Abstract
Mesencephalic astrocyte-derived neurotrophic factor (MANF), otherwise named Arginine-Rich, Mutated in Early-stage Tumors (ARMET), is a secretory endoplasmic reticulum stress (ERS) protein that is widely expressed in mammalian tissues. To date, little is known about the distribution and expression of MANF in the retina and optic nerve (ON). Therefore, we studied the expression and distribution of MANF in the ON and retina by real-time PCR, immunofluorescence staining and western blotting. Results from rat and mouse were highly consistent in the retina. MANF was detected in both tissues in rat, wherein it was principally localized to the ganglion cell layer (GCL), followed by the inner nuclear layer (INL). The MANF protein levels in the rat retina were 3.33-fold higher than in the rat ON. Additionally, MANF was robustly expressed by retinal ganglion cells (RGCs) in the human retina. In human ON, MANF was partially co-localized with glial fibrillary acidic protein (GFAP), suggesting that it was not restricted to astrocytes. In vitro studies confirmed that MANF could be robustly expressed in RGCs and was found principally within the cytoplasm. Hypoxia can stimulate up-regulation by of MANF expression over time, suggesting that MANF may play a vital role in the functional regulation of RGCs both in health and disease. We believe that the present study improves our understanding of the distribution and expression of MANF in the retina and ON and could help in further analysis of its interact and correlate with the relevant ophthalmic diseases.
Highlights
Mesencephalic astrocyte-derived neurotrophic factor (MANF), known as Arginine-Rich, Mutated in Early-stage Tumors (ARMET), is a secreted endoplasmic reticulum stress (ERS) protein (Apostolou et al, 2008)
Retinal and retinal ganglion cells (RGCs) samples incubated with secondary antibodies without primary antibody displayed little or no autofluorescence (Supplementary Figure 2)
Western blotting was used to find out if MANF could be expressed in the rodent retina and optic nerve (ON)
Summary
Mesencephalic astrocyte-derived neurotrophic factor (MANF), known as Arginine-Rich, Mutated in Early-stage Tumors (ARMET), is a secreted endoplasmic reticulum stress (ERS) protein (Apostolou et al, 2008). It was first isolated from a rat mesencephalic type-1 astrocyte cell line. The widespread expression of MANF together with its regulation by cellular insults and evolutionary conserved nature implies that it likely has important roles in many tissue types, including the eyes, and could present a potentially novel therapeutic modality for the treatment of hypoxic-ischemic or neurodegenerative diseases of the retina. We applied a combination of real-time PCR, immunofluorescence staining and western blotting to determine the expression and distribution of MANF in the retina and optic nerve (ON)
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