Lasting effects of prenatal exposure to Cannabis in the retina of the offspring: an experimental study in mice

Abstract

Background: Prenatal exposure to Cannabis is a worldwide growing problem. Although retina is part of the central nervous system, the impact of maternal Cannabis use on the retinal development and its postnatal consequences remains unknown. As the prenatal period is potentially sensitive in the normal development of the retina, we hypothesized that recreational use of Cannabis during pregnancy may alter retina structure in the offspring. To test this, we developed a murine model that mimics human exposure in terms of dose and use.

Methods: Pregnant BalbC mice were exposed daily for 5 min to Cannabis smoke (0.2 g of Cannabis) or filtered air, from gestational day 5 to 18 (N = 10/group). After weaning period, pups were separated and examined weekly. On days 60, 120, 200, and 360 after birth, 10 pups from each group were randomly selected for Spectral Domain Optical Coherence Tomography (SD-OCT) analysis of the retina. All retina layers were measured and inner, outer, and total retina thickness were calculated. Other 37 mice from both groups were sacrificed on days 20, 60, and 360 for retinal stereology (total volume of the retina and volume fraction of each retinal layer) and light microscopy. Means and standard deviations were calculated and MANOVA was performed.

Results: The retina of animals which mother was exposed to Cannabis during gestation was 17% thinner on day 120 (young adult) than controls (P = 0.003) due to 21% thinning of the outer retina (P = 0.001). The offspring of mice from the exposed group presented thickening of the IS/OS in comparison to controls on day 200 (P < 0.001). In the volumetric analyzes by retinal stereology, the exposed mice presented transitory increase of the IS/OS total volume and volume fraction on day 60 (young adult) compared to controls (P = 0.008 and P = 0.035, respectively). On light microscopy, exposed mice presented thickening of the IS/OS on day 360 (adult) compared to controls (P = 0.03).

Conclusion: Gestational exposure to Cannabis smoke may cause structural changes in the retina of the offspring that return to normal on mice adulthood. These experimental evidences suggest that children and young adults whose mothers smoked Cannabis during pregnancy may require earlier and more frequent clinical care than the non-exposed population.

Keywords: Cannabis; Mice; Microscopy; Optical coherence; Prenatal exposure delayed effects; Retina; Tomography.

Fig. 1

Total retina thickness on SD-OCT of the offspring of pregnant mice exposed to Cannabis smoke or filtered air during pregnancy (mean ± standard deviation). *P = 0.003 (MANOVA, 95% CI: 251–263, age*exposition)

Fig. 2

Inner and outer retina thickness on SD-OCT of the offspring of pregnant mice exposed to Cannabis smoke or filtered air during pregnancy (mean ± standard deviation). Inner retina included nerve fiber layer, ganglion cell layer (NFL + GCL), inner plexiform layer (IPL), and inner nuclear layer (INL). Outer retina included outer plexiform layer (OPL), outer nuclear layer (ONL), photoreceptors inner segments and outer segments junction (IS/OS), and retinal pigment epithelium (RPE). *P = 0.001 (MANOVA, 95% CI: 117–126, age*exposition)

Fig. 3

Retinal layers thickness on SD-OCT of the offspring of pregnant mice exposed to Cannabis smoke or filtered air during pregnancy (mean ± standard deviation).*P = 0.0001 (MANOVA, 95% CI: 47–54, age*exposition)

Fig. 4

Mean total volume of each retina layer under stereology of the offspring of pregnant mice exposed to Cannabis smoke or filtered air during pregnancy. *P = 0.008 (MANOVA, 95% CI: 0.8–1.5, age*exposition)

Fig. 5

Mean volume fraction of each retina layer under stereology of the offspring of pregnant mice exposed to Cannabis smoke or filtered air during pregnancy. *P = 0.035 (MANOVA, 95% CI: 10.6–17.4, age*exposition) **P = 0.005 (MANOVA, 95% CI: 22.8–26.4, sex*exposition)

Fig. 6

Retina layers thickness under light microscopy of the offspring of pregnant mice exposed to Cannabis smoke or filtered air during pregnancy (mean ± standard deviation). *P = 0.03 (MANOVA, 95% CI: 55–78, age*exposition)