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Biomedical subjects

K S Ha

Publications and source records attributed to K S Ha.

44 records · Page 3Linked to original sources
PubMed1994

Lysophosphatidic acid activation of phosphatidylcholine-hydrolysing phospholipase D and actin polymerization by a pertussis toxin-sensitive mechanism.

Incubation of IIC9 fibroblasts with lysophosphatidic acid (LPA) induced an increase in the amount of filamentous actin (F-actin), which was concentration-dependent with a maximal effect at 100 ng/ml. Phosphatidic acid (PA) also produced a concentration-dependent increase of F-actin, but it was less potent than LPA. The LPA-induced increase in F-actin was rapid and sustained for at least 60 min. LPA rapidly increased the levels of PA and choline, with maximal increases at 5 min and 30 s respectively. LPA also caused a monophasic increase in diacylglycerol (DAG) which lagged behind the increases in PA and choline. LPA stimulated phosphatidylbutanol formation in the presence of butanol and produced a small increase in inositol phosphates that was much less than that induced by alpha-thrombin. Pretreatment of cells with pertussis toxin (PTX) caused greater than 50% inhibition of the LPA-stimulated increases in PA, DAG and choline. PTX increased the LPA concentration required to induce half-maximal actin polymerization by about 10-fold. PTX caused a similar shift in the dose-response curve for LPA-induced PA formation. These results suggest that LPA induces an increase in PA by activating a phosphatidylcholine-hydrolysing phospholipase D via a PTX-sensitive G-protein and that the increase in PA is involved in the activation of actin polymerization.

Actins↗
PubMed1993

Differential translocation of protein kinase C isozymes by thrombin and platelet-derived growth factor. A possible function for phosphatidylcholine-derived diacylglycerol.

The translocation of protein kinase C (PKC) from the cytosolic to the particulate fraction in IIC9 fibroblasts has been studied to define the functions of 1,2-diacylglycerol (DAG) derived from the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) and phosphatidylcholine (PC). alpha-Thrombin caused a biphasic change in DAG, with two peaks at 15-60 s and 5-15 min, derived from PIP2 and PC, respectively, while platelet-derived growth factor (PDGF) induced a monophasic DAG increase from PC at 5-15 min. alpha-Thrombin also induced a rapid, but transient, increase of inositol 1,4,5-trisphosphate and cytosolic Ca2+, whereas PDGF did not. Three PKC isozymes, alpha, epsilon, and zeta, were identified by Western blotting in IIC9 cells and were mainly localized in the cytosol. A fraction of cytosolic PKC alpha was rapidly translocated by alpha-thrombin at 15 s, but its membrane association was lost within 1 min. PKC epsilon was also rapidly translocated; however, its membrane association was sustained for almost 60 min. PKC zeta was not translocated by alpha-thrombin or phorbol 12-myristate 13-acetate. PDGF translocated PKC epsilon at 5 min but had little effect at 15 s and did not translocate PKC alpha or zeta. Incubation with Bacillus cereus PC- or phosphatidylinositol-specific phospholipase C, which increased DAG but not phosphatidic acid, stimulated translocation of PKC epsilon, but not PKC alpha or zeta. Addition of chelators to inhibit the rise in intracellular Ca2+ largely blocked PKC alpha translocation induced by alpha-thrombin but had no effect on PKC epsilon translocation. Addition of ionomycin allowed alpha-thrombin to induce PKC alpha translocation at 5 min. PKC alpha translocation was mimicked by 1,2-dioctanoylglycerol plus ionomycin, but not by either alone. On the other hand, PKC epsilon was translocated by the DAG alone. These results support the conclusion that PIP2 hydrolysis activates both PKC alpha and epsilon at 15 s, whereas PC hydrolysis activates only PKC epsilon at 5 min. The differential activation at 5 min can be attributed to the failure of PC hydrolysis to increase Ca2+ and not to a difference in the molecular species of DAG derived from the phospholipids.

Animals↗
PubMed1993

Activation of actin polymerization by phosphatidic acid derived from phosphatidylcholine in IIC9 fibroblasts.

alpha-Thrombin induced a change in the cell morphology of IIC9 fibroblasts from a semiround to an elongated form, accompanied by an increase in stress fibers. Incubation of the cells with phospholipase D (PLD) from Streptomyces chromofuscus and exogenous phosphatidic acid (PA) caused similar morphological changes, whereas platelet-derived growth factor (PDGF) and phorbol 12-myristate 13-acetate (PMA) induced different changes, e.g., disruption of stress fibers and cell rounding. alpha-Thrombin, PDGF, and exogenous PLD increased PA by 20-40%, and PMA produced a smaller increase. alpha-Thrombin and exogenous PLD produced rapid increases in the amount of filamentous actin (F-actin) that were sustained for at least 60 min. However, PDGF produced a transient increase of F-actin at 1 min and PMA caused no significant change. Dioctanoylglycerol was ineffective except at 50 micrograms/ml. Phospholipase C from Bacillus cereus, which increased diacylglycerol (DAG) but not PA, did not change F-actin content. Down-regulation of protein kinase C (PKC) did not block actin polymerization induced by alpha-thrombin. H-7 was also ineffective. Exogenous PA activated actin polymerization with a significant effect at 0.01 microgram/ml and a maximal increase at 1 microgram/ml. No other phospholipids tested, including polyphosphoinositides, significantly activated actin polymerization. PDGF partially inhibited PA-induced actin polymerization after an initial increase at 1 min. PMA completely or largely blocked actin polymerization induced by PA or PLD. These results show that PC-derived PA, but not DAG or PKC, activates actin polymerization in IIC9 fibroblasts, and indicate that PDGF and PMA have inhibitory effects on PA-induced actin polymerization.

Actins↗
PubMed1992

Biphasic changes in the level and composition of Dunaliella salina plasma membrane diacylglycerols following hypoosmotic shock.

Hypoosmotic shock has been shown to trigger an immediate and selective increase of plasma membrane diacylglycerols (DAG) in the green alga Dunaliella salina, coinciding with an approximately equivalent loss of phosphatidylinositol 4,5-bisphosphate from this membrane [Ha, K.S., & Thompson, G.A., Jr. (1991) Plant Physiol. 97, 921-927]. Following a slight decline in amount, DAG levels of the plasma membrane resumed their rise by 2 min after the shock and by 40 min had achieved a maximum concentration equivalent to 230% of DAG levels in unstressed cells. This second, more sustained increase of plasma membrane DAG was matched by a DAG increase in the microsome-enriched cytoplasmic membrane fraction, commencing at 2 min and peaking at 140% of control values. The changing pattern of DAG molecular species produced in the plasma membrane during the early phases of hypoosmotic stress was compatible with their derivation from phospholipase C hydrolysis of inositol phospholipids and phosphatidylcholine. From 8 min following hypoosmotic shock, as relatively larger scale DAG accumulations developed in the cytoplasmic membranes, the molecular species composition changed to reflect a marked increase in de novo synthesis of sn-1-oleoyl, sn-2-palmitoylglycerol, and dioleoylglycerol. The former molecular species appears to be synthesized in the chloroplast while the latter is produced in the endoplasmic reticulum. The radioisotope labeling data with Na2(14)CO3 confirmed that the biphasic formation of DAG triggered by hypoosmotic shock culminates in a large-scale de novo synthesis of DAG. This is the first clear evidence for de novo synthesis as a source of DAG following PIP2-mediated signaling.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Membrane↗
PubMed1991

Diacylglycerol Metabolism in the Green Alga Dunaliella salina under Osmotic Stress : Possible Role of Diacylglycerols in Phospholipase C-Mediated Signal Transduction.

The sn-1,2-diacylglycerol (DAG) content and molecular species composition of Dunaliella salina whole cells and cell fractions were measured by complementary high performance liquid chromatography and gas chromatography techniques. At 4.2 nanomoles per 100 nanomoles lipid phosphorus, the whole cell DAG level was high in comparison with most animal tissues. The DAG concentration was highest in the microsome-enriched fraction, followed by that in the chloroplast and in the plasma membrane fractions. The predominant DAG molecular species in all cell fractions contained oleic (18:1), linoleic (18:2), or linolenic (18:3) acid in the sn-1 position and palmitate (16:0) in the sn-2 position. Recent studies have raised the possibility of DAG serving a signal transducing function in osmotically stressed D. salina cells. During the first 30 seconds following hypoosmotic shock, there was a 40% increase in the plasma membrane DAG content, whereas the DAG content of the microsome-enriched fraction was unchanged. On a nanomole per 100 nanomoles phospholipid basis, the rise in plasma membrane DAG nearly matched the previously reported (KJ Einspahr, TC Peeler, GA Thompson Jr [1988] J Biol Chem 263: 5775-5779) transient fall in phosphatidylinositol 4,5-bisphosphate. Furthermore, 18:1/16:0 DAG, one of the major plasma membrane DAG molecular species increasing in amount after hypoosmotic shock, was the characteristic molecular species of plasma membrane phosphatidylinositol, phosphatidylinositol 4-phosphate, and phosphatidylinositol 4,5-bisphosphate, but no other lipid of that membrane. Evidence was found for a rise in 16:0/18:2 and 16:0/18:3 DAG as well following hypoosmotic shock. This pattern suggested that phosphatidylcholine hydrolysis also contributed to the stress-induced production of DAG in the D. salina plasma membrane. The extent of the sudden DAG increase was sufficient to consider it a potential second messenger in phospholipase C-mediated signal transduction.

Journal Article↗
PubMed1983

Childhood non-Hodgkin's lymphoma developing from recurrent neck mass.

Non-Hodgkin's lymphoma (NHL) in an 8-year-old boy developing from recurrent neck mass during 15-months history is presented. Two biopsies carried out during this period were interpreted as reactive change. The third biopsy was diagnosed as NHL. He died of the systemic disease 19 months after the first admission. Clinical, histological and immunocytochemical investigations for the three biopsy materials revealed that the first and the second biopsies were reactive, and the third biopsy was neoplastic, B cell lymphoma. Known etiological factors causing lymphoreticular disorders are reviewed by the literatures, and absence of these factors in the present case is described.

B-Lymphocytes↗
PubMed1981

A new approach to detection of heterozygotes for adenosine deaminase deficiency: a hypothetical method.

In a second and third families with ADA deficiency found in Japan, we tried a new approach to evaluate heterozygote detection. This is based on the hypothesis that ADA activity of red blood cell is the quantitative sum of the activities of ADA proteins expressed by two allelic genes at the ADA autosomal locus, and that these activities are not changed by the gene transmission from parents to children. We have detected red blood cell-ADA activities expressed by the one normal allelic gene in heterozygotes (including parents and paternal or maternal grandfather or grandmother) and from these values have determined combinations for the pair of ADA activities expressed by the two allelic genes of other family members. These combinations were consistently made in all relatives examined in the two families, and we conclude that several members of each family who were judged to have nil activity in the combinations were heterozygotes for ADA deficiency.

Adenosine Deaminase↗