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Comparative localization of cystathionine beta synthases and cystathionine gamma lyase in canine, non-human primate and human retina.
Exp Eye Res. 2019 04; 181:72-84.EE

Abstract

Chronic exposure of the retina to light and high concentrations of polyunsaturated fatty acid in photoreceptor cells make this tissue susceptible to oxidative damage. As retinal degenerative diseases are associated with photoreceptor degeneration, the antioxidant activity of both hydrogen sulfide (H2S) and glutathione (GSH) may play an important role in ameliorating disease progression. H2S production is driven by cystathionine-γ-lyase (CSE) and cystathionine-β-synthase (CBS), the key enzymes that also drive transsulfuration pathway (TSP) necessary for GSH production. As it is currently unclear whether localized production of either H2S or GSH contributes to retinal homeostasis, we undertook a comparative analysis of CBS and CSE expression in canine, non-human primate (NHP) and human retinas to determine if these antioxidants could play a regulatory role in age-related or disease-associated retinal degeneration. Retinas from normal dogs, NHPs and humans were used for the study. Laser capture microdissection (LCM) was performed to isolate individual layers of the canine retina and analyze CBS and CSE gene expression by qRT-PCR. Immunohistochemistry and western blotting were performed for CBS and CSE labeling and protein expression in dog, NHP, and human retina, respectively. Using qRT-PCR, western blot, and immunohistochemistry (IHC), we showed that CBS and CSE are expressed in the canine, NHP, and human retina. IHC results from canine retina demonstrated increased expression levels of CBS but not CSE with post-developmental aging. IHC results also showed non-overlapping localization of both proteins with CBS presenting in rods, amacrine, horizontal, and nerve fiber cell layers while CSE was expressed by RPE, cones and Mϋller cells. Finally, we demonstrated that these enzymes localized to all three layers of canine, NHP and human retina: photoreceptors, outer plexiform layer (OPL) and notably in the ganglion cells layer/nerve fiber layer (GCL/NFL). QRT-PCR performed using RNA extracted from tissues isolated from these cell layers using laser capture microdissection (LCM) confirmed that each of CBS and CSE are expressed equally in these three layers. Together, these findings reveal that CSE and CBS are expressed in the retina, thereby supporting further studies to determine the role of H2S and these proteins in oxidative stress and apoptosis in retinal degenerative diseases.

Authors+Show Affiliations

Department of Clinical Sciences and Advanced Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.Department of Clinical Sciences and Advanced Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.Department of Clinical Sciences and Advanced Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.F. M. Kirby Center for Molecular Ophthalmology, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA, USA.Department of Clinical Sciences and Advanced Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA. Electronic address: gda@vet.upenn.edu.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

30653965

Citation

Badiei, Alireza, et al. "Comparative Localization of Cystathionine Beta Synthases and Cystathionine Gamma Lyase in Canine, Non-human Primate and Human Retina." Experimental Eye Research, vol. 181, 2019, pp. 72-84.
Badiei A, Sudharsan R, Santana E, et al. Comparative localization of cystathionine beta synthases and cystathionine gamma lyase in canine, non-human primate and human retina. Exp Eye Res. 2019;181:72-84.
Badiei, A., Sudharsan, R., Santana, E., Dunaief, J. L., & Aguirre, G. D. (2019). Comparative localization of cystathionine beta synthases and cystathionine gamma lyase in canine, non-human primate and human retina. Experimental Eye Research, 181, 72-84. https://doi.org/10.1016/j.exer.2019.01.007
Badiei A, et al. Comparative Localization of Cystathionine Beta Synthases and Cystathionine Gamma Lyase in Canine, Non-human Primate and Human Retina. Exp Eye Res. 2019;181:72-84. PubMed PMID: 30653965.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparative localization of cystathionine beta synthases and cystathionine gamma lyase in canine, non-human primate and human retina. AU - Badiei,Alireza, AU - Sudharsan,Raghavi, AU - Santana,Evelyn, AU - Dunaief,Joshua L, AU - Aguirre,Gustavo D, Y1 - 2019/01/14/ PY - 2018/11/16/received PY - 2018/12/18/revised PY - 2019/01/03/accepted PY - 2019/1/18/pubmed PY - 2020/2/14/medline PY - 2019/1/18/entrez KW - Cystathionine-β-synthase (CBS) KW - Cystathionine-γ-lyase (CSE) KW - Dog KW - Human KW - Retina SP - 72 EP - 84 JF - Experimental eye research JO - Exp Eye Res VL - 181 N2 - Chronic exposure of the retina to light and high concentrations of polyunsaturated fatty acid in photoreceptor cells make this tissue susceptible to oxidative damage. As retinal degenerative diseases are associated with photoreceptor degeneration, the antioxidant activity of both hydrogen sulfide (H2S) and glutathione (GSH) may play an important role in ameliorating disease progression. H2S production is driven by cystathionine-γ-lyase (CSE) and cystathionine-β-synthase (CBS), the key enzymes that also drive transsulfuration pathway (TSP) necessary for GSH production. As it is currently unclear whether localized production of either H2S or GSH contributes to retinal homeostasis, we undertook a comparative analysis of CBS and CSE expression in canine, non-human primate (NHP) and human retinas to determine if these antioxidants could play a regulatory role in age-related or disease-associated retinal degeneration. Retinas from normal dogs, NHPs and humans were used for the study. Laser capture microdissection (LCM) was performed to isolate individual layers of the canine retina and analyze CBS and CSE gene expression by qRT-PCR. Immunohistochemistry and western blotting were performed for CBS and CSE labeling and protein expression in dog, NHP, and human retina, respectively. Using qRT-PCR, western blot, and immunohistochemistry (IHC), we showed that CBS and CSE are expressed in the canine, NHP, and human retina. IHC results from canine retina demonstrated increased expression levels of CBS but not CSE with post-developmental aging. IHC results also showed non-overlapping localization of both proteins with CBS presenting in rods, amacrine, horizontal, and nerve fiber cell layers while CSE was expressed by RPE, cones and Mϋller cells. Finally, we demonstrated that these enzymes localized to all three layers of canine, NHP and human retina: photoreceptors, outer plexiform layer (OPL) and notably in the ganglion cells layer/nerve fiber layer (GCL/NFL). QRT-PCR performed using RNA extracted from tissues isolated from these cell layers using laser capture microdissection (LCM) confirmed that each of CBS and CSE are expressed equally in these three layers. Together, these findings reveal that CSE and CBS are expressed in the retina, thereby supporting further studies to determine the role of H2S and these proteins in oxidative stress and apoptosis in retinal degenerative diseases. SN - 1096-0007 UR - https://www.unboundmedicine.com/medline/citation/30653965/Comparative_localization_of_cystathionine_beta_synthases_and_cystathionine_gamma_lyase_in_canine_non_human_primate_and_human_retina_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0014-4835(18)30839-X DB - PRIME DP - Unbound Medicine ER -