 |
||||
| 2007 JASN IMPACT FACTOR 7.111 | HOME AUTHOR INFO EDITORIAL BOARD SUBSCRIBE FEEDBACK ALERTS HELP | |||
| CURRENT ISSUE | ARCHIVES | JASN Express | ONLINE SUBMISSION | |
|
||||
| 2007 JASN IMPACT FACTOR 7.111 | HOME AUTHOR INFO EDITORIAL BOARD SUBSCRIBE FEEDBACK ALERTS HELP | |||
| CURRENT ISSUE | ARCHIVES | JASN Express | ONLINE SUBMISSION | |
REGULAR ARTICLES |
*
Department of Renal Pathology, Institute of Nephrology, Faculty of
Medicine, Niigata University, Niigata, Japan
Department of Medicine (II), Faculty of Medicine, Niigata University,
Niigata, Japan
Department of Molecular Biology, The Scripps Research Institute, La Jolla,
California.
Correspondence to Dr. Tadashi Yamamoto, Department of Renal Pathology, Institute of Nephrology, Faculty of Medicine, Niigata University, Asahimachidori 1-757, Niigata 951-8510, Japan. Phone: +81 25 227 2152; Fax: +81 25 227 0768; E-mail: tdsymmt{at}med.niigata-u.ac.jp
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Because the kidney has not yet yielded evidence of olfactomedin-related glycoprotein, we found the expression of this molecule in rat glomerulus by using ribonuclease protection assay. Additionally, B-type mRNA variants of this gene were prominently expressed in the renal glomeruli and the translation product in the Golgi apparatus of podocytes, suggesting a role of this glycoprotein in these cells. The unique expression may also indicate that the glycoprotein is a potential new marker for podocytes.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Ribonuclease Protection Assay
The ribonuclease protection assay has been described in detail
(9,10).
Plasmids containing the cDNA inserts were linearized with EcoRI to
prepare templates providing antisense cRNA probes. cRNA probes were labeled
with [
-32P]-UTP by in vitro transcription using T3
RNA polymerase (Promega, Madison, WI), and specific radioactivity was adjusted
to 1 x 105 cpm/µl in hybridization buffer (80% formamide,
40 mM 1,4-piperazinediethanesulfonic acid, 0.4 M NaCl, 1 mM
ethylenediaminetetra-acetic acid). Aliquots (10 µg) of total cellular RNA
from rat systemic organs were hybridized with 1 x 105 cpm of
cRNA probes at 48°C for 16 h. Unhybridized probes were digested with
ribonuclease A (1.0 µg/ml) and T1 (100 U/ml) mixture at 30°C for 1 h,
and then digested with proteinase K (0.5 mg/ml) at 37°C for 30 min. After
phenol/chloroform extraction, hybridized probes were precipitated with
ethanol, denatured at 85°C, and electrophoresed on 6% polyacrylamide gels.
The dried gels were exposed to x-ray films (Fuji Photo Film Co., Kanagawa,
Japan).
For screening genes predominantly expressed in rat glomerulus, plasmids isolated from the rat glomerulus cDNA library were linearized with EcoRI for preparation of templates, and radiolabeled probes were synthesized using three to five templates in combination as described above. Total cellular RNA samples isolated from rat glomeruli, cortex, and medulla were incubated with 1 x 105 cpm of the cRNA probe mixture at 48°C for 16 h. Then, unhybridized probes were digested with RNase ONETM ribonuclease (Promega) (10 U/ml) at 30°C for 1 h, then treated with 0.2% sodium dodecyl sulfate (SDS) to inactivate the RNase before precipitation with ethanol. After gel electrophoresis and autoradiography, the template sets that gave bands predominantly in the glomerular RNA lanes were selected and then individual template was used for cRNA probe synthesis to identify glomerulus-specific clones by ribonuclease protection assay. These clones were sequenced by an automated DNA sequencer (Perkin Elmer Japan, Urayasu, Japan).
Antibody and cDNA
Antibody against the olfactomedin-related glycoprotein was obtained by
immunizing rabbits with synthetic peptide corresponding to 39 amino acids of
the M region sequence of the olfactomedin-related glycoprotein
(TQRDLQYVEKMENQMKGLESKFRQVEESHKQHLARQFKG). The antibody's specificity for
olfactomedin-related glycoprotein was previously confirmed by Western blot
analysis (5).
AMY (40-766) cDNA and ß-coatomer (ß-COP) protein cDNA (3066-3256), which was also isolated during the search for glomerulus-rich genes in the rat glomerulus cDNA library, were subcloned into pGEM3 vector and pGEM3Z vector (Promega), respectively. Rat glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA of 123-bp fragment was subcloned into pGEM4Z vector as described previously (11).
Western Blot Analysis
Rat tissues (brain, lung, eye, and glomeruli) were homogenized in buffer
(0.16 M NaCl, 11 mM sodium phosphate, 1 mM phenylmethylsulfonyl fluoride, 10
mg/ml leupeptin, 10 mM pepstatin A, pH 7.4) on ice with a Potter-type
homogenizer. The homogenate was centrifuged for 10 min at 600 x
g to remove large debris, and the resulting supernatant was
solubilized in 125 mM Tris-HCl, pH 6.8, ß-mercaptoethanol, 4.6% SDS, and
0.1% glycerol and boiled for 5 min. The sample of ~50 µg of protein
each was loaded on SDS-polyacrylamide gels (4 to 20% gradient gel), and the
bands were transferred to polyvinylidene difluoride membranes by
electroblotting. The membranes were preincubated with 10% nonfat milk
overnight, incubated with the anti-olfactomedin-related glycoprotein (1:2000
dilution) or anti-ß-COP antibody (1:500 dilution, G-2279, Sigma Aldrich
Japan, Tokyo, Japan) overnight, and washed in 0.05% Tween 20 phosphatebuffered
saline (PBS). Then, they were incubated with a horseradish peroxidase-labeled
second antibody (1:200 dilution; EnVision, DAKO Japan, Kyoto, Japan), and the
immunoreactivity was visualized by an enhanced chemiluminescence detection
system (Amersham Pharmacia Biotech, Tokyo, Japan).
Immunofluorescence Microscopy
Several organs obtained from adult Wistar-Kyoto rats and kidneys from
neonatal (day 4) rats were quick-frozen in n-hexane at -70°C and
cryosectioned at 4 µm thickness. The sections were fixed in acetone at
4°C for 5 min and incubated with the anti-olfactomedin-related
glycoprotein antibody (1:500 diluted) at 4°C overnight. They were then
incubated with FITC-conjugated goat anti-rabbit IgG (Seikagaku Kogyo Co.,
Tokyo, Japan) for 30 min at 37°C. The kidney sections were doubly stained
with tetramethyl rhodamine isothiocyanate (TRITC)-conjugated rabbit anti-rat
basement membrane antibody for 30 min at 37°C to outline the glomerular
basement membrane. These sections were examined under a Vanox AH-2 microscope
(Olympus, Tokyo, Japan).
To examine stereologic localization of the olfactomedin-related glycoprotein, laser scanning confocal microscopy was also done. The kidneys of adult rats were perfusion-fixed with periodate-lysine-paraformaldehyde fixative, and glomeruli isolated by a sieving method were further fixed by immersion in the same fixative for 10 min. After washing three times with PBS, the tissues were permeabilized with 0.3% Triton-X 100 in PBS for 10 min. They were then washed with PBS and incubated with the anti-olfactomedin-related glycoprotein rabbit antibody and anti-vimentin mouse monoclonal antibody (Boehringer Mannheim, Indianapolis, IN) overnight. After rinsing in PBS, the glomeruli were incubated with TRITC-labeled goat antimouse IgG (Cooper Biomedical, Malvern, PA) and FITC-labeled goat anti-rabbit IgG for 2 h. After washing in PBS, the samples were mounted on slides in buffered glycerin and observed under a confocal laser microscope (Bio-Rad Laboratories, Hercules, CA).
The presence of olfactomedin-related glycoprotein was also examined in glomerular epithelial cells in culture by immunofluorescence microscopy, using the anti-olfactomedin-related glycoprotein antibody. Glomeruli were isolated from rat kidneys and cultivated for 5 d on type I collagen-coated Lab-Tek glass slides (Miles Scientific, Naperville, IL) in RPMI 1640 supplemented with 5% fetal bovine serum, penicillin (100 U/ml), and streptomycin (100 mg/ml). The outgrown cells were examined by immunofluorescence microscopy using anti-rat synaptopodin antibody (anti-pp44, kindly provided by Dr. P. Mundel, Albert Einstein College of Medicine, Bronx, NY), as described previously (12,13).
Immunoelectron Microscopy
To seek the olfactomedin-related glycoprotein at an ultrastructural level
in the glomerulus, rat kidneys were fixed with
periodate-lysine-paraformaldehyde fixative by perfusion and immersion, and the
tissue blocks were embedded in glycol methacrylate resin. Ultrathin sections
of the resin-embedded tissues were collected on nickel grids and incubated
with 5% normal goat serum for 1 h and then with the anti-olfactomedin-related
glycoprotein antibody or normal rabbit serum (1:1000 diluted) overnight. After
washing in PBS, the sections were incubated with gold (15 nm)-labeled
anti-rabbit IgG (Amersham Pharmacia Biotech) for 2 h. These sections were
washed in PBS and distilled water, post-fixed with 2.5% glutaraldehyde, and
then counterstained with 2% aqueous uranyl acetate and 1% lead citrate for
observation by electron microscopy (Hitachi, Ibaragi, Japan).
Computer Analyses
Comparisons to known sequences were performed by BLAST or FASTA on the
Internet server. Alignment of protein sequences was achieved with the GAP
program (BESTFIT) (14).
Secondary structure analysis for coiled-coil motifs was conducted with the
software program "COILS"
(15).
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
To determine which variants of the olfactomedin-related glycoprotein mRNA appeared in the glomerulus, a ribonuclease protection assay was performed using the AMY (40-766) cDNA as a template. The AMY antisense cRNA probe was expected to hybridize with AMY, AMZ, BMY, and BMZ mRNA with its counterparts for AMY (726 nt), AM (570 nt), MY (462 nt), and M (306 nt), respectively (Figure 1B). Accordingly, four distinct bands corresponding to these four variants were intensely detected in the brain (Figure 1C, lane 1). In contrast, the glomeruli yielded only two bands corresponding to BMY and BMZ, which were comparable in intensity to those in the brain (Figure 1C, lane 2); however, AMY and AMZ mRNA variants were not expressed. These two BMY and BMZ mRNA variants were also apparent, but faint in the lung and eye (Figure 1C, lanes 7 and 8).
Western blot analysis confirmed translation of the mRNA in these tissues (Figure 2). The anti-olfactomedin-related glyco-protein antibody reacted to four major bands of ~81, ~70, ~29, and ~24 kD in the brain extract; two bands of ~56 and ~29 kD in the eye; and two bands of ~63 and ~29 kD in the lung. In contrast, an intense band of ~68 kD and a faint band of ~30 kD were visualized in the glomerular extract. Extraction of Golgi apparatus components was verified by detection of ß-COP in all of these samples.
|
Localization of Olfactomedin-Related Glycoprotein
In the kidney, olfactomedin-related glycoprotein was predominantly
immunolocalized in the glomeruli in a pattern of several large patches
(Figure 3A). The glycoprotein
was localized predominantly in the cytoplasmic bodies of podocytes, but was
not found in glomerular mesangial cells, parietal epithelial cells of Bowman's
capsule, or tubular epithelial cells at a higher magnification
(Figure 3B). Occasionally, a
single small dot of weak staining was observed in the cytoplasm of each
endothelial cell, both in the glomeruli and peritubular capillaries.
|
Laser scanning confocal microscopy revealed conspicuous staining of irregularly shaped structures in the juxtanuclear cytoplasms of podocytes and significant, but much less, staining in cytoplasms of glomerular endothelial cells (Figure 3, C and D). The olfactomedin-related glycoprotein was particularly clear in cytoplasmic bodies of podocytes in contrast to vimentin staining in their primary processes.
In the cytoplasms of neurons at ubiquitous sites in the brain, intense immunofluorescence staining produced a speckled or patchy pattern (Figure 3, E and F). Besides the distinct speckled staining within neurons, ambiguous faint immunostaining was associated in and around the neurons. Immunostaining was also observed at perinuclear cytoplasms of ciliary, corneal, and iris epithelial cells in the eye and arteriolar and alveolar capillary endothelial cells and bronchiolar epithelial cells in the lung (Figure 3, G and H).
The subcellular sites of the glycoprotein were then determined by immunogold electron microscopy. A heavy labeling of gold particles was found within the cytoplasms of podocytes, exclusively in the well-developed Golgi apparatus (Figure 4, A and B). However, only background labeling was observed in other sites such as the cytoplasmic membrane, rough endoplasmic reticulum (ER), primary processes, or foot processes of the podocytes. When the anti-olfactomedin-related glycoprotein antibody was replaced with normal rabbit serum, no Golgi apparatus of podocytes was labeled. In glomerular or peritubular endothelial cells, the Golgi apparatus had only a sparse sprinkling of gold particles (Figure 4C); however, no significant labeling was observed in the Golgi apparatus of other types of cells such as tubular epithelial cells in the kidney.
|
In the neonatal kidney, most of the cells in the commashaped and S-shaped bodies and some of the cells around these bodies were weakly positive for the olfactomedin-related glycoprotein (Figure 5, A and B). However, the staining in the kidney disappeared, leaving the staining in podocytes in the capillary loop stage (Figure 5C). During the maturing stage, the podocyte staining became more intense and larger in size (Figure 5D).
|
Two different phenotypes of cells were outgrown from rat glomeruli in culture; large cells of irregular shape and small cells of polygonal cobblestone-like appearance. The former cells were intensely stained with mouse monoclonal antibody against synaptopodin and also with the anti-olfactomedin-related glycoprotein antibody in a patchy pattern at juxtanuclear regions (Figure 5E). The latter cells were negative for synaptopodin and positive very faintly for the olfactomedin-related glycoprotein (Figure 5F).
Tissue Specific mRNA Expression of Olfactomedin-Related
Glycoprotein
To examine whether expression of the olfactomedin-related glycoprotein was
selective in the Golgi apparatus of particular tissues or simply correlated
with the size or number of Golgi apparatus, the mRNA expression in several
tissues was compared with that of ß-COP, which is a ubiquitous
Golgi-specific marker (Figure
6). The expression of olfactomedin-related glycoprotein mRNA was
extremely high in the brain and the glomerulus and was negligible in renal
cortex, medulla, liver, and small intestine. In contrast, all of these tissues
had nearly equal amounts of ß-COP mRNA expression, although it was
relatively high in the glomerulus.
|
Structural Feature of Deduced Olfactomedin-Related Glycoprotein
Computer analysis of the glycoprotein's deduced amino acid sequence
revealed the following characteristics. (1) Its Z region had homology
with the carboxy terminus of bullfrog olfactomedin, in which 33% of amino
acids (59 of 178) are identical and an additional 33 amino acids (33 of 178)
are conservative substitutions, yielding 54% sequence similarity
(5). The C-terminal half of the
olfactomedin-related glycoprotein also showed homology with the human
trabecular meshwork-inducible glucocorticoid response protein (TIGR), the
product of a candidate gene responsible for juvenile open angle glaucoma
(16). In this comparison,
41.3% of amino acids (107 of 259) were identical to each other and an
additional 28 amino acids were conservative substitutions, yielding 52.5%
sequence similarity (Figure
7A). (2) The olfactomedin-related glycoprotein also had
some similarity (~20% identity, ~40% similarity) with several
cytoskeleton-related proteins including various myosin heavy chains
(Figure 7B), NuMA
(17), Rad 50
(18), and
giantin/macrogolgin/GCP372
(19,20,21,22).
These similarities were marginal and variable depending on the search
algorithm used (FASTA and BLAST). (3) Secondary structure analysis of
the olfactomedin-related glycoprotein using the program "COILS"
revealed that the protein had three predicted coiled-coil domains in the M and
Z regions (Figure 7C).
Coiled-coil domains are overlapped on regions that were similar to myosin
heavy chains.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
In the brain, four mRNA variants (AMY, AMZ, BMY, and BMZ) are transcribed from a single gene (D2Sutle) by differential promoter utilization to generate A or B and alternative splicing to generate Y or Z (5). However, we found that only two of the variants, BMY and BMZ, were expressed in the glomerulus, eye, and lung, and no expression of AMY or AMZ mRNA was detectable in these tissues. The expression of AMY and AMZ in the brain may be attributable to the presence of a PIT-1 element in the A region of the gene, which is a binding element for a pituitary-specific transcription factor (5,28,29). With Western blot analysis, we also identified four distinct bands corresponding to transcripts from the four mRNA variants in the brain: an ~81-kD band for BMZ, an ~70-kD band for AMZ, an ~29-kD band for BMY, and an ~24-kD band for AMY. The two bands of ~63 kD and ~29 kD in the lung or ~54 kD and ~29 kD in the eye are predicted as isoforms translated from BMZ and BMY mRNA variants, respectively. However, an intense single band of ~68 kD was detected with an additional minor band of ~30 kD in glomeruli by Western blot analysis. Because the ribonuclease protection assay clearly demonstrated two mRNA variants of BMY and BMZ in glomeruli, the ~68-kD band was regarded as a molecule translated from BMZ variant and the ~30-kD band from BMY variant. Several explanations have been proposed to account for this striking difference in the density of these two bands. BMY translation might be restricted in the glomerulus by unknown reason. BMY isoform may be translated with the same amount of BMZ, but it is rapidly fragmented in glomerular cells or it is released from them. Because the difference in density of these two BMY and BMZ isoforms was not striking in other tissues (brain, eye, and lung), translation of these isoforms may also take place equally in the glomerulus but the fate after translation may not be identical. By immunofluorescence microscopy, the ambiguous faint immunostaining besides the distinct intense Golgi staining in neurons as shown in Figure 3 may indicate a possible difference in destination of Z and Y isoforms in the brain. However, such ambiguous staining was undetectable in the glomeruli. This observation may also suggest that the BMY isoform is translated in the glomeruli but released into its ultrafiltrate or metabolized rapidly. If it was secreted, it might be possible to affect on podocytes or other nephron epithelia cells present downstream in an autocrine or paracrine manner, although the function of this molecule is unknown. The variations in gel mobility of BMZ and BMY glycoproteins in each tissue were presumably due to differences in glycosylational modifications.
The present study clearly demonstrates localization of the olfactomedin-related glycoprotein in the Golgi apparatus of renal podocytes. In contrast, a mouse homologue of the rat olfactomedin-related glycoprotein, termed pancortin, has been reported to be present in the ER of cortical neurons in the brain immunoelectron microscopy using an antibody against the M region (6). The amino acid sequence homology between rat and murine AMZ or BMZ variants is quite high with substitution of a few amino acids. The destination of the four isoforms may be different from each other after the synthesis of this protein in ER. The long isoforms AMZ and BMZ have the sequence Ser-Asp-Glu-Leu (SDEL) at the carboxy terminus (5), and the sequence may be a functional ER retention signal in Plasmodium falciparum (30,31,32,33). Additionally, the tetrapeptide Lys-Asp-Glu-Leu (KDEL) or analogous sequence at the carboxy terminus has been considered an ER retention signal (33). The similarity between KDEL (the ER retention signal) and SDEL located at the carboxy terminus of AMZ and BMZ isoforms of the olfactomedin-related glycoprotein may indicate that these isoforms remain inside the ER. However, we were unable to detect them in the ER of rat podocytes by immuno-electron microscopy. Moreover, we readily colocalized giantin (Golgi apparatus marker) with the olfactomedin-related glycoprotein in rat podocytes or brain neurons with immunofluorescence microscopy (data not shown). Furthermore, we identified both AMZ and BMZ isoforms predominantly in the Golgi-enriched fraction isolated from a rat brain homogenate by sucrose density ultracentrifugation (unpublished data). These data indicate that the long isoforms of the glycoprotein probably occupy the same sites in the Golgi apparatus of podocytes and neurons in rats. The SDEL sequence at the carboxy terminus of the long isoforms AMZ and BMZ may be an ER retention signal in mice, but may play a role in their retention in Golgi complex in rats.
The Golgi complex, as the main factory for protein processing, distributes processed proteins, lipids, and polysaccharide products to multiple destinations. The cytoplasms of podocytes contain several huge Golgi apparatuses, and the well-developed Golgi apparatus may be an indication of the great capacity of these cells to modify glycoconjugation (4). Glycoconjugation is achieved in a cell-specific manner and is finely regulated spatio-temporally in the central nervous system (34). Therefore, the high level of olfactomedin-related glycoprotein expressed in podocytes may be associated simply with a number or size of Golgi apparatuses or may relate to a specific activity of the podocytes in rats. These issues remain to be determined, however. Two of our results suggest that the olfactomedin-related glycoprotein is expressed in a cell-specific manner. First, the expression of this glycoprotein mRNA was not comparable to that of ß-COP, a ubiquitous Golgi protein. Second, the olfactomedin-related glycoprotein was found in podocytes but not tubular epithelial cells of rat kidneys, although the Golgi apparatus is well developed in the tubular epithelial cells. This cellular specificity may also be true in the glomerular cell culture. At least two types of epithelial cells have been reported in the culture; large irregularly shaped cells and polygonal ones with cobblestone-like appearance. The former cells are considered to retain the phenotypes of podocytes in vivo, and the latter ones retain the phenotypes of parietal epithelial cells of Bowman's capsule because the large cells of irregular shape exhibit the podocyte-specific markers podocalyxin and synaptopodin, but the polygonal cells do not (13,35). In addition, the polygonal cells are rarely cultivated from rat glomeruli without Bowman's capsule, but from those with Bowman's capsule. Our results revealed the intense staining for the olfactomedin-related glycoprotein in the former cells and very faint staining in the latter, supporting the idea of the cellar specificity of this molecule. The cell-specific expression is accomplished during the development of rat kidneys. In the comma-shaped and S-shaped body stages, most of the cells were weakly positive for the glycoprotein. The podocyte-specific expression became obvious in the capillary loop stage, while podocytes differentiate to lose mitotic activity.
Recently, antibodies directed against the Golgi complex have been reported in the sera of patients with systemic lupus erythematosus (36), Sjögren's syndrome (37,38) and other systemic rheumatic diseases (39,40,41), idiopathic cerebellar ataxia (42), paraneoplastic cerebellar degeneration (43), and viral infections, including those by Epstein—Barr virus (44) and HIV (45). As the target antigens, several Golgi complex proteins including golgins-95 and - 160 (46), giantin/macrogolgin/GCP372 (19,20,21,22), and golgin-245 (47) have been identified. These Golgi complex autoantigens apparently share significant sequence similarity to several cytoskeleton-related proteins including desmin, myosin proteins (myosin heavy chain and tropomyosin), kinesin (48), and 150-kD dynein-associated polypeptide (49), and they also bear coiled-coil domains in their secondary structure. The functions and pathogenic potential of these proteins are unclear, but their structural features predict their potential participation in the transport of vesicles from the ER to the Golgi complex or within the Golgi stack. The secondary structure of three coiled-coil domains was predicted in the M and Z regions of the olfactomedin-related glycoprotein as the other Golgi autoantigens. The olfactomedin-related glycoprotein has regions that share weak sequence similarity to a variety of proteins with predicted coiled-coil domains: myosin heavy-chains, NuMA involved in nuclear structure, spindle assembly and nuclear reformation (17), Rad50 involved in the double-strand break, formation and repair of DNA (18), and giantin/macrogolgin/GCP372, which is a Golgi resident protein (19,20,21,22). The olfactomedin-related glycoprotein had no significant sequence similarity to Golgi glycosylation enzymes or other Golgi complex proteins. Therefore, the structural similarity of olfactomedin-related glycoprotein to the Golgi autoantigens may indicate that the glycoprotein is a structural component of Golgi apparatus as a cytoskeleton-related entity.
The present study demonstrated that the olfactomedin-related glycoprotein was present primarily in the Golgi apparatus of podocytes and neurons in rats. The predominant expression of this glycoprotein in these cells suggests its probable contribution to biosynthesis and posttranslational modification of products in their Golgi apparatuses or to the structural maintenance of the Golgi complexes.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
This article has been cited by other articles:
 |
|
 |
|
  E. Bertelli, M. Regoli, L. Fonzi, R. Occhini, S. Mannucci, L. Ermini, and P. Toti Nestin Expression in Adult and Developing Human Kidney J. Histochem. Cytochem., April 1, 2007; 55(4): 411 - 421. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Malyukova, H.-S. Lee, R. N. Fariss, and S. I. Tomarev Mutated Mouse and Human Myocilins Have Similar Properties and Do Not Block General Secretory Pathway Invest. Ophthalmol. Vis. Sci., January 1, 2006; 47(1): 206 - 212. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Grinchuk, Z. Kozmik, X. Wu, and S. Tomarev The Optimedin Gene Is a Downstream Target of Pax6 J. Biol. Chem., October 21, 2005; 280(42): 35228 - 35237. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. M. Hardy, E. A. Hoffman, P. Gonzalez, B. S. McKay, and W. D. Stamer Extracellular Trafficking of Myocilin in Human Trabecular Meshwork Cells J. Biol. Chem., August 12, 2005; 280(32): 28917 - 28926. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Torrado, R. Trivedi, R. Zinovieva, I. Karavanova, and S. I. Tomarev Optimedin: a novel olfactomedin-related protein that interacts with myocilin Hum. Mol. Genet., May 16, 2002; 11(11): 1291 - 1301. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
HOME
CURRENT ISSUE
ARCHIVES
JASN Express
ONLINE SUBMISSION
AUTHOR INFO
EDITORIAL BOARD SUBSCRIBE FEEDBACK ALERTS HELP |
