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Effect of the PGD2-DP signaling pathway on primary cultured rat hippocampal neuron injury caused by aluminum overload.

Ma J, Yang Q, Wei Y, Yang Y, Ji C, Hu X, Mai S, Kuang S, Tian X, Luo Y, Liang G, Yang J - Sci Rep (2016)

Bottom Line: BW245C reduced the Ca(2+) fluorescence intensity and protected the neurons.DK-PGD2 increased the intensity of Ca(2+) fluorescence, while CAY10471 had the opposite effect.In conclusion, contrary to the effect of DP2, the PGD2-DP1 signaling pathway protects against the primary cultured rat hippocampal neuronal injury caused by aluminum overload.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Chongqing Medical University, the Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing 400016, China.

ABSTRACT
In the present study, the agonists and antagonists of DP receptor were used to examine whether the PGD2-DP signaling pathway affects neuronal function. Primary cultured hippocampal neuron was prepared and treated with aluminum maltolate (100 μM) to establish the neuronal damage model. PGD2 and cAMP content was detected by ELISA. L-PGDS and DPs mRNA and protein expression were measured by RT-PCR and Western blotting, respectively. The aluminium-load neuron was treated with the DP1 agonist BW245C, the DP1 antagonist BWA868C, the DP2 agonist DK-PGD2, and the DP2 antagonist CAY10471, respectively. Neuronal pathomorphology was observed using H-E staining. The cell viability and the lactate dehydrogenase leakage rates of neurons were measured with MTT and LDH kit, respectively. Ca(2+) level was detected by Fluo-3/AM. In the model group, the MTT values obviously decreased; LDH leakage rates and PGD2 content increased significantly; L-PGDS, DP1 mRNA and protein expressions increased, and DP2 level decreased. BW245C reduced the Ca(2+) fluorescence intensity and protected the neurons. DK-PGD2 increased the intensity of Ca(2+) fluorescence, while CAY10471 had the opposite effect. In conclusion, contrary to the effect of DP2, the PGD2-DP1 signaling pathway protects against the primary cultured rat hippocampal neuronal injury caused by aluminum overload.

No MeSH data available.


Related in: MedlinePlus

The morphology of primary cultured hippocampal neurons and NSE staining of neurons.(a1,b1,c1) Representative microscopic photographs show the morphology of hippocampal neurons after inoculation for 1d, 3d and 7d. Cultured 7 days later, adjacent neurons had developed to mutual cross-linked cell. Sections were pictured at 200× power. (a2,b2,c2) NSE staining showed that more than 95% of positive neurons existed. Sections were pictured at 100×, 200×, 400×, respectively.
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f1: The morphology of primary cultured hippocampal neurons and NSE staining of neurons.(a1,b1,c1) Representative microscopic photographs show the morphology of hippocampal neurons after inoculation for 1d, 3d and 7d. Cultured 7 days later, adjacent neurons had developed to mutual cross-linked cell. Sections were pictured at 200× power. (a2,b2,c2) NSE staining showed that more than 95% of positive neurons existed. Sections were pictured at 100×, 200×, 400×, respectively.

Mentions: Observed under inverted microscope, the neuronal soma was plumped with short processes after cultured 1 day. The neurons appeared oval or vertebral after 3 days, and they had become stereoscopic. For 7 days, the cultured neuronal bodies enlarged and the nervous process eruption and extension into reticulation. The purity of the hippocampus neurons was detected by neuron specific enolase (NSE) after cultured for 7 d. The bodies and axons of the positive cells were stained to be brown by NSE and nuclei were stained to be blue after counterstained with hematoxylin. One hundred cells were selected randomly to evaluate the number and the percentage of positive cells. Our results showed that there were more than 95% of positive cells (Fig. 1).


Effect of the PGD2-DP signaling pathway on primary cultured rat hippocampal neuron injury caused by aluminum overload.

Ma J, Yang Q, Wei Y, Yang Y, Ji C, Hu X, Mai S, Kuang S, Tian X, Luo Y, Liang G, Yang J - Sci Rep (2016)

The morphology of primary cultured hippocampal neurons and NSE staining of neurons.(a1,b1,c1) Representative microscopic photographs show the morphology of hippocampal neurons after inoculation for 1d, 3d and 7d. Cultured 7 days later, adjacent neurons had developed to mutual cross-linked cell. Sections were pictured at 200× power. (a2,b2,c2) NSE staining showed that more than 95% of positive neurons existed. Sections were pictured at 100×, 200×, 400×, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835855&req=5

f1: The morphology of primary cultured hippocampal neurons and NSE staining of neurons.(a1,b1,c1) Representative microscopic photographs show the morphology of hippocampal neurons after inoculation for 1d, 3d and 7d. Cultured 7 days later, adjacent neurons had developed to mutual cross-linked cell. Sections were pictured at 200× power. (a2,b2,c2) NSE staining showed that more than 95% of positive neurons existed. Sections were pictured at 100×, 200×, 400×, respectively.
Mentions: Observed under inverted microscope, the neuronal soma was plumped with short processes after cultured 1 day. The neurons appeared oval or vertebral after 3 days, and they had become stereoscopic. For 7 days, the cultured neuronal bodies enlarged and the nervous process eruption and extension into reticulation. The purity of the hippocampus neurons was detected by neuron specific enolase (NSE) after cultured for 7 d. The bodies and axons of the positive cells were stained to be brown by NSE and nuclei were stained to be blue after counterstained with hematoxylin. One hundred cells were selected randomly to evaluate the number and the percentage of positive cells. Our results showed that there were more than 95% of positive cells (Fig. 1).

Bottom Line: BW245C reduced the Ca(2+) fluorescence intensity and protected the neurons.DK-PGD2 increased the intensity of Ca(2+) fluorescence, while CAY10471 had the opposite effect.In conclusion, contrary to the effect of DP2, the PGD2-DP1 signaling pathway protects against the primary cultured rat hippocampal neuronal injury caused by aluminum overload.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Chongqing Medical University, the Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing 400016, China.

ABSTRACT
In the present study, the agonists and antagonists of DP receptor were used to examine whether the PGD2-DP signaling pathway affects neuronal function. Primary cultured hippocampal neuron was prepared and treated with aluminum maltolate (100 μM) to establish the neuronal damage model. PGD2 and cAMP content was detected by ELISA. L-PGDS and DPs mRNA and protein expression were measured by RT-PCR and Western blotting, respectively. The aluminium-load neuron was treated with the DP1 agonist BW245C, the DP1 antagonist BWA868C, the DP2 agonist DK-PGD2, and the DP2 antagonist CAY10471, respectively. Neuronal pathomorphology was observed using H-E staining. The cell viability and the lactate dehydrogenase leakage rates of neurons were measured with MTT and LDH kit, respectively. Ca(2+) level was detected by Fluo-3/AM. In the model group, the MTT values obviously decreased; LDH leakage rates and PGD2 content increased significantly; L-PGDS, DP1 mRNA and protein expressions increased, and DP2 level decreased. BW245C reduced the Ca(2+) fluorescence intensity and protected the neurons. DK-PGD2 increased the intensity of Ca(2+) fluorescence, while CAY10471 had the opposite effect. In conclusion, contrary to the effect of DP2, the PGD2-DP1 signaling pathway protects against the primary cultured rat hippocampal neuronal injury caused by aluminum overload.

No MeSH data available.


Related in: MedlinePlus