595
2003
eng
doctoralthesis
1
2003-11-10
--
2003-11-05
Systematic analysis of genes expressed in the retinal pigment epithelium (RPE) and identification of candidates for genetic susceptibility to age-related macular degeneration (AMD)
Age related macular degeneration (AMD) is the leading cause of visual impairment in the elderly and the major cause of blindness in the developed world. To date, the molecular mechanisms underlying the disease are not well understood although in recent years a primary involvement of the retinal pigment epithelium (RPE) has become evident. The aim of the present study is to systematically analyse genes which are differentially expressed in the RPE, and to assess their possible association with mechanisms and pathways likely to be related to retinal disease, in particular AMD. Towards this goal, 2379 expressed sequence tags (ESTs) were established from an inhouse generated RPE cDNA library. This library was constructed by using the suppression subtraction hybridization (SSH) technique which normalises redundant sequences and ensures enrichment of rare transcripts. In a first phase, 1002 ESTs were sequenced and subjected to comprehensive alignment with public nucleotide and protein databases. A search of the 1002 ESTs against the human genome draft sequence yielded 168 known genes, 51 predicted genes, 15 unknown transcripts and 41 clones with no significant similarity. Reverse Northern blot hybridization was performed for 318 EST clusters to identify abundantly expressed genes in the RPE and to prioritize subsequent analyses. Representative clones were spotted onto a nylon membrane and hybridized with cDNA probes of driver (heart and liver) and tester (RPE) used in the cDNA library construction. Subsequently, 107 EST clusters were subjected to Northern blot hybridizations. These analyses identified 7 RPE-specific, 3 retina-specific, 7 RPE/retina-specific, and 7 tissue restricted transcripts, while 29 EST clusters were ubiquitously expressed, and evaluation was not possible for another 54 EST clusters. Of the 24 transcripts with specific or restricted expression, 16 clones were selected for further characterization. The predicted gene MGC2477 and 2 novel isoforms of the human transient receptor potential cation channel, subfamily M, member 3 (TRPM3) were cloned and further described in detail. In addition, polymorphic variations for these 2 genes as well as for the human MT-Protocadherin gene were determined. For MGC2477, 15 single nucleotide polymorphisms (SNPs) were identified, with 13 having a frequency of the minor allele greater than 20%. 10 of the 15 SNPs have not been reported in so far in public SNP repertoires. Partial assessment of the TRPM3 gene yielded 35 SNPs. Of these, 30 (85.7%) were highly frequent (0.17-0.5%), and 14 (40%) were novel. The MT-Protocadherin gene revealed 35 SNPs, including 28 (80%) with high frequency of the minor allele. 23 (65.7%) were novel SNPs. These SNPs will be used to construct the most common haplotypes. These will be used in case/control association studies in 400 AMD patients and 200 ethnically and aged matched controls to assess a possible contribution of these genes in the etiology of AMD.
Die altersabhängige Makuladegeneration (AMD) ist die häufigste Ursache von gravierenden Einschränkungen des Sehvermögens im fortgeschrittenen Lebensalter. In den Industriestaaten ist die AMD zudem die Hauptursache für Altersblindheit. Die molekularen Mechanismen, die zur Entstehung der AMD führen, sind bisher nur unzureichend bekannt. In den letzten Jahren hat es sich jedoch herausgestellt, dass das retinale Pigmentepithel (RPE) eine primäre Rolle in der Pathogenese der AMD spielt. Ziel dieser Arbeit war die systematische Analyse von Genen, welche im RPE differentiell exprimiert werden. Entsprechende Kandidatengene sollten auf deren mögliche Beteiligung an der Entstehung von Erkrankungen der Retina, insbesondere der AMD, untersucht werden. Zunächst wurden 2379 ESTs aus einer innerhalb der Arbeitsgruppe generierten RPE cDNA Bibliothek definiert. Die dazu verwendete cDNA Bibliothek wurde durch die Suppressions- Subtraktions Hybridisierungs-Technik (SSH) konstruiert. Diese Technik gestattet eine Normalisierung gegenüber redundanten Sequenzen und begünstigt gleichzeitig die Anreicherung von seltenen Transkripten. In einer ersten Phase wurden 1002 ESTs sequenziert und einer umfassenden bioinformatischen Analyse mit Hilfe der verfügbaren DNA- und Protein Datenbanken unterzogen. Der Vergleich der 1002 ESTs mit der Draft Sequenz des menschlichen Genoms ergab den Hinweis auf 168 bereits bekannte Gene, 51 mögliche Gene, 15 völlig unbekannte Transkripte und 41 nicht weiter zuordenbare cDNA Klone. 318 EST Cluster wurden einer reversen Northen-Blot Analyse unterzogen um hochexprimierte Gene zu identifizieren und damit Prioritäten für die weiteren Analysen zu setzen. Im Rahmen der Northern-Analyse wurden repräsentative Klone von 107 EST-Klustern mit cDNA Sonden der ursprünglichen cDNA-Bibliothek hybridisiert. Als Ergebnis dieser Analyse fanden sich 7 RPE-spezifische, 3 Retina-spezifische, 7 sowohl RPE- als auch Retinaspezifische sowie 7 auf einzelne Gewebe limitierte Transkripte. 29 EST Cluster erwiesen sich als ubiquitär exprimiert, und 54 Kluster konnten nicht näher zugeordnet werden. Von den 24 Transkripten mit spezifischer oder zumindest begrenzter Expression wurden 16 Klone zur weiteren Charakterisierung ausgewählt. Aus diesen Material wurden im Rahmen dieser Arbeit das Kandidatengen MGC2477 sowie 2 neue Isoformen des menschlichen TRPM3-Gens kloniert und näher charakterisiert. Weiterhin wurden polymorphe Varianten dieser beiden Isoformen und des menschlichen MTProtocadherin- Gens definiert. Im Gen MGC2477 wurden 15 SNPs identifiziert, wovon die Allelhäufigkeit des selteneren Allels bei 13 der SNPs über 20% lag. Für 10 der insgesamt 15 vii SNPs dieses Gens fanden sich bisher keine Einträge in den entprechenden Datenbanken. Die SNP-Suche wurde auch für das TRPM3-Gen durchgeführt und ergab 35 SNPs, wovon 30 (85,7%) als hochfrequent eingestuft werden konnten. 14 dieser 35 SNPs waren bisher nicht in den Datenbanken verzeichnet. Beim MT-Protocadherin-Gen fanden sich ebenfalls 35 SNPs, wobei 80% eine hohe Frequenz des selteneren Allels aufwiesen. In diesem Fall handelte es sich bei 23 der insgesamt 35 SNPs um bisher unbekannte Allele. Diese SNPs bilden den Ausgangspunkt zur Konstruktion der häufigsten Haplotypen der genannten Gene. Mit der Charakterisierung der Einzel-Nukleotid Polymorphismen der Kandidatengene wurde die Grundlage zur Durchführung von Fall/Kontrollstudien gelegt, in deren Rahmen die Bedeutung der jeweiligen Kandidatengene in der Pathogense der AMD untersucht werden kann.
urn:nbn:de:bvb:20-opus-7053
705
X119225
Faisal Mirghani Abdel Rahman
deu
swd
Senile Makuladegeneration / Pigmentepithel / Genexpression
eng
uncontrolled
RPE
eng
uncontrolled
AMD
eng
uncontrolled
RPE specific genes
Biowissenschaften; Biologie
open_access
Institut für Humangenetik
Theodor-Boveri-Institut für Biowissenschaften
Universität Würzburg
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/595/RPE_project.pdf
27122
2022
eng
5
11
article
1
--
2022-04-28
--
Cisplatin-induced reproductive toxicity and oxidative stress: ameliorative effect of kinetin
Cisplatin is a commonly used chemotherapeutic agent; however, its potential side effects, including gonadotoxicity and infertility, are a critical problem. Oxidative stress has been implicated in the pathogenesis of cisplatin-induced testicular dysfunction. We investigated whether kinetin use at different concentrations could alleviate gonadal injury associated with cisplatin treatment, with an exploration of the involvement of its antioxidant capacity. Kinetin was administered in different doses of 0.25, 0.5, and 1 mg/kg, alone or along with cisplatin for 10 days. Cisplatin toxicity was induced via a single IP dose of 7 mg/kg on day four. In a dose-dependent manner, concomitant administration of kinetin with cisplatin significantly restored testicular oxidative stress parameters, corrected the distorted sperm quality parameters and histopathological changes, enhanced levels of serum testosterone and testicular StAR protein expression, as well as reduced the up-regulation of testicular TNF-α, IL-1β, Il-6, and caspase-3, caused by cisplatin. It is worth noting that the testicular protective effect of the highest kinetin dose was comparable/more potent and significantly higher than the effects of vitamin C and the lowest kinetin dose, respectively. Overall, these data indicate that kinetin may offer a promising approach for alleviating cisplatin-induced reproductive toxicity and organ damage, via ameliorating oxidative stress and reducing inflammation and apoptosis.
Antioxidants
2076-3921
10.3390/antiox11050863
urn:nbn:de:bvb:20-opus-271223
2022-05-09T06:54:26+00:00
sword
swordwue
attachment; filename=deposit.zip
8a2724b8ddc2c3935ce284b68c429946
Antioxidants (2022) 11:5, 863. doi:10.3390/antiox11050863
CC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International
Rania Abdel-Latif
Moustafa Fathy
Hend Ali Anwar
Muhammad Naseem
Thomas Dandekar
Eman M. Othman
eng
uncontrolled
cytokinins
eng
uncontrolled
kinetin
eng
uncontrolled
cisplatin
eng
uncontrolled
reproductive toxicity
Biowissenschaften; Biologie
open_access
Theodor-Boveri-Institut für Biowissenschaften
Import
Förderzeitraum 2022
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/27122/antioxidants-11-00863-v2.pdf
28512
2018
eng
1
19
article
1
--
2018-01-11
--
Protein activity of the \(Fusarium\) \(fujikuroi\) rhodopsins CarO and OpsA and their relation to fungus−plant interaction
Fungi possess diverse photosensory proteins that allow them to perceive different light wavelengths and to adapt to changing light conditions in their environment. The biological and physiological roles of the green light-sensing rhodopsins in fungi are not yet resolved. The rice plant pathogen Fusarium fujikuroi exhibits two different rhodopsins, CarO and OpsA. CarO was previously characterized as a light-driven proton pump. We further analyzed the pumping behavior of CarO by patch-clamp experiments. Our data show that CarO pumping activity is strongly augmented in the presence of the plant hormone indole-3-acetic acid and in sodium acetate, in a dose-dependent manner under slightly acidic conditions. By contrast, under these and other tested conditions, the Neurospora rhodopsin (NR)-like rhodopsin OpsA did not exhibit any pump activity. Basic local alignment search tool (BLAST) searches in the genomes of ascomycetes revealed the occurrence of rhodopsin-encoding genes mainly in phyto-associated or phytopathogenic fungi, suggesting a possible correlation of the presence of rhodopsins with fungal ecology. In accordance, rice plants infected with a CarO-deficient F. fujikuroi strain showed more severe bakanae symptoms than the reference strain, indicating a potential role of the CarO rhodopsin in the regulation of plant infection by this fungus.
International Journal of Molecular Sciences
1422-0067
10.3390/ijms19010215
urn:nbn:de:bvb:20-opus-285125
2022-09-05T19:03:22+00:00
sword
swordwue
attachment; filename=deposit.zip
b6a93a82ad902c57d24bf47e49dd32ba
International Journal of Molecular Sciences (2018) 19:1, 215. https://doi.org/10.3390/ijms19010215
CC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International
Alexander Adam
Stephan Deimel
Javier Pardo-Medina
Jorge García-Martínez
Tilen Konte
M. Carmen Limón
Javier Avalos
Ulrich Terpitz
eng
uncontrolled
fungal rhodopsins
eng
uncontrolled
CarO
eng
uncontrolled
OpsA
eng
uncontrolled
Fusarium fujikuroi
eng
uncontrolled
Oryza sativa
eng
uncontrolled
rice–plant infection
eng
uncontrolled
green light perception
eng
uncontrolled
indole-3-acetic acid (IAA)
eng
uncontrolled
bakanae
eng
uncontrolled
patch-clamp
Biowissenschaften; Biologie
open_access
Theodor-Boveri-Institut für Biowissenschaften
Import
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/28512/ijms-19-00215.pdf
7266
1991
eng
bookpart
1
2013-09-03
--
--
Genetic factors in tumour formation: The melanoma-inducing gene of Xiphophorus
No abstract available.
8067
urn:nbn:de:bvb:20-opus-86388
In: Molecular biology intumour research / ed. Olaf-Georg Issinger. - Heidelberg : Hüthig, 1991. - S. 79-94. - (BioTechForum 5: Adv. Molecular Genetics ; 3)
Deutsches Urheberrecht
D. Adam
A. Schartl
S. Andexinger
S. Hölter
B. Wilde
Manfred Schartl
deu
swd
Humangenetik
deu
swd
Tumor
deu
swd
Entstehung
Chemie und zugeordnete Wissenschaften
open_access
Theodor-Boveri-Institut für Biowissenschaften
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/7266/Schartl_7266.pdf
5936
1988
eng
article
1
2012-06-25
--
--
RFLP for an EGF-receptor related gene associated with the melanoma oncogene locus of Xiphophorus maculatus
No abstract available
urn:nbn:de:bvb:20-opus-61822
6182
In: Nucleic Acids Research (1988 ) 16, 7212.
Deutsches Urheberrecht
D. Adam
J. Wittbrodt
A. Telling
Manfred Schartl
deu
swd
Physiologische Chemie
Chemie und zugeordnete Wissenschaften
open_access
Theodor-Boveri-Institut für Biowissenschaften
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/5936/Schartl42.pdf
5337
1993
eng
article
1
2011-12-07
--
--
Tumor suppression in Xiphophorus by an accidentally acquired promoter
Melanoma formation in the teleost Xiphophorus is caused by a dominant genetic locus, Tu. This locus includes the Xmrk oncogene, which encodes a receptor tyrosine kinase. Tumor induction is. suppressed in wild-type fish by a tumor suppressor locus, R. Molecular genetic analyses revealed that the Tu locus emerged by nonhomologaus recombination of the Xmrk proto-oncogene with a previously uncharacterized sequence, D. This event generated an additional copy of Xmrk with a new promoter. Suppression of the new Xmrk promoter by R in parental fish and its deregulation in hybrids explain the genetics of melanoma formation in Xiphophorus.
urn:nbn:de:bvb:20-opus-61630
6163
In: Science (1993) , 259, 816-819
Deutsches Urheberrecht
Dieter Adam
Nicola Dimitrijevic
Manfred Schartl
deu
swd
Physiologische Chemie
Chemie und zugeordnete Wissenschaften
open_access
Theodor-Boveri-Institut für Biowissenschaften
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/5337/Schartl11.pdf
7193
1991
eng
article
1
2013-08-22
--
--
Transcriptional activation of the melanoma inducing Xmrk oncogene in Xiphophorus
The melanoma inducing locus of Xiphophorus encodes a tumorigenic version of a novel putative receptor tyrosine kinase (Xmrk). To elucidate the mechanism of oncogenic activation of Xmrk, we compared the structure and expression of two oncogenic loci with the corresponding proto-oncogene. Only minor structural alterations were found to be specific for the oncogenic Xmrk genes. Marked overexpression of the oncogene transcripts in melanoma, which are approximately 1 kb shorter than the proto-oncogene transcript, correlates with the malignancy of the tumors. The tumor transcripts are derived from an alternative transcription start site that is used only in the oncogenic loci. Thus, oncogenic activation of the melanoma inducing Xmrk gene appears primarily to be due to novel transcriptional control and overexpression.
8033
urn:nbn:de:bvb:20-opus-87584
In: Oncogene, 1991, 6, S. 73-80
Deutsches Urheberrecht
Dieter Adam
Winfried Maueler
Manfred Schartl
deu
swd
Schwertkärpfling
deu
swd
Onkogen
deu
swd
Melanom
Chemie und zugeordnete Wissenschaften
open_access
Theodor-Boveri-Institut für Biowissenschaften
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/7193/Schartl_7193.pdf
11938
2014
eng
e104337
8
9
article
1
2015-09-29
--
--
Evaluation of a New Recombinant Oncolytic Vaccinia Virus Strain GLV-5b451 for Feline Mammary Carcinoma Therapy
Virotherapy on the basis of oncolytic vaccinia virus (VACV) infection is a promising approach for cancer therapy. In this study we describe the establishment of a new preclinical model of feline mammary carcinoma (FMC) using a recently established cancer cell line, DT09/06. In addition, we evaluated a recombinant vaccinia virus strain, GLV-5b451, expressing the anti-vascular endothelial growth factor (VEGF) single-chain antibody (scAb) GLAF-2 as an oncolytic agent against FMC. Cell culture data demonstrate that GLV-5b451 virus efficiently infected, replicated in and destroyed DT09/06 cancer cells. In the selected xenografts of FMC, a single systemic administration of GLV-5b451 led to significant inhibition of tumor growth in comparison to untreated tumor-bearing mice. Furthermore, tumor-specific virus infection led to overproduction of functional scAb GLAF-2, which caused drastic reduction of intratumoral VEGF levels and inhibition of angiogenesis.
In summary, here we have shown, for the first time, that the vaccinia virus strains and especially GLV-5b451 have great potential for effective treatment of FMC in animal model.
PLoS ONE
10.1371/journal.pone.0104337
urn:nbn:de:bvb:20-opus-119387
PLoS ONE 9(8): e104337. doi:10.1371/journal.pone.0104337
Marion Adelfinger
Ivaylo Gentschev
Julio Grimm de Guibert
Stephanie Weibel
Johanna Langbein-Laugwitz
Barbara Härtl
Hugo Murua Escobar
Ingo Nolte
Nanhai G. Chen
Richard J. Aguilar
Yong A. Yu
Qian Zhang
Alexa Frentzen
Aladar A. Szalay
eng
uncontrolled
antibodies
eng
uncontrolled
cancer treatment
eng
uncontrolled
carcinomas
eng
uncontrolled
vaccinia virus
eng
uncontrolled
oncolytic viruses
eng
uncontrolled
viral replication
eng
uncontrolled
cell cultures
eng
uncontrolled
enzyme-linked immunoassays
Krankheiten
open_access
Institut für Molekulare Infektionsbiologie
Theodor-Boveri-Institut für Biowissenschaften
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/11938/033_Adelfinger_PLOS_ONE.pdf
12648
2015
eng
2
13
article
1
2016-01-31
--
--
Molecular cloning and expression analysis of dmrt1 and sox9 during gonad development and male reproductive cycle in the lambari fish, Astyanax altiparanae
Background
The dmrt1 and sox9 genes have a well conserved function related to testis formation in vertebrates, and the group of fish presents a great diversity of species and reproductive mechanisms. The lambari fish (Astyanax altiparanae) is an important Neotropical species, where studies on molecular level of sex determination and gonad maturation are scarce.
Methods
Here, we employed molecular cloning techniques to analyze the cDNA sequences of the dmrt1 and sox9 genes, and describe the expression pattern of those genes during development and the male reproductive cycle by qRT-PCR, and related to histology of the gonad.
Results
Phylogenetic analyses of predicted amino acid sequences of dmrt1 and sox9 clustered A. altiparanae in the Ostariophysi group, which is consistent with the morphological phylogeny of this species. Studies of the gonad development revealed that ovary formation occurred at 58 days after hatching (dah), 2 weeks earlier than testis formation. Expression studies of sox9 and dmrt1 in different tissues of adult males and females and during development revealed specific expression in the testis, indicating that both genes also have a male-specific role in the adult. During the period of gonad sex differentiation, dmrt1 seems to have a more significant role than sox9. During the male reproductive cycle dmrt1 and sox9 are down-regulated after spermiation, indicating a role of these genes in spermatogenesis.
Conclusions
For the first time the dmrt1 and sox9 were cloned in a Characiformes species. We show that both genes have a conserved structure and expression, evidencing their role in sex determination, sex differentiation and the male reproductive cycle in A. altiparanae. These findings contribute to a better understanding of the molecular mechanisms of sex determination and differentiation in fish.
Reproductive Biology and Endocrinology
10.1186/1477-7827-13-2
urn:nbn:de:bvb:20-opus-126486
Reproductive Biology and Endocrinology 2015, 13:2. DOI:10.1186/1477-7827-13-2
Mateus C. Adolfi
Ana C. O. Carreira
Lázaro W. O. Jesus
Jan Bogerd
Rejane M. Funes
Manfred Schartl
Mari C. Sogayar
Maria I. Borella
eng
uncontrolled
spermatogenesis
eng
uncontrolled
SOX9
eng
uncontrolled
DMRT1
eng
uncontrolled
sex differentiation
eng
uncontrolled
teleostei
Humanphysiologie
open_access
Theodor-Boveri-Institut für Biowissenschaften
Förderzeitraum 2015
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/12648/Adolfi_ReproductiveBiology_1477-7827-13-2.pdf
26567
2021
eng
1
11
article
1
2022-04-05
--
--
A duplicated copy of id2b is an unusual sex-determining candidate gene on the Y chromosome of arapaima (Arapaima gigas)
Arapaima gigas is one of the largest freshwater fish species of high ecological and economic importance. Overfishing and habitat destruction are severe threats to the remaining wild populations. By incorporating a chromosomal Hi-C contact map, we improved the arapaima genome assembly to chromosome-level, revealing an unexpected high degree of chromosome rearrangements during evolution of the bonytongues (Osteoglossiformes). Combining this new assembly with pool-sequencing of male and female genomes, we identified id2bbY, a duplicated copy of the inhibitor of DNA binding 2b (id2b) gene on the Y chromosome as candidate male sex-determining gene. A PCR-test for id2bbY was developed, demonstrating that this gene is a reliable male-specific marker for genotyping. Expression analyses showed that this gene is expressed in juvenile male gonads. Its paralog, id2ba, exhibits a male-biased expression in immature gonads. Transcriptome analyses and protein structure predictions confirm id2bbY as a prime candidate for the master sex-determiner. Acting through the TGF beta signaling pathway, id2bbY from arapaima would provide the first evidence for a link of this family of transcriptional regulators to sex determination. Our study broadens our current understanding about the evolution of sex determination genetic networks and provide a tool for improving arapaima aquaculture for commercial and conservation purposes.
Scientific Reports
10.1038/s41598-021-01066-z
urn:nbn:de:bvb:20-opus-265672
publish
Scientific Reports (2021) 11:1, 21544. https://doi.org/10.1038/s41598-021-01066-z
false
true
CC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International
Mateus C. Adolfi
Kang Du
Susanne Kneitz
Cédric Cabau
Margot Zahm
Christophe Klopp
Romain Feron
Rômulo V. Paixão
Eduardo S. Varela
Fernanda L. de Almeida
Marcos A. de Oliveira
Rafael H. Nóbrega
Céline Lopez-Roques
Carole Iampietro
Jérôme Lluch
Werner Kloas
Sven Wuertz
Fabian Schaefer
Matthias Stöck
Yann Guiguen
Manfred Schartl
eng
uncontrolled
evolutionary genetics
eng
uncontrolled
genetic markers
eng
uncontrolled
genome
Medizin und Gesundheit
open_access
Theodor-Boveri-Institut für Biowissenschaften
Förderzeitraum 2021
Comprehensive Cancer Center Mainfranken
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/26567/s41598-021-01066-z.pdf