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Activation of Neuropeptide Y Receptors Modulates Retinal Ganglion Cell Physiology and Exerts Neuroprotective Actions In Vitro.

Martins J, Elvas F, Brudzewsky D, Martins T, Kolomiets B, Tralhão P, Gøtzsche CR, Cavadas C, Castelo-Branco M, Woldbye DP, Picaud S, Santiago AR, Ambrósio AF - ASN Neuro (2015)

Bottom Line: We found that NPY attenuated the increase in the [Ca2+]i triggered by glutamate mainly via Y1 receptor activation.Using in vitro cultures of rat retinal explants exposed to NMDA, we found that NPY pretreatment prevented NMDA-induced cell death.In conclusion, we found modulatory effects of NPY application that for the first time were detected at the level of RGCs.

View Article: PubMed Central - PubMed

Affiliation: Institute for Biomedical Imaging and Life Sciences (IBILI), Faculty of Medicine, University of Coimbra, 3004-548 Coimbra, Portugal CNC.IBILI, University of Coimbra, 3004-548 Coimbra, Portugal.

No MeSH data available.


Related in: MedlinePlus

Expression of NPY and NPY receptors in RGCs. (a) Detection of NPY and NPY receptors (Y1, Y2, Y4, and Y5) mRNA expression by RT-PCR in purified RGCs. RGCs were isolated from the retinas of either pups or young adult Long Evans and Wistar rats. DNA ladder. (b) NPY and NPY receptor immunoreactivity (ir) in a purified RGC culture obtained from Wistar pups (a–e, red). RGCs were stained with antiBrn3a (RGC marker, green) and nuclei with DAPI (blue). Scale bar: 20 µm. (c) NPY-ir and NPY receptor-ir in retinal slices obtained from young adult Wistar rats (a–e, red). RGCs were stained with the RGC marker Brn3a (green) and nuclei with DAPI (blue). NPY-ir was mainly detected in strata 1, 3, and 5 of IPL (arrows; a). Y1-ir was detected in distal and proximal INL, strata 2 and 4 of IPL, and RGCs (arrows; b). Y2-ir was detected in INL (c). Y4-ir was detected in INL and GCL (d). and Y5-ir was detected in Müller cells (e). (d) Y5-ir (red) is colocalized with Müller cells (green) in retinal slices of young adult Wistar rats. Müller cells were identified by vimentin-ir (green). Nuclei were stained with DAPI (blue). GCL = ganglion cell layer; IPL = inner plexiform layer; INL = inner nuclear layer; OPL = outer plexiform layer; ONL = outer nuclear layer. Scale bar: 50 µm.
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fig1-1759091415598292: Expression of NPY and NPY receptors in RGCs. (a) Detection of NPY and NPY receptors (Y1, Y2, Y4, and Y5) mRNA expression by RT-PCR in purified RGCs. RGCs were isolated from the retinas of either pups or young adult Long Evans and Wistar rats. DNA ladder. (b) NPY and NPY receptor immunoreactivity (ir) in a purified RGC culture obtained from Wistar pups (a–e, red). RGCs were stained with antiBrn3a (RGC marker, green) and nuclei with DAPI (blue). Scale bar: 20 µm. (c) NPY-ir and NPY receptor-ir in retinal slices obtained from young adult Wistar rats (a–e, red). RGCs were stained with the RGC marker Brn3a (green) and nuclei with DAPI (blue). NPY-ir was mainly detected in strata 1, 3, and 5 of IPL (arrows; a). Y1-ir was detected in distal and proximal INL, strata 2 and 4 of IPL, and RGCs (arrows; b). Y2-ir was detected in INL (c). Y4-ir was detected in INL and GCL (d). and Y5-ir was detected in Müller cells (e). (d) Y5-ir (red) is colocalized with Müller cells (green) in retinal slices of young adult Wistar rats. Müller cells were identified by vimentin-ir (green). Nuclei were stained with DAPI (blue). GCL = ganglion cell layer; IPL = inner plexiform layer; INL = inner nuclear layer; OPL = outer plexiform layer; ONL = outer nuclear layer. Scale bar: 50 µm.

Mentions: The presence of NPY and NPY receptors (Y1, Y2, Y4, and Y5) was assessed in purified rat RGCs. Total RNA was extracted immediately after purification. To assess the presence of mRNA encoding for NPY and NPY receptors (Y1, Y2, Y4, and Y5) by RT-PCR, we used cDNA from RGCs isolated from the retinas of P3 to P4 pups and 8 weeks old adult rats of two different strains: Wistar and Long Evans (Figure 1(a)). We found that mRNA for both NPY and NPY receptors (Y1, Y2, Y4, and Y5) was detected in purified RGCs from both rat strains and at both ages.Figure 1.


Activation of Neuropeptide Y Receptors Modulates Retinal Ganglion Cell Physiology and Exerts Neuroprotective Actions In Vitro.

Martins J, Elvas F, Brudzewsky D, Martins T, Kolomiets B, Tralhão P, Gøtzsche CR, Cavadas C, Castelo-Branco M, Woldbye DP, Picaud S, Santiago AR, Ambrósio AF - ASN Neuro (2015)

Expression of NPY and NPY receptors in RGCs. (a) Detection of NPY and NPY receptors (Y1, Y2, Y4, and Y5) mRNA expression by RT-PCR in purified RGCs. RGCs were isolated from the retinas of either pups or young adult Long Evans and Wistar rats. DNA ladder. (b) NPY and NPY receptor immunoreactivity (ir) in a purified RGC culture obtained from Wistar pups (a–e, red). RGCs were stained with antiBrn3a (RGC marker, green) and nuclei with DAPI (blue). Scale bar: 20 µm. (c) NPY-ir and NPY receptor-ir in retinal slices obtained from young adult Wistar rats (a–e, red). RGCs were stained with the RGC marker Brn3a (green) and nuclei with DAPI (blue). NPY-ir was mainly detected in strata 1, 3, and 5 of IPL (arrows; a). Y1-ir was detected in distal and proximal INL, strata 2 and 4 of IPL, and RGCs (arrows; b). Y2-ir was detected in INL (c). Y4-ir was detected in INL and GCL (d). and Y5-ir was detected in Müller cells (e). (d) Y5-ir (red) is colocalized with Müller cells (green) in retinal slices of young adult Wistar rats. Müller cells were identified by vimentin-ir (green). Nuclei were stained with DAPI (blue). GCL = ganglion cell layer; IPL = inner plexiform layer; INL = inner nuclear layer; OPL = outer plexiform layer; ONL = outer nuclear layer. Scale bar: 50 µm.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4552225&req=5

fig1-1759091415598292: Expression of NPY and NPY receptors in RGCs. (a) Detection of NPY and NPY receptors (Y1, Y2, Y4, and Y5) mRNA expression by RT-PCR in purified RGCs. RGCs were isolated from the retinas of either pups or young adult Long Evans and Wistar rats. DNA ladder. (b) NPY and NPY receptor immunoreactivity (ir) in a purified RGC culture obtained from Wistar pups (a–e, red). RGCs were stained with antiBrn3a (RGC marker, green) and nuclei with DAPI (blue). Scale bar: 20 µm. (c) NPY-ir and NPY receptor-ir in retinal slices obtained from young adult Wistar rats (a–e, red). RGCs were stained with the RGC marker Brn3a (green) and nuclei with DAPI (blue). NPY-ir was mainly detected in strata 1, 3, and 5 of IPL (arrows; a). Y1-ir was detected in distal and proximal INL, strata 2 and 4 of IPL, and RGCs (arrows; b). Y2-ir was detected in INL (c). Y4-ir was detected in INL and GCL (d). and Y5-ir was detected in Müller cells (e). (d) Y5-ir (red) is colocalized with Müller cells (green) in retinal slices of young adult Wistar rats. Müller cells were identified by vimentin-ir (green). Nuclei were stained with DAPI (blue). GCL = ganglion cell layer; IPL = inner plexiform layer; INL = inner nuclear layer; OPL = outer plexiform layer; ONL = outer nuclear layer. Scale bar: 50 µm.
Mentions: The presence of NPY and NPY receptors (Y1, Y2, Y4, and Y5) was assessed in purified rat RGCs. Total RNA was extracted immediately after purification. To assess the presence of mRNA encoding for NPY and NPY receptors (Y1, Y2, Y4, and Y5) by RT-PCR, we used cDNA from RGCs isolated from the retinas of P3 to P4 pups and 8 weeks old adult rats of two different strains: Wistar and Long Evans (Figure 1(a)). We found that mRNA for both NPY and NPY receptors (Y1, Y2, Y4, and Y5) was detected in purified RGCs from both rat strains and at both ages.Figure 1.

Bottom Line: We found that NPY attenuated the increase in the [Ca2+]i triggered by glutamate mainly via Y1 receptor activation.Using in vitro cultures of rat retinal explants exposed to NMDA, we found that NPY pretreatment prevented NMDA-induced cell death.In conclusion, we found modulatory effects of NPY application that for the first time were detected at the level of RGCs.

View Article: PubMed Central - PubMed

Affiliation: Institute for Biomedical Imaging and Life Sciences (IBILI), Faculty of Medicine, University of Coimbra, 3004-548 Coimbra, Portugal CNC.IBILI, University of Coimbra, 3004-548 Coimbra, Portugal.

No MeSH data available.


Related in: MedlinePlus