Protein Kinase C (αand β) Immunoreactivity in Rabbit and Rat Retina: Effect of Phorbol Esters and Transmitter Agonists on Immunoreactivity and the Translocation of the Enzyme from Cytosolic to Membrane Compartments

N. N. Osborne, N. J. Broyden, N. L. Barnett, N. J. Morris

Research output: Contribution to journalArticleResearchpeer-review

48 Citations (Scopus)

Abstract

Using a monoclonal antibody against protein kinase C (PKC) that recognises the isoenzymes α, βI, and βII, positive immunoreactivity was observed throughout the cytoplasm of bipolar cells in both rat and rabbit retinas. PKC immunoreactivity was also associated with the outer segment of photoreceptors in the rabbit retina and presumed amacrine cells in the rat retina. The PKC immunoreactivity in the retina was unaffected in content or localisation in rats kept in continuous dark or light conditions over a period of 6 days. The localisation of PKC immunoreactivity in retinas was similar in 6‐day‐old, 16 day‐old, or adult rabbits. However, the content of PKC was lowest at the youngest stage and highest in the adult rabbit retinas. Of the two active phorbol esters studied, only phorbol 12, 13‐dibutyrate (PDbut) at a concentration of 1 μM caused the PKC immunoreactivity in rabbit retina bipolar cells to be „transported” from the perikarya towards the axonal terminal processes. Biochemical analyses showed that most of the cytosolic PKC was translocated to the membrane compartment following such treatment. The other phorbol ester, phorbol 12‐myristate 13‐acetate, even at a concentration of 10 μM did not cause a similar transport of PKC immunoreactivity in the bipolar cells, although a partial translocation of the enzyme could be followed biochemically. Both the translocation and transport of PKC by PDbut could be reversed by simply incubating the retinas in physiological solution for 60 min. The „transport” and translocation processes were not obviously affected by the transport inhibitor colchicine or by known PKC inhibitors such as staurosporine, H‐7, sphingosine, or polymyxin B. In addition, agonists known to stimulate inositol phosphates in the retina, viz., carbachol, noradrenaline, and quisqualate, or 4‐aminopyridine did not cause a translocation or „transport” of PKC as observed for the phorbol esters.

Original languageEnglish
Pages (from-to)594-604
Number of pages11
JournalJournal of Neurochemistry
Volume57
Issue number2
DOIs
Publication statusPublished - 1 Jan 1991
Externally publishedYes

Fingerprint

Phorbol Esters
Protein Kinase C
Retina
Rats
Transmitters
Rabbits
Membranes
Enzymes
Carrier Proteins
Quisqualic Acid
Amacrine Cells
Polymyxin B
Sphingosine
Inositol Phosphates
Staurosporine
Protein C Inhibitor
Colchicine
Carbachol
Protein Kinase Inhibitors
Isoenzymes

Cite this

@article{6d089344b2c846fea7e50f549f66e954,
title = "Protein Kinase C (αand β) Immunoreactivity in Rabbit and Rat Retina: Effect of Phorbol Esters and Transmitter Agonists on Immunoreactivity and the Translocation of the Enzyme from Cytosolic to Membrane Compartments",
abstract = "Using a monoclonal antibody against protein kinase C (PKC) that recognises the isoenzymes α, βI, and βII, positive immunoreactivity was observed throughout the cytoplasm of bipolar cells in both rat and rabbit retinas. PKC immunoreactivity was also associated with the outer segment of photoreceptors in the rabbit retina and presumed amacrine cells in the rat retina. The PKC immunoreactivity in the retina was unaffected in content or localisation in rats kept in continuous dark or light conditions over a period of 6 days. The localisation of PKC immunoreactivity in retinas was similar in 6‐day‐old, 16 day‐old, or adult rabbits. However, the content of PKC was lowest at the youngest stage and highest in the adult rabbit retinas. Of the two active phorbol esters studied, only phorbol 12, 13‐dibutyrate (PDbut) at a concentration of 1 μM caused the PKC immunoreactivity in rabbit retina bipolar cells to be „transported” from the perikarya towards the axonal terminal processes. Biochemical analyses showed that most of the cytosolic PKC was translocated to the membrane compartment following such treatment. The other phorbol ester, phorbol 12‐myristate 13‐acetate, even at a concentration of 10 μM did not cause a similar transport of PKC immunoreactivity in the bipolar cells, although a partial translocation of the enzyme could be followed biochemically. Both the translocation and transport of PKC by PDbut could be reversed by simply incubating the retinas in physiological solution for 60 min. The „transport” and translocation processes were not obviously affected by the transport inhibitor colchicine or by known PKC inhibitors such as staurosporine, H‐7, sphingosine, or polymyxin B. In addition, agonists known to stimulate inositol phosphates in the retina, viz., carbachol, noradrenaline, and quisqualate, or 4‐aminopyridine did not cause a translocation or „transport” of PKC as observed for the phorbol esters.",
author = "Osborne, {N. N.} and Broyden, {N. J.} and Barnett, {N. L.} and Morris, {N. J.}",
year = "1991",
month = "1",
day = "1",
doi = "10.1111/j.1471-4159.1991.tb03790.x",
language = "English",
volume = "57",
pages = "594--604",
journal = "Journal of Neurochemistry",
issn = "0022-3042",
publisher = "Wiley-Blackwell",
number = "2",

}

Protein Kinase C (αand β) Immunoreactivity in Rabbit and Rat Retina : Effect of Phorbol Esters and Transmitter Agonists on Immunoreactivity and the Translocation of the Enzyme from Cytosolic to Membrane Compartments. / Osborne, N. N.; Broyden, N. J.; Barnett, N. L.; Morris, N. J.

In: Journal of Neurochemistry, Vol. 57, No. 2, 01.01.1991, p. 594-604.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Protein Kinase C (αand β) Immunoreactivity in Rabbit and Rat Retina

T2 - Effect of Phorbol Esters and Transmitter Agonists on Immunoreactivity and the Translocation of the Enzyme from Cytosolic to Membrane Compartments

AU - Osborne, N. N.

AU - Broyden, N. J.

AU - Barnett, N. L.

AU - Morris, N. J.

PY - 1991/1/1

Y1 - 1991/1/1

N2 - Using a monoclonal antibody against protein kinase C (PKC) that recognises the isoenzymes α, βI, and βII, positive immunoreactivity was observed throughout the cytoplasm of bipolar cells in both rat and rabbit retinas. PKC immunoreactivity was also associated with the outer segment of photoreceptors in the rabbit retina and presumed amacrine cells in the rat retina. The PKC immunoreactivity in the retina was unaffected in content or localisation in rats kept in continuous dark or light conditions over a period of 6 days. The localisation of PKC immunoreactivity in retinas was similar in 6‐day‐old, 16 day‐old, or adult rabbits. However, the content of PKC was lowest at the youngest stage and highest in the adult rabbit retinas. Of the two active phorbol esters studied, only phorbol 12, 13‐dibutyrate (PDbut) at a concentration of 1 μM caused the PKC immunoreactivity in rabbit retina bipolar cells to be „transported” from the perikarya towards the axonal terminal processes. Biochemical analyses showed that most of the cytosolic PKC was translocated to the membrane compartment following such treatment. The other phorbol ester, phorbol 12‐myristate 13‐acetate, even at a concentration of 10 μM did not cause a similar transport of PKC immunoreactivity in the bipolar cells, although a partial translocation of the enzyme could be followed biochemically. Both the translocation and transport of PKC by PDbut could be reversed by simply incubating the retinas in physiological solution for 60 min. The „transport” and translocation processes were not obviously affected by the transport inhibitor colchicine or by known PKC inhibitors such as staurosporine, H‐7, sphingosine, or polymyxin B. In addition, agonists known to stimulate inositol phosphates in the retina, viz., carbachol, noradrenaline, and quisqualate, or 4‐aminopyridine did not cause a translocation or „transport” of PKC as observed for the phorbol esters.

AB - Using a monoclonal antibody against protein kinase C (PKC) that recognises the isoenzymes α, βI, and βII, positive immunoreactivity was observed throughout the cytoplasm of bipolar cells in both rat and rabbit retinas. PKC immunoreactivity was also associated with the outer segment of photoreceptors in the rabbit retina and presumed amacrine cells in the rat retina. The PKC immunoreactivity in the retina was unaffected in content or localisation in rats kept in continuous dark or light conditions over a period of 6 days. The localisation of PKC immunoreactivity in retinas was similar in 6‐day‐old, 16 day‐old, or adult rabbits. However, the content of PKC was lowest at the youngest stage and highest in the adult rabbit retinas. Of the two active phorbol esters studied, only phorbol 12, 13‐dibutyrate (PDbut) at a concentration of 1 μM caused the PKC immunoreactivity in rabbit retina bipolar cells to be „transported” from the perikarya towards the axonal terminal processes. Biochemical analyses showed that most of the cytosolic PKC was translocated to the membrane compartment following such treatment. The other phorbol ester, phorbol 12‐myristate 13‐acetate, even at a concentration of 10 μM did not cause a similar transport of PKC immunoreactivity in the bipolar cells, although a partial translocation of the enzyme could be followed biochemically. Both the translocation and transport of PKC by PDbut could be reversed by simply incubating the retinas in physiological solution for 60 min. The „transport” and translocation processes were not obviously affected by the transport inhibitor colchicine or by known PKC inhibitors such as staurosporine, H‐7, sphingosine, or polymyxin B. In addition, agonists known to stimulate inositol phosphates in the retina, viz., carbachol, noradrenaline, and quisqualate, or 4‐aminopyridine did not cause a translocation or „transport” of PKC as observed for the phorbol esters.

UR - http://www.scopus.com/inward/record.url?scp=0025738263&partnerID=8YFLogxK

U2 - 10.1111/j.1471-4159.1991.tb03790.x

DO - 10.1111/j.1471-4159.1991.tb03790.x

M3 - Article

VL - 57

SP - 594

EP - 604

JO - Journal of Neurochemistry

JF - Journal of Neurochemistry

SN - 0022-3042

IS - 2

ER -