From the Department of Nutritional Sciences and
Toxicology, University of California-Berkeley, Berkeley, California
94720 and the § Department of Nutritional Sciences,
University of Wisconsin, Madison, Wisconsin 53706
Received for publication, November 8, 2002, and in revised form, December 13, 2002
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ABSTRACT |
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This study describes cDNA cloning and
characterization of mouse RALDH4. The 2.3-kb cDNA encodes an
aldehyde dehydrogenase of 487 amino acid residues, about two-orders of
magnitude more active in vitro with
9-cis-retinal than with all-trans-retinal. RALDH4 recognizes as substrate 9-cis-retinal generated in
transfected cells by the short-chain dehydrogenases CRAD1, CRAD3, or
RDH1, to reconstitute a path of 9-cis-retinoic acid
biosynthesis in situ. Northern blot analysis showed
expression of RALDH4 mRNA in adult mouse liver and kidney. In
situ hybridization revealed expression of RALDH4 in liver on
embryo day 14.5, in adult hepatocytes, and kidney cortex.
Immunohistochemistry confirmed RALDH4 expression in hepatocytes and
showed that hepatocytes also express RALDH1, RALDH2, and RALDH3. Kidney
expresses the RALDH4 protein primarily in the proximal and distal
convoluted tubules of the cortex but not in the glomeruli or the
medulla. Kidney expresses RALDH2 in the proximal convoluted tubules of
the cortex but not in the distal convoluted tubules or glomeruli.
Kidney expresses RALDH1 and RALDH2 in the medulla. The enzymatic
characteristics of RALDH4, its expression in fetal liver, and its
unique expression pattern in adult kidney compared with RALDH1, -2, and
-3 suggest that it could meet specific needs for
9-cis-retinoic acid biosynthesis.
All-trans-retinoic acid
(atRA)1 induces a variety of
biological responses by modulating gene expression to control
differentiation or entry into apoptosis of diverse cell types in
numerous organs (1-3). atRA seems to function as the major RAR ligand
in vivo and satisfies all known retinoid endocrine functions
(4-6). 9cRA has been identified as an RXR ligand in cells treated with
atRA and serves as a high-affinity ligand for both RAR and RXR in
vitro (7, 8). RXR forms heterodimers with numerous nuclear
receptors, including RAR, vitamin D receptor (VDR), and peroxisome
proliferator-activated receptor (PPAR). The heterodimers seem to
require only the non-RXR ligands for maximum activation, if present in
sufficient concentrations. Nevertheless, 9cRA increases heterodimer
activation in vitro when non-RXR ligand concentrations fall
below maximum. 9cRA and its analogues may affect disease progression,
including noninsulin-dependent diabetes mellitus and
specific cancers (9, 10). If endogenous 9cRA has important functions
in vivo, its biosynthesis likely would be controlled.
Understanding control would require detailed knowledge of the enzymes
that catalyze 9cRA biosynthesis and their expression loci.
RA biosynthesis from retinol proceeds via two reactions with
retinal as an intermediate (11). Only one mouse SDR, RDH1, functions
efficiently with all-trans-retinol, but at least three mouse
SDR, RDH1, CRAD1, and CRAD3, function efficiently with
9-cis-retinol (12-15). Four different RALDH have
been identified that convert all-trans- or
9-cis-retinal into RA. All belong to the ALDH superfamily, which consists of at least 86 eukaryotic members (16). ALDH1A1 (human
ALDH1 (17, 18), mouse AHD2 (19), rat RALDH1 (20-23)) catalyzes
conversion of all-trans- and 9-cis-retinal into
atRA and 9cRA with similar efficiencies
(Vm/K0.5) (24). ALDH1A2
(human ALDH11 (25), mouse/rat RALDH2 (26, 27)) also catalyzes
dehydrogenation of all-trans- and 9-cis-retinal
into RA but functions about 4-fold less efficiently with
9-cis- than with all-trans-retinal (11, 24).
ALDH1A3 (mouse RALDH3 (28)) catalyzes dehydrogenation of
all-trans-retinal efficiently but not that of
9-cis-retinal.2 A
fourth candidate, the human ALDH12 (not the same enzyme as Here, we report cDNA cloning and characterization of a new mouse
ALDH, RALDH4, and its mRNA and protein expression patterns in the
adult and embryo. We reconstituted 9cRA biosynthesis in intact cells to
compare the activity of mRALDH4 to the other two mouse RALDH (RALDH1
and RALDH2) that recognize 9-cis-retinal and to assess
ability of the three RALDH to generate 9cRA from
9-cis-retinal produced in intact cells by the retinol
dehydrogenases mCRAD1, mCRAD3, and mRDH1. We found that RALDH4
functions in intact cells to generate 9cRA when co-expressed with
CRAD1, CRAD2, or RDH1 and has a unique expression pattern in kidney
compared with the other RALDH.
Cloning of Mouse RALDH4 cDNA--
The expressed sequence
tag clone AI504872 (ATCC, IMAGE 987012) in GenBankTM
shows significant homology with human ALDH12 (31). We sequenced the
entire insert and generated the complete 3'-untranslated region with two rounds of RACE using mouse e17 Marathon-Ready cDNA
(Clontech). First-round PCR was done with a program
of 3 min/94 °C, 30 s/94 °C, 30 s/55 °C, and 90 s/72 °C for
35 cycles and finally 72 °C/10 min. Nested PCR was done using the
same program except with 25 cycles. The gene-specific primers for the
two rounds of RACE-PCR were 5'-TGTCAGTGCAGTCAACAGAG-3' (nt 1561-1580)
and 5'-CTCAATGACCACTGACGGTA-3' (nt 1628-1647) of the final cDNA.
An additional 5'-untranslated region sequence was amplified by 5'-RACE
using the GeneRacer kit (Invitrogen) and mouse liver
poly(A+) RNA (Clontech) with the primer
5'-AGAGACTGCTCCAGTACATC-3' (nt 282-301 of the final cDNA),
following the manufacturer's instructions. PCR products were cloned
into pGEN-T (Promega) for sequencing.
Northern Blot--
Mouse multiple tissue and embryo Northern
blots were purchased from Clontech. A 304-base pair
probe corresponding to nt 1343-1646 of the cDNA sequence was
amplified by PCR, labeled with 32P using the RadPrime DNA
labeling system (Invitrogen), and hybridized according to the
manufacturer's protocol. Prehybridization and hybridization were done
in 10 ml of ExpressHyb (Clontech) solution at
68 °C for 30 min and 1 h, respectively. The blot was washed four times in 2× SSC with 0.05% SDS at room temperature for 30 min,
followed by two washes in 0.1× SSC with 0.1% SDS at 68 °C for 30 min. We stripped the RALDH4 probe and reprobed the blot with a
Enzyme Assays--
The coding region of mRALDH4 was amplified
with primers (restriction sites are underlined),
5'-CGGGATCCGGCACTTTCCTAACATGGCT-3' and
5'-CGGAATTCCACAGCAGCTGTAGGATGAT-3', and was cloned into the BamHI/EcoRI sites of pcDNA3 (Invitrogen) to
create pcDNA3/mRALDH4. pcDNA3/mRALDH4 was transfected into
CHO-K1 cells with LipofectAMINE reagent (Invitrogen). Cells transfected
with pcDNA3 were used as control. 24 h after transfection,
cells were sonicated in 20 mM Hepes, 150 mM
KCl, 1 mM EDTA, 10% sucrose, 2 mM
dithiothreitol, pH 7.5. The 800 × g supernatants
(10-min spins) were the source of protein for enzyme assays done at
37 °C in 0.5 ml of 20 mM Hepes, pH 8.5, 150 mM KCl, 1 mM EDTA, 2 mM
dithiothreitol, and 2 mM NAD+. Retinoids were
added in 2 µl of Me2SO. RA was quantified by high
performance liquid chromatography (29).
Reconstitution in Intact Cells--
The day before transfection
with LipofectAMINE 2000 (Invitrogen), CHO-K1 cells were seeded into
6-well plates. 24 h after transfection, the medium was replaced
with fresh medium (1 ml/well) containing 1 µM
9-cis-retinol. After 1 h of incubation, cells and
medium were extracted to quantify 9cRA.
In Situ Hybridization--
The same probe used for Northern blot
hybridization was cloned into pGEM-T. The direction of insertion was
determined by DNA sequencing. The construct was digested with
SpeI and transcribed from the T7 promoter to generate the
antisense probe or was digested with SacII and transcribed
from the SP6 promoter to generate the sense probe. Adult liver and
kidney samples were harvested from 4-week-old male C57BL/6J mice
(Jackson Laboratory). NIH Swiss mouse embryos were obtained from Harlan
Sprague-Dawley (Indianapolis, IN). The morning of plug detection was
designated e0.5. In situ hybridization was done on
paraformaldehyde-fixed, paraffin-embedded tissue sections using
35S-labeled sense and antisense riboprobes as described
(28, 32). Tissue sections were counterstained with propidium iodide
after development. Images were obtained under dark-field and
epifluorescent illumination.
Western Blot and Immunohistochemistry--
Rabbit antibodies
were raised against peptides designed from mouse RALDH4 (residues
406-419, DSEEEVITRANSVR), RALDH1 (residues 434-455, TKDLDKAITVSS),
RALDH2 (residues 482-493, REYSEVKTVTVK), and RALDH3 (residues
446-459, KNLDKALKLAAALE). Each was purified by affinity chromatography
using peptide coupled to
N-hydroxysuccinimide-activated-Sepharose 4 (Amersham
Biosciences). For Western blot analyses, ~5 µg of protein from
CHO cell lysates were run on 10% SDS-PAGE. The samples were
transferred onto a nitrocellulose membrane using a Trans-Blot S.D.
semi-dry transfer cell (Bio-Rad). Nonspecific binding was blocked with
5% nonfat milk. The membrane was then incubated with 0.1 µg/µl
primary antibody for 1 h at room temperature. Signals were
detected by chemiluminescence reagent plus (PerkinElmer Life Sciences)
with horseradish peroxidase-conjugated goat anti-rabbit antibody
(Sigma). Immunohistochemistry was done with the rabbit ABC staining
system following the manufacturer's instructions (Santa Cruz
Biotechnology, Inc.). Adult mouse tissue sections were obtained from
Histo-Tec Laboratory (Hayward, CA). Slides were de-paraffined using
Citrisolv (Fisher) and rehydrated with serial dilutions of aqueous
ethanol. Antigens were unmasked by a 10-min treatment with 0.1% pepsin
in HCl buffer, pH 2.5. Endogenous peroxidase activity was quenched by a
10-min incubation with 1% H2O2. Slides were
incubated with 1.5% blocking serum at room temperature for 1 h,
then with 0.5 µg/µl primary antibody for 30 min at room temperature
or 4 °C overnight, and finally with biotinylated secondary antibody
at room temperature for 30 min. Signals were detected by incubation
with biotinylated horse radish peroxidase together with avidin and were
stained with peroxidase substrate. Negative controls were obtained by
omitting the primary antibody. Sections were counterstained with hematoxylin.
cDNA Cloning of Mouse RALDH4--
Amplification by RACE of the
5'- and 3'-untranslated region of the expressed sequence tag clone
IMAGE 987012 produced a cDNA of 2.3 kb that included 50 nucleotides
upstream of the ATG start site, continued through the polyadenylation
signal, and concluded with 22 adenosine residues (data not shown). The
deduced amino acid sequence showed high identify with a human RALDH
(90%) and with the partial sequence of a rat RALDH (91%) and
contained all 23 invariant residues of the ALDH superfamily (Fig.
1). The mouse ALDH, designated RALDH4,
shows no more than 50% amino acid sequence similarity with mouse
RALDH1, -2, and -3 (Table I)
(19-23).
Enzymatic Properties--
Initial assays were done with 10 µM substrate and the 800 × g supernatant
of transfected CHO cells. mRALDH4 had high enzymatic activity with
9-cis-retinal (2.5 ± 0.2 nmol/min/mg protein;
means ± S.D., n = 3), lower activity with
13-cis-retinal (0.6 ± 0.06), and very low activity
with all-trans-retinal (0.03 ± 0.002). Kinetic constants were determined in two independent assays under initial velocity conditions with the most efficient substrate,
9-cis-retinal. The average Km value was
2.3 ± 0.3 µM (± S.E.) and the average
Vmax value was 3.4 ± 0.1 nmol/min/mg (Fig.
2, top). Disulfiram and citral
inhibited mRALDH4 activity with IC50 values of 5.3 and 31 µM, respectively (Fig. 2, bottom).
Acetaldehyde inhibited weakly (IC50 value = 9.6 mM).
mRALDH4 functioned in reconstituted paths of 9cRA biosynthesis in
intact cells, with 9-cis-retinal generated in
situ by mCRAD1, mCRAD3, or mRDH1 (Fig.
3). CRAD1 and CRAD3 were about equivalent in generating 9-cis-retinal, as predicted by their kinetic
constants (13, 14), whereas RDH1 was less efficient, consistent with its greater efficiency with all-trans-retinol than
9-cis-retinol (15). RALDH4 was less efficient than RALDH1 or
RALDH2 in generating 9cRA from 9-cis-retinal produced by
CRAD1. Western blots showed that differences in protein expression did
not contribute markedly to the differences in activities.
mRNA Expression of Mouse RALDH4--
Northern blot showed
intense expression of mRALDH4 mRNA in liver and kidney (Fig.
4). No signals were detected in six other tissues screened. Northern blot also showed expression at e15 and e17
but no signal at e7 or e11, indicating initiation of RALDH4 expression
after e11 and on or before e15.
In Situ Hybridization--
No mRALDH4 mRNA signal was detected
in the e10.5 embryo, consistent with the Northern blot data (Fig.
5). At e14.5, expression was detected in
liver but not in kidney or elsewhere except for low but consistent
signals in some blood cell populations (data not shown). In the
4-week-old male mouse, RALDH4 mRNA was expressed throughout the
liver and intensely in the kidney cortex (Fig. 6).
Expression of RALDH Protein in Adult Liver and Kidney--
To test
whether the anti-peptide antibodies were specific, Western blot
analyses were done with lysates of CHO cells transfected with each
RALDH construct. Each RALDH was detected as a ~54-kDa band, and each
antibody reacted specifically with its corresponding enzyme (Fig.
7).
Immunohistochemical staining of adult mouse liver sections showed that
many but not all hepatocytes throughout the liver express the four
RALDH (Fig. 8). No pattern emerged for
RALDH expression in any particular liver section. Frequent signals for
RALDH1 were detected in cells with lipid droplets (Fig. 8, panel
1, arrow). Signals for RALDH2 and 3 were much less
frequent in cells with lipid droplets, and signals for RALDH4 did
appear in cells with lipid droplets. The epithelial cells of the bile
ducts and the endothelial cells of blood vessels showed no specific
signal for any of the four enzymes. Negative controls (no primary
antibody) showed no staining (Fig. 8C).
Immunohistochemistry showed that kidney expresses RALDH4 primarily in
the cortex in the proximal and distal convoluted tubules, overlapping
with its mRNA expression (Fig.
9A). RALDH4 was not detected
in glomeruli (Fig. 9, B and C). RALDH1 produced
weak signals close to background in the cortex (Fig. 9D) but
showed strong signals in the medulla, localized to the straight
segments of loops of Henle (Fig. 9F), collecting tubules
(Fig. 9G), and thin segments of loops of Henle (Fig.
9H). RALDH2 was detected in the cortex in proximal
convoluted tubules but not in distal convoluted tubules or glomeruli
(Fig. 9E). RALDH2 also was detected in the medulla in
collecting tubules (Fig. 9I) and straight segments of loops
of Henle (Fig. 9J). Negative controls (no primary antibody) showed no signals (Fig. 9K). No RALDH3 signals were detected
in kidney.
This report shows that a fourth mouse ALDH, RALDH4, recognizes
retinoids as substrates. Mouse RALDH4 likely represents an ortholog of
human ALDH123 because the two
share 90% amino acid identity, have similar Km values for 9-cis-retinal (3.2 µM for ALDH12),
do not catalyze all-trans-retinal dehydrogenation well, and
are expressed intensely in the adult only in liver and kidney (31).
Characterization of mouse RALDH4 extends insight into retinoid
metabolism by demonstrating its generation of 9cRA in intact cells in
cooperation with any one of three mouse SDR (CRAD1, CRAD3, RDH1) and by
showing that its mRNA and protein expression patterns differ from
the other RALDH. In contrast to RALDH4, the e14.5 mouse liver expresses RALDH1 mRNA weakly and does not express RALDH2 or RALDH3 mRNA (33, 34). This early intense expression of RALDH4 in fetal liver
suggests a function in fetal liver development. Expression of RALDH4 in
liver and some blood cells at e14.5 suggests a potential contribution
to fetal hemopoiesis. This is significant because mouse embryos that
lack RXR RALDH1 accounts for at least 90% of the all-trans-retinal
and 9-cis-retinal dehydrogenase activities in adult
rat/mouse liver and kidney (18, 19, 24). RALDH2, -3, and -4 contribute the remaining activity, without apparent domination by any
one. Multiple RALDH expressed in hepatocytes, albeit not necessarily in
the same hepatocytes, may allow differential regulation or may reflect the status of the cell cycle or differentiation state. Distinctive functions for each isozyme are suggested by the observation that adult
mouse kidney cortex expresses RALDH2 and -4, whereas the medulla
expresses RALDH1 and -2. Adult mouse kidney expresses RDH1 protein in
both the cortex and medulla, which could provide 9-cis-retinal for 9cRA production by
RALDH.4 The lack of a protein
signal for RALDH3 in kidney contrasts with detection of RALDH3 mRNA
in the collecting ducts of the renal papilla in fetal and adult mouse
(34). It is not clear whether this reflects a technical problem or weak
translation of RALDH3 mRNA and/or rapid turnover of RALDH3 protein
in adult kidney.
Citral inhibits retinal metabolism catalyzed by rat RALDH1 and -2 and
chick RALDH3 with IC50 values of 1, 12, and 3.5 µM, respectively, compared with the IC50
value of 25 µM with mRALDH4 (20, 23, 28). Areas affected
morphologically by citral have been considered loci of RA biosynthesis
and signaling, even though no data indicates that citral inhibits RALDH
and/or any ALDH specifically. The aldehyde functional group of citral
could impair actions of many proteins through Schiff's-base formation
unrelated to active site binding. Acetaldehyde inhibition of all RALDH
suggests potential impairment of RA generation through two mechanisms,
competition at active sites and acetylation of lysine residues.
In summary, this study describes cDNA cloning and characterization
of mouse RALDH4, an enzyme active with endogenously generated 9-cis-retinal in intact cells. RALDH4 has different
expression during embryonic development and in the adult kidney
compared with three other mouse RALDH. RALDH4 can function with any one of CRAD1, CRAD3, or RDH1 to reconstitute a path of 9cRA biosynthesis in
cells. These data are consistent with a distinct contribution of RALDH4
to retinoid metabolism.
INTRODUCTION
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ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
1-pyrroline-5-carboxylate, which also has been
designated ALDH12/ALDH8A1 (29, 30)), has considerable activity with
9-cis-retinal but negligible activity with
all-trans-retinal (31). It is the only known ALDH more
efficient for 9-cis-retinal than for
all-trans-retinal.
MATERIALS AND METHODS
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ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
-actin probe (Clontech) using the same protocol.
Blots were exposed to Kodak X-OMAT LS film overnight.
RESULTS
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
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Fig. 1.
Amino acid sequences of mouse and rat RALDH4
and the human ortholog. Underlining indicates residues
that differ in each species. Boldface identifies the 23 invariant residues in the ALDH superfamily. The box shows
the peptide used to generate antibodies.
Amino acid sequence comparisons of mouse RALDH
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Fig. 2.
Enzymatic properties of mouse RALDH4 with
9-cis-retinal. Top panel, kinetic data
represent means ± S.D. of triplicates from a representative
experiment. Bottom panel, inhibition by disulfiram
(filled squares), citral (filled circles), and
acetaldehyde (open circles). Inhibition data are averages of
duplicates with 3 µM substrate; inhibitors were added in
5 µl of ethanol. Kinetic and inhibitor assays were done for 10 min
with 4 µg of protein and were analyzed by nonlinear regression using
GraftPad Prism 3.0.
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Fig. 3.
Contribution of mRALDH4 to 9cRA acid
biosynthesis in intact cells. Top panel, CHO cells were
transfected with pcDNA3/RALDH4 (0, 0.25, 0.5, and 1 µg,
left to right) and 0.5 µg of one of three vectors that
express SDR, which generate 9-cis-retinal from
9-cis-retinol. A, pcDNA3/CRAD1 (open
bars). B, pcDNA3/CRAD3 (striped bars);
C, pcDNA3/RDH1 (filled bars). Bottom
panel, in a separate experiment, CHO cells were transfected with
pcDNA3/CRAD1 (0, 0.25, 0.5, and 1 µg) and 0.5 µg of
pcDNA3/RALDH1 (D, open bars),
pcDNA3/RALDH2 (E, striped bars),
pcDNA3/RALDH4 (F, filled bars; 0.25, 0.5, and
1 µg vector only). Data are pmol 9cRA generated per well in 1 h
of incubation from 1 µM 9-cis-retinol
(means ± S.D., n = 3). Western blots are shown
below each graph. The top rows show expression
from the varied vectors; the bottom rows show expression
from the fixed vectors.
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Fig. 4.
mRNA expression of mRALDH4 in adult and
embryo. Adult tissues screened included: heart (1);
brain (2); spleen (3); lung (4); liver
(5); skeletal muscle (6); kidney (7);
testis (8). Embryo samples assayed included: e7
(9); e11 (10); e15 (11); e17
(12). Blots were reprobed with -actin (lower
panels).
- or
-Actin account for the faster-migrating
bands.
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Fig. 5.
In situ hybridization of RALDH4 in
mouse embryo. Silver grain signals represent
hybridization of antisense cRNA against RALDH4; red signals
represent nuclei stained by propidium iodide. Sense cRNA showed
no signals (data not shown). A, RALDH4 signal was not
detected within the heart or liver primordia of the e10.5 mouse.
B, view of e14.5 embryo thoracic region showing low but
specific expression of RALDH4 in liver. No signal was detected in lung,
heart, mesentery, or other tissues. White arrows indicate
liver. Magnifications are 10× for panel A and 4× for
panel B.
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Fig. 6.
In situ hybridization of RALDH4 in
adult mouse liver and kidney. Hybridization signals are
represented by silver grains; nuclei were visualized with
propidium iodide. A, antisense cRNA reveals robust RALDH4
transcript levels throughout liver. B, sense cRNA does not
show significant hybridization in liver. C, antisense cRNA
shows strong RALDH4 expression within the kidney cortex and
low-to-absent expression within the medulla. D, sense cRNA
does not show significant hybridization in adult kidney. Magnification
is 4× in all panels.
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Fig. 7.
Analysis of anti-RALDH antibodies.
Western blots were done with each anti-serum (Ab) on lysates
of CHO cells (5 µg of protein) transfected with pcDNA3
(m) or a pcDNA3/RALDH. Bands migrated ~54 kDa.
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Fig. 8.
RALDH protein expression in adult mouse
liver. Immunohistochemistry was done on adult mouse liver with
anti-sera raised against: RALDH1 (1); RALDH2 (2);
RALDH3 (3); RALDH4 (4); negative control
(C). The arrow in panel 1 points to
lipid-engorged cells that express RALDH1. The arrowheads in
panels 2-4 show RALDH expression in hepatocytes, whereas the
arrows point to hepatocytes that do not express RALDH.
Panels have the same magnification; the scale bar represents
100 µm.
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Fig. 9.
RALDH protein expression in adult mouse
kidney. Immunohistochemistry was done with anti-sera against
RALDH4 (A-C) at different magnifications, showing cortex
and medulla (A) and cortex (B and C).
D, RALDH1 in cortex; E, RALDH2 in cortex;
F-H, RALDH1 in medulla; I and J,
RALDH2 in medulla. Panel K shows a negative control (no
primary antibody). Panels C-K have the same magnification.
Scale bars represent 100 µm.
DISCUSSION
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ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
have decreased erythropoietin expression and impaired
erythropoiesis between e10.25 and e14.5 (35). These observations may
provide insight into the mechanism for the synergistic effects of
vitamin A and iron supplementation on erythropoiesis and relief of
nutritional anemia relative to iron supplementation alone (36).
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FOOTNOTES |
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* This work was supported in part by National Institutes of Health Grant DK47839.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AF510322.
¶ To whom correspondence should be addressed: Dept. of Nutritional Sciences and Toxicology, University of California-Berkeley, 119 Morgan Hall, MC 3104, Berkeley, CA 94720. Tel.: 510-642-0809; Fax: 510-642-0535; E-mail: jna@uclink4.berkeley.edu.
Published, JBC Papers in Press, January 7, 2003, DOI 10.1074/jbc.M211417200
2 Min Lin and J. L. Napoli, unpublished data.
3
The standardized ALDH nomenclature approved by
HUGO has named 17 human ALDH genes, but the human ALDH with
9-cis-retinal dehydrogenase activity reported in Ref. 31 as
ALDH12 was not among them. The trivial name ALDH12 has been assigned to
1-pyrroline-5-carboxylase (16).
4 Na Bien-Ly and J. L. Napoli, unpublished data.
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ABBREVIATIONS |
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The abbreviations used are: atRA, all-trans-retinoic acid; CRAD, cis-retinoid/androgen dehydrogenase; e, mouse embryo day; RACE, rapid amplification of cDNA ends; RALDH, retinal dehydrogenase(s); 9cRA, 9-cis-retinoic acid; RAR, retinoic acid receptor(s); mRDH1, mouse retinol dehydrogenase type 1; RXR, retinoid X receptor(s); SDR, short-chain dehydrogenase(s)/reductase(s); CHO, Chinese hamster ovary.
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1. | Chytil, F., and Haq, R. (1990) Crit. Rev. Eukaryot. Gene Expr. 1, 61-73[Medline] [Order article via Infotrieve] |
2. | Festus, L., Davies, P. J., and Piacentini, M. (1991) Eur. J. Cell Biol. 56, 170-177[Medline] [Order article via Infotrieve] |
3. | Lohnes, D., Dierich, A., Ghyselinck, N., Kastner, P., Lampron, C., LeMeur, M., Lufkin, T., Mendelsohn, C., Nakshatri, H., and Chambon, P. (1992) J. Cell Sci. 16, 69-76 |
4. |
Chambon, P.
(1996)
FASEB J.
10,
940-954 |
5. | Glass, C. K., Rosenfeld, M. G., Rose, D. W., Kurokawa, R., Kamei, Y., Xu, L., Torchia, J., Ogliastro, M. H., and Westin, S. (1997) Biochem. Soc. Trans. 25, 602-605[Medline] [Order article via Infotrieve] |
6. | Renaud, J. P., and Moras, D. (2000) Cell. Mol. Life Sci. 57, 1748-1769[Medline] [Order article via Infotrieve] |
7. | Heyman, R. A., Mangelsdorf, D. J., Dyck, J. A., Stein, R. B., Eichele, G., Evans, R. M., and Thaller, C. (1992) Cell 68, 397-406[Medline] [Order article via Infotrieve] |
8. | Levin, A. A., Sturzenbecker, L. J., Kazmer, S., Bosakowski, T., Huselton, C., Allenby, G., Speck, J., Kratzeisen, C., Rosenberger, M., and Lovey, A. (1992) Nature 355, 359-361[CrossRef][Medline] [Order article via Infotrieve] |
9. | Nagpal, S., and Chandraratna, R. A. (2000) Curr. Pharm. Des. 6, 919-931[Medline] [Order article via Infotrieve] |
10. | Altucci, L., and Gronemeyer, H. (2001) Nature Reviews 1, 181-193[CrossRef][Medline] [Order article via Infotrieve] |
11. | Napoli, J. L. (1999) Biochim. Biophys. Acta 1440, 139-162[Medline] [Order article via Infotrieve] |
12. | Napoli, J. L. (2001) Mol. Cell. Endocrinol. 171, 103-109[CrossRef][Medline] [Order article via Infotrieve] |
13. |
Chai, X.,
Zhai, Y.,
and Napoli, J. L.
(1997)
J. Biol. Chem.
272,
33125-33131 |
14. | Zhuang, R., Lin, M., and Napoli, J. L. (2002) Biochemistry 41, 3477-3483[CrossRef][Medline] [Order article via Infotrieve] |
15. |
Zhang, M.,
Chen, W.,
Smith, S. M.,
and Napoli, J. L.
(2001)
J. Biol. Chem.
276,
44083-44090 |
16. | Sophos, N. A., Pappa, A., Ziegler, T. L., and Vasiliou, V. (2001) Chem.-Biol. Inter. 130-132, 323-337 |
17. | Dockham, P. A., Lee, M. O., and Sladek, N. E. (1992) Biochem. Pharmacol. 43, 2453-2469[CrossRef][Medline] [Order article via Infotrieve] |
18. | Yoshida, A., Hsu, L. C., and Yanagawa, Y. (1993) Adv. Exp. Med. Biol. 328, 37-44[Medline] [Order article via Infotrieve] |
19. | Lee, M. O., Manthey, C. L., and Sladek, N. E. (1991) Biochem. Pharmacol. 42, 1279-1285[CrossRef][Medline] [Order article via Infotrieve] |
20. |
Posch, K. C.,
Burns, R. D.,
and Napoli, J. L.
(1992)
J. Biol. Chem.
267,
19676-19682 |
21. | Penzes, P., Wang, X., and Napoli, J. L. (1997) Gene 191, 167-172[CrossRef][Medline] [Order article via Infotrieve] |
22. | Penzes, P., Wang, X., and Napoli, J. L. (1997) Biochim. Biophys. Acta 1342, 175-181[Medline] [Order article via Infotrieve] |
23. | Bhat, P. V., Labrecque, J., Boutin, J. M., Lacroix, A., and Yoshida, A. (1995) Gene 166, 303-306[CrossRef][Medline] [Order article via Infotrieve] |
24. | El Akawi, Z., and Napoli, J. L. (1994) Biochemistry 33, 1938-1943[Medline] [Order article via Infotrieve] |
25. | Yoshida, A., Rzhetsky, A., Hsu, L. C., and Chang, C. (1998) Eur. J. Biochem. 251, 549-557[Abstract] |
26. |
Wang, X.,
Penzes, P.,
and Napoli, J. L.
(1996)
J. Biol. Chem.
271,
16288-16293 |
27. | Zhao, D., McCaffery, P., Ivins, K. J., Neve, R. L., Hogan, P., and Dräger, U. (1996) Eur. J. Biochem. 240, 5-22 |
28. |
Grun, F.,
Hirose, Y.,
Kawauchi, S.,
Ogura, T.,
and Umesono, K.
(2000)
J. Biol. Chem.
275,
41210-41218 |
29. |
Hu, C.,
Lin, W.-W.,
Cassandra, O.,
and Valle, D.
(1999)
J. Biol. Chem.
274,
6754-6762 |
30. | Vasiliou, V., Bairoch, A., Tipton, K. F., and Nebert, D. W. (1999) Pharmacogenetics 9, 421-434[Medline] [Order article via Infotrieve] |
31. |
Lin, M.,
and Napoli, J. L.
(2000)
J. Biol. Chem.
275,
40106-40112 |
32. | Wilkinson, D. G. (1998) In Situ Hybridization , 2nd Ed. , Oxford University Press, New York |
33. | Niederreither, K., McCaffery, P., Dräger, U. C., Chambon, P., and Dollé, P. (1997) Mech. Dev. 62, 67-78[CrossRef][Medline] [Order article via Infotrieve] |
34. | Niederreither, K., Fraulab, V., Garnier, J. M., Chambon, P., and Dollé, P. (2002) Mech. Dev. 110, 165-171[CrossRef][Medline] [Order article via Infotrieve] |
35. |
Makita, T.,
Hernandez-Hoyos, G.,
Chen, T. H-P.,
Wu, H.,
Rothenberg, E. V.,
and Sucov, H. M.
(2001)
Genes Dev.
15,
889-901 |
36. | Suharno, D., West, C. E., Muhilal, K. D., and Jautvast, J. G. (1993) Lancet 342, 1325-1328[CrossRef][Medline] [Order article via Infotrieve] |