Itaru Kojima

A novel action of activin A: stimulation of insulin secretion in rat pancreatic islets.

The present study was conducted to examine an action of activin A on insulin secretion from rat pancreatic islets. In a batch incubation system, activin A stimulated insulin secretion in a dose-dependent manner at concentrations higher than 1 nM. Furthermore, activin A greatly potentiated glucose-induced insulin release. When islets were perifused with 1 nM activin A, insulin secretion was barely affected in this system. However, the insulin response to 16.7 mM glucose was greatly enhanced. Both the first and second phases of insulin response were enhanced by 1 nM activin A. These results indicate that, in addition to its known actions on pituitary-gonadal and hematopoietic systems, activin A modulates the function of pancreatic islets and stimulates insulin secretion.

Activin-A: A modulator of multiple types of anterior pituitary cells

When primary culture of rat pituitary cells were incubated with 1 nM activin-A for more than 24 hrs, activin-A significantly increased intracellular content of FSH without affecting the control of LH. Pretreatment of the cells with activin-A also enhanced LHRH-induced FSH release without affecting LH release. Furthermore, pretreatment of the cells with activin-A significantly reduced both GRF-mediated GH release and TRH-mediated PRL release. However, activin-A did not affect the response of ACTH and TSH to their releasing hormones. These results indicate that, in addition to the known action on gonadotrophs, activin-A also modifies the function of somatotrophs and lactotrophs.

Involvement of endogenous digitalis-like substance in genesis of deoxycorticosterone-salt hypertension

In an attempt to evaluate the role of endogenous digitalis-like substance in the genesis of deoxycorticosterone (DOCA) -salt hypertension, the effects of intravenous anti-digoxin antibody on the blood pressure response were observed in male Wistar rats that underwent heminephrectomy followed by treatment with DOCA and saline. Administration of anti-digoxin antibody caused a marked decrease in the blood pressure which continued for about an hour. Such a change in the blood pressure was not observed in pertinent control animals. Thus, it seems that endogenous digitalis-like substance plays an important role in the genesis of DOCA-salt hypertension.

Pertussis toxin blocks angiotensin II-induced calcium influx but not inositol trisphosphate production in adrenal glomerulosa cell

Involvement of GTP‐binding proteins in angiotensin II‐induced mobilization of calcium has been examined in adrenal glomerulosa cells by using pertussis toxin. Pretreatment of glomerulosa cells with pertussis toxin abolishes angiotensin II‐induced calcium influx without blocking inositol trisphosphate production. These results suggest a role of the pertussis toxin‐sensitive GTP‐binding protein in transducing angiotensin II‐receptor occupancy into opening of calcium channel.

Evidence that type II insulin-like growth factor receptor is coupled to calcium gating system

In competent Balb/c 3T3 cells primed with epidermal growth factor (primed competent cells), insulin-like growth factor-II (IGF-II) stimulated calcium influx in a concentration dependent manner with the ED50 of 450 pM. When receptor-bound [125I]IGF-II was cross-linked by use of disuccinimidyl suberate, a 240 K-Da protein was radiolabeled. Excess amount of unlabeled IGF-II inhibited the affinity-labeling of the 240 K-Da protein. To further examine whether IGF-II stimulates calcium influx by acting on the type II IGF receptor, we employed polyclonal antibody raised against rat type II IGF receptor, R-II-PABl. This antibody immunoprecipitated the type II IGF receptor and inhibited IGF-II binding in Balb/c 3T3 cell membrane without affecting IGF-I binding. In primed competent cells, R-II-PABl elicited an agonistic action in stimulating [3H]thymidine incorporation. Under the same condition, R-II-PABl elicited a marked stimulation of calcium influx. These results suggest that, in Balb/c 3T3 cells, 1) relatively low concentrations of IGF-II act mainly on the type II IGF receptor; 2) the type II IGF receptor is coupled to a calcium gating system; and 3) binding of a ligand to the type II IGF receptor leads to the stimulation of DNA synthesis.

Insulin-like Growth Factor II Stimulates Calcium Influx in Competent Balb/c 3T3 Cells Primed with Epidermal Growth Factor

Insulin-like growth factors (IGFs) are potent mitogens in mammalian cells (Zapfet al., 1981). The mode of action of IGFs has been extensively studied in Balb/c 3T3 cells. Thus, IGFs promote cell cycle progression specifically in competent Balb/c 3T3 cells while IGFs have essentially no effect in Go-arrested cells (Pledgeret al., 1977; Stileset al., 1979). To promote cell cycle progression, IGFs should be exposed to competent cells continuously throughout the Giphase (Pledgeret al., 1977). Hence, it is presumed that IGFs may generate a continuous mitogenic signal. This observation together with the fact that extracellular calcium is required for cell cycle progression have led to the consideration that calcium influx may be a mitogenic signal of IGFs.

Circulating digitalis-like substance is increased in doca-salt hypertension

Blood pressure and digitalis-like substance were measured in the plasma of control, salt-treated, and DOCA-salt treated rats. Blood pressure in DOCA-salt treated rats was significantly higher than that of either control or salt-treated animals. Digitalis-like activity was measured by two methods, radioimmunoassay for digoxin, and a receptor binding assay employing a rat brain synaptosomal membrane fraction. Digoxin-like immunoreactivity in plasma was not detected in either control or salt-treated rats, but was detected in DOCA-salt treated rats. Receptor binding activity in salt-treated rats was slightly but significantly higher than that of control rats. In DOCA-salt treated rats, receptor binding activity was significantly higher than that of salt-treated rats. Partial purification of the digitalis-like substance in plasma was performed by gel filtration using Sephadex G-25. Two peaks containing digoxin-like immunoreactivity were observed. Receptor binding activity, as well as Na+-K+ ATPase inhibitory activity, was detected only in the second peak, in which approximately 70% of the digoxin-like immunoreactivity was eluted. These results indicate that a circulating digitalis-like substance is increased in DOCA-salt hypertension.

Dual effect of activin a on cell growth in Balbc 3T3 cells

Effects of activin A on cell growth were studied in Balb/c 3T3 cells. When incubated with serum, activin A inhibited serum-induced increase in DNA synthesis in a concentration-dependent manner. Activin A also inhibited serum-induced increase in cell number. When added in quiescent cells, activin A did not affect competence-inducing activity of PDGF. Activin A by itself had a small competence-inducing activity. In contrast, when added in competent cells, activin A inhibited progression activity of platelet-poor plasma. These results indicate that activin A has dual action on cell proliferation in Balb/c 3T3 cells.

Transient Ca2+-channel current characterized by a low-threshold voltage in zona glomerulosa cells of rat adrenal cortex

Voltage-gated Ca2+-current was identified in single isolated cells of the zona glomerulosa of adrenal cortex and its properties were studied by the “tight-seal” whole cell recording technique. The Ca2+-channel current was dissected from the net current by dialyzing the cells with CsCl. The identified Ca2+-current was found to be activated by a relatively small depolarization only when the cells were held at a large negative holding potential, but it was inactivated within 10–30 ms. The time course of activation and inactivation was voltage-dependent and became faster when the amplitude of depolarization was increased. The transmembrane potential of the glomerulosa cells was highly sensitive to [K+]e, the slope of the potential change per tenfold change in [K+]e being 48 mV. An increase in [K+]e from 4.7 to 10 mM induce a membrane depolarization by 15 mV, which was sufficient to cause the membrane to reach the threshold potential (−60 mV) for activation of the Ca2+-current at physiological concentration of [Ca2+]e (2.5 mM −CaCl2). The observed properties of the Ca2+-current and K+-dependence of the membrane potential may give reasonable explanation for the mechanism of Ca2+-uptake and consequent aldosterone secretion induced by a small increase in [K+]e, which is known to be one of the major stimulations for aldosterone secretion.

Evidence for two distinct voltage-gated calcium channel currents in bovine adrenal glomerulosa cells

We analyzed inward Ca2+ currents in single bovine adrenal glomerulosa cell using whole-cell patch clamp techniques. Two types of voltage-gated Ca2+ channel currents were identified. One was a transient (T) type which decayed within 100 ms, characterized by a low threshold voltage (about -70 mv) similar to that seen in rat adrenal glomerulosa cells (Matsunaga, H. et al. (1987) Pflügers Arch. 408, 351-355.) Another was a long-lasting (L) type which shows a more positive threshold potential. The present results suggest that while T type Ca2+ channels may explain initial calcium influx in response to an elevation in extracellular K+, L type Ca2+ channels may allow sustained calcium influx which is necessary for sustained aldosterone secretion.