May 2005
Volume 46, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2005
Properties of the Tight Junction Relevant for Fluid Transport as Studied With Modulators
Author Affiliations & Notes
  • L. Ma
    Ophthalmology, Columbia University, New York, NY
  • K. Kuang
    Ophthalmology, Columbia University, New York, NY
  • P. Iserovich
    Ophthalmology, Columbia University, New York, NY
  • J. Li
    Ophthalmology, Columbia University, New York, NY
  • J. Fischbarg
    Ophthalmology, Columbia University, New York, NY
  • Footnotes
    Commercial Relationships  L. Ma, None; K. Kuang, None; P. Iserovich, None; J. Li, None; J. Fischbarg, None.
  • Footnotes
    Support  NIH Grant EY 06178, and RPB
Investigative Ophthalmology & Visual Science May 2005, Vol.46, 2206. doi:
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      L. Ma, K. Kuang, P. Iserovich, J. Li, J. Fischbarg; Properties of the Tight Junction Relevant for Fluid Transport as Studied With Modulators . Invest. Ophthalmol. Vis. Sci. 2005;46(13):2206.

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Abstract
 
Abstract:
 

 

According to a recent hypothesis, the corneal endothelium removes water from the corneal stroma through the paracellular route via electro–osmotic coupling taking place in the tight junctions (TJ). Hence, we investigated the relationship between TJ electrical resistance and endothelial fluid transport.

 

 

ATP, dBcAMP, palmitoyl carnitine (PC), ouabain and poly–L–lysine (PLL) were used to modulate corneal endothelial fluid transport and TJ. We used the standard deepithelialized rabbit corneal preparation to determine fluid transport (FT). The corneal thickness was determined using an automated specular reflection (Dikstein–Maurice) microscope. Reagents were added when corneal thickness had stabilized after mounting (about 60 minutes). Transendothelial resistance (TER) was determined using cultured rabbit corneal endothelial cells (CRCEC) grown to full confluence for two weeks on 24 mm permeable membrane inserts.

 

 

The rates (um/h) of corneal stroma swelling and the TER (% of initial value) after 60 minutes of drugs application were:

 

 

 

(1)

 

If fluid transport is paracellular, opening of that route would increase FT. This is what we observe when using dBcAMP and ATP: fluid transport increases, and TER decreases.

 

(2)

 

If electro–osmosis is the driving force, changing the TJ zeta potential by treatment with the polycation PLL would disrupt electro–osmosis without significant effect on TER. This is what we observe.

 

(3)

 

If TJs are however disrupted (opened in an unspecific manner), one would expect both decreased TER and decreased FT, as we also observe.

 

 
Keywords: cornea: endothelium • cornea: basic science • cell adhesions/cell junctions 
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