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Masaru Inatani, Hidenobu Tanihara, Atsuhiko Oohira, Megumi Honjo, Noriaki Kido, Yoshihito Honda; Spatiotemporal Expression Patterns of 6B4 Proteoglycan/Phosphacan in the Developing Rat Retina. Invest. Ophthalmol. Vis. Sci. 2000;41(7):1990-1997.
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purpose. To investigate expression of 6B4 proteoglycan/phosphacan, the major
constituent of chondroitin sulfate proteoglycan and a possible
modulator of neural network formation in the developing central nervous
system, in developing rat retina.
methods. Changes in expression and localization of 6B4 proteoglycan in
developing rat retina were investigated by reverse
transcription–initiated polymerase chain reaction (RT–PCR),
immunohistochemistry, and immunoblot analysis.
results. Semiquantitative RT–PCR revealed that mRNA expression of 6B4
proteoglycan in retinas peaked at postnatal day 14 (P14) and then
decreased at P42. Immunohistochemical analyses using MAb 6B4, a
monoclonal antibody against 6B4 proteoglycan, revealed faint
immunoreactivity in the inner aspects of the retina at embryonal day 16
(E16). At birth, weak immunoreactivity was present in the nerve fiber
layer (NFL) and inner plexiform layer (IPL). At P7 and P14, the NFL,
IPL, and outer plexiform layer (OPL) stained intensely, but the
ganglion cell layer (GCL) remained unstained. Between P21 and P42,
immunoreactivity in the NFL and IPL weakened slightly. Immunoblot
analyses showed a MAb 6B4 immunopositive band in the retinal soluble
fraction treated with chondroitinase ABC. The amount of the
immunopositive band increased rapidly as retinal development proceeded.
Surprisingly, a significant amount of the immunopositive band was
present in the retina even before digestion with chondroitinase ABC,
indicating that at least part of 6B4 proteoglycan in rat retina exists
in a non-proteoglycan form.
conclusions. The existence of 6B4 proteoglycan/phosphacan was thus
demonstrated in rat retina, although some biochemical parameters were
different from those of the 6B4 proteoglycan seen in
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