An in vitro organ culture system with developing mouse palates was improved to characterize the cleft palate (CP)-inducing potential of chemicals and underlying mechanisms. Palatal explants collected from gestation day 12 mouse fetuses were cultured with various concentrations of teratogens and examined for palatal development after 48 and 72 h of culture to assess effects of the chemicals on growth and/or fusion of palatal shelves. When the explants were exposed to diphenylhydantoin or 5-fluorouracil, palatal growth was inhibited in a concentration-dependent manner at 48 h. Suppression of the expression of proliferative cell nuclear antigen revealed poor cell proliferation. At 72 h, the incidence of explants with CP was significantly increased in the high-dose groups, suggesting that CP induction is mainly attributable to inhibition of palatal growth. By contrast, retinoic acid and hydrocortisone significantly lowered the rates of fused palates at 72 h in all treated groups, while they exhibited no effects on palatal growth at 48 h even at the highest concentration. Because no apoptosis was found in the epithelial cells at the tip of these palates, these chemicals are suggested to inhibit palatal fusion process by preventing apoptosis.

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PubMedDate not supplied

Modulation by Zn2+ of GABA responses in bipolar cells of the mouse retina.

The localization of endogenous Zn2+ in the mouse retina was examined histochemically and the inhibitory action of Zn2+ on GABA-induced responses was studied in bipolar cells isolated from the mouse retina. Accumulation of endogenous Zn2+ was detected in photoreceptors, bipolar, and/or amacrine cells by either the bromopyridylazo-diethylaminophenol method or the dithizone method. Under whole-cell recording conditions, GABA induced a Cl- current in isolated bipolar cells. The current consisted of two components. The first component was inhibited completely by application of 100 microM bicuculline, suggesting that this is a GABA(A)-receptor mediated current. The second component was inhibited completely by 100 microM 3-aminopropyl-(methyl)-phosphinic acid, suggesting that this is a GABA(C)-receptor mediated current. GABA(C) receptors were present at a higher density on the axon terminal than on dendrites. Zn2+ inhibited both GABA(A) and GABA(C) receptors. GABA(C) receptors were more susceptible to Zn2+; the IC50 for the GABA(A) receptor was 67.4 microM and that for the GABA(C) receptor was 1.9 microM. These results suggest that Zn2+ modulates the inhibitory interaction between amacrine and bipolar cells, particularly that mediated by the GABA(C) receptor.

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PubMedDate not supplied

GABA-mediated component in the feedback response of turtle retinal cones.

The negative feedback from horizontal cells to cone photoreceptors contributes to the formation of the receptive-field surround in cone photoreceptors. Recently, studies on the modulation of voltage-gated Ca(2+) currents in cone photoreceptors have led to great progress in our understanding of the mechanism of horizontal-cone feedback. Another highly probable hypothesis is that GABA mediates this feedback. This hypothesis is supported by the facts that cone photoreceptors respond to GABA and that horizontal cells release GABA. However, GABA-mediated synaptic inputs from horizontal cells to cone photoreceptors have not been demonstrated. In the present study, we examined whether cone photoreceptors receive GABAergic inputs from horizontal cells using a slice patch technique in the turtle retina. When 1 mM of GABA was applied to the cone photoreceptors, GABA-induced currents were activated. GABA-induced currents reversed their polarity at the equilibrium potential of Cl-. The application of 30 microM of SR95531, an antagonist of GABAA receptors, alone did not produce any change in the holding currents. When 200 microM of pentobarbital was introduced to potentiate the GABAergic inputs to the cone photoreceptors, however, the inhibitory action of SR95531 on GABAergic inputs became detectable. The amplitude of the GABAergic inputs, potentiated by pentobarbital, increased when the horizontal cells were depolarized by the application of 20 microM of kainate, while the amplitude decreased when the horizontal cells were hyperpolarized by the application of 10 microM of CNQX. When the cone photoreceptors were voltage clamped at a potential at which the voltage-gated Ca(2+) current was inactive, horizontal-cone feedback was not observed. However, the horizontal-cone feedback became detectable when the GABAergic inputs to the cone photoreceptors were potentiated by pentobarbital. We concluded that the contribution of GABAergic inputs from horizontal cells to cone pedicles in the formation of the receptive-field surround in cone photoreceptors is very limited but that the modulation of voltage-gated Ca(2+) currents in cone photoreceptors is a physiologically relevant mechanism for horizontal-cone feedback.

6-Cyano-7-nitroquinoxaline-2,3-dione↗