The consequences of selectively knocking down key metabolic genes in Muller cells of the mouse retina

Weiyong Shen, So-Ra Lee, Michelle X. Yam, Nigel L. Barnett, Ashish Easow Mathai, Rui Zhang, Ling Zhu, James Hurley, Jianhai Du, Pankaj Seth, Yoshio Hirabayashi, Shigeki Furuya, Mark C. Gillies

Research output: Contribution to journalMeeting AbstractResearchpeer-review

Abstract

Purpose : Derangement of glucose metabolism is a potential cause of photoreceptor degeneration. We have previously shown that selectively ablating Müller cells leads to photoreceptor degeneration, indicating that Müller cells are critical for their survival (Shen et al. J Neurosci 2012). This project aimed to study the consequences of selectively knocking down key metabolic genes in Müller cells of the mouse retina.

Methods : Transgenic mice for inducible Müller cell-specific gene targeting (Rlbp1-CreER mice) were crossed with mice carrying floxed metabolic genes including insulin receptor (IR), hexokinase 2 (Hk2), lactate dehydrogenase A (LDH-A), pyruvate dehydrogenase E1α (PDH-E1α) and phosphoglycerate dehydrogenase (PHGDH). Rlbp1-CreER mice crossed with Cre reporter mice were used as controls. Changes in target gene expression and the rod and cone photoreceptors were studied by immunostaining and Western blots. Retinal function was assessed by dark-adapted flash electroretinography (ERG).

Results : Double label immunofluorescent staining indicated that Müller cells expressed IR, Hk2 and PHGDH but not LDH-A and PDH-E1α. Knocking down IR, HK2 and PHGDH in Müller cells resulted in photoreceptor degeneration 3-4 weeks after tamoxifen-induced Cre expression, as evidenced by the loss of cone photoreceptor apical processes after staining retinas with fluorescently labelled peanut agglutinin and antibodies against blue- and red-green opsin. These changes were accompanied by impaired retinal function as measured by ERG. We also found that knocking down PHGDH appeared to produce more profound effects on photoreceptor degeneration compared with knocking down IR and Hk2 in Müller cells. We did not observe any retinal abnormalities after crossing Rlbp1-CreER mice with transgenic lines carrying floxed LDH-A and PDH-E1α genes.

Conclusions : Our data suggest that Müller cells may use glucose through the upstream of aerobic glycolysis and the serine/glycine metabolic pathway to support photoreceptors in the mammalian retina.
Original languageEnglish
Article number1492
Number of pages3
JournalInvestigative Ophthalmology and Visual Science
Volume59
Issue number9
Publication statusPublished - Jul 2018
Externally publishedYes
EventAnnual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO) - Honolulu
Duration: 29 Apr 20183 May 2018

Cite this

Shen, Weiyong ; Lee, So-Ra ; Yam, Michelle X. ; Barnett, Nigel L. ; Mathai, Ashish Easow ; Zhang, Rui ; Zhu, Ling ; Hurley, James ; Du, Jianhai ; Seth, Pankaj ; Hirabayashi, Yoshio ; Furuya, Shigeki ; Gillies, Mark C. / The consequences of selectively knocking down key metabolic genes in Muller cells of the mouse retina. In: Investigative Ophthalmology and Visual Science. 2018 ; Vol. 59, No. 9.
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title = "The consequences of selectively knocking down key metabolic genes in Muller cells of the mouse retina",
abstract = "Purpose : Derangement of glucose metabolism is a potential cause of photoreceptor degeneration. We have previously shown that selectively ablating M{\"u}ller cells leads to photoreceptor degeneration, indicating that M{\"u}ller cells are critical for their survival (Shen et al. J Neurosci 2012). This project aimed to study the consequences of selectively knocking down key metabolic genes in M{\"u}ller cells of the mouse retina.Methods : Transgenic mice for inducible M{\"u}ller cell-specific gene targeting (Rlbp1-CreER mice) were crossed with mice carrying floxed metabolic genes including insulin receptor (IR), hexokinase 2 (Hk2), lactate dehydrogenase A (LDH-A), pyruvate dehydrogenase E1α (PDH-E1α) and phosphoglycerate dehydrogenase (PHGDH). Rlbp1-CreER mice crossed with Cre reporter mice were used as controls. Changes in target gene expression and the rod and cone photoreceptors were studied by immunostaining and Western blots. Retinal function was assessed by dark-adapted flash electroretinography (ERG).Results : Double label immunofluorescent staining indicated that M{\"u}ller cells expressed IR, Hk2 and PHGDH but not LDH-A and PDH-E1α. Knocking down IR, HK2 and PHGDH in M{\"u}ller cells resulted in photoreceptor degeneration 3-4 weeks after tamoxifen-induced Cre expression, as evidenced by the loss of cone photoreceptor apical processes after staining retinas with fluorescently labelled peanut agglutinin and antibodies against blue- and red-green opsin. These changes were accompanied by impaired retinal function as measured by ERG. We also found that knocking down PHGDH appeared to produce more profound effects on photoreceptor degeneration compared with knocking down IR and Hk2 in M{\"u}ller cells. We did not observe any retinal abnormalities after crossing Rlbp1-CreER mice with transgenic lines carrying floxed LDH-A and PDH-E1α genes.Conclusions : Our data suggest that M{\"u}ller cells may use glucose through the upstream of aerobic glycolysis and the serine/glycine metabolic pathway to support photoreceptors in the mammalian retina.",
author = "Weiyong Shen and So-Ra Lee and Yam, {Michelle X.} and Barnett, {Nigel L.} and Mathai, {Ashish Easow} and Rui Zhang and Ling Zhu and James Hurley and Jianhai Du and Pankaj Seth and Yoshio Hirabayashi and Shigeki Furuya and Gillies, {Mark C.}",
year = "2018",
month = "7",
language = "English",
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journal = "Investigative Ophthalmology",
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Shen, W, Lee, S-R, Yam, MX, Barnett, NL, Mathai, AE, Zhang, R, Zhu, L, Hurley, J, Du, J, Seth, P, Hirabayashi, Y, Furuya, S & Gillies, MC 2018, 'The consequences of selectively knocking down key metabolic genes in Muller cells of the mouse retina', Investigative Ophthalmology and Visual Science, vol. 59, no. 9, 1492.

The consequences of selectively knocking down key metabolic genes in Muller cells of the mouse retina. / Shen, Weiyong; Lee, So-Ra; Yam, Michelle X.; Barnett, Nigel L.; Mathai, Ashish Easow; Zhang, Rui; Zhu, Ling; Hurley, James; Du, Jianhai; Seth, Pankaj; Hirabayashi, Yoshio; Furuya, Shigeki; Gillies, Mark C.

In: Investigative Ophthalmology and Visual Science, Vol. 59, No. 9, 1492, 07.2018.

Research output: Contribution to journalMeeting AbstractResearchpeer-review

TY - JOUR

T1 - The consequences of selectively knocking down key metabolic genes in Muller cells of the mouse retina

AU - Shen, Weiyong

AU - Lee, So-Ra

AU - Yam, Michelle X.

AU - Barnett, Nigel L.

AU - Mathai, Ashish Easow

AU - Zhang, Rui

AU - Zhu, Ling

AU - Hurley, James

AU - Du, Jianhai

AU - Seth, Pankaj

AU - Hirabayashi, Yoshio

AU - Furuya, Shigeki

AU - Gillies, Mark C.

PY - 2018/7

Y1 - 2018/7

N2 - Purpose : Derangement of glucose metabolism is a potential cause of photoreceptor degeneration. We have previously shown that selectively ablating Müller cells leads to photoreceptor degeneration, indicating that Müller cells are critical for their survival (Shen et al. J Neurosci 2012). This project aimed to study the consequences of selectively knocking down key metabolic genes in Müller cells of the mouse retina.Methods : Transgenic mice for inducible Müller cell-specific gene targeting (Rlbp1-CreER mice) were crossed with mice carrying floxed metabolic genes including insulin receptor (IR), hexokinase 2 (Hk2), lactate dehydrogenase A (LDH-A), pyruvate dehydrogenase E1α (PDH-E1α) and phosphoglycerate dehydrogenase (PHGDH). Rlbp1-CreER mice crossed with Cre reporter mice were used as controls. Changes in target gene expression and the rod and cone photoreceptors were studied by immunostaining and Western blots. Retinal function was assessed by dark-adapted flash electroretinography (ERG).Results : Double label immunofluorescent staining indicated that Müller cells expressed IR, Hk2 and PHGDH but not LDH-A and PDH-E1α. Knocking down IR, HK2 and PHGDH in Müller cells resulted in photoreceptor degeneration 3-4 weeks after tamoxifen-induced Cre expression, as evidenced by the loss of cone photoreceptor apical processes after staining retinas with fluorescently labelled peanut agglutinin and antibodies against blue- and red-green opsin. These changes were accompanied by impaired retinal function as measured by ERG. We also found that knocking down PHGDH appeared to produce more profound effects on photoreceptor degeneration compared with knocking down IR and Hk2 in Müller cells. We did not observe any retinal abnormalities after crossing Rlbp1-CreER mice with transgenic lines carrying floxed LDH-A and PDH-E1α genes.Conclusions : Our data suggest that Müller cells may use glucose through the upstream of aerobic glycolysis and the serine/glycine metabolic pathway to support photoreceptors in the mammalian retina.

AB - Purpose : Derangement of glucose metabolism is a potential cause of photoreceptor degeneration. We have previously shown that selectively ablating Müller cells leads to photoreceptor degeneration, indicating that Müller cells are critical for their survival (Shen et al. J Neurosci 2012). This project aimed to study the consequences of selectively knocking down key metabolic genes in Müller cells of the mouse retina.Methods : Transgenic mice for inducible Müller cell-specific gene targeting (Rlbp1-CreER mice) were crossed with mice carrying floxed metabolic genes including insulin receptor (IR), hexokinase 2 (Hk2), lactate dehydrogenase A (LDH-A), pyruvate dehydrogenase E1α (PDH-E1α) and phosphoglycerate dehydrogenase (PHGDH). Rlbp1-CreER mice crossed with Cre reporter mice were used as controls. Changes in target gene expression and the rod and cone photoreceptors were studied by immunostaining and Western blots. Retinal function was assessed by dark-adapted flash electroretinography (ERG).Results : Double label immunofluorescent staining indicated that Müller cells expressed IR, Hk2 and PHGDH but not LDH-A and PDH-E1α. Knocking down IR, HK2 and PHGDH in Müller cells resulted in photoreceptor degeneration 3-4 weeks after tamoxifen-induced Cre expression, as evidenced by the loss of cone photoreceptor apical processes after staining retinas with fluorescently labelled peanut agglutinin and antibodies against blue- and red-green opsin. These changes were accompanied by impaired retinal function as measured by ERG. We also found that knocking down PHGDH appeared to produce more profound effects on photoreceptor degeneration compared with knocking down IR and Hk2 in Müller cells. We did not observe any retinal abnormalities after crossing Rlbp1-CreER mice with transgenic lines carrying floxed LDH-A and PDH-E1α genes.Conclusions : Our data suggest that Müller cells may use glucose through the upstream of aerobic glycolysis and the serine/glycine metabolic pathway to support photoreceptors in the mammalian retina.

M3 - Meeting Abstract

VL - 59

JO - Investigative Ophthalmology

JF - Investigative Ophthalmology

SN - 0146-0404

IS - 9

M1 - 1492

ER -