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Biochemical and Molecular Actions of Nutrients

Vitamin A Deficiency Alters the Structure and Collagen IV Composition of Rat Renal Basement Membranes¹

M. Pilar Marín, Guillermo Esteban-Pretel, Ruth Alonso, Yoshikazu Sado^*, Teresa Barber, Jaime Renau-Piqueras and Joaquín Timoneda²

Departamento de Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad de Valencia, Valencia, Spain; ^* Division of Immunology, Shigei Medical Research Institute, Okayama, Japan; and Centro de Investigación, Hospital "La Fe," Valencia, Spain

²To whom correspondence should be addressed. E-mail: Joaquin.Timoneda{at}uv.es.

Retinoids can modulate the expression of extracellular matrix (ECM) proteins with variable results depending on other contributing factors. Because changes in these proteins may alter the composition and impair the function of specialized ECM structures such as basement membranes (BMs), we studied the effects of vitamin A deficiency on renal BMs during the growing period. Newborn male rats were fed a vitamin A-deficient (VAD) diet for 50 d. The ultrastructure of renal BMs was analyzed by electron microscopy. Total collagen IV, the different (IV) chains, matrix degrading metalloproteinases (MMP), and tissue inhibitors of metalloproteinases (TIMP) were quantified by immunocytochemistry and/or Western blotting. Tumor necrosis factor- and interleukin-1ß were measured by ELISA. Semiquantitative RT-PCR was used for determining the steady-state levels for each (IV) chain mRNA. VAD renal BMs showed an irregular thickening, particularly tubular BM. The total collagen IV content was increased, but there was a differential expression of the collagen IV chains. The protein amounts for 1(IV), 4(IV), and 5(IV) were similarly increased, whereas 2(IV) and 3(IV) were decreased. The levels of mRNA for each collagen IV chain changed in parallel with those of the corresponding protein. Both MMP2 and MMP9 were diminished, but no change was detected in TIMP1 or TIMP2. Our data indicate that nutritional VAD leads to alterations in the structure of renal BMs and to quantitative and qualitative variations in its collagen IV composition. These changes may be a factor predisposing to or resulting in kidney malfunction and renal disease.

KEY WORDS: • collagen IV • vitamin A • basement membranes • rat kidney • metalloproteinase

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