Korean J Ophthalmol.  2005 Dec;19(4):288-292. 10.3341/kjo.2005.19.4.288.

Delay of Photoreceptor Cell Degeneration in rd Mice by Systemically Administered Phenyl-N-tert-butylnitrone

• Kim JH
• Kim JH
• Yu YS
• Jeong SM
• Kim KW

• KMID: 754441
• DOI: http://doi.org/10.3341/kjo.2005.19.4.288

Abstract

PURPOSE
To study the effect of systemic administration of phenyl-N-tert-butylnitrone (PBN) on the degeneration of photoreceptor cells in rd mice. METHODS: PBN was injected intraperitoneally into FVB/rd mice on postnatal days (P) 5 to 14 (group A), and P10 to 18 (group B). At days P14, 16, 18, 20 and 27, morphological changes and apoptosis were analyzed by staining with hematoxylin and eosin or DAPI. The effect of PBN on apoptosis was analyzed in retinal pigment epithelial (RPE) cells by the measurement of caspase-3 activity. RESULTS: In control and group B mice, the outer nuclear layer (ONL) of the retina was composed of 8-10 rows at P12, and rapidly decreased to one row at P18. In group A mice, the ONL was preserved with 5-7 rows at P18, and decreased to one row at P22. PBN inhibited caspase-3 activity in cultured RPE cells. CONCLUSIONS: PBN delayed, but did not block, the degeneration of photoreceptor cells in rd mice. PBN may exert its inhibitory effect during the early phase of photoreceptor cell degeneration.

Keyword

Blood retinal barrier; Caspase-3; Delay; Phenyl-N-tert-butylnitrone; rd mice

MeSH Terms

Retinal Degeneration/*drug therapy/metabolism/pathology
Pigment Epithelium of Eye/drug effects/metabolism/pathology
Photoreceptors, Vertebrate/drug effects/metabolism/*pathology
Nitrogen Oxides/*administration & dosage/pharmacokinetics/therapeutic use
Neuroprotective Agents/*administration & dosage/pharmacokinetics/therapeutic use
Mice
Male
Injections, Intraperitoneal
Free Radical Scavengers/*administration & dosage/pharmacokinetics/therapeutic use
Follow-Up Studies
Female
Enzyme Precursors/metabolism
Disease Models, Animal
Cells, Cultured
Caspases/metabolism
Caspase 3
Apoptosis/drug effects
Animals

Figure

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  Fig. 1. PBN concentration in sensory retina after intraperitoneal injection. The concentration of PBN in the sensory retina was measured 1, 27 3, and 6 hours after intraperitoneal injection using HPLC.

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  Fig. 2. ONL preservation following Intraperitoneal Injection of PBN. (A) PBN injection from P5 to P14 (group A) delayed the loss of cells in the ONL. H & E staining (X400) (P: postnatal day; White arrow: ONL); (B) The number of photoreceptor nuclei per column in the ONL at days P12, 14, 16, 18, 20 and 27 was as follows; (Control/ Group A/ Group B): P12, 15.2i0.84/16.6 i0.89/15.6i0.55; P14, 7.8i1.3/14.4i1.14/8.2i0.84; P16, 1.4 i0.55/10.8i0.84/3.2i1.09; P187 1i0/7.4i1.14/1.6i0.55; P20, 0.8i0.45/1iO/O.8i0.45; P27, 0.8i0.45/0.8i0.45/0.8i0.45. In group A, C611 loss was significantly blocked by PBN from P14 to 18, compared to control and group B (∗p<0.05)

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  Fig. 3. DAPI staining. Strong fluorescent spots, indicating apoptotic bodies, were observed primarily in the ONL. Apoptosis is occurring in the rows of OLN cells maintained in group A at day P18. (X400 (P: postnatal day; Black arrow: ONL).

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  Fig. 4. Caspase—3 activity inhibition by PBN in ARPE cells were treated with 120 NM H202 for 24 hrs in the presence or absence of 1 mM PBN. PBN inhibited the increase in caspase—3 activity that occurs in response to H202.

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