Hisashi Koda

Involvement of EP3 subtype of prostaglandin E receptors in PGE2-induced enhancement of the bradykinin response of nociceptors.

Prostaglandin E2 augments bradykinin-induced discharges of polymodal receptors as studied in vitro preparations. The antagonist and agonists for three subtypes of EP receptors were used to determine which subtype is involved in this phenomenon. The agonist for EP3 (M&B28767) simulated the PGE2-induced effect but not for EP2 (butaprost). The antagonist for EP1 (AH6809) did not suppress the effect. These findings indicate the involvement of the EP3 receptor subtype in the effect.

Excitation and sensitization of the heat response induced by a phorbol ester in canine visceral polymodal receptors studied in vitro

To clarify the possible involvement of protein kinase (PK) C activation in the excitation and sensitization of polymodal receptors (PMRs), the effects of phorbol 12,13-dibutyrate (PDBu) on PMRs were studied in canine testis-spermatic nerve preparations in vitro. Application of PDBu (10(-7), 10(-6), AND 10(-5) M) for 5 min evoked a significant increase in the ongoing activity of the PMRs within 15 min. PDBu (10(-8) to 10(-5) M) significantly augmented the subsequent heat responses of the PMRs. Staurosporine (10(-6) M), a PK inhibitor, attenuated the effect of PDBu on heat responses. These data suggest that activation of PKC contributes to the activities of PMRs.

Potentiation and suppression of the histamine response by raising and lowering the temperature in canine visceral polymodal receptors in vitro.

It is well known that itch and inflammatory pain are enhanced when tissue is warmed, while they are suppressed when tissue is cooled. To see whether these changed sensations are based on the changed response of sensory receptors, the temperature dependency of the excitation of polymodal receptors induced by histamine, which plays an important role both in itch and inflammatory pain, was studied. Single nerve activities of polymodal receptors were recorded from canine testis-spermatic nerve preparations in vitro. Raising the temperature from 34 to 40 degrees C, a temperature below the threshold for the heat response of polymodal receptors, facilitated the histamine-induced nerve discharge to 268% of that at 34 degrees C, while lowering the temperature to 28 degrees C decreased it to 25%. Facilitation of the histamine response was also observed in the noxious temperature range (48 and 51 degrees C). These results suggest that the potentiation of the histamine-induced sensation by increasing the tissue temperature, as well as its suppression by lowering tissue temperature, can be explained by a temperature-dependent response of peripheral sensory receptors to histamine. However, the suppression of itch by noxious heat reported by Bickford (Bickford, R.G., Experiments relating to the itch sensation, its peripheral mechanism, and central pathways, Clin. Sci. Incorp. Heart, 3 (1937) 377-386) cannot be explained by the noxious heat-induced facilitation of the peripheral receptor response reported in this paper.

Evidence that protein kinase C activation is involved in the excitatory and facilitatory effects of bradykinin on canine visceral nociceptors in vitro

The involvement of protein kinase (PK) C activation in the effects of bradykinin (BK) on peripheral nociceptors, polymodal receptors, was examined using canine testis-spermatic nerve preparations in vitro. Phorbol 12,13-dibutyrate 0.1 microM, which activates PKC, suppressed the BK-induced excitation when applied for 3-5 min prior to BK application, but facilitated it when applied simultaneously with BK. Neither effect was induced by an inactive phorbol ester, 4alpha-phorbol 12, 13-didecanoate, demonstrating that both effects were mediated through the activation of PKC. In addition, staurosporine 1 microM, a PK inhibitor, suppressed both BK-induced excitation and facilitation of the heat response of testicular polymodal receptors without influencing on-going activities and the heat response itself. These results suggest that PKC activation is involved in the excitatory and facilitatory effects of BK on peripheral nociceptors.

Possible contribution of protein kinase C in the effects of histamine on the visceral nociceptor activities in vitro.

To clarify the possible contribution of protein kinase C activation in histamine-induced excitation and sensitization of the heat response of testicular polymodal receptors, the effects of staurosporine, a protein kinase C inhibitor, and phorbol 12,13-dibutyrate, a protein kinase C activating phorbol ester, were studied in visceral polymodal receptors. Single polymodal receptor activities were recorded in vitro from testis-spermatic nerve preparations obtained from deeply anesthetized dogs (pentobarbital sodium, 30 mg/kg, i.v.). Histamine (10 microM)-induced excitation and facilitation of the heat response of polymodal receptors were both suppressed by staurosporine (1 microM), suggesting that activation of protein kinase C is involved in both these effects of histamine. Application of phorbol 12,13-dibutyrate (0.1 microM) mixed with histamine increased the histamine-induced excitation, whereas a 5 min application of phorbol 12,13-dibutyrate before histamine suppressed it. These results suggest that phorbol 12,13-dibutyrate-activated protein kinase C has inactivation as well as activation effects on the intracellular cascade connected to histamine receptors, and that the former has a slower time course.

Augmenting effects of cyclic AMP on the heat response of canine testicular polymodal receptors

It has long been thought that the augmenting effects of E series prostaglandins (PG) are mediated by cyclic AMP (cAMP). In clarifying the validity of this hypothesis, we studied the effects of increases in intracellular cAMP on the heat response of testicular polymodal receptors. Recordings were obtained from testis-spermatic nerve preparations excised from dogs anesthetized with pentobarbital (30 mg/kg, i.v.). Induction of increases in intracellular cAMP by either forskolin, an adenylyl cyclase activator, or a mixture of dibutyryl cAMP (10(-4) M) and 3-isobutyl-1-methyl xanthine (10(-4) M) significantly and reversibly augmented the heat response. These results support the notion that cAMP is involved in the augmenting effect of PG E2 on the heat response of nociceptors.

Opposite effects of increased intracellular cyclic AMP on the heat and bradykinin responses of canine visceral polymodal receptors in vitro

To clarify the validity of the long standing hypothesis that effects of E series prostaglandin (PG)S are mediated by cyclic AMP (cAMP), we studied the effects of increases in intracellular cAMP on the heat and bradykinin responses of testicular polymodal receptors. Polymodal receptor activities were recorded in vitro from testis-spermatic nerve preparations excised from dogs anesthetized with pentobarbital (30 mg/kg, i.v.). Increases in intracellular cAMP induced by either forskolin (5 or 10 microM), an adenylyl cyclase activator, or a mixture of dibutyryl cAMP (20-100 microM), a membrane permeable cAMP analog, and 3-isobutyl-1-methyl xanthine (20-100 microM), an inhibitor of the cAMP degrading enzyme, significantly augmented the response to heat (42-48 degrees C). In contrast, these substances failed to facilitate the response to bradykinin (0.1 or 1 microM) and instead suppressed it. Dideoxyforskolin (10 microM), an inactive analog of forskolin, had no effects on both the heat and bradykinin responses. These results demonstrate that an increase in intracellular cAMP induces opposite effects on the heat and bradykinin responses. Possible involvement of intracellular cAMP in the facilitatory effects of PGE2 on both responses was discussed in connection with the PGE receptor subtypes involved in the sensitization of the bradykinin and heat responses.

Influence of histamine on the bradykinin response of canine testicular polymodal receptors in vitro

Effects of histamine on the testicular polymodal receptors were studied in vitro using canine testisspermatic nerve preparations. Histamine induced distinct increase in the discharge rate in 6 out of 17 units tested (high responders), while it only weakly excited the remaining 11 units (low responders). The bradykinin response of low responders tended to be slightly facilitated by pretreatment with histamine, while that of high responders tended to be suppressed. Significant correlation was observed between the magnitude of histamine-induced discharges and the magnitude of change in the bradykinin responses.

Histamine-induced sensitization of the heat response of canine visceral polymodal receptors.

In elucidating the possible involvement of histamine (His) in hyperalgesia to heat, the effects of His on the heat response of testicular polymodal receptors were studied in vitro in canine testis-spermatic nerve preparations. His (100 and 1000 microM) induced discharges > 0.5 impulses/s (high-responders) in about one third of units tested. His (> or = 10 microM) facilitated the heat response irrespective of whether His alone induced substantial excitation, and the magnitude of facilitation at 100 microM did not differ between high- and low-responders (His-induced discharges < or = 0.5 impulses/s). Conduction velocity of high-responders at 100 microM (2.2 +/- 0.9 m/s, n = 5) was slower on average than that of low-responders (12.9 +/- 1.6 m/s, n = 9). The sensitizing effect of His reported in this paper suggests that His plays a role in hyperalgesia.

Possible involvement of the EP2 receptor subtype in PGE2-induced enhancement of the heat response of nociceptors

Prostaglandin E2 augments bradykinin- and heat-induced discharges of polymodal receptors as studied in vitro preparations. Our previous study revealed the involvement of the EP3 receptor subtype in the PGE2 induced enhancement of the BK response [Brain Res. 632 (1993) 321-324]. The agonist for EP2 (butaprost; 10(-8) M) significantly augmented heat responses, but did not augment the BK responses at concentrations from 10(-8) to 10(-5) M; however, the agonist for EP3 (M&B28767) or EP1 (17-phenyl-trinor-PGE2) at 10(-7) M did not affect the heat responses. These findings indicate the involvement of the EP2 receptor subtype in the augmenting effect of PGE2 on heat responses.