Related Experiment Videos
Differential expression of nitric oxide synthase isoforms in form-deprived chick eyes
Purpose:
To clarify whether nitric oxide synthase (NOS) is involved in development of myopia, we examined the influence of form deprivation on the expressions of NOS isoform mRNA.
Methods:
NOS isoform cDNAs were amplified from total RNA extracted from control and 7-day-form-deprived chick retina-RPE (retinal pigment epithelium)-choroid, using competitive RT-PCR (reverse-transcription polymerase chain reaction). Each NOS isoform protein was also analyzed by Western blotting and immunohistochemistry.
Result:
Expression of inducible NOS (iNOS) mRNA was highest in the control chick retina-RPE-choroid, followed by the expression of brain NOS (bNOS) mRNA. Expression of endothelial NOS (eNOS) mRNA was faint. The iNOS protein level, however, was only slightly higher than the levels of the bNOS and eNOS proteins and was found mainly in the outer part of the photoreceptor layer and inner and outer parts of RPE and choroid. bNOS alone was found in the outer nuclear layer. Although form deprivation reduced the iNOS and bNOS mRNA expressions, only the iNOS protein showed significant reduction.
Conclusion:
All three NOS isoforms were expressed in chick retina-RPE-choroid. Predominant expression of iNOS, instead of bNOS and eNOS, suggested the existence of ocular tissue-specific regulation of the iNOS gene. In addition to differences in expression level, bNOS displayed regional differential expression. Moreover, only iNOS was reduced in response to form deprivation. It is suggested that NOS isoforms may be differentially involved in the mechanisms regulating the posterior eye tissues, including myopic eye growth.
Insights
Nitric oxide synthase (NOS) isoforms are present in chick eyes. Form deprivation reduced inducible NOS (iNOS) protein, suggesting a role in myopia development.
Area of Science:
- Ophthalmology
- Molecular Biology
- Neuroscience
Background:
- Nitric oxide synthase (NOS) plays a role in various physiological processes.
- The involvement of specific NOS isoforms in eye development and myopia is not fully understood.
Purpose of the Study:
- To investigate the role of nitric oxide synthase (NOS) isoforms in myopia development.
- To examine the influence of form deprivation on NOS isoform expression in chick ocular tissues.
Main Methods:
- Competitive RT-PCR was used to analyze NOS isoform mRNA expression in control and form-deprived chick retina-RPE-choroid.
- Western blotting and immunohistochemistry were employed to assess NOS isoform protein levels and localization.
Main Results:
- All three NOS isoforms (iNOS, bNOS, eNOS) were detected in chick retina-RPE-choroid.
- Inducible NOS (iNOS) mRNA and protein were predominantly expressed.
- Form deprivation led to a significant reduction in iNOS protein levels, while mRNA levels of iNOS and bNOS were reduced.
Conclusions:
- The expression of NOS isoforms in ocular tissues suggests differential roles in eye physiology.
- The reduction in iNOS protein following form deprivation indicates its potential involvement in the regulation of myopic eye growth.