The localization of PGE2 receptor subtypes in rat retinal cultures and the neuroprotective effect of the EP2 agonist butaprost.
It is concluded from immunohistochemical that all four types of prostaglandin-E(2) (PGE(2)) (EP1, EP2, EP3 and EP4) receptors are associated with specific cell-types in primary rat retinal cultures. Analysis specifically of EP2 receptor immunoreactivity shows it to coexist with some neurones expressing Thy-1 and calbindin immunoreactivities as well as with vimentin-positive Müller cells. Moreover, exposure of cultures to the EP2 specific agonist butaprost (100 nM) for a period of 24h results in a generation of cAMP thus providing support for the functionality of EP2 receptors. Cell survival was significantly affected in cultures where the serum concentration was reduced from 10 to 1% for 24h. This was reflected by a reduction in the number of GABA-positive neurons and an elevation of released lactate dehydrogenase (LDH) into the culture medium. Moreover, a number of cells displayed a clear generation of reactive oxygen species (ROS) and a staining for the breakdown of DNA by the TUNEL procedure as an indicator for apoptosis. These negative effects were attenuated when butaprost (100 nM) was present during the serum reduction and 30 min before the insult. The present studies provide evidence to show that all PGE(2) receptor types exist in the retina of rat pups, remain functional when the retinal cells are cultured and that specific activation of EP2 receptors with butaprost can attenuate a detrimental insult caused by insufficient serum that may occur in situ by reduced trophic support.
Andrade da Costa
Belmira Lara da Silveira
BL
Departamento de Fisiologia e Farmacologia, Centro de Ciências Biológicas (CCB), CCB-UFPE, Av. Prof. Moraes Rego s/n, Cidade Universitária, 50670901 Recife, PE, Brazil.
Kang
Kui Dong
KD
Rittenhouse
Kay D
KD
Osborne
Neville N
NN
D000527:Alprostadil / Q000031:analogs & derivatives* / Q000097:blood / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000187:drug effects; D064026:Calbindins; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects; D015232:Dinoprostone / Q000819:agonists* / Q000378:metabolism; D020287:In Situ Nick-End Labeling; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009457:Neuroglia / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018696:Neuroprotective Agents / Q000097:blood / Q000494:pharmacology*; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D018078:Receptors, Prostaglandin E / Q000819:agonists* / Q000378:metabolism; D012160:Retina / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D064030:S100 Calcium Binding Protein G / Q000378:metabolism; D018800:Thy-1 Antigens / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D064026:Calbindins; D016895:Culture Media, Serum-Free; D018696:Neuroprotective Agents; D017382:Reactive Oxygen Species; D018078:Receptors, Prostaglandin E; D064030:S100 Calcium Binding Protein G; D018800:Thy-1 Antigens; D014746:Vimentin; D005680:gamma-Aminobutyric Acid; D000242:Cyclic AMP; D007770:L-Lactate Dehydrogenase; D000527:Alprostadil; C048491:butaprost; D015232:Dinoprostone