Limits...
Differential alterations in the expression of neurotransmitter receptors in inner retina following loss of photoreceptors in rd1 mouse.

Srivastava P, Sinha-Mahapatra SK, Ghosh A, Srivastava I, Dhingra NK - PLoS ONE (2015)

Bottom Line: Synaptic activity in the excitatory bipolar cells and the predominantly inhibitory amacrine cells is enhanced.We found significantly higher levels of AMPA, glycine and GABAa receptors, but lower levels of GABAc receptors in rd1 mouse than in wild-type.We found that the levels of GABAa receptors in inner plexiform layer of wild-type and rd1 mice were similar, whereas those in outer plexiform layer and inner nuclear layer combined were higher in rd1 mouse.

View Article: PubMed Central - PubMed

Affiliation: National Brain Research Centre, Manesar (Gurgaon) Haryana, India.

ABSTRACT
Loss of photoreceptors leads to significant remodeling in inner retina of rd1 mouse, a widely used model of retinal degeneration. Several morphological and physiological alterations occur in the second- and third-order retinal neurons. Synaptic activity in the excitatory bipolar cells and the predominantly inhibitory amacrine cells is enhanced. Retinal ganglion cells (RGCs) exhibit hyperactivity and aberrant spiking pattern, which adversely affects the quality of signals they can carry to the brain. To further understand the pathophysiology of retinal degeneration, and how it may lead to aberrant spiking in RGCs, we asked how loss of photoreceptors affects some of the neurotransmitter receptors in rd1 mouse. Using Western blotting, we measured the levels of several neurotransmitter receptors in adult rd1 mouse retina. We found significantly higher levels of AMPA, glycine and GABAa receptors, but lower levels of GABAc receptors in rd1 mouse than in wild-type. Since GABAa receptor is expressed in several retinal layers, we employed quantitative immunohistochemistry to measure GABAa receptor levels in specific retinal layers. We found that the levels of GABAa receptors in inner plexiform layer of wild-type and rd1 mice were similar, whereas those in outer plexiform layer and inner nuclear layer combined were higher in rd1 mouse. Specifically, we found that the number of GABAa-immunoreactive somas in the inner nuclear layer of rd1 mouse retina was significantly higher than in wild-type. These findings provide further insights into neurochemical remodeling in the inner retina of rd1 mouse, and how it might lead to oscillatory activity in RGCs.

No MeSH data available.


Related in: MedlinePlus

GABAcR-ρ2 levels were downregulated in rd1 mouse retina.A) Representative blots of GABAcR-ρ2 and β-tubulin in adult wild-type and rd1 mouse retinas. Black vertical line indicates that the two bands were not adjacent. B) Levels of GABAcR-ρ2 (mean±SD) were significantly lower in rd1 mouse retina than in wild-type (n = 8). ** p<0.005.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4383516&req=5

pone.0123896.g004: GABAcR-ρ2 levels were downregulated in rd1 mouse retina.A) Representative blots of GABAcR-ρ2 and β-tubulin in adult wild-type and rd1 mouse retinas. Black vertical line indicates that the two bands were not adjacent. B) Levels of GABAcR-ρ2 (mean±SD) were significantly lower in rd1 mouse retina than in wild-type (n = 8). ** p<0.005.

Mentions: The goat polyclonal GABAcR antibody recognizes ρ2 subunit of the receptor and detects a single band at 50 kDa in mouse brain (manufacturer’s data). In mouse retina, this antibody produced a single band near 57 kDa in our hands, probably representing a specific GABAc isoform (manufacture’s data/ emails; http://www.uniprot.org/uniprot/P47742; see Fig 4A). This band disappeared when we preincubated the antibody with the relevant peptide antigen (sc-21343P; Santa Cruz). β-Tubulin antibody was used as loading control for Western blotting. This antibody detects a single band at 55 kDa which was used for normalization when quantifying the proteins.


Differential alterations in the expression of neurotransmitter receptors in inner retina following loss of photoreceptors in rd1 mouse.

Srivastava P, Sinha-Mahapatra SK, Ghosh A, Srivastava I, Dhingra NK - PLoS ONE (2015)

GABAcR-ρ2 levels were downregulated in rd1 mouse retina.A) Representative blots of GABAcR-ρ2 and β-tubulin in adult wild-type and rd1 mouse retinas. Black vertical line indicates that the two bands were not adjacent. B) Levels of GABAcR-ρ2 (mean±SD) were significantly lower in rd1 mouse retina than in wild-type (n = 8). ** p<0.005.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4383516&req=5

pone.0123896.g004: GABAcR-ρ2 levels were downregulated in rd1 mouse retina.A) Representative blots of GABAcR-ρ2 and β-tubulin in adult wild-type and rd1 mouse retinas. Black vertical line indicates that the two bands were not adjacent. B) Levels of GABAcR-ρ2 (mean±SD) were significantly lower in rd1 mouse retina than in wild-type (n = 8). ** p<0.005.
Mentions: The goat polyclonal GABAcR antibody recognizes ρ2 subunit of the receptor and detects a single band at 50 kDa in mouse brain (manufacturer’s data). In mouse retina, this antibody produced a single band near 57 kDa in our hands, probably representing a specific GABAc isoform (manufacture’s data/ emails; http://www.uniprot.org/uniprot/P47742; see Fig 4A). This band disappeared when we preincubated the antibody with the relevant peptide antigen (sc-21343P; Santa Cruz). β-Tubulin antibody was used as loading control for Western blotting. This antibody detects a single band at 55 kDa which was used for normalization when quantifying the proteins.

Bottom Line: Synaptic activity in the excitatory bipolar cells and the predominantly inhibitory amacrine cells is enhanced.We found significantly higher levels of AMPA, glycine and GABAa receptors, but lower levels of GABAc receptors in rd1 mouse than in wild-type.We found that the levels of GABAa receptors in inner plexiform layer of wild-type and rd1 mice were similar, whereas those in outer plexiform layer and inner nuclear layer combined were higher in rd1 mouse.

View Article: PubMed Central - PubMed

Affiliation: National Brain Research Centre, Manesar (Gurgaon) Haryana, India.

ABSTRACT
Loss of photoreceptors leads to significant remodeling in inner retina of rd1 mouse, a widely used model of retinal degeneration. Several morphological and physiological alterations occur in the second- and third-order retinal neurons. Synaptic activity in the excitatory bipolar cells and the predominantly inhibitory amacrine cells is enhanced. Retinal ganglion cells (RGCs) exhibit hyperactivity and aberrant spiking pattern, which adversely affects the quality of signals they can carry to the brain. To further understand the pathophysiology of retinal degeneration, and how it may lead to aberrant spiking in RGCs, we asked how loss of photoreceptors affects some of the neurotransmitter receptors in rd1 mouse. Using Western blotting, we measured the levels of several neurotransmitter receptors in adult rd1 mouse retina. We found significantly higher levels of AMPA, glycine and GABAa receptors, but lower levels of GABAc receptors in rd1 mouse than in wild-type. Since GABAa receptor is expressed in several retinal layers, we employed quantitative immunohistochemistry to measure GABAa receptor levels in specific retinal layers. We found that the levels of GABAa receptors in inner plexiform layer of wild-type and rd1 mice were similar, whereas those in outer plexiform layer and inner nuclear layer combined were higher in rd1 mouse. Specifically, we found that the number of GABAa-immunoreactive somas in the inner nuclear layer of rd1 mouse retina was significantly higher than in wild-type. These findings provide further insights into neurochemical remodeling in the inner retina of rd1 mouse, and how it might lead to oscillatory activity in RGCs.

No MeSH data available.


Related in: MedlinePlus