The zebrafish mutants for the V-ATPase subunits d, ac45, E, H and c and their variable pigment dilution phenotype
© Ramos-Balderas et al.; licensee BioMed Central Ltd. 2013
Received: 1 September 2012
Accepted: 23 January 2013
Published: 2 February 2013
The V-ATPase is a proton pump that creates an acidic medium, necessary for lysosome function and vesicular traffic. It is also essential for several developmental processes. Many enzymes, like the V-ATPase, are assemblies of multiple subunits, in which each one performs a specific function required to achieve full activity. In the zebrafish V-ATPase 15 different subunits form this multimeric complex and mutations in any of these subunits induce hypopigmentation or pigment dilution phenotype. We have previously found variability in the pigment dilution phenotype among five of the V-ATPase zebrafish mutants. This work presents additional information about such differences and is an update from a previous report.
We describe the variable phenotype severity observed among zebrafish V-ATPase pigment dilution mutants studying mRNA expression levels from their corresponding genes. At the same time we carried out phylogenetic analysis for this genes.
Based in the similarities between different pigment dilution mutants we suggest that there is an essential role for V-ATPases in melanosome biogenesis and melanocyte survival. Neither variable expression levels for the different V-ATPase subunits studied here or the presence of duplicated genes seems to account for the variable phenotype severity from this group of mutants. We believe there are some similarities between the pigment dilution phenotype from zebrafish V-ATPase insertional mutants and pigment mutants obtained in a chemical screening (“Tubingen pigmentation mutants”). As for some of these “Tubingen mutants” the mutated gene has not been found we suggest that mutations in V-ATPase genes may be inducing their defects.
KeywordsZebrafish Insertional mutants V-ATPase Subunits Pigment dilution
Zebrafish pigment dilution mutants are the consequence of mutations in genes involved in vesicular traffic or endo-lysosomal function [1, 2] and therefore they are tools to understand how melanosomes and other organelles are formed. This report is an update from data previously published by us, and others, about five V-ATPase mutants with a pigment dilution phenotype [3, 4].
A large-scale retroviral insertional mutagenesis generated about 400 zebrafish mutants, which are currently at the “Zebrafish International Resource Center” and are used by several research groups to study different aspects of developmental biology and cell differentiation [5–8]. At the same time some of these insertional mutants are models for human diseases like oculocutaneous albinism , cancer [9, 10], liver and kidney diseases [11, 12] and neuroblastoma . In total 16 insertional mutants have pigment defects and for 7 of these, insertions have been found tightly linked to subunits from the V-ATPase complex [8, 14]. V-ATPase mutants have hypopigmentation or pigment dilution as the more visible phenotype  but also have retinal defects and some other physiological problems [4, 15]. Since V-ATPase function has been found to be essential for melanosome biogenesis it was suggested that the pigment dilution phenotype is the consequence of a reduction in the number of melanosomes, which may affect melanocyte survival [3, 4], the same is true for mutations in other genes also required for melanosome biogenesis [1, 16]. Melanosomes are the organelles where melanin synthesis is carried out in the melanocyte (pigment cell) .
The V-ATPase complex is oriented in the membrane in such a way that it pumps protons from the cytoplasm out of the cell or to the interior of certain organelles. V-ATPase function is essential for secretion, lysosome function, vesicular traffic and phagocytosis . V-ATPases are also important during development in invertebrates [19, 20] and vertebrates . Its function is required for specific processes like acquiring left-right asymmetry [22, 23], biliary function , bone formation [25, 26] and neural system development [27, 28]. The vacuolar ATPase complex from vertebrates is composed of at least 15 subunits arranged in two functional domains: V1 with subunits A, B, C, D, E, F, G and H and V0 with subunits a, c, c”/b and e. Both domains have a combined molecular weight around 750 kDa. V-ATPases also have two accessory subunits known as the membrane proteins Ac45 and Ap2/M8-9. The V1 sector is peripheral to the membrane being the site for ATP hydrolysis and V0 domain is a multimeric membrane proton channel. V1 and V0 are connected by a stalk, which is formed by the V1 subunits C, E, G and H in the configuration CE2G2H. The assembly, interactions and function for each of the V-ATPase subunits have been reviewed extensively [18, 29, 30].
The seven known V-ATPase insertional mutants are: atp6vod1 hi2188bTg (V0-d1), atp6ap1b hi112Tg (V0-ac45b), atp6v1e1b hi577aTg (V1-E1b), atp6v1h hi923Tg (V1-H), atp6v0ca hi1207Tg (V0-ca), atp6v1f hi1988Tg (V1-F) and atp6ap2 hi3681Tg (V0-ap2) (there are several alleles for each of these mutants, which are either insertional or chemically induced). Three more insertional mutants for V-ATPase subunits were recently obtained, which are: atp6v1ba la013065Tg (V1-B), atp6v1d la013933Tg (V1-D) and atp6v0a1a la015092Tg (V0-a1a) . Furthermore, knockdown morpholino experiments have also been carried out for V-ATPase subunits genes atp6v0a1a, atp6v0b, atp6v0ca, atp6v0cb and atp6v1a. This means that for 12 out of the 15 V-ATPase subunits there is a mutation or knock down experiment (the exceptions are V1-C, V1-G and V0-e subunits). Even though each mutant or morphant may have its own characteristics, pigment dilution is a common characteristic when V-ATPase function is affected.
In this work we present additional information about five of the V-ATPase insertional mutants and their genes. These are subunits V0-d1 (gene atp6vod1), V0-ac45b (gene atp6ap1b), V1-E1b (gene atp6v1e1b), V1-H (gene atp6v1h) and V0-ca (gene atp6v0ca).
Materials and methods
Fish husbandry and genotyping
All protocols carried out in animals were approved by the Office of Laboratory Animal Welfare (OLAW) from the National Institutes of Health (NIH) at the US, approval #A5281-01. Wild type zebrafish (strain TAB-14) and the five V-ATPase insertional mutant strains used in this work were obtained as a kind gift from Professor Nancy Hopkins of the Massachusetts Institute of Technology. Zebrafish embryos were obtained by natural crosses and then keep at 28.5°C, they were staged according to Kimmel et al. . Adult zebrafish were maintained in a recirculation system (Aquatic Habitats) with constant pH, temperature and dark–light cycles . Food consisted of harvested nauplii larvae mixed with macerated TetraminPro (Tetra). We genotyped fin clips to identify carriers by PCR as described before .
Photography and dark adaptation assay
Live zebrafish larvae, WT and mutants, were placed in 3% methylcellulose (SIGMA) in excavated slides and oriented with a pin. No anesthetic was used because melanocyte appearance may be affected in the presence of tricaine. Photographs were taken in a StemiSV11 Zeiss Stereomicroscope using a Sony DSC-F707 camera attached to an adapter for microscopes (Martin Microscope Co.). Images were taken in the same light conditions for all larvae fish, photos were enhanced using Adobe Photoshop, however previous to any modification all images in the same figure were placed side by side in the same layer and only then enhanced together to avoid biased comparisons. The dark adaptation assay was carried out in 5 dpf larvae as described before . In total 12 zebrafish larvae were used in this analysis, 1 WT and 5 mutants for each experiment, the assay was made in duplicate.
RT-PCR and semi-quantitative RT-PCR
Total RNA was obtained using the Trizol method (Invitrogen), 60 WT zebrafish embryos were collected for each developmental stage, placed over ice for 20 min and then transferred directly to Trizol. For adult organ RNA extraction 12 adult fish were dissected, these were of 8 to 12 months old and both male and female were used indistinctly. Adult zebrafish were euthanized first in ice cold 0.025% in tricaine (3-aminobenzoic acid methyl ester from Sigma) as recommended . TURBO DNase (Ambion) was added in the reaction mix to eliminate residual genomic DNA. Using 2 μg of total RNA we prepared first strand cDNA with oligo-dT primers and superscript III Reverse Transcriptase (Invitrogen). We always included a control without superscript III in order to confirm that prepared cDNA was free of genomic DNA as contaminant. In particular in brain and liver tissue from zebrafish it is difficult to eliminate genomic DNA and required double trizol extraction and repeated TURBO DNase treatments. Routinely RT-PCR was performed using a Taq enzyme and reagents from SIGMA. All experiments were repeated at least two times. The primers used to amplify actin were 5′- CATCAGCATGGCTTCTGCTCTGTATGG-3′ and 5′- GACTTGTCAGTGTACAGAGACACCCTG-3′. The primers used to amplify V1-d1 (gene atp6vod1) were 5′- GATTTGGATGAGATGAACATTGAGA-3′ and 5′- CACAATGTTCCGGCACTCTTGCTCC-3′. The primers used to amplify V0-ac45b (gene atp6ap1b) were 5′- GGAACCAGGCTGATCTAGCAAGCA-3′ and 5′-TCACGCTGAAGCTCTGGATCTGAA-3′. The primers used to amplify V1-E1b (gene atp6v1e1b) were 5′- CCAAACAGGGCGACTGGTATTTCA-3′ and 5′- GCCGACGTCCAGAAACAGATCAAG-3′. The primers used to amplify V1-H (gene atp6v1h) were 5′-TGTGGCTTCCAGCTGCAGTATCAG-3′ and 5′- ACTCCATTCCAGGCGTCCAGACTT-3′. The primers used to amplify the subunit V0-ca (gene atp6v0ca) were 5′-ACTCAGAGTCTAAACCTGCTAGACTG-3′ and 5′ GAGCCCCAGCACCTCTGCAAAGATC-3′. All RT-PCR products were in the range of 400 to 600 bp in size. An MJ-Research PTC-200 thermal cycler was used (25 cycles only for most reactions). RT-PCR was carried out with 2 μl of cDNA (25 μl final volume) but when semi-quantitative RT-PCR was performed the cDNA source was diluted 1:100 and 1:1000.
Multiple alignments and phylogenetic tree building
Multiple alignments and Phylogenetic tree building were carried out using MacVector12. Protein sequences were used for multiple alignments and were obtained, by BLAST searches, from public databases (Ensembl and NCBI). Multiple alignments were made using ClustalW using the BLOSUM matrix with open gap penalty of 10%, extended gap penalty of 0.2% while the delay divergence was 30%. Phylogenetic trees were built with the Neighbor Joining method in the Best Tree mode with a systematic tie breaking.
Variable phenotype in the V-ATPase insertional mutants
Variability in the phenotype severity could also be observed in developmental delay, because the mutants for V1-H and V0-ca subunits, have the shortest body size and were more affected in general development (see Additional file 1: Figure S1). As a further test for variability in phenotype severity we performed a dark adaptation assay in the V-ATPase mutants. When larvae are exposed to a dark environment melanosomes become broadly distributed and the melanocytes look expanded, this is consequence of a physiological hormonal response that involves the retina and the pituitary gland . We observed that zebrafish V0-d1, V0-ac45b mutants expand their melanocytes but V1-E1b, V1-H and V0-ca mutants did not present a comparable response (see Additional file 2: Figure S2). It is worth mentioning that while carrying out this experiment we notice that, in all the mutants, melanin spots remain unresponsive to the dark adaptation assay, supporting the idea that melanin spots are the consequence of melanocyte degeneration.
mRNA expression for the V0-d1, V0-ac45b, V1-E1b, V1-H and V0-ca V-ATPase subunits
Gene duplications in some V-ATPase subunits
Zebrafish mutants used in this study, their phenotype severity and duplicated genes
We described here that there is a variable severity in pigment dilution and developmental delay between the five V-ATPase mutants even though these mutants do not possess functional mRNA for the affected genes . Variability in phenotype severity suggest that V-ATPase function is not equally impaired in all these mutants and that each subunit may play its functional role only in some developmental stages, only in some organs or only in some physiological conditions. It also reveals how plastic is the functionality of the V-ATPase and how complex is regulation could be.
We find similarities between the zebrafish five V-ATPase insertional mutants and the Tubingen VI.C mutants (previously published by others); both groups of mutants show melanin spots that collect ventrolaterally to the ear (Figure 2), in the hindgut and in clusters in the dorsal stripe (not shown). Mutants in the class VI.C that were originally found in the Tubingen screening are delayed fade (dfd), fade out (fad), fading vision (fdv) and Quasimodo (qam) . Only the fdv mutation has been positionally cloned and it was found to be the consequence of a premature stop codon halfway the silva/pmela gene, which codes for a protein required for intraluminal fibril formation required for melanosome maturation . Due to the similarities between the V-ATPase mutants and the Tubingen class VI.C mutants we believe some of the yet uncloned pigment dilution mutants (dfd, fad or qam) may harbor mutations in V-ATPase subunits. The fad mutant shows retinal degeneration , which could also be observed in some of the insertional V-ATPase mutants . It was reported that the chemically induced zebrafish mutant a82 has a pigment dilution phenotype and therefore was a candidate for mutations in a V-ATPase subunit, so it was tested by complementation crosses against several V-ATPase mutants and found to be an allele of V0-ac45b .
V-ATPase function must be essential for melanosome biogenesis, the lack of such organelles cause the pigment dilution phenotype and may also induce melanocyte degradation, which is observed in the form of melanin spots in the zebrafish pigment dilution mutants [1, 2, 38]. It has been shown that pre-melanosomes are more acidic than mature melanosomes, which suggests they could have high requirements for V-ATPase activity, furthermore the V-ATPase subunit V0-a3 has been physically found in melanosomes from mouse melanocytes .
We also described that the variable pigment dilution phenotype, in this mutant collection, could not be explained by differences in mRNA expression of the analyzed V-ATPase subunits and although some of these subunits have duplicated genes, it does not seem to exist any functional redundancy that lessen the phenotypic severity observed, as it has been shown for other genes . With the sole exception of V0-a45b, in which, the phenotype is among the less severe and has a duplicated V0-a45a that may be compensating for the lack of V0-a45b, however this will need confirmation. It is common that zebrafish mutants with specific phenotypes are induced by mutations that affect the expression of transcription factors, ligands or receptors . This collection of pigment dilution zebrafish mutants shows that there is a new set of models with a very specific phenotype named “pigment dilution” induced when melanosome biogenesis is affected, and as a group could be used to get a better understanding about melanosome biogenesis and how multimeric enzymes achieve staggering levels of functional plasticity.
Tubingen-AB-14 zebrafish strain
Polymerase chain reaction
V1-B, V1-C, V1-D, V1-E1b, V1-F, V1-G, V1-H: Subunits A B, C, D, E1b, F, G H from the V1 domain of the V-ATPase
V0-ca, V0-cb, V0-c”/b, V0-d1, V0-e: Subunits a1a ca, cb, c”/b, d1 and e from the V0 domain of the V-ATPase.
The authors are grateful to Dr. Nancy Hopkins from the Massachusetts Institute of Technology for providing the zebrafish strains used in this work. We thank Vladimir Pelcastre for fish husbandry and breeding and Maggie Brunner for reviewing and editing the manuscript. Funding for this work was provided by the CONACYT grant 166046 and as well for the PAPIIT-UNAM grant IN208512.
- Maldonado E, Hernandez F, Lozano C, Castro ME, Navarro RE: The zebrafish mutant vps18 as a model for vesicle-traffic related hypopigmentation diseases. Pigment Cell Res. 2006, 19 (4): 315-326. 10.1111/j.1600-0749.2006.00320.x.PubMedView ArticleGoogle Scholar
- Schonthaler HB, Fleisch VC, Biehlmaier O, Makhankov Y, Rinner O, Bahadori R, Geisler R, Schwarz H, Neuhauss SC, Dahm R: The zebrafish mutant lbk/vam6 resembles human multisystemic disorders caused by aberrant trafficking of endosomal vesicles. Development. 2008, 135 (2): 387-399.PubMedView ArticleGoogle Scholar
- Navarro RE, Ramos-Balderas JL, Guerrero I, Pelcastre V, Maldonado E: Pigment dilution mutants from fish models with connection to lysosome-related organelles and vesicular traffic genes. Zebrafish. 2008, 5 (4): 309-318. 10.1089/zeb.2008.0549.PubMedView ArticleGoogle Scholar
- Nuckels RJ, Ng A, Darland T, Gross JM: The vacuolar-ATPase complex regulates retinoblast proliferation and survival, photoreceptor morphogenesis and pigmentation in the zebrafish eye. Invest Ophtalmol Vis. 2009, 50 (2): 893-905.View ArticleGoogle Scholar
- Amsterdam A, Varshney GK, Burgess SM: Retroviral-mediated insertional mutagenesis in zebrafish. Methods Cell Biol. 2011, 104: 59-82.PubMedPubMed CentralView ArticleGoogle Scholar
- Barresi MJ, Burton S, Dipietrantonio K, Amsterdam A, Hopkins N, Karlstrom RO: Essential genes for astroglial development and axon pathfinding during zebrafish embryogenesis. Dev Dyn. 2010, 239 (10): 2603-2618. 10.1002/dvdy.22393.PubMedPubMed CentralView ArticleGoogle Scholar
- Carney TJ, von der Hardt S, Sonntag C, Amsterdam A, Topczewski J, Hopkins N, Hammerschmidt M: Inactivation of serine protease Matriptase1a by its inhibitor Hai1 is required for epithelial integrity of the zebrafish epidermis. Development. 2007, 134 (19): 3461-3471. 10.1242/dev.004556.PubMedView ArticleGoogle Scholar
- Golling G, Amsterdam A, Sun Z, Antonelli M, Maldonado E, Chen W, Burgess S, Haldi M, Artzt K, Farrington S, et al: Insertional mutagenesis in zebrafish rapidly identifies genes essential for early vertebrate development. Nat Genet. 2002, 31 (2): 135-140. 10.1038/ng896.PubMedView ArticleGoogle Scholar
- Amsterdam A, Sadler KC, Lai K, Farrington S, Bronson RT, Lees JA, Hopkins N: Many ribosomal protein genes are cancer genes in zebrafish. PLoS Biol. 2004, 2 (5): E139-10.1371/journal.pbio.0020139.PubMedPubMed CentralView ArticleGoogle Scholar
- Lai K, Amsterdam A, Farrington S, Bronson RT, Hopkins N, Lees JA: Many ribosomal protein mutations are associated with growth impairment and tumor predisposition in zebrafish. Dev Dyn. 2009, 238 (1): 76-85. 10.1002/dvdy.21815.PubMedPubMed CentralView ArticleGoogle Scholar
- Sadler KC, Amsterdam A, Soroka C, Boyer J, Hopkins N: A genetic screen in zebrafish identifies the mutants vps18, nf2 and foie gras as models of liver disease. Development. 2005, 132 (15): 3561-3572. 10.1242/dev.01918.PubMedView ArticleGoogle Scholar
- Sun Z, Hopkins N: vhnf1, the MODY5 and familial GCKD-associated gene, regulates regional specification of the zebrafish gut, pronephros, and hindbrain. Genes Dev. 2001, 15 (23): 3217-3229. 10.1101/gad946701.PubMedPubMed CentralView ArticleGoogle Scholar
- Amsterdam A, Lai K, Komisarczuk AZ, Becker TS, Bronson RT, Hopkins N, Lees JA: Zebrafish Hagoromo mutants up-regulate fgf8 postembryonically and develop neuroblastoma. Mol Cancer Res: MCR. 2009, 7 (6): 841-850. 10.1158/1541-7786.MCR-08-0555.PubMedPubMed CentralView ArticleGoogle Scholar
- Amsterdam A, Nissen RM, Sun Z, Swindell EC, Farrington S, Hopkins N: Identification of 315 genes essential for early zebrafish development. Proc Natl Acad Sci USA. 2004, 101 (35): 12792-12797. 10.1073/pnas.0403929101.PubMedPubMed CentralView ArticleGoogle Scholar
- Gross JM, Perkins BD, Amsterdam A, Egana A, Darland T, Matsui JI, Sciascia S, Hopkins N, Dowling JE: Identification of zebrafish insertional mutants with defects in visual system development and function. Genetics. 2005, 170 (1): 245-261. 10.1534/genetics.104.039727.PubMedPubMed CentralView ArticleGoogle Scholar
- Schonthaler HB, Lampert JM, von Lintig J, Schwarz H, Geisler R, Neuhauss SC: A mutation in the silver gene leads to defects in melanosome biogenesis and alterations in the visual system in the zebrafish mutant fading vision. Dev Biol. 2005, 284 (2): 421-436. 10.1016/j.ydbio.2005.06.001.PubMedView ArticleGoogle Scholar
- Raposo G, Marks MS: Melanosomes–dark organelles enlighten endosomal membrane transport. Nat Rev Mol Cell Biol. 2007, 8 (10): 786-797. 10.1038/nrm2258.PubMedPubMed CentralView ArticleGoogle Scholar
- Ma B, Xiang Y, An L: Structural bases of physiological functions and roles of the vacuolar H(+)-ATPase. Cell Signal. 2011, 23 (8): 1244-1256. 10.1016/j.cellsig.2011.03.003.PubMedView ArticleGoogle Scholar
- Allan AK, Du J, Davies SA, Dow JA: Genome-wide survey of V-ATPase genes in Drosophila reveals a conserved renal phenotype for lethal alleles. Physiol Genomics. 2005, 22 (2): 128-138. 10.1152/physiolgenomics.00233.2004.PubMedView ArticleGoogle Scholar
- Oka T, Futai M: Requirement of V-ATPase for ovulation and embryogenesis in Caenorhabditis elegans. J Biol Chem. 2000, 275 (38): 29556-29561. 10.1074/jbc.M002756200.PubMedView ArticleGoogle Scholar
- Schoonderwoert VT, Martens GJ: Targeted disruption of the mouse gene encoding the V-ATPase accessory subunit Ac45. Mol Membr Biol. 2002, 19 (1): 67-71. 10.1080/09687680110112910.PubMedView ArticleGoogle Scholar
- Adams DS, Robinson KR, Fukumoto T, Yuan S, Albertson RC, Yelick P, Kuo L, McSweeney M, Levin M: Early, H+-V-ATPase-dependent proton flux is necessary for consistent left-right patterning of non-mammalian vertebrates. Development. 2006, 133 (9): 1657-1671. 10.1242/dev.02341.PubMedPubMed CentralView ArticleGoogle Scholar
- Chen Y, Wu B, Xu L, Li H, Xia J, Yin W, Li Z, Shi D, Li S, Lin S, et al: A SNX10/V-ATPase pathway regulates ciliogenesis in vitro and in vivo. Cell Res. 2012, 22 (2): 333-345. 10.1038/cr.2011.134.PubMedPubMed CentralView ArticleGoogle Scholar
- EauClaire SF, Cui S, Ma L, Matous J, Marlow FL, Gupta T, Burgess HA, Abrams EW, Kapp LD, Granato M, et al: Mutations in vacuolar H+ -ATPase subunits lead to biliary developmental defects in zebrafish. Dev Biol. 2012, 365 (2): 434-444. 10.1016/j.ydbio.2012.03.009.PubMedPubMed CentralView ArticleGoogle Scholar
- Qin A, Cheng TS, Lin Z, Pavlos NJ, Jiang Q, Xu J, Dai KR, Zheng MH: Versatile roles of V-ATPases accessory subunit Ac45 in osteoclast formation and function. PLoS One. 2011, 6 (11): e27155-10.1371/journal.pone.0027155.PubMedPubMed CentralView ArticleGoogle Scholar
- Qin A, Cheng TS, Pavlos NJ, Lin Z, Dai KR, Zheng MH: V-ATPases in osteoclasts: Structure, function and potential inhibitors of bone resorption. Int J Biochem Cell Biol. 2012, 44 (9): 1422-1435. 10.1016/j.biocel.2012.05.014.PubMedView ArticleGoogle Scholar
- Chung AY, Kim MJ, Kim D, Bang S, Hwang SW, Lim CS, Lee S, Park HC, Huh TL: Neuron-specific expression of atp6v0c2 in zebrafish CNS. Dev Dyn. 2010, 239 (9): 2501-2508. 10.1002/dvdy.22383.PubMedView ArticleGoogle Scholar
- Peri F, Nusslein-Volhard C: Live imaging of neuronal degradation by microglia reveals a role for v0-ATPase a1 in phagosomal fusion in vivo. Cell. 2008, 133 (5): 916-927. 10.1016/j.cell.2008.04.037.PubMedView ArticleGoogle Scholar
- Marshansky V, Futai M: The V-type H(+)-ATPase in vesicular trafficking: targeting, regulation and function. Curr Opin Cell Biol. 2008, 20 (4): 415-426. 10.1016/j.ceb.2008.03.015.PubMedView ArticleGoogle Scholar
- Muench SP, Trinick J, Harrison MA: Structural divergence of the rotary ATPases. Q Rev Biophys. 2011, 44 (3): 311-356. 10.1017/S0033583510000338.PubMedView ArticleGoogle Scholar
- Wang D, Jao LE, Zheng N, Dolan K, Ivey J, Zonies S, Wu X, Wu K, Yang H, Meng Q, et al: Efficient genome-wide mutagenesis of zebrafish genes by retroviral insertions. Proc Natl Acad Sci USA. 2007, 104 (30): 12428-12433. 10.1073/pnas.0705502104.PubMedPubMed CentralView ArticleGoogle Scholar
- Lee J, Willer JR, Willer GB, Smith K, Gregg RG, Gross JM: Zebrafish blowout provides genetic evidence for Patched1-mediated negative regulation of Hedgehog signaling within the proximal optic vesicle of the vertebrate eye. Dev Biol. 2008, 319 (1): 10-22. 10.1016/j.ydbio.2008.03.035.PubMedPubMed CentralView ArticleGoogle Scholar
- Pickart MA, Sivasubbu S, Nielsen AL, Shriram S, King RA, Ekker SC: Functional genomics tools for the analysis of zebrafish pigment. Pigment Cell Res. 2004, 17 (5): 461-470. 10.1111/j.1600-0749.2004.00189.x.PubMedView ArticleGoogle Scholar
- Horng JL, Lin LY, Huang CJ, Katoh F, Kaneko T, Hwang PP: Knockdown of V-ATPase subunit A (atp6v1a) impairs acid secretion and ion balance in zebrafish (Danio rerio). Am J Physiol Regul Integr Comp Physiol. 2007, 292 (5): R2068-2076. 10.1152/ajpregu.00578.2006.PubMedView ArticleGoogle Scholar
- Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF: Stages of embryonic development of the zebrafish. Dev Dyn. 1995, 203 (3): 253-310. 10.1002/aja.1002030302.PubMedView ArticleGoogle Scholar
- Trevarrow B: Zebrafish facilities for small and large laboratories. Methods Cell Biol. 2004, 77: 565-591.PubMedView ArticleGoogle Scholar
- Wilson JM, Bunte RM, Carty AJ: Evaluation of rapid cooling and tricaine methanesulfonate (MS222) as methods of euthanasia in zebrafish (Danio rerio). J Am Assoc Lab Anim Sci: JAALAS. 2009, 48 (6): 785-789.PubMedPubMed CentralGoogle Scholar
- Kelsh RN, Brand M, Jiang YJ, Heisenberg CP, Lin S, Haffter P, Odenthal J, Mullins MC, van Eeden FJ, Furutani-Seiki M, et al: Zebrafish pigmentation mutations and the processes of neural crest development. Development. 1996, 123: 369-389.PubMedGoogle Scholar
- Ho MN, Hirata R, Umemoto N, Ohya Y, Takatsuki A, Stevens TH, Anraku Y: VMA13 encodes a 54-kDa vacuolar H(+)-ATPase subunit required for activity but not assembly of the enzyme complex in Saccharomyces cerevisiae. J Biol Chem. 1993, 268 (24): 18286-18292.PubMedGoogle Scholar
- Wilkens S, Inoue T, Forgac M: Three-dimensional structure of the vacuolar ATPase. Localization of subunit H by difference imaging and chemical cross-linking. J Biol Chem. 2004, 279 (40): 41942-41949. 10.1074/jbc.M407821200.PubMedView ArticleGoogle Scholar
- Kay JN, Finger-Baier KC, Roeser T, Staub W, Baier H: Retinal ganglion cell genesis requires lakritz, a Zebrafish atonal Homolog. Neuron. 2001, 30 (3): 725-736. 10.1016/S0896-6273(01)00312-9.PubMedView ArticleGoogle Scholar
- El Far O, Seagar M: A role for V-ATPase subunits in synaptic vesicle fusion?. J Neurochem. 2011, 117 (4): 603-612.PubMedGoogle Scholar
- Peters C, Bayer MJ, Buhler S, Andersen JS, Mann M, Mayer A: Trans-complex formation by proteolipid channels in the terminal phase of membrane fusion. Nature. 2001, 409 (6820): 581-588. 10.1038/35054500.PubMedView ArticleGoogle Scholar
- Dougan ST, Warga RM, Kane DA, Schier AF, Talbot WS: The role of the zebrafish nodal-related genes squint and cyclops in patterning of mesendoderm. Development. 2003, 130 (9): 1837-1851. 10.1242/dev.00400.PubMedView ArticleGoogle Scholar
- Nishi T, Kawasaki-Nishi S, Forgac M: Expression and function of the mouse V-ATPase d subunit isoforms. J Biol Chem. 2003, 278 (47): 46396-46402. 10.1074/jbc.M303924200.PubMedView ArticleGoogle Scholar
- Jansen EJ, Hafmans TG, Martens GJ: V-ATPase-mediated granular acidification is regulated by the V-ATPase accessory subunit Ac45 in POMC-producing cells. Mol Biol Cell. 2010, 21 (19): 3330-3339. 10.1091/mbc.E10-04-0274.PubMedPubMed CentralView ArticleGoogle Scholar
- Miranda KC, Karet FE, Brown D: An extended nomenclature for mammalian V-ATPase subunit genes and splice variants. PLoS One. 2010, 5 (3): e9531-10.1371/journal.pone.0009531.PubMedPubMed CentralView ArticleGoogle Scholar
- Bahadori R, Rinner O, Schonthaler HB, Biehlmaier O, Makhankov YV, Rao P, Jagadeeswaran P, Neuhauss SC: The Zebrafish fade out mutant: a novel genetic model for Hermansky-Pudlak syndrome. Invest Ophthalmol Vis Sci. 2006, 47 (10): 4523-4531. 10.1167/iovs.05-1596.PubMedView ArticleGoogle Scholar
- Tabata H, Kawamura N, Sun-Wada GH, Wada Y: Vacuolar-type H(+)-ATPase with the a3 isoform is the proton pump on premature melanosomes. Cell Tissue Res. 2008, 332 (3): 447-460. 10.1007/s00441-008-0597-5.PubMedView ArticleGoogle Scholar
- Patton EE, Zon LI: The art and design of genetic screens: zebrafish. Nat Rev Genet. 2001, 2 (12): 956-966. 10.1038/35103567.PubMedView ArticleGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.