Mao C, Agca C, Mocko-Strand J, Wang J, Ullrich-Lüter E, Pan P, Wang S, Arnone M, Frishman L, Klein W (2016) Substituting mouse transcription factor Pou4f2 with a sea urchin orthologue restores retinal ganglion cell development. Proc Biol Sci 283:20152978. doi: 10.1098/rspb.2015.2978 PMID: 26962139

Summary: Pou4f2 is Pou domain transcription factor that is essential for the development of retinal ganglion cells (RGCs) in the vertebrate retina. The sea urchin genome contains SpPou4f1/2, a distant orthologue of Pou4f2, but they have no obvious eyes and their photoreceptors are located around their tube feet disc. Scientists replaced genomic Pou4f2 with an SpPou4f1/2 cDNA to see if SpPou4f1/2 could support RGC development in mice. Mice expressing SpPou4f1/2 developed retinas that looked like wild-type mice. Immunolabeling of retinas with a 1:1000 dilution of Anti-Melanopsin (Cat. #AB-N39) showed the presence of many well-bundled axons emanating from SpPou4f1/2-expressing RGCs. Electroretinogram recordings from these mice indicate that their RGCs are functionally active. These results suggest that there is a high degree of functional conservation between the two genes despite more than 540 million years of divergence from the common ancestor of mice and sea urchins.

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Alenciks E, Frazier K, Porter A, Fraley G (2015) Immunolesions of melanopsin receptive neurons attenuates the hormonal reproductive axis in the adult but has no effect on growth in immature Peking ducks. Neuroscience 2015 Abstracts 613.05/R20. Society for Neuroscience, Chicago IL.

Summary: Several light sensitive receptors have been described in the avian brain that are thought to regulate the reproductive axis independently from the eyes and pineal gland. Recently, our lab has described the presence of 3 photoneuroendocrine systems in the Pekin duck: rhodopsin, opsin 5, & melanopsin. We set out to test the hypothesis that melanopsin receptive neurons are necessary to maintain seasonal reproductive status along with growth and development in the Pekin drake. To accomplish these goals we first investigated 50-week-old Pekin drakes that were housed in the aviary at Hope College under long day length (18 hrs lights on) conditions in floor pens. To specifically lesion melanopsin-receptive neurons, 3 μl of an anti-melanopsin-saporin conjugate (MSAP, 100 ng/ul) was injected into the lateral ventricle (n = 10). Control drakes were injected with 3 ul of equimolar unconjugated anti-melanopsin and saporin (SAP, n = 10). The drakes were returned to the aviary after complete recovery. Reproductive behaviors were analyzed weekly in a test pen with adult hens. After 4 weeks, birds were euthanized and body weights were measured, and brains, pituitaries, and testes collected and stored for analyses. To test melanopsin’s effect on immature ducks the same surgery was performed on a group of 10 day old ducks (n= 10). Ducks were weighed weekly starting at 3 days of age. After a final weight was obtained at 50 days of age, ducks were euthanized and a blood sample was collected and sent out for an avian panel. Mature MSAP-treated drakes had significantly (p< 0.001) reduced relative teste weights compared to SAP controls. qRT-PCR analyses (n= 3 per treatment) of anterior pituitary showed a significant reduction (p< 0.001) in both LH-beta and FSH mRNA’s. Immunoctyochemical analyses (n= 3 per treatment) showed a significant reduction in melanopsin and GnRH-immunoreactivities. Immature drake BW did not differ significantly between MSAP and SAP animals at any of the measured days. The data appeared to drift toward significance near the end of the sampling period (p = 0.297). Blood panel results revealed no significant differences between MSAP and SAP animals in any CBC component. Serum glutamic-oxaloacetic transaminase (SGOT) (p= 0.022) and creatine phosphokinase (CPK) values were significantly elevated (p = 0.006) in MSAP animals compared to controls. Although melanopsin neurons in the PMM appear to have an important role in adult drakes, their importance in the growth of immature ducks is still unclear. However, these data underscore the importance of the photoneuroendocrine system in maintaining the reproductive axis along with growth and development in seasonally breeding birds.

Related Products: Melanopsin-SAP (Cat. #IT-44), Saporin (Cat. #PR-01)

Buhr E, Yue W, Ren X, Jiang Z, Liao H, Mei X, Vemaraju S, Nguyen M, Reed R, Lang R, Yau K, Van Gelder R (2015) Neuropsin (OPN5)-mediated photoentrainment of local circadian oscillators in mammalian retina and cornea. Proc Natl Acad Sci U S A 112:13093-13098. doi: 10.1073/pnas.1516259112 PMID: 26392540

Summary: Circadian clocks are found in most mammalian tissues. These clocks are synchronized by the suprachiasmatic nuclei (SCN) in the brain. The local clock found in the retina does not require rods, cones, intrinsically photosensitive retinal ganglion cells, or the SCN. In order to determine what photopigments are responsible for local retinal photoentrainment, the authors used a candidate gene approach. For immunohistochemical studies on flat mount retinas they used a melanopsin antibody (Cat. #AB-N38) at a 1:1000 dilution. The data indicate that OPN5, also known as neuropsin, has a light-sensing function and is involved in retinal photoentrainment.

Related Products: Melanopsin Rabbit Polyclonal (Cat. #AB-N38)

Riazifar H, Sun G, Wang X, Rupp A, Vemaraju S, Ross-Cisneros F, Lang R, Sadun A, Hattar S, Guan M, Huang T (2015) Phenotypic and functional characterization of Bst+/- mouse retina. Dis Model Mech 8:969-976. doi: 10.1242/dmm.018176 PMID: 26035379

Summary: The belly spot and tail mutant mouse strain was first reported on in 1976. Among other phenotypic changes, it carries ocular mutations including retinal colobomas, reduced retinal ganglion cells (RGCs), and axon misrouting. In order to assess the use of this strain as a murine model for stem cell therapies of retinal degenerative diseases the authors performed a number of characterization experiments including electron microscopy, immunohistochemistry, testing of circadian rhythms, and morphological studies. Some of the immunohistochemistry was done using Anti-Melanopsin (Cat. #AB-N38) at a 1:5000 dilution.

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El-Danaf R, Huberman A (2015) Characteristic patterns of dendritic remodeling in early-stage glaucoma: evidence from genetically identified retinal ganglion cell types. J Neurosci 35:2329-2343. doi: 10.1523/JNEUROSCI.1419-14.2015 PMID: 25673829

Summary: The loss of retinal ganglion cells (RGC) is the second-most common cause of blindness worldwide. Using several mouse transgenic cell lines, the authors investigated the changes that occur on the establishment of elevated ocular pressure. Anti-melanopsin (Cat. #AB-N39) at 1:1000 was used to illuminate the morphology of the M1 intrinsically photosensitive RGC.

Related Products: Melanopsin Rabbit Polyclonal, affinity-purified (Cat. #AB-N39)

Fraley GS (2014) Immunolesions of melanopsin receptive neurons in the adult Pekin drake attenuates the hormonal reproductive axis. Neuroscience 2014 Abstracts 543.01. Society for Neuroscience, Washington, DC.

Summary: Several light sensitive receptors have been described in the avian brain that are thought to regulate the reproductive axis independently from the eyes and pineal gland. Recently, my lab has described the presence of three of these photoneuroendocrine systems in the Pekin duck: opsin, opsin 5, and melanopsin. I set out to test the hypothesis that melanopsin receptive neurons are necessary to maintain seasonal reproductive status in the Pekin drake. To accomplish this, 50-week-old Pekin drakes were housed in the aviary at Hope College under long day length (18 hrs lights on) conditions in floor pens (5 drakes per pen). To specifically lesion melanopsin-receptive neurons, drakes were anethestized (8 mg/kg Propofol, IV), given analgesics (2 mg/kg ketfen, SC) skin incised and a trephine hole drilled 10 mm caudal to bony orbits and 1 mm to the left of midline. A 33 gauge stainless steel needle attached to a Hamilton syringe was lowered stereotactically 3.5 mm ventral to dura into the lateral ventricle. Three microliters of an anti-melanopsin-saporin conjugate (MSAP, 100 ng/ul) was injected into the lateral ventricle (n = 10). Control drakes were injected with 3 ul of equimolar unconjugated anti-melanopsin and saporin (SAP, n = 10). The incision was closed with VetBond, and the drakes returned to the aviary after complete recovery from anesthesia. After 4 weeks, birds were euthanized (400 mg/kg FatalPlus, IP) and body weight measured, and brains, pituitaries, and testes collected and stored for analyses. MSAP-treated drakes had significantly (p < 0.001) reduced relative teste weights compared to SAP controls. qRT-PCR analyses (n = 5 per treatment) of anterior pituitary showed a significant reduction (p < 0.001) in both LH-beta and FSH mRNA’s. Immunoctyochemical analyses (n = 5 per treatment) showed a significant reduction in melanopsin and GnRH-immunoreactivities. These data underscore the importance of the photoneuroendocrine system in maintaining the reproductive axis in seasonally breeding birds.

Related Products: Melanopsin-SAP (Cat. #IT-44)

Mao C, Li H, Zhang Z, Kiyama T, Panda S, Hattar S, Ribelayga C, Mills S, Wang S (2014) T-box transcription regulator Tbr2 is essential for the formation and maintenance of Opn4/melanopsin-expressing intrinsically photosensitive retinal ganglion cells. J Neurosci 34:13083-13095. doi: 10.1523/JNEUROSCI.1027-14.2014 PMID: 25253855

Summary: Opsin 4/melanopsin-expressing intrinsically photosensitive retinal ganglion cells (ipRGCs) are responsible for controlling non-image-forming visual functions in the retina. The findings show that opsin 4 is only expressed in Tbr2-positive ipRGCs, no ipRGCs are found if Tbr2 is deleted before RGC specialization, and most ipRGCs are eliminated when Tbr2 is deleted from established ipRGCs. An antibody against melanopsin (Cat. #AB-N39) was used at a 1:1000 dilution for immunohistochemical analyses.

Related Products: Melanopsin Rabbit Polyclonal, affinity-purified (Cat. #AB-N39)

Joly S, Guzik-Kornacka A, Schwab M, Pernet V (2014) New mouse retinal stroke model reveals direction-selective circuit damage linked to permanent optokinetic response loss. Invest Ophthalmol Vis Sci 55:4476-4489. doi: 10.1167/iovs.14-14521 PMID: 24970264

Summary: The authors used a mouse model of ‘retinal stroke’ to better delineate the optokinetic response deficits at the cellular level. Damage was found in the processes of starburst amacrine cells (SACs), and to a lesser extent, the dendrites. Anti-melanopsin (Cat. #AB-N38) at 1:2500 was used for immunohistochemistry.

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Ren C, Luan L, Wui-Man Lau B, Huang X, Yang J, Zhou Y, Wu X, Gao J, Pickard GE, So KF, Pu M (2013) Direct retino-raphe projection alters serotonergic tone and affective behavior. Neuropsychopharmacology 38(7):1163-1175. doi: 10.1038/npp.2013.35

Summary: Although recent work has shown that some intrinsically photosensitive retinal ganglion cells (ipRGCs) are responsible for processing nonimage-forming visual functions, it is unclear whether the ipRGCs or conventional RGCs modulate affective behavior. The authors injected 2 μg of melanopsin-SAP (Cat. #IT-44) into each eye of gerbils, or biotinylated CTB monoclonal antibody coupled to Streptavidin-ZAP (Cat. #IT-27). The data suggest that retino-raphe signals modulate dorsal raphe nucleus serotonergic tone and affective behavior.

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Estevez ME, Fogerson PM, Ilardi MC, Borghuis BG, Chan E, Weng S, Auferkorte ON, Demb JB, Berson DM (2012) Form and function of the M4 cell, an intrinsically photosensitive retinal ganglion cell type contributing to geniculocortical vision. J Neurosci 32(39):13608-13620. doi: 10.1523/JNEUROSCI.1422-12.2012 PMID: 23015450

Summary: Intrinsically photosensitive retinal ganglion cells (ipRGCs) are cells that contain the photopigment melanopsin. In this work the authors extensively characterize the M4 ipRGCs. A melanopsin antibody (Cat. #AB-N38) at a 1:10,000 dilution was used to determine the presence of melanopsin by immunohistochemistry.

Related Products: Melanopsin Rabbit Polyclonal (Cat. #AB-N38)