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Tracy Nguyen

Publications and source records attributed to Tracy Nguyen.

5 recordsLinked to original sources
PubMed2005

The G alpha(o/i)-coupled cannabinoid receptor-mediated neurite outgrowth involves Rap regulation of Src and Stat3.

The study of the signaling pathways regulating neurite outgrowth in culture is important because of their potential role in neuronal differentiation in vivo. We have previously shown that the G alpha(o/i)-coupled CB1 cannabinoid receptor (CB1R) activates Rap1 to induce neurite outgrowth. G alpha(o/i) also activates the Src-Stat3 pathway. Here, we studied the relationship between the G alpha(o/i)-Rap1 and Src-Stat3 pathways and the role of these signaling pathways in CB1R-mediated neurite outgrowth in Neuro-2A cells. The CB1 agonist HU-210 induced pertussis toxin-sensitive Src and Stat3 phosphorylation. Dominant negative (DN) mutants of Src and Stat3 blocked CB1R-induced neurite outgrowth. Constitutively active Rap 1B and Ral-activated Src and CB1R-induced Src phosphorylation was inhibited by Rap1-DN and Ral-DN, indicating that both Rap1 and Ral mediate downstream signaling from G alpha(o/i) for Src activation. Rap1-activated Ral and Ral-DN blocked Rap-induced Src phosphorylation. G alpha(o)-induced Stat3 activation was blocked by Ral-DN, whereas v-Src-induced Stat3 activation was not inhibited by Ral-DN, indicating that the CB1R, through G alpha(o), mediates the sequential activation of Rap1 to Ral to Src to Stat3 in Neuro-2A cells. Downstream of Src, the CB1R also activated Rac1 and JNK, which enhanced CBR1-mediated Stat3 activation. Rac-DN blocked CB1R-induced activation of JNK. Pharmacological inhibition of JNK blocked Src and CB1R activation of Stat3, indicating that Rac and JNK are also involved in CB1R-mediated neurite outgrowth. Overall, this study demonstrated that G alpha(o/i)-coupled CB1R triggers neurite outgrowth in Neuro-2A through the activation of a signaling network containing two pathways that bifurcate at Src and converge at Stat3.

Animals↗
PubMed2005

Cannabinoid receptor-induced neurite outgrowth is mediated by Rap1 activation through G(alpha)o/i-triggered proteasomal degradation of Rap1GAPII.

The G(alpha)o/i-coupled CB1 cannabionoid receptor induces neurite outgrowth in Neuro-2A cells. The mechanisms of signaling through G(alpha)o/i to induce neurite outgrowth were studied. The expression of G(alpha)o/i reduces the stability of its direct interactor protein, Rap1GAPII, by targeting it for ubiquitination and proteasomal degradation. This results in the activation of Rap1. G(alpha)o/i-induced activation of endogenous Rap1 in Neuro-2A cells is blocked by the proteasomal inhibitor lactacystin. G(alpha)o/i stimulates neurite outgrowth that is blocked by the expression of dominant negative Rap1. Expression of Rap1GAPII also blocks the G(alpha)o/i-induced neurite outgrowth and treatment with proteasomal inhibitors potentiates this inhibition. The endogenous G(alpha)o/i-coupled cannabinoid (CB1) receptor in Neuro-2A cells stimulates the degradation of Rap1GAPII; activation of Rap1 and treatment with pertussis toxin or lactacystin blocks these effects. The CB1 receptor-stimulated neurite outgrowth is blocked by treatment with pertussis toxin, small interfering RNA for Rap, lactacystin, and expression of Rap1GAPII. Thus, the G(alpha)o/i-coupled cannabinoid receptor, by regulating the proteasomal degradation of Rap1GAPII, activates Rap1 to induce neurite outgrowth.

Amino Acid Sequence↗
PubMed2003

Variability in hypoxia-induced corneal swelling is associated with variability in corneal metabolism and endothelial function.

PURPOSE: To determine if the variability in contact lens-induced corneal swelling is associated with variability in corneal oxygen consumption (Q(C)) or corneal endothelial function. METHODS: Corneal swelling was induced in 30 non-contact lens wearers by 2 hours of closed-eye contact lens wear, using thick (oxygen transmission [Dk/t] = 4.0 x 10-9) and thin (Dk/t = 12 x 10-9) hydrogel lenses of identical design. Following the induction of swelling, corneal thickness was continually measured by pachymetry until open-eye steady-state (OESS) thickness was achieved. The percentage of recovery per hour was calculated as a measure of endothelial function. Tear oxygen tension (PO(2)) beneath the hydrogels was measured in the open eye and after 5 minutes of eye closure to obtain estimates of Q(C). A change in corneal pH during eye closure while wearing the hydrogels was used as a measure of hypoxic acidosis. Associations between corneal swelling and endothelial cell density or corneal epithelial thickness were also tested. RESULTS: There were modest but significant (P < 0.05) correlations between thick-lens corneal swelling and thick-lens closed-eye PO(2) (r = -0.36); thin lens corneal swelling and thin-lens closed-eye and open-eye PO(2) (r = -0.40 and -0.39, respectively). Corneal swelling also increases with increasing Q(C) (r = + 0.29 to + 0.33). Corneal swelling was associated with the decrease in pH during lens wear (r = + 0.30 and + 0.20 for thick and thin lenses, respectively). Thick- and thin-lens corneal swelling was significantly associated with percentage of recovery per hour (r = -0.40 and -0.34, respectively). Multiple regression analysis of corneal swelling with PO(2) and percentage of recovery per hour suggested an additive effect, however the effects were not significant. There was a modest inverse association between corneal swelling and OESS, however there were no associations between corneal swelling and endothelial cell density or epithelial thickness. CONCLUSIONS: The variability in contact lens-induced corneal swelling is associated with both corneal metabolic activity (Q(C) and pH) and endothelial function (percentage of recovery per hour). Our interpretation is that individuals with larger Q(C) produce more lactic acid (i.e., more swelling) whereas stronger endothelial function resists swelling. The modest correlations, however, suggest that other factors also are involved in explaining the phenomenon of corneal swelling.

Acidosis↗
PubMed2002

Estimation of human corneal oxygen consumption by noninvasive measurement of tear oxygen tension while wearing hydrogel lenses.

PURPOSE: To devise a procedure for direct estimation of corneal oxygen consumption in human subjects. METHODS: Tear oxygen tension (PO2) was measured at the posterior surface of two standard hydrogel contact lenses (38% water, 0.2 and 0.06 mm thick, oxygen transmissibility [Dk/t] = 4.2 and 14 x 10(-9) cm x mL O2/mL x sec x torr) and one newly available hydrogel-silicone polymer lens (Dk/t = 99 x 10(-9)). The oxygen-sensitive dye, Pd-meso-tetra (4-carboxyphenyl) porphine, bound to bovine serum albumin, was incubated with the lenses overnight. The lenses, coated with the protein-dye complex, were placed on four subjects' eyes, and tear PO2 was measured in the open eye and after 5 minutes of eye closure, using a time-domain phosphorescence measurement system. Given the tear PO2, lens Dk/t, and corneal thickness, oxygen consumption (Q(C), in mL O2/cm(3) x sec) could be calculated from established oxygen diffusion models. RESULTS: Protein-dye complex bound to the lens surface enabled reporting of tear PO2 for long periods. As expected, estimated tear PO2 was higher in subjects wearing lenses with higher Dk/t: mean open-eye PO2 = 30.6 +/- 3.1 and 8.1 +/- 1.3 torr for the thin and thick hydrogel lenses, respectively, and 97.6 +/- 22.9 torr for the hydrogel-silicone lens. After 5 minutes of eye closure, tear PO2 was significantly reduced and reached a new steady state in approximately 20 seconds after eye opening. Fitting a single exponential model to the data and extrapolating to t = 0 provided an estimate of PO2 under the closed lid for the thin hydrogel (PO2 = 7 +/- 2.3 torr) and the hydrogel-silicone lens (PO2 = 22.6 +/- 4 torr). After 5 minutes of eye closure with the thick hydrogel lens, tear PO2 remained constant for approximately 10 seconds after eye opening (mean PO2 = 3.9 +/- 0.7) before increasing to a new steady state. This delay could be accounted for by the time needed for oxygen to diffuse to the posterior surface of the lens. Calculated Q(C) ranged from 2.2 x 10(-4) to 3.7 x 10(-6) mL O2/cm(3) x sec) at the highest and lowest PO2s, respectively, and is comparable to previous in vitro and in vivo estimates. CONCLUSIONS: Tear PO2 behind hydrogel lenses can be measured in human subjects using the phosphorescence of the porphyrin-protein complex bound to the lens surface. The method is simple, fast, reliable, and noninvasive, allowing quick and direct estimates of Q(C). In addition to contact lens wear, this method should be useful for examining the effects of disease, surgery, or topical drugs on the corneal oxygen consumption rate.

Adult↗