Anat Cell Biol.  2022 Sep;55(3):311-319. 10.5115/acb.22.034.

Effect of caffeine on genes expressions of developing retinas in the chick model

• Lekchaoum T
• Buddawong A
• Ahi S
• Chandee N
• Weerachatyanukul W
• Asuvapongpatana S

• KMID: 2533357
• DOI: http://doi.org/10.5115/acb.22.034

Abstract

It has been reported that overconsumption of caffeine during pregnancy leads to a deleterious effect within the nervous tissues during embryonic development. In this study, we further extrapolated the effect of caffeine in the developing retinas, which is known to be one of the most sensitive tissues in chick embryos. Morphological changes of retinal thickness and organization of neuroretinal epithelium were monitored using three gene markers, Atoh7, FoxN4, and Lim1. Upon treating with a single dose of caffeine (15 µmol at embryonic day 1 [E1]), relative thicknesses of developing retinas (particularly of E7 and E9) were significantly altered. Among the three genes studied, the expression pattern of Atoh7 was notably altered while those of FoxN4, and Lim1 mRNA showed only a slight change in these developing retinas. Quantitative polymerase chain reaction results supported the most notable changes of Atoh7 but not FoxN4, and Lim1 gene in the developing retinas, particularly at E7. The effect of caffeine towards other organs during development should be extrapolated and the awareness of its intensive consumption should be raised.

Keyword

Caffeine; Chick embryo; Retina; Epithelium; Retinal ganglion cells

Figure

• 

  Fig. 1 Histology of developing retinas of control and caffeine-treated embryos at the stages of E3–E9. The paraffin sections were stained with hematoxylin and eosin and the images were acquired at low magnification (bar 200 µm, A–H) and high magnification (bar 100 µm, I–P) to demonstrate cellular organization in retinas. NBL, neuroblastic layer; pGCL, presumptive ganglion cell layer; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer. *Delaminated IPL.

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  Fig. 2 Localization of the three retinal marker genes in developing retinas of the control and caffeinetreated embryos. Note the apparent changes in the staining pattern and intensity of Atoh7 gene in E7 and E9 retinas of control (A–D) and caffeine-treated animals (E–H), while those of FoxN4 (I–P), and Lim1 (Q–X) are minimally changed. Bars in E3 column=100 μm; the E5, E7, E9 column=50 μm. NBL, neuroblastic layer; pGCL, presumptive ganglion cell layer; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer.

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  Fig. 3 Quantitative analysis of the three retinal marker genes in embryos treated with a single dose of caffeine compared with the control embryos. Total of RNA of the whole eyes were collected from E7 and E9 embryos and the expression levels of Atoh7 (A), FoxN4 (B), and Lim1 (C) were quantitated by real-time polymerase chain reaction (PCR). The data were calculated from 2–δδCq quantitative PCR values of caffeine-treated and control embryos and expressed as mean±standard error of log2 fold changes of the triplicated.

Reference

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