydrogen peroxide (H2O2) treatment, which is known to oxidize the thiols, these effects of nitric oxide donors were probably due to modification of thiol groups present in a GAPDH molecule. On the other hand, DTT posttreatment after the treatment of GAPDH with SNP, SNAP, or H2O2 did not completely restore the modified thiols and the inhibited enzyme activity. DTT posttreatment after the 30-min-treatment with these agents restored free thiols to 14 in all treatments. In the case of SNAP treatment, all 4 active sites were restored and enzyme activity reached more than 80% of the control activity, but in two other cases one active site remained modified and enzyme activity was restored to about only 20%. Therefore, all 4 free thiols in the active site seem to be very important for full enzyme activity. DTT posttreatment in the presence of sodium arsenite, which is known to reduce sulfenic acid to thiol, almost completely restored both thiol groups and enzyme activity. These findings suggest that nitric oxide inhibits GAPDH activity by modifications of the thiols which are essential for this activity, and that the modification includes formation of sulfenic acid, which is not restored by DTT. S-nitrosylation, which is one type of thiol modification by NO, occurred when GAPDH was treated with SNAP but not SNP. Analysis of thiol modification showed that SNAP preferentially nitrosylated the active site thiols, the nitrosylation of which fully disappeared by DTT posttreatment. It seems that SNAP nitrosylates the active site thiols of GAPDH to prevent these thiols from oxidizing to sulfenic acid.

Binding Sites↗

Tyrosine kinase involvement in apamin-sensitive inhibitory responses of rat distal colon.

1. It has been suggested that pituitary adenylate cyclase activating peptide (PACAP) may be involved in the non-adrenergic, non-cholinergic (NANC) inhibitory response of longitudinal muscle of rat distal colon. In this study, we have investigated the intracellular mechanism of PACAP-induced relaxation in this muscle. 2. PACAP induced an apamin-sensitive relaxation of the longitudinal muscle. The tyrosine kinase inhibitors genistein at 10 microM and tyrphostin 25 at 30 microM, but not the cyclic AMP-dependent protein kinase inhibitor Rp-8-bromoadenosine-3',5'-cyclic monophosphorothioate at 30 microM significantly inhibited the PACAP-induced relaxation to 60% and 25% of control values, respectively. PACAP did not increase the cyclic AMP content of the muscle. 3. Tyrphostin 25 at 10 microM significantly inhibited the relaxation of longitudinal muscle induced by electrical field stimulation (EFS), to 50% of control values. Apamin at 1 microM, an antagonist of small conductance Ca2+-activated K+ channels, also inhibited the relaxation, to 42 % of control values. The inhibitory effects of tyrphostin 25 and apamin were not additive (44 % of control values). 4. PACAP induced an apamin-sensitive, slow hyperpolarization of the cell membrane of the muscle. Tyrphostin 25 at 3 microM inhibited this PACAP-induced hyperpolarization. Tyrphostin 25 at 10 microM and genistein at 10 microM inhibited the apamin-sensitive inhibitory junction potentials induced by a single pulse of EFS. 5. The PACAP-induced relaxation of longitudinal muscle occurred with a concomitant decrease in intracellular Ca2+ levels ([Ca2+]i). Tyrphostin 25 at 10 microM and apamin at 1 microM abolished these PACAP-induced responses. 6. From these findings it is suggested that the activation of tyrosine kinase is involved in PACAP-induced relaxation of longitudinal muscle from rat distal colon, 'upstream of' the activation of apamin-sensitive K+ channels.

8-Bromo Cyclic Adenosine Monophosphate↗

Identification of segments VI and VII of the liver based on the ramification patterns of the intrahepatic portal and hepatic veins.

We describe the pattern of intrahepatic vessel ramification in the right posterior hepatic sector in a population of 197 adults. Each specimen was dissected from its visceral (inferior) surface in order to demonstrate variations in the distribution of the portal vein branches to the hepatic segments of the right lobe, especially to segments VI (S6) and VII (S7) as described by Couinaud. We also examine whether three hepatic veins, i.e., the right hepatic vein (RHV), middle hepatic vein (MHV), and the short hepatic vein (SHV), aid the identification of segmental portal branches in the lower posterior sector. Four major patterns of branching of the posterior sectorial trunk of the portal vein system are described. In group A (32.0%) a single posterior trunk formed an arch-like pattern sending multiple branches to S6 and S7 (P6 and P7). We named the multiple branches to the apparent S6 the inferoposterior portal branches. It was difficult to identify which of these branches were equivalent to P6. In group B (27.9%), the posterior sectorial trunk bifurcated to form P6 and P7. In most of the specimens in this group, therefore, we were able clearly to identify both S6 and S7 based on the portal vein system. In group C (6.6%), the trunk trifurcated to form P6, P7, and an intermediate branch, which supplied both segments or a gray zone between them. Group D (33.5%) included variations of the anterior segmental branches, and in specimens of this group, the anteromedial border of the sector was difficult to identify. Notably, the three-dimensional interdigitating topographical relationship of the hepatic veins and the portal branches was not evident in the lower posterior sector, since tributaries of the RHV and the portal branches followed similar courses and paralleled each other in the region and since the territory of the SHV was usually restricted to the superficial parenchyma near the inferior surface. In group A, tributaries of the RHV/SHV (>3 mm in diameter) passed between the inferoposterior portal branches in only 22.2%/14.3% of the specimens. Thus the hepatic veins often did not reveal which of the multiple inferoposterior branches was P6. Moreover, in the subset of Group B in which the segments were identified based on the portal vein ramification, tributaries of the RHV/SHV (>3 mm in diameter) showed the intersegmental interdigitating arrangement in only 32.0%/6.0% of the specimens. In addition, a thick tributary of the MHV, sometimes arising from S6, did not run along, but penetrated the S5/S6 border plane from the lateral to the medial side. Therefore, the three hepatic veins (RHV, SHV, MHV) often did not aid the identification of the liver segments in the region. Consequently, the less than ideal combinations of irregular configurations of the portal and hepatic venous systems suggest that the right posterior segments cannot be conclusively identified anatomically in 30-40% of cases. Other means of identification, such as the conventional proportional manner (the upper and lower halves of the posterior sector roughly correspond to S6 and S7) may be required.

Adult↗

Vascular anatomy of the pancreaticoduodenal region: A review.

Vascular anatomy of the pancreaticoduodenal region has been the subject of numerous studies. However, several essential areas of confusion remain in interpretation of the vascular configuration. We note and discuss three key points in relation to this confusion: (1) a missing vascular arcade, (2) a rearrangement of the arcade by collateral and/or transverse vessels, and (3) a solitary vessel without an accompanying comites vein or artery. In addition, we consider that different interpretations as well as varying reported incidences depend on different "thresholds" when observations are made. Consideration of new aspects of vascular anatomy of the pancreaticoduodenal region is required for further improvement of surgical procedures. In terms of the selection of lymph node resection procedure, we discuss mainly the inferior arterial origin. Special attention should be paid to the ligation of inferior arteries because of the high incidence of the common trunk formation of the upper jejunal and inferior pancreaticoduodenal arteries. With regard to duodenum-preserving pancreatic head resection for benign tumors, our observations are introduced in view of either arterial or venous configuration. First, a communicating artery between the anterior and posterior arterial arcades is noted because of its possible critical role in blood supply to the papilla of Vater. Second, a venous drainage route from the duodenum to the retroperitoneal space in "normal" specimens is described.

Dissection↗

Mediators of nonadrenergic, noncholinergic relaxation in longitudinal muscle of the intestine of ICR mice.

Mediators of nonadrenergic, noncholinergic (NANC) relaxation in longitudinal muscle of several regions of ICR mouse intestine were studied. An inhibitor of synthesis of nitric oxide, N(G)-nitro-L-arginine (L-NOARG) at 10 microM significantly inhibited NANC relaxations induced by electrical field stimulation (EFS) in the jejunum, ileum, and the proximal and distal colon. Especially in the ileum extent of the inhibition was more than 80%. An antagonist of vasoactive intestinal peptide (VIP) receptors, VIP(10-28) at 3 microM partially inhibited the EFS induced relaxations in the jejunum and proximal colon, but very slightly in the distal colon and had no effect in the ileum. An antagonist of pituitary adenylate cyclase activating peptide (PACAP) receptor, PACAP(6-38) at 3 microM partially inhibited the EFS-induced relaxations in the proximal and distal colon, but not in the jejunum and ileum. Totals of the percentages of relaxant components mediated by nitric oxide, VIP and PACAP in every region are roughly equal to a hundred percent. In another series of experiments, EFS-induced relaxations were almost completely inhibited by the treatment of the segments with L-NOARG and VIP(10-28) in the jejunum, with L-NOARG, VIP(10-28) and PACAP(6-38) in the proximal colon, and with L NOARG and PACAP(6-38) in the distal colon. The present results suggest that nitric oxide solely mediates the relaxation of longitudinal muscle of the ileum of ICR mice, whereas nitric oxide and VIP co-mediate it in the jejunum, nitric oxide, VIP and PACAP in the proximal colon, and nitric oxide and PACAP in the distal colon.

Animals↗

Configuration of hepatic veins in the right surgical lobe of the human liver with special reference to their complementary territorial relationships: morphometric analysis of controlled specimens with clearly defined portal segmentation.

The configurations of hepatic veins, particularly the complementary territorial relationships between the veins, in the right surgical lobes of 156 human livers, the segments of which were identified clearly according to the portal ramification, were studied. In order to assess the functional roles of the vessels, we compared the diameter values of the vessels in the upper region of the lobe under the assumption that this parameter corresponds very closely to the actual blood flow volume through the vessel. The right hepatic vein (RHV) sometimes (17%) developed poorly and drained neither segment V (S5) nor VI (S6); instead, the inferomedial part of the S6 was often (26%) drained by the middle hepatic vein (MHV). However, most thick short hepatic veins (SHVs) did not drain S6 specifically instead of a poorly-developed RHV, but usually drained both S6 and segment VII (S7), irrespective of the configurations of other veins. Sometimes (8%), the RHV, rather than the MHV, drained a large part of S5. Overall, in the lower region of the lobe, the RHV, SHV and MHV showed complementary venous drainage relationships. Furthermore, the RHV usually (75%) ran not between S5 and S6 but through S6, corresponding to its usual territory, and could not be regarded as the intersectorial landmark in the lower region. Although various names have been given to the thick venous tributaries in the upper region of the right lobe, we systematically classified these tributaries into 5 types of "right superior radicles" (RSR), according to their topographical relationships with the right superior portal branches (P7 and P8), RHV, MHV, SHVs and their tributaries. When just thick RSRs were considered, the 5 types (anterior, posterior, lateral, medial and intersegmental) were observed in 90%, 88%, 89%, 38% and 34%, respectively. Notably, all but the intersegmntal RSRs almost always corresponded to a specific segmental territory (S7 or segment VIII (S8) and had specific terminals (at the RHV or MHV), suggesting that these 4 RSRs were proper segmental or intrasegmental veins, whereas, the 5th type showed a different configuration, i.e., an intersegmental vein that drained both S7 and S8, ran along and above the RHV and merged with it. Our morphometric examination of the upper region revealed that the sizes of P7 and P8 increased or decreased simultaneously, and suggested that the RSRs played limited roles in venous return. Rather than the RSRs, the SHVs appeared to drain the overflow in excess of the venous return capacity of the RSRs. Despite their limitations and possible complementary relationships with some veins, however, all the superior veins tended to developed evenly well or poorly. Overall, the venous return did not seem to converge into a single particular vein, but was carried away by multiple superior veins: the RHV, MHV, SHVs, and 5 RSRs.

Adult↗

[Combination chemotherapy of continuous infusion 5-FU and daily low-dose CDDP in colorectal carcinoma].

Continuous intravenous infusion (CVI) of 5-fluorouracil (5-FU) plus daily low-dose cisplatin (CDDP) was evaluated in 59 patients with advanced and unresectable colorectal carcinoma. The procedure consisted of 5-FU 320 mg/m2/day, CVI and CDDP 2.5-3.5 mg/m2/day, infused for one hour five times a week (low-dose FP therapy) was performed for at least four weeks. The rates of complete and partial response, along with side effects were studied. Also, the effect of low-dose FP therapy on the prognosis of patients with colorectal carcinoma was investigated. The rate of complete and partial response was 57.1%. The frequency of severe side effects (grade 3 and 4) was 10.5%, and no death from side effects was noted. The effect of low-dose FP therapy on the prognosis of advanced and unresectable colorectal carcinoma was studied. It is suggested that this chemotherapy might contribute to the survival of patients with colorectal carcinoma. We designed the chemotherapy of intermittent administration (day by day) of 5-FU 600 mg/m2/day and daily CDDP 2.5-3.5 mg/m2/day (intermittent FP) in order to decrease the rate of side effects and their severity. No cases with grade 3 or 4 side effects were experienced, but the response rate was mostly similar to that of low-dose FP therapy. We concluded that low-dose FP and intermittent FP therapy might be fairly effective for advanced and unresectable colorectal carcinoma.

Antineoplastic Combined Chemotherapy Protocols↗

Inhibition of skeletal muscle sarcoplasmic reticulum Ca2+-ATPase by nitric oxide.

The effects of nitric oxide on the activities of thapsigargin-sensitive sarcoplasmic reticulum Ca2+-ATPase (SERCA) and Ca2+ uptake by sarcoplasmic reticulum (SR) membranes prepared from white skeletal muscle of rabbit femoral muscle were studied. Pretreatment of the SR preparations with nitric oxide at concentrations of up to 250 microM for 1 min decreased the SERCA activity concentration dependently, and also decreased their Ca2+ uptake. Both these effects of nitric oxide were reversible. Inhibitors of guanylyl cyclase and protein kinase G (PKG) had no significant effect on the nitric oxide-induced inhibitions of SERCA and Ca2+ uptake. Moreover, dithiothreitol did not reverse the inhibitory effects of nitric oxide on SERCA and Ca2+ uptake. These findings suggest that nitric oxide inhibits SERCA, mainly SERCA 1, of rabbit femoral skeletal muscle by an action independent of the cyclic GMP-PKG system or oxidation of thiols, and probably by a direct action on SERCA protein.

Aminoquinolines↗

Nonadrenergic, noncholinergic relaxation mediated by nitric oxide with concomitant change in Ca2+ level in rectal circular muscle of rats.

The mediators of nonadrenergic, noncholinergic (NANC) relaxation of the circular muscle of rat rectum were examined in vitro. In the circular muscle of rat rectum, NG-nitro-L-arginine (L-NOARG) at 10 microM did not affect electrical field stimulation-induced relaxation but at 100 microM it inhibited electrical field stimulation-induced relaxation by about 75% and 1-mM L-arginine reversed the inhibition. Exogenous nitric oxide (NO) (1-10 microM) concentration dependently relaxed the circular muscle. Electrical field stimulation increased the cyclic GMP content of the circular muscle to about twice its resting level. L-NOARG, even at 10 microM, completely inhibited the electrical field stimulation-induced elevation of cyclic GMP content. However, L-arginine at 1 mM did not reverse the inhibition in cyclic GMP content. Inhibitory junction potentials (i.j.ps) induced by electrical field stimulation in the circular muscle cells were not affected by L-NOARG, 100 microM. Apamin ( < or = microM) did not affect the electrical field stimulation-induced relaxation, but almost completely inhibited electrical field stimulation-induced i.j.ps. NO (0.3-10 microM) induced relaxation of the circular muscle with a concomitant decrease in intracellular Ca2+ level ([Ca2+]i). Abundant immunoreactivity of NO synthase was found in the circular muscle layer, in addition to myenteric and submucosal plexus. The results suggest that NO induces NANC relaxation with a concomitant change in [Ca2+]i in the circular muscle of rat rectum. However, the involvement of changes in cyclic GMP level and in membrane potentials in the mechanism was not shown in the present experimental conditions.

Animals↗

Pranidipine, a new 1,4-dihydropyridine calcium channel blocker, enhances cyclic GMP-independent nitric oxide-induced relaxation of the rat aorta.

Pranidipine, a new calcium channel blocker, prolonged endothelium-dependent relaxation induced by acetylcholine in an aortic ring preparation, contracted with prostaglandin F2alpha. This action was not shared by amlodipine. The effect was not modified by indomethacin, suggesting that the action of pranidipine does not involve prostanoid metabolism. N(G)-nitro-L-arginine completely prevented the action of Pranidipine. The drug affected neither nitric oxide (NO) synthase activity nor the level of cyclic GMP in the vessel. Pranidipine did not affect the sensitivity of the contractile proteins to calcium. Pranidipine also did not alter cyclic GMP-induced relaxation in alpha-toxin-skinned vascular preparations. Pranidipine also prolonged glyceryl trinitrate-induced relaxation in the endothelium denuded rat aorta. Furthermore, pranidipine enhanced relaxation of the aorta induced by glyceryl trinitrate even in the presence of methylene blue, a guanylyl cyclase inhibitor. This action was not modified by iberiotoxin or by charybdotoxin, two inhibitors of the calcium-activated potassium channel. The results strongly suggest that pranidipine enhances cyclic GMP-independent NO-induced relaxation of smooth muscle by a mechanism other than through NO-induced hyperpolarization. These effects were in direct contrast to amlodipine, another new 1,4-dihydropyridine calcium antagonist.

Amlodipine↗

Relationship between muscarinic autoinhibition and the inhibitory effect of morphine on acetylcholine release from myenteric plexus of guinea pig ileum.

The relationship between muscarinic autoinhibition and the inhibitory effect of morphine on acetylcholine (ACh) release was investigated in a longitudinal muscle with myenteric plexus (LMMP) preparation of guinea pig ileum. Morphine (10 microM) inhibited spontaneous and evoked ACh release by electrical field stimulation (EFS) at 1 Hz but not at 10 Hz. Atropine (1 microM) did not affect the resting ACh release, but it significantly increased EFS-evoked release, suggesting activation of muscarinic autoreceptors by ACh released during EFS. Only when the autoinhibition was weakened by atropine, morphine exhibited an inhibitory effect on the EFS-evoked release at 10 Hz. Bethanechol (300 microM) inhibited the EFS-evoked release at 1 Hz but not 10 Hz, suggesting that muscarinic autoreceptors are partially or almost fully activated at 1 or 10 Hz stimulation, respectively. After bethanechol treatment, morphine did not exhibit its inhibitory effect on the EFS-evoked release at 1 Hz. Naloxone (1 microM) increased spontaneous and EFS-evoked ACh release at 1 Hz but not at 10 Hz. Following treatment with atropine, naloxone also increased ACh release at 10-Hz stimulation. These results suggest that morphine and an endogenous opioid inhibit ACh release from LMMP preparations when muscarinic autoinhibition mechanism does not fully work. This inhibitory effect of morphine is discussed in relation to the calcium sensitivity of the preparations in ACh release.

Acetylcholine↗

Relationship between inhibitory effect of endogenous opioid via mu-receptors and muscarinic autoinhibition in acetylcholine release from myenteric plexus of guinea pig ileum.

Relationship between activation of opioid receptors and muscarinic autoinhibition in acetylcholine (ACh) release from the myenteric plexus was studied in longitudinal muscle myenteric plexus (LMMP) preparations of guinea pig ileum. A mu-receptor agonist, [D-Ala2, N-Me-Phe4, Gly5-ol] enkephalin (DAMGO), at a concentration of 1 microM inhibited the ACh release evoked by electrical field stimulation (EFS) at 1 Hz but not at 10 Hz. After the muscarinic autoreceptors were blocked with atropine (1 microM), DAMGO inhibited EFS-evoked ACh release also at 10 Hz. After the autoreceptors were potently activated with muscarine (200 microM), the inhibitory effect of DAMGO at 1 Hz was abolished. A kappa-receptor agonist, U-50,488, at 1 microM inhibited the EFS-evoked ACh release both at 1 and 10 Hz. U-50,488 inhibited ACh release regardless of the presence of atropine or muscarine. A delta-agonist, enkephalin [D-PEN2.5] (PDPDE), did not show any significant effect. On the other hand, a selective mu-receptor antagonist, cyprodime, increased ACh release evoked by EFS at 1 Hz, but not at 10 Hz. After the autoreceptors were blocked, cyprodime increased EFS-evoked ACh release also at 10 Hz. The selective kappa-receptor antagonist, nor-binaltorphimine, did not affect ACh release in the absence or presence of atropine. The results suggest that endogenous opioid(s) inhibits ACh release by activating mu-, but not kappa- and delta-receptors in the LMMP of guinea pig ileum and that the inhibitory effect of endogenous opioid(s) in the ACh release is important when muscarinic autoinhibition mechanism does not fully work.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

Decrease in participation of nitric oxide in nonadrenergic, noncholinergic relaxation of rat intestine with age.

Participation of nitric oxide in the electrical field stimulation-induced nonadrenergic, noncholinergic (NANC) relaxation in various intestinal regions was studied in 2- to 50-week-old Wistar rats. In the jejunum of 2-week-old rats, the extent of the nitric oxide-mediated component of the relaxation of longitudinal muscle was approximately 60-70%, whereas the component was 40-50% in 4-week-old rats and was absent in 8- and 50-week-old rats. Thus, nitric oxide seems to be the most important mediator at young ages but its significance is lost with age. The same tendency as that in the jejunum was also shown in longitudinal muscle of the ileum, proximal and distal colon, and rectum. The tendency was also shown in the circular muscle of the rectum. Sensitivity of the longitudinal muscle of the jejunum and proximal colon to exogenously added nitric oxide was high in younger rats. Immunoreactive structures for nitric oxide synthase were observed in the circular muscle layer of the rectum. The population of the structures was denser in 4-week-old than that in 50-week-old. The results suggest that NANC relaxation in every region of the intestine at 2-week-old is almost solely mediated by nitric oxide, and its significance as an inhibitory mediator gradually or rapidly decreases with age.

Adrenergic Agents↗

Inhibitory effect of endomorphin-1 and -2 on acetylcholine release from myenteric plexus of guinea pig ileum.

Endomorphin-1 and -2, putative endogenous ligands for the mu-opioid receptor, inhibited acetylcholine (ACh) release evoked by electrical field stimulation (EFS) at 1 Hz, which partially activates muscarinic autoreceptors, but not at 10 Hz, which fully activates muscarinic autoreceptors, in longitudinal muscle with the myenteric plexus (LMMP) preparations of guinea pig ileum. After blockade of autoinhibition by atropine, the peptides also inhibited EFS-evoked ACh release at 10 Hz. The inhibitory effects on ACh release were abolished by the mu-opioid antagonist cyprodime. These results suggest that endomorphin-1 and -2 inhibit ACh release from LMMP preparations of guinea pig ileum and that the mechanism of the inhibition must have a component in common with muscarinic autoinhibition.

Acetylcholine↗

Carboxy-terminal regions of the sarcoplasmic/endoplasmic reticulum Ca(2+)- and the Na+/K(+)-ATPases control their K+ sensitivity.

The Na+,K(+)-ATPase and the sarcoplasmic/endoplasmic reticulum Ca(2+)-(SERCA-) ATPase belong to a family of P-type ATPases that undergo a cycle of conformational changes between the phosphorylated and dephosphorylated stages in an ion-specific manner. The ouabain-inhibitable Na+,K(+)-ATPase activity requires Na+ and K+. On the other hand, the Ca(2+)-dependent and thapsigargin-inhibitable activity of the SERCA-ATPase does not depend upon Na+ and K+ for its basal activity. However, the SERCA-ATPase and Ca(2+)-transport activities can be further activated either by K+ in a two-step fashion with high (ED50 approximately 20 mM) and low affinity (ED50 approximately 70 mM) or by Na+ in a one-step fashion with an ED50 value of approximately 50 mM. A chimera, in which the carboxy-terminal region (Leu861-COOH) of the Na+,K(+)-ATPase alpha 1 subunit replaced the corresponding region (Ser830-COOH) of the SERCA1-ATPase, lacked the low-affinity K+ activation of the SERCA-ATPase but displayed a higher-affinity (ED50 < 10 mM) activation by K+, similar to that of the Na+,K(+)-ATPase, whereas activation by Na+ was not affected. The replacement of the large cytosolic loop (Gly354-Lys712) and the amino-terminal regions (Met1-Asp162) of the SERCA1-ATPase with the corresponding portions of the Na+,K(+)-ATPase alpha 1 subunit did not affect the sensitivity of the SERCA-ATPase activity to K+. Thus, the carboxy-terminal regions of both the SERCA1 and the Na+,K(+)-ATPase alpha 1 subunit are critical for K+ sensitivity. Analysis of additional (Ca2+/Na+,K+)-ATPase chimeras demonstrated that the carboxy-terminal 102 amino acids (Phe920-Tyr1021) of the Na+/K(+)-ATPase alpha 1 subunit are sufficient to shift the K+ affinity for activation of the SERCA-ATPase without the beta subunit. No change in the two-step activation of SERCA-ATPase by K+ was seen when residues Thr871-Thr898 of the SERCA1-ATPase were replaced with residues Asn894-Ala919 of the Na+,K(+)-ATPase alpha 1 subunit, a region known to bind the Na+,K(+)-ATPase beta subunit [Lemas, M. V., et al. (1994) J. Biol. Chem. 269, 8255-8259]. Thus, the Na+,K(+)-ATPase subunit-assembly domain and the K(+)-sensitive region are distinct within the carboxy-terminal 161 amino acids of the Na+,K(+)-ATPase.

Animals↗

[Inhibitory neuronal control of smooth muscle activity of the gastrointestinal tract].

Recent findings suggest that nonadrenergic inhibitory responses of the smooth muscle of the gastrointestinal tract is mediated by nitric oxide and some intestinal peptides including VIP, PACAP and CGRP. Although nitric oxide was suggested to mediate the nonadrenergic relaxation in various regions of gastrointestinal tracts of many species, further careful studies revealed that nitric oxide participates in the relaxation in restricted regions, not throughout the tract. It was also found that the peptides work in extremely restricted regions of the tract. Importance of the role of nitric oxide in the relaxation varies with different regions, strains and species. Moreover, it significantly decreases with the age of the rat, especially between 4-8 weeks of age. Inhibitors of Ca(2+)-activated K+ channels inhibited the relaxation in almost all regions examined in rats, although the magnitude of the inhibition varied from region to region. The intracellular action mechanism(s) of nitric oxide was discussed in relation to changes in cyclic GMP level, intracellular Ca2+ level and membrane potentials of the smooth muscle cells.

Aging↗

Nonadrenergic, noncholinergic relaxation in longitudinal muscle of rat jejunum.

The mediators of nonadrenergic, noncholinergic (NANC) relaxation in the longitudinal muscle of rat jejunum were studied in vitro. Electrical field stimulation (EFS) of segments of rat jejunum induced a rapid transient relaxation followed by a subsequent contraction in the presence of atropine and guanethidine. NG-Nitro-L-arginine (L-NOARG, 10 microM) inhibited the EFS-induced NANC relaxation by about 25%, and L-arginine (1 mM) completely reversed this inhibition. Exogenously added nitric oxide (0.1-10 microM) induced relaxation of the segment. Treatment of the segment with alpha-chymotrypsin resulted in about 50% inhibition of the EFS-induced relaxation. Several peptide candidates for the mediator of NANC relaxation were examined by using selective antagonists of their receptors or by a receptor-desensitization method. Results indicated that vasoactive intestinal peptide, pituitary adenylate cyclase activating peptide, peptide histidine isoleucine, atrial natriuretic peptide and neurotensin are not associated with NANC relaxation of the segments. On the other hand, apamin at 1 microM inhibited the EFS-induced relaxation by 74%. Inhibitory effects of L-NOARG and, apamin or alpha-chymotrypsin treatment on the EFS-induced relaxation were additive and almost complete. Exogenous nitric oxide-induced relaxation was not affected by apamin. Inhibitory junction potentials (i.j.p.'s) were recorded from longitudinal muscle cells of rat jejunum. Apamin at 200 nM abolished i.j.p.'s induced by two pulses of EFS. These results suggest that NANC relaxation in longitudinal muscle of rat jejunum involves two independent components: one is a nitric oxide-mediated minor component, and the other is an unknown substance-mediated apamin-sensitive major component that is inhibited by alpha-chymotrypsin treatment.

Adrenergic Fibers↗