Electrolyte Blood Press.  2007 Dec;5(2):75-88. 10.5049/EBP.2007.5.2.75.

Immunolocalization of Protein Kinase C Isoenzymes alpha, betaI, betaII and gamma in Adult and Developing Rat Kidney

• Kim WY
• Lee GS
• Kim YH
• Park EY
• Hwang JS
• Kim H
• Kim J

• KMID: 2052289
• DOI: http://doi.org/10.5049/EBP.2007.5.2.75

Abstract

Protein kinase C (PKC) plays an important role not only in signal transduction mechanisms in various biological processes, but also in the regulation of growth and differentiation during development. We studied the classical PKC alpha, betaI, betaII and gamma, with regard to their expression in adult and developing rat kidney. PKCalpha appeared in the ureteric bud at embryonic day (E) 16, and the proximal and distal anlage at E18. After birth, the immunoreactivity of PKCalpha gradually decreased. In adult, PKCalpha was expressed intensely in the connecting tubule (CNT), the collecting ducts (CD) and the renal corpuscle, and weakly in the proximal and distal tubules. PKCbetaI appeared in the ureteric bud at E16, and the proximal anlage at E18. After birth, the immunoreactivity of PKCbetaI gradually disappeared from the CD and proximal tubule. In adult, PKCbetaI was expressed in the intercalated cells of the CNT and cortical CD, the proximal straight tubule, and the renal corpuscle. PKCbII appeared in distal anlage at E18, and increased markedly after birth. In the CD, PKCbetaII immunoreactivity appeared after birth. In adult, PKCbetaII was expressed in the distal tubule, the CNT and the CD. The immunoreactivity for PKCgamma appeared only in the proximal anlage at E18, and increased temporally around the time of birth. However, no immunoreactivity for PKCgamma was observed in adult rat kidney. These results indicate that classical PKC isoforms appear to play a role in the regulation of various renal functions and differentiation within specific functional units of the uriniferous tubule in rat kidney.

MeSH Terms

Adult*
Animals
Biological Processes
Humans
Kidney*
Parturition
Protein Isoforms
Protein Kinase C beta
Protein Kinase C*
Protein Kinases*
Rats*
Signal Transduction
Ureter
Protein Isoforms
Protein Kinase C
Protein Kinases

Figure

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  Fig. 1 Differential interference contrast (DIC) micrographs of wax sections illustrating immunostaining for protein kinase C α (PKCα) in the cortical labyrinth (A), medullary ray (B), initial part of the inner medulla (C), and the terminal part of the inner medulla (D) of adult rat kidneys. PKCα immunostaining is observed in the cytoplasm of mesangial cells (arrows) and the parietal epithelium (arrowhead) of renal corpuscles, intercalated cells (open arrows) of the connecting tubule (CNT) and cortical collecting duct (CCD), and on the basolateral plasma membrane of the CNT cells and the principal cells of the collecting duct. Note the PKCα-negative intercalated cells (open arrowheads) in the initial part of the inner medullary collecting duct (IMCDi). IMCDt, terminal part of the IMCD. Magnifications: A-D, ×400.

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  Fig. 2 DIC micrographs of wax sections illustrating immunostaining for PKCα in 16- (A), 18- (C & F), and 20-day-old (G) fetal kidneys, and 3- (D & H) and 7-day-old (B, E & I) pups. protein kinase C α (PKCα) appeared in the ureteric buds (UB) at 16 days of gestation (A) and in the proximal and distal anlage (stars) at 18 days of gestation (C). B. Note that the PKCα-positive tubular profiles (arrows), which are newly formed proximal and distal tubules, are located only in the subcapsular region in 7-day-old pups, whereas PKCα immunoreactivity is decreased in the mature tubules such as the proximal convoluted tubule (PCT) and connecting tubule (CNT) cells located in the inner cortex. C-E, Note the PKCα immunoreactivity in the mesangial cells (arrows) and parietal epithelium (arrowheads) of the developing renal corpuscle. F-G. Immunoreactivity for PKCα in the basolateral plasma membrane of inner medullary collecting duct (IMCD) cells gradually decreased during development. Note the disappearance of apical PKC expression in the intercalated cells (arrows) of the medullary collecting duct (MCD) after birth. Magnifications: A, ×200; B, ×200; C-I, ×528.

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  Fig. 3 DIC micrographs of wax sections illustrating protein kinase C βI (PKCβI) immunostaining in the cortical labyrinth (A), medullary rays (B), outer stripe of the outer medulla (C), inner stripe of the outer medulla (D), and initial part of the inner medulla (E) of the adult rat kidney. PKCβI immunoreactivity is strong in the intercalated cells (open arrows) of the connecting tubules (CNT) and cortical collecting ducts (CCD), moderate in the proximal straight tubules (PST), and weak in the mesangial cells (arrows) and parietal epithelium of the renal corpuscle. In the inner medulla, PKCβI immunostaining is observed only in the descending thin limb (stars) of the Loop of Henle. There is no immunoreactivity in the CNT cells, principal cells of the CCD, outer (OMCDo) and inner stripe of the outer medullary collecting duct (OMCDi), and initial inner medullary collecting duct (IMCDi). Note that there is no immunoreactivity in the intercalated cells (open arrowheads) of the OMCD and IMCDi. Magnifications: A-E, ×400.

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  Fig. 4 DIC micrographs of wax sections illustrating immunostaining for protein kinase C βI (PKCβI) in 16- (A), 18- (B) and 20-day-old (E) fetal kidneys, and 3-(F) and 7-day-old (C, D & G) pups. PKCβI appeared in the ureteric buds (UB) at 16 days of gestation and in the proximal anlage (open arrows) at 18 days of gestation. PKCβI immunostaining appeared in the differentiating proximal tubules (open arrows) in the fetal stage and disappeared from the mature proximal tubule after birth. PKCβI-positive proximal anlage are observed in the subcapsular region until 7 days after birth. Note the PKCβI immunostaining in the mesangial cells (arrows) and parietal epithelium (arrowheads) of the developing renal corpuscle in D. Immunoreactivity for PKCβI in the medullary collecting duct (MCD) cells gradually disappeared after birth (E-G). IMCD, inner medullary collecting duct. Magnifications: A, ×200; B, ×264; C & D-G, ×528.

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  Fig. 5 DIC micrographs of 1-mm-thick plastic sections illustrating immunostaining for protein kinase C βII (PKCβII) in the cortex (A), medullary rays (B), the border between the outer and inner medulla (C), and the terminal part of the inner medulla (D) of adult rat kidney. Immunoreactivity for PKCβII is well localized on the basolateral plasma membrane of distal convoluted tubule (DCT) cells, connecting tubule (CNT) cells, thick ascending limb (TAL) cells, and principal cells throughout the collecting duct. Note that there is no immunoreactivity for PKCβII in both the type A intercalated cells (arrow) and type B intercalated cells (arrowhead) in the cortical collecting duct (CCD). There is weak basolateral PKCβII labeling in the proximal convoluted tubule (PCT). The open arrow indicates a PKCβII-negative intercalated cell in the CNT. ATL, ascending thin limb; G, glomerulus; OMCDi, inner stripe of the outer medullary collecting duct; IMCDi, initial part of the inner medullary collecting duct; IMCDt, terminal part of the IMCD. Magnifications: A, ×368; B, ×640; C & D, ×320.

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  Fig. 6 DIC micrographs of wax sections illustrating immunostaining for protein kinase C βII (PKCβII) in 18-day-old fetuses (A), and 1- (B), 3- (C), and 7-day-old (D) pups. A. PKCβII immunoreactivity appeared in the distal tubules (asterisks) at 18 days of gestation. B-C. Note a marked increase in PKCβII immunostaining in the distal tubules (asterisks) after birth. Insets demonstrating PKCβII-negative medullary collecting ducts (MCD) from an 18-day-old fetus (A) and a 1-day-old pup (B). In the thick ascending limb (TAL), PKCβII immunoreactivity appeared around the time of birth. P, proximal tubule; S, S-shaped body; III, stage III renal corpuscle; V, renal vesicle. Magnifications: A-D, ×200; insets, ×528.

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  Fig. 7 Light micrographs of wax sections illustrating immunostaining for protein kinase C γ (PKCγ) in adult rat kidneys. There is very weak immunostaining for PKCγ only in the cytoplasm of proximal convoluted tubules (PCT) and proximal straight tubules (PST). Co, cortex; DCT, distal convoluted tubule; OMCDi, inner stripe part of outer medullary collecting duct; OMCDo, outer stripe part of OMCD; OSOM, outer stripe of outer medulla; ISOM, inner stripe of outer medulla; TAL, thick ascending limb. Magnifications: A-C, ×330.

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  Fig. 8 Light micrographs of wax sections illustrating immunostaining for protein kinase C γ (PKCγ) in kidneys from 20-day-old fetuses (A & B), and 1- (C & D), 3- (E), and 21-day-old (F) pups. PKCγ immunoreactivity is highly expressed in the proximal tubules (PT) during development and markedly decreased in the PT from 21 days after birth. B and D are higer magnification area indicate by rectangular in A and C, respectively. DT, distal tubule; PCT, proximal convoluted tubule; PST, proximal straight tubule. Magnifications: A, C, E & F, ×83; B & D, ×330.

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  Fig. 9 Changes in immunoreactivity of classical protein kinase C isoforms in uriniferous tubules during renal development in the rat. RC, renal corpuscle; PT, proximal tubule; DT, distal tubule; CD, collecting duct.

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