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Pain related channels are differentially expressed in neuronal and non-neuronal cells of glabrous skin of fabry knockout male mice.

Lakomá J, Rimondini R, Donadio V, Liguori R, Caprini M - PLoS ONE (2014)

Bottom Line: Additionally, KO mice showed a decreased and scattered pattern of neuronal terminations consistent with the reduction in neuronal terminations in skin biopsies of patients with small fiber neuropathies.At the molecular level KO animals showed an increase in the expression of TRPV1 and Nav1.8, and a decrease in the expression of TRPM8.Notably, these alterations are observed in young animals.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Human and General Physiology, Department of Pharmacy and Biotechnology (FaBiT), University of Bologna, Bologna, Italy.

ABSTRACT
Fabry disease (FD) is one of the X-linked lysosomal storage disorders caused by deficient functioning of the alpha-galactosidase A (α-GalA) enzyme. The α-GalA deficiency leads to multi-systemic clinical manifestations caused by the preferential accumulation of globotriaosylceramide in the endothelium and vascular smooth muscles. A hallmark symptom of FD patients is peripheral pain that appears in the early stage of the disease. Pain in FD patients is a peripheral small-fiber idiopathic neuropathy, with intra-epidermal fiber density and integrity being used for diagnosing FD in humans. However, the molecular correlates underlying pain sensation in FD remain elusive. Here, we have employed the α-GalA gene KO mouse as a model of FD in rodents to investigate molecular changes in their peripheral nervous system that may account for their algesic symptoms. The α-GalA mice display neuropathic pain as evidenced by thermal hyperalgesia and mechanical allodynia, with histological analyses showing alterations in cutaneous innervation. Additionally, KO mice showed a decreased and scattered pattern of neuronal terminations consistent with the reduction in neuronal terminations in skin biopsies of patients with small fiber neuropathies. At the molecular level KO animals showed an increase in the expression of TRPV1 and Nav1.8, and a decrease in the expression of TRPM8. Notably, these alterations are observed in young animals. Taken together, our findings imply that the α-GalA KO mouse is a good model in which to study the peripheral small fiber neuropathy exhibited by FD patients, and provides molecular evidence for a hyperexcitability of small nociceptors in FD.

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Detection of ENFs in α-GalA KO males frontal paws.The DAPI immunostaining of 50 µm floating sagittal mice frontal paw section with marked region of interest (A). The immunohistochemistry of α-GalA KO males (n = 3) revealed the scattered expression of PGP9.5 (red) - specific marker of neuronal terminations in the epidermis of frontal paw skin in comparison to their WT controls (n = 3). The dermis and epidermis border was distinguished by staining for Collagen IV (green) and visually determined by dotted lines. Paw epidermal PGP9.5 positive fibers showed morphological abnormality such as fragmentation in α-GalA KO males, whereas the epidermal fibers showed a more regular morphology in WT males (white arrows) (B). Scale bar represents 100 µm. Numerical analysis of neuronal fibers terminations showed significant decrease (about 50%; p = 0.0161) in α-GalA KO males in comparison to WT (C).
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pone-0108641-g003: Detection of ENFs in α-GalA KO males frontal paws.The DAPI immunostaining of 50 µm floating sagittal mice frontal paw section with marked region of interest (A). The immunohistochemistry of α-GalA KO males (n = 3) revealed the scattered expression of PGP9.5 (red) - specific marker of neuronal terminations in the epidermis of frontal paw skin in comparison to their WT controls (n = 3). The dermis and epidermis border was distinguished by staining for Collagen IV (green) and visually determined by dotted lines. Paw epidermal PGP9.5 positive fibers showed morphological abnormality such as fragmentation in α-GalA KO males, whereas the epidermal fibers showed a more regular morphology in WT males (white arrows) (B). Scale bar represents 100 µm. Numerical analysis of neuronal fibers terminations showed significant decrease (about 50%; p = 0.0161) in α-GalA KO males in comparison to WT (C).

Mentions: The epidermis of mammalian non-hairy (glabrous) skin is densely innervated by nociceptor endings that carry harmful chemical, mechanical and temperature stimuli [22], [23]. The immunohistochemistry of floating longitudinal sections of frontal mice paw revealed that neuronal fibers in dermis of glabrous skin were present in lower abundance in α-GalA(−/0) individuals compared to control mice. Notably, density of the ENFs was markedly decreased in KO mice compared to WT (37% less PGP 9.5 positive fibers in KO (5.11 fibers/mm) in comparison to WT (9.58 fibers/mm) (Figure 3p = 0.0161, n = 3 animals for WT, n = 3 animals for KO). Moreover, in the case of α-GalA(−/0) mice, we found morphological abnormalities such as nerve fibers swelling and fragmentations (Figure 3, white arrows). Interestingly, the reduction of PGP 9.5 immunostaining was not limited to the epidermis but also involved the sub-epidermis plexus.


Pain related channels are differentially expressed in neuronal and non-neuronal cells of glabrous skin of fabry knockout male mice.

Lakomá J, Rimondini R, Donadio V, Liguori R, Caprini M - PLoS ONE (2014)

Detection of ENFs in α-GalA KO males frontal paws.The DAPI immunostaining of 50 µm floating sagittal mice frontal paw section with marked region of interest (A). The immunohistochemistry of α-GalA KO males (n = 3) revealed the scattered expression of PGP9.5 (red) - specific marker of neuronal terminations in the epidermis of frontal paw skin in comparison to their WT controls (n = 3). The dermis and epidermis border was distinguished by staining for Collagen IV (green) and visually determined by dotted lines. Paw epidermal PGP9.5 positive fibers showed morphological abnormality such as fragmentation in α-GalA KO males, whereas the epidermal fibers showed a more regular morphology in WT males (white arrows) (B). Scale bar represents 100 µm. Numerical analysis of neuronal fibers terminations showed significant decrease (about 50%; p = 0.0161) in α-GalA KO males in comparison to WT (C).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4206276&req=5

pone-0108641-g003: Detection of ENFs in α-GalA KO males frontal paws.The DAPI immunostaining of 50 µm floating sagittal mice frontal paw section with marked region of interest (A). The immunohistochemistry of α-GalA KO males (n = 3) revealed the scattered expression of PGP9.5 (red) - specific marker of neuronal terminations in the epidermis of frontal paw skin in comparison to their WT controls (n = 3). The dermis and epidermis border was distinguished by staining for Collagen IV (green) and visually determined by dotted lines. Paw epidermal PGP9.5 positive fibers showed morphological abnormality such as fragmentation in α-GalA KO males, whereas the epidermal fibers showed a more regular morphology in WT males (white arrows) (B). Scale bar represents 100 µm. Numerical analysis of neuronal fibers terminations showed significant decrease (about 50%; p = 0.0161) in α-GalA KO males in comparison to WT (C).
Mentions: The epidermis of mammalian non-hairy (glabrous) skin is densely innervated by nociceptor endings that carry harmful chemical, mechanical and temperature stimuli [22], [23]. The immunohistochemistry of floating longitudinal sections of frontal mice paw revealed that neuronal fibers in dermis of glabrous skin were present in lower abundance in α-GalA(−/0) individuals compared to control mice. Notably, density of the ENFs was markedly decreased in KO mice compared to WT (37% less PGP 9.5 positive fibers in KO (5.11 fibers/mm) in comparison to WT (9.58 fibers/mm) (Figure 3p = 0.0161, n = 3 animals for WT, n = 3 animals for KO). Moreover, in the case of α-GalA(−/0) mice, we found morphological abnormalities such as nerve fibers swelling and fragmentations (Figure 3, white arrows). Interestingly, the reduction of PGP 9.5 immunostaining was not limited to the epidermis but also involved the sub-epidermis plexus.

Bottom Line: Additionally, KO mice showed a decreased and scattered pattern of neuronal terminations consistent with the reduction in neuronal terminations in skin biopsies of patients with small fiber neuropathies.At the molecular level KO animals showed an increase in the expression of TRPV1 and Nav1.8, and a decrease in the expression of TRPM8.Notably, these alterations are observed in young animals.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Human and General Physiology, Department of Pharmacy and Biotechnology (FaBiT), University of Bologna, Bologna, Italy.

ABSTRACT
Fabry disease (FD) is one of the X-linked lysosomal storage disorders caused by deficient functioning of the alpha-galactosidase A (α-GalA) enzyme. The α-GalA deficiency leads to multi-systemic clinical manifestations caused by the preferential accumulation of globotriaosylceramide in the endothelium and vascular smooth muscles. A hallmark symptom of FD patients is peripheral pain that appears in the early stage of the disease. Pain in FD patients is a peripheral small-fiber idiopathic neuropathy, with intra-epidermal fiber density and integrity being used for diagnosing FD in humans. However, the molecular correlates underlying pain sensation in FD remain elusive. Here, we have employed the α-GalA gene KO mouse as a model of FD in rodents to investigate molecular changes in their peripheral nervous system that may account for their algesic symptoms. The α-GalA mice display neuropathic pain as evidenced by thermal hyperalgesia and mechanical allodynia, with histological analyses showing alterations in cutaneous innervation. Additionally, KO mice showed a decreased and scattered pattern of neuronal terminations consistent with the reduction in neuronal terminations in skin biopsies of patients with small fiber neuropathies. At the molecular level KO animals showed an increase in the expression of TRPV1 and Nav1.8, and a decrease in the expression of TRPM8. Notably, these alterations are observed in young animals. Taken together, our findings imply that the α-GalA KO mouse is a good model in which to study the peripheral small fiber neuropathy exhibited by FD patients, and provides molecular evidence for a hyperexcitability of small nociceptors in FD.

Show MeSH
Related in: MedlinePlus