Subcellular localization of renal kallikrein by ultrastructural immunocytochemistry

C. P. Vio; C. D. Figueroa

doi:10.1038/ki.1985.115

Subcellular localization of renal kallikrein by ultrastructural immunocytochemistry

C. P. Vio, C. D. Figueroa

Universidad Austral de Chile

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84 Scopus citations

Abstract

The subcellular distribution of immunoreactive kallikrein was described in the rat nephron using ultrastructural immunocytochemistry. The renal tissue was fixed with a mixture of buffered picric acid-paraformaldehyde-glutaraldehyde and immunostained with the peroxidase-antiperoxidase method for the electron microscope with the following steps: (1) antikallikrein antiserum, (2) anti-IgG serum, (3) peroxidase-antiperoxidase complex, (4) 3-3' diaminobenzidine-H₂O₂, and (5) post-staining with osmium tetroxide. Preabsorption of the primary antiserum with purified rat urinary kallikrein and substitution with normal serum were used as controls. As we have described previously, kallikrein was present exclusively in the connecting tubule cell of the distal nephron. Subcellularly, kallikrein was distributed in luminal membranes, basal membranes, rough edoplasmic reticulum, Golgi apparatus, and vesicles. The immunoreactive vesicles were present in the proximity of the Golgi apparatus and in the cytoplasm in the way between the Golgi and the luminal and basal plasma membranes. No immunostaining was observed in other subcellular components of the connecting tubule cell or in the other type of cell. With the description of kallikrein in subcellular organelles involved in the synthesis, processing, and transport of glycoproteins, we have advanced an hypothetical intracellular processing pathway for renal kallikrein.

Original language	English
Pages (from-to)	36-42
Number of pages	7
Journal	Kidney International
Volume	28
Issue number	1
DOIs	https://doi.org/10.1038/ki.1985.115
State	Published - 1985
Externally published	Yes

Bibliographical note

Funding Information:
Portions of this work were presented at the Kinin '84 meeting in Savannah, Georgia, USA, October, 1984. This work was supported by Grant RS-82-32 from the Universidad Austral de Chile, DirecciOn de InvestigaciOn y Desarrollo, and by Grant 1119/83 from CONICYT. The authors acknowledge Professor Italo Caorsi, M.D., for his comments on this work, and thank Mr. L. Delannoy and Mr. C. Lizama for technical assistance, and Mrs. R. Guzmán for editorial assistance.

ASJC Scopus subject areas

Nephrology

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10.1038/ki.1985.115

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title = "Subcellular localization of renal kallikrein by ultrastructural immunocytochemistry",

abstract = "The subcellular distribution of immunoreactive kallikrein was described in the rat nephron using ultrastructural immunocytochemistry. The renal tissue was fixed with a mixture of buffered picric acid-paraformaldehyde-glutaraldehyde and immunostained with the peroxidase-antiperoxidase method for the electron microscope with the following steps: (1) antikallikrein antiserum, (2) anti-IgG serum, (3) peroxidase-antiperoxidase complex, (4) 3-3' diaminobenzidine-H2O2, and (5) post-staining with osmium tetroxide. Preabsorption of the primary antiserum with purified rat urinary kallikrein and substitution with normal serum were used as controls. As we have described previously, kallikrein was present exclusively in the connecting tubule cell of the distal nephron. Subcellularly, kallikrein was distributed in luminal membranes, basal membranes, rough edoplasmic reticulum, Golgi apparatus, and vesicles. The immunoreactive vesicles were present in the proximity of the Golgi apparatus and in the cytoplasm in the way between the Golgi and the luminal and basal plasma membranes. No immunostaining was observed in other subcellular components of the connecting tubule cell or in the other type of cell. With the description of kallikrein in subcellular organelles involved in the synthesis, processing, and transport of glycoproteins, we have advanced an hypothetical intracellular processing pathway for renal kallikrein.",

author = "Vio, {C. P.} and Figueroa, {C. D.}",

note = "Funding Information: Portions of this work were presented at the Kinin '84 meeting in Savannah, Georgia, USA, October, 1984. This work was supported by Grant RS-82-32 from the Universidad Austral de Chile, DirecciOn de InvestigaciOn y Desarrollo, and by Grant 1119/83 from CONICYT. The authors acknowledge Professor Italo Caorsi, M.D., for his comments on this work, and thank Mr. L. Delannoy and Mr. C. Lizama for technical assistance, and Mrs. R. Guzm{\'a}n for editorial assistance.",

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N1 - Funding Information: Portions of this work were presented at the Kinin '84 meeting in Savannah, Georgia, USA, October, 1984. This work was supported by Grant RS-82-32 from the Universidad Austral de Chile, DirecciOn de InvestigaciOn y Desarrollo, and by Grant 1119/83 from CONICYT. The authors acknowledge Professor Italo Caorsi, M.D., for his comments on this work, and thank Mr. L. Delannoy and Mr. C. Lizama for technical assistance, and Mrs. R. Guzmán for editorial assistance.

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N2 - The subcellular distribution of immunoreactive kallikrein was described in the rat nephron using ultrastructural immunocytochemistry. The renal tissue was fixed with a mixture of buffered picric acid-paraformaldehyde-glutaraldehyde and immunostained with the peroxidase-antiperoxidase method for the electron microscope with the following steps: (1) antikallikrein antiserum, (2) anti-IgG serum, (3) peroxidase-antiperoxidase complex, (4) 3-3' diaminobenzidine-H2O2, and (5) post-staining with osmium tetroxide. Preabsorption of the primary antiserum with purified rat urinary kallikrein and substitution with normal serum were used as controls. As we have described previously, kallikrein was present exclusively in the connecting tubule cell of the distal nephron. Subcellularly, kallikrein was distributed in luminal membranes, basal membranes, rough edoplasmic reticulum, Golgi apparatus, and vesicles. The immunoreactive vesicles were present in the proximity of the Golgi apparatus and in the cytoplasm in the way between the Golgi and the luminal and basal plasma membranes. No immunostaining was observed in other subcellular components of the connecting tubule cell or in the other type of cell. With the description of kallikrein in subcellular organelles involved in the synthesis, processing, and transport of glycoproteins, we have advanced an hypothetical intracellular processing pathway for renal kallikrein.

AB - The subcellular distribution of immunoreactive kallikrein was described in the rat nephron using ultrastructural immunocytochemistry. The renal tissue was fixed with a mixture of buffered picric acid-paraformaldehyde-glutaraldehyde and immunostained with the peroxidase-antiperoxidase method for the electron microscope with the following steps: (1) antikallikrein antiserum, (2) anti-IgG serum, (3) peroxidase-antiperoxidase complex, (4) 3-3' diaminobenzidine-H2O2, and (5) post-staining with osmium tetroxide. Preabsorption of the primary antiserum with purified rat urinary kallikrein and substitution with normal serum were used as controls. As we have described previously, kallikrein was present exclusively in the connecting tubule cell of the distal nephron. Subcellularly, kallikrein was distributed in luminal membranes, basal membranes, rough edoplasmic reticulum, Golgi apparatus, and vesicles. The immunoreactive vesicles were present in the proximity of the Golgi apparatus and in the cytoplasm in the way between the Golgi and the luminal and basal plasma membranes. No immunostaining was observed in other subcellular components of the connecting tubule cell or in the other type of cell. With the description of kallikrein in subcellular organelles involved in the synthesis, processing, and transport of glycoproteins, we have advanced an hypothetical intracellular processing pathway for renal kallikrein.

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Subcellular localization of renal kallikrein by ultrastructural immunocytochemistry

Abstract

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