Simultaneous quantification of retinol, retinal, and retinoic acid isomers by high-performance liquid chromatography with a simple gradiation

Most opsins selectively bind 11-cis retinal as a chromophore to form a photosensitive pigment, which underlies various physiological functions, such as vision and circadian photoentrainment. Recently, opsin 3 (Opn3), originally called encephalopsin or panopsin, and its homologs were identified in various tissues including brain, eye, and liver in both vertebrates and invertebrates, including human. Because Opn3s are mainly expressed in tissues that are not considered to contain sufficient amounts of 11-cis retinal to form pigments, the photopigment formation ability of Opn3 has been of interest. Here, we report the successful expression of Opn3 homologs, pufferfish teleost multiple tissue opsin (PufTMT) and mosquito Opn3 (MosOpn3) and show that these proteins formed functional photopigments with 11-cis and 9-cis retinals. The PufTMT-and MosOpn3-based pigments have absorption maxima in the blue-to-green region and exhibit a bistable nature. These Opn3 homolog-based pigments activate G i -type and G o -type G proteins light dependently, indicating that they potentially serve as light-sensitive G i /G o -coupled receptors. We also demonstrated that mammalian cultured cells transfected with the MosOpn3 or PufTMT became light sensitive without the addition of 11-cis retinal and the photosensitivity retained after the continuous light exposure, showing a reusable pigment formation with retinal endogenously contained in culture medium. Interestingly, we found that the MosOpn3 also acts as a light sensor when constituted with 13-cis retinal, a ubiquitously present retinal isomer. Our findings suggest that homologs of vertebrate Opn3 might function as photoreceptors in various tissues; furthermore, these Opn3s, particularly the mosquito homolog, could provide a promising optogenetic tool for regulating cAMP-related G protein-coupled receptor signalings.rhodopsin | phototransduction | nonvisual photoreception | opsin diversity M ost opsins bind 11-cis retinal as a chromophore to form a photosensitive pigment (opsin-based pigment) that serves as a light-sensitive G protein-coupled receptor (GPCR). The 11-cis to all-trans isomerization of the chromophore in an opsinbased pigment upon light absorption triggers G protein activation (1-4), and the photoreception of the opsin-based pigment initiates vision and nonvisual functions, including circadian entrainment and pupil responses. Several thousand opsins have been identified, and they are phylogenetically classified into eight groups based on the members that have existed early in animal evolution (5). The phylogenetic classification of each opsin also roughly corresponds to its molecular function. The G t -coupled opsin group contains vertebrate visual pigments, which activate transducin (G t )-type G proteins, and some nonvisual pigments, such as pinopsin, VA-opsin, parapinopsin, and parietopsin (1-4). The G q -coupled opsin group is composed of the visual pigments from many invertebrates, such as insects and cephalopods, and the vertebrate opsin 4 (Opn4) homologs (melanops...

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“…S4, and Abs. at 360nm (14-17) and their translated protein could be functional by binding to retinoids in the serum (20). As shown in Fig.…”

Section: Resultsmentioning

confidence: 94%

“…AB753162 and AB753163). addition, the invertebrate Opn3 homolog acts as a photopigment when bound to 13-cis retinal, which is thermally equilibrated with all-trans retinal and, therefore, ubiquitously present in animals (19,20). These results suggest that Opn3 homolog-expressing tissues may be photosensitive.…”

mentioning

confidence: 90%

Homologs of vertebrate Opn3 potentially serve as a light sensor in nonphotoreceptive tissue

Koyanagi

Takada

Nagata

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

144

212

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“…Apparently, an abnormal retinoid status is related to carcinogenesis. Therefore, it is not surprising that the administration of retinoid prevents the development of squamous cell carcinoma in the skin, oral cavity, and lung in animal models (Moon et al 1994). Notably, some synthetic retinoids were reported to be effective in preventing the development of esophageal cancer from severe esophageal squamous dysplasia (Han 1993).…”

Section: Retinoids and Cancermentioning

confidence: 99%

“…It allowed us to quantify them up to 2.5 ng/ml. The details are described in the article of Miyagi et al 2001.…”

Section: Feature Of An Nad Dependent In Situ Ra Supply System In Rat mentioning

confidence: 99%

The Interaction Between the Metabolism of Retinol and Ethanol in Esophageal Mucosa – A Possible Mechanism of Esophageal Cancer in Alcoholics

Yokoyama¹,

Yokoyama²,

Hibi³

2012

Esophageal Cancer - Cell and Molecular Biology, Biomarkers, Nutrition and Treatment

Self Cite

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“…The chemical stability of tretinoin in dermatological lotions and hydrogels [11] and in hydrophilic gels [12] has been studied by simpler HPLC methods, but these methods do not report analysis of isotretinoin. HPLC separation methods for determination of tretinoin in ovules [13] and in solution [14] have been briefly described, and methods for analysis in biological samples have been reported [15,16]. The photostability of tretinoin lotion in the presence and absence of additives such as UV sunscreen, cyclodextrin and surfactants used an HPLC method with a gradient mobile phase composed of methanol-water-acetic acid mixture [17].…”

Section: Introductionmentioning

confidence: 99%

A rapid HPLC method for simultaneous determination of tretinoin and isotretinoin in dermatological formulations

Tashtoush

Jacobson

2007

Journal of Pharmaceutical and Biomedical Analysis

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Simultaneous quantification of retinol, retinal, and retinoic acid isomers by high-performance liquid chromatography with a simple gradiation

Cited by 32 publications

References 4 publications

Homologs of vertebrate Opn3 potentially serve as a light sensor in nonphotoreceptive tissue

Homologs of vertebrate Opn3 potentially serve as a light sensor in nonphotoreceptive tissue

The Interaction Between the Metabolism of Retinol and Ethanol in Esophageal Mucosa – A Possible Mechanism of Esophageal Cancer in Alcoholics

A rapid HPLC method for simultaneous determination of tretinoin and isotretinoin in dermatological formulations

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