Differential Expression of Melanopsin Isoforms Opn4L and Opn4S during Postnatal Development of the Mouse Retina

Photosensitive retinal ganglion cells (pRGCs) respond to light from birth and represent the earliest known light detection system to develop in the mouse retina. A number of morphologically and functionally distinct subtypes of pRGCs have been described in the adult retina, and have been linked to different physiological roles. We have previously identified two distinct isoforms of mouse melanopsin, Opn4L and Opn4S, which are generated by alternate splicing of the Opn4 locus. These isoforms are differentially expressed in pRGC subtypes of the adult mouse retina, with both Opn4L and Opn4S detected in M1 type pRGCs, and only Opn4L detected in M2 type pRGCs. Here we investigate the developmental expression of Opn4L and Opn4S and show a differential profile of expression during postnatal development. Opn4S mRNA is detected at relatively constant levels throughout postnatal development, with levels of Opn4S protein showing a marked increase between P0 and P3, and then increasing progressively over time until adult levels are reached by P10. By contrast, levels of Opn4L mRNA and protein are low at birth and show a marked increase at P14 and P30 compared to earlier time points. We suggest that these differing profiles of expression are associated with the functional maturation of M1 and M2 subtypes of pRGCs. Based upon our data, Opn4S expressing M1 type pRGCs mature first and are the dominant pRGC subtype in the neonate retina, whereas increased expression of Opn4L and the maturation of M2 type pRGCs occurs later, between P10 and P14, at a similar time to the maturation of rod and cone photoreceptors. We suggest that the distinct functions associated with these cell types will develop at different times during postnatal development

Neuropsin (Opn5): a novel opsin identified in mammalian neural tissue.

We have cloned and characterised the expression of a new opsin gene, neuropsin (Opn5), in mice and humans. Neuropsin comprises seven exons on mouse chromosome 17. Its deduced protein sequence suggests a polypeptide of 377 amino acids in mice (354 in humans), with many structural features common to all opsins, including a lysine in the seventh transmembrane domain required to form a Schiff base link with retinaldehyde. Neuropsin shares 25-30% amino acid identity with all known opsins, making it the founding member of a new opsin family. It is expressed in the eye, brain, testis and spinal cord

How blind are they? Phototactic responses in stygobiont diving beetles (Coleoptera: Dytiscidae) from calcrete aquifers of Western Australia

Subterranean water beetles endemic to groundwater calcretes of Western Australia exhibit convergent traits typical of troglomorphic arthropods, including loss of eyes, pigmentation and wings. As these dytiscid species are estimated to have been isolated underground in permanent darkness for over three million years, it is predicted that they will completely lack phototactic responses. We tested this hypothesis by analysing the behaviour of six subterranean beetle species within an observational arena with dark and light hemispheres. Scan samples at 1 min intervals and total time spent on each hemisphere were recorded over a 20 min period, testing at least 15 individuals per species. We quantified behaviour as an index (dark ratio) so that individual species in this, and future, studies can be consistently compared. Results analysed as both categorical and absolute proportion of time spent in each hemisphere suggest negative phototaxis in Paroster macrosturtensis. The remaining five species did not display any preference for either light or dark hemispheres. These results raise the possibility that some ancestral Paroster species may have exhibited negative phototactic behaviour prior to subterranean colonization. The retention of such a behavioural trait in lightless environments could represent the maintenance for some unknown pleiotropic function. Alternatively, it is possible that insufficient time has passed for neutral processes to render photoreception genes and phototactic behaviours non-functional. Our study adds to a growing body of evidence that implies highly troglomorphic animals may have evolved from ancestral species that exhibited negative phototaxis as a preadaptation to living in permanent darkness

Expression of opsin genes early in ocular development of humans and mice.

We have compared the onsets of expression of the classical visual opsins with those of the non-rod, non-cone opsins in foetal and post-natal eye tissue from mice and humans. Mouse Rgr-opsin, peropsin, encephalopsin and melanopsin are all expressed in foetal development by E11.5, unlike the murine rod and cone opsins that exhibit post-natal expression, e.g. P1 for ultraviolet cone opsin and P5 for rod opsin. Human non-rod, non-cone opsins are also all expressed early, by 8.6 weeks post-conception. The implications of these observations are discussed with regard to the possible functions of these opsins at early stages of ocular development