scholarly journals The rhodopsin-retinochrome system for retinal re-isomerization predates the origin of cephalopod eyes

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Oliver Vöcking ◽  
Lucas Leclère ◽  
Harald Hausen
Keyword(s):  
In Situ Hybridization ◽  
Photoreceptor Cells ◽  
Visual Cycle ◽  
Phylogenetic Distribution ◽  
Enzymatic Process ◽  
Antibody Staining ◽  
The Individual ◽  
Functional Components ◽  
Visual Rhodopsin

Abstract Background The process of photoreception in most animals depends on the light induced isomerization of the chromophore retinal, bound to rhodopsin. To re-use retinal, the all-trans-retinal form needs to be re-isomerized to 11-cis-retinal, which can be achieved in different ways. In vertebrates, this mostly includes a stepwise enzymatic process called the visual cycle. The best studied re-isomerization system in protostomes is the rhodopsin-retinochrome system of cephalopods, which consists of rhodopsin, the photoisomerase retinochrome and the protein RALBP functioning as shuttle for retinal. In this study we investigate the expression of the rhodopsin-retinochrome system and functional components of the vertebrate visual cycle in a polyplacophoran mollusk, Leptochiton asellus, and examine the phylogenetic distribution of the individual components in other protostome animals. Results Tree-based orthology assignments revealed that orthologs of the cephalopod retinochrome and RALBP are present in mollusks outside of cephalopods. By mining our dataset for vertebrate visual cycle components, we also found orthologs of the retinoid binding protein RLBP1, in polyplacophoran mollusks, cephalopods and a phoronid. In situ hybridization and antibody staining revealed that L. asellus retinochrome is co-expressed in the larval chiton photoreceptor cells (PRCs) with the visual rhodopsin, RALBP and RLBP1. In addition, multiple retinal dehydrogenases are expressed in the PRCs, which might also contribute to the rhodopsin-retinochrome system. Conclusions We conclude that the rhodopsin-retinochrome system is a common feature of mollusk PRCs and predates the origin of cephalopod eyes. Our results show that this system has to be extended by adding further components, which surprisingly, are shared with vertebrates.

scholarly journals Selected In Situ Hybridization Methods: Principles and Application

Molecules ◽  
2021 ◽  
Vol 26 (13) ◽  
pp. 3874
Author(s):  
Dominika Veselinyová ◽  
Jana Mašlanková ◽  
Katarina Kalinová ◽  
Helena Mičková ◽  
Mária Mareková ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Imaging Techniques ◽  
Cell Communication ◽  
Probe Design ◽  
Rapid Progress ◽  
In Situ Techniques ◽  
Different Types ◽  
Laboratory Procedures ◽  
The Individual

We are experiencing rapid progress in all types of imaging techniques used in the detection of various numbers and types of mutation. In situ hybridization (ISH) is the primary technique for the discovery of mutation agents, which are presented in a variety of cells. The ability of DNA to complementary bind is one of the main principles in every method used in ISH. From the first use of in situ techniques, scientists paid attention to the improvement of the probe design and detection, to enhance the fluorescent signal intensity and inhibition of cross-hybrid presence. This article discusses the individual types and modifications, and is focused on explaining the principles and limitations of ISH division on different types of probes. The article describes a design of probes for individual types of in situ hybridization (ISH), as well as the gradual combination of several laboratory procedures to achieve the highest possible sensitivity and to prevent undesirable events accompanying hybridization. The article also informs about applications of the methodology, in practice and in research, to detect cell to cell communication and principles of gene silencing, process of oncogenesis, and many other unknown processes taking place in organisms at the DNA/RNA level.

Variation in highly repetitive DNA composition of heterochromatin in rye studied by fluorescence in situ hybridization

Genome ◽  
1995 ◽  
Vol 38 (6) ◽  
pp. 1061-1069 ◽  
Author(s):  
A. Cuadrado ◽  
N. Jouve ◽  
C. Ceoloni
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Dna ◽  
Dna Sequences ◽  
5S Rdna ◽  
Individual Identification ◽  
Highly Repetitive Dna ◽  
The Individual

The molecular characterization of heterochromatin in six lines of rye has been performed using fluorescence in situ hybridization (FISH). The highly repetitive rye DNA sequences pSc 119.2, pSc74, and pSc34, and the probes pTa71 and pSc794 containing the 25S–5.8S–18S rDNA (NOR) and the 5S rDNA multigene families, respectively, were used. This allowed the individual identification of all seven rye chromosomes and most chromosome arms in all lines. All varieties showed similar but not identical patterns. A standard in situ hybridization map was constructed following the nomenclature system recommended for C-bands. All FISH sites observed appeared to correspond well with C-band locations, but not all C-banding sites coincided with hybridization sites of the repetitive DNA probes used. Quantitative and qualitative differences between different varieties were found for in situ hybridization response at corresponding sites. Variation between plants and even between homologous chromosomes of the same plant was found in open-pollinated lines. In inbred lines, the in situ pattern of the homologues was practically identical and no variation between plants was detected. The observed quantitative and qualitative differences are consistent with a corresponding variation for C-bands detected both within and between cultivars.Key words: fluorescence in situ hybridization, repetitive DNA, rye, Secale cereale, polymorphism.

scholarly journals The effect of avidin-biotin interactions in detection systems for in situ hybridization.

1992 ◽  
Vol 40 (1) ◽  
pp. 135-141 ◽  
Author(s):  
E J Speel ◽  
B Schutte ◽  
F C Ramaekers ◽  
A H Hopman
Keyword(s):  
In Situ Hybridization ◽  
Network Formation ◽  
Transitional Cell ◽  
Fluorescein Isothiocyanate ◽  
Staining Technique ◽  
Dna Probe ◽  
Detection Systems ◽  
Amplification Method ◽  
The Individual

The effect of avidin-biotin interactions in several detection systems for the non-radioactive in situ hybridization (ISH) technique was studied in a model system using a transitional cell carcinoma line and a biotinylated DNA probe. We performed fluorescence ISH to unravel the individual steps in a sensitive and frequently used amplification method which makes use of the alternating cytochemical detection layers of fluorescein isothiocyanate-conjugated avidin (AvFITC) and biotinylated goat anti-avidin (BioGAA) antibodies to detect the hybridized and biotinylated probe. Our experiments revealed that BioGAA antibodies bind with their antigen binding sites and not with their biotin moieties to avidin molecules that have already interacted with the DNA probe. The probable working mechanism of this amplification method is presented in a model. Furthermore, we used a peroxidase staining technique to compare with each other the sensitivity of several other detection systems in which avidin-biotin interactions play an important role, e.g., the avidin-biotinylated peroxidase complex (ABC) system. The experiments show that avidin molecules can not be efficiently used to interconnect two biotinylated molecular layers, since their introduction leads to firmly closed cytochemical networks. Such a closed network is already formed between the hybridized and biotinylated DNA probe and a first detection layer of avidin molecules, as appears from the finding that biotinylated molecules could hardly be coupled to these avidin molecules in a following detection layer. Therefore, the results presented here provide us with new insight into the molecular basis of cytochemical network formation. This will enable us to choose the proper procedures for increasing the sensitivity of ISH detection systems.

scholarly journals X-gal Staining of Drosophila Embryos Compatible with Antibody Staining or In Situ Hybridization

BioTechniques ◽  
1998 ◽  
Vol 24 (6) ◽  
pp. 918-922 ◽  
Author(s):  
Ming-Tsan Su ◽  
Krista Golden ◽  
Rolf Bodmer
Keyword(s):  
In Situ Hybridization ◽  
Antibody Staining ◽  
Drosophila Embryos

Extrarenal tissue distribution of CHIP28 water channels by in situ hybridization and antibody staining

1994 ◽  
Vol 266 (4) ◽  
pp. C893-C903 ◽  
Author(s):  
H. Hasegawa ◽  
S. C. Lian ◽  
W. E. Finkbeiner ◽  
A. S. Verkman
Keyword(s):  
In Situ Hybridization ◽  
Epithelial Cells ◽  
Tissue Distribution ◽  
Choroid Plexus ◽  
Ciliary Body ◽  
Fluid Transport ◽  
Pancreatic Acini ◽  
Antibody Staining ◽  
Strong Hybridization

This study is an extension of in situ hybridization experiments showing expression of mRNA encoding CHIP28 in selected epithelial or endothelia in spleen, colon, lung, and eye (H. Hasegawa, R. Zhang, A. Dohrman, and A. S. Verkman. Am. J. Physiol. 264 (Cell Physiol. 33): C237-C245, 1993). Additional tissues from rat were screened by in situ hybridization, and tissues from rat and humans were stained with a polyclonal anti-CHIP28 antibody. Northern blot showed the 2.8-kilobase mRNA encoding CHIP28 in kidney, lung, and heart. In situ hybridization showed strong hybridization in epithelial cells in choroid plexus, iris, ciliary body, and lens and in epithelial and subepithelial layers of trachea. Except for colonic crypts, specific hybridization was not observed in the gastrointestinal tract, liver, thyroid gland, and muscle. Immunoblot of tissues from exsanguinated rats showed immunoreactive CHIP28 protein in kidney, lung, trachea, and heart. In fixed frozen rat and/or human tissues, the anti-CHIP28 antibody stained epithelial cells in kidney proximal tubule and thin limb of Henle, lung alveolus, bronchial mucosa and glands, choroid plexus, ciliary body, iris, lens surface, colonic crypt, sweat gland, pancreatic acini, gallbladder epithelium, and placental syncytial trophoblast cells. Endothelial cells were stained in many tissues. These studies indicate a wide and selective CHIP28 tissue distribution, suggesting an important role for CHIP28 in fluid transport. The absence of CHIP28 in many nonrenal membranes believed to be water permeable suggests the existence of non-CHIP28 water transporters.

scholarly journals Specific detection of neuronal cell bodies: in situ hybridization with a biotin-labeled neurofilament cDNA probe.

1986 ◽  
Vol 34 (7) ◽  
pp. 923-926 ◽  
Author(s):  
P Liesi ◽  
J P Julien ◽  
P Vilja ◽  
F Grosveld ◽  
L Rechardt
Keyword(s):  
In Situ Hybridization ◽  
Cdna Probe ◽  
Neuronal Cell ◽  
Dna Probe ◽  
Specific Detection ◽  
Neurofilament Protein ◽  
Antibody Staining ◽  
Neuronal Cell Bodies ◽  
Specific Mrna

We have used a biotinylated, 300-nucleotide cDNA probe which encodes the 68,000 MW neurofilament protein to detect neurofilament-specific mRNA in situ. The neurofilament message specifically demonstrates the neuronal cell bodies, in contrast to the usual antibody staining which detects their neurites. The hybridization is detected only in neuronal structures. Consequently, detection of the biotinylated neurofilament DNA probe by silver-intensified streptavidin-gold can be specifically used to identify neuronal cell bodies.

scholarly journals ANTIBODY STAINING AND IN SITU HYBRIDIZATION REVEALS THAT A SUBSET OF NEURONS IN THE RABBIT NEONATAL BRAIN PRODUCE INSULIN

1987 ◽  
Vol 21 (4) ◽  
pp. 221A-221A
Author(s):  
Ruben Schechter ◽  
Lynn Karyckl ◽  
Farouk Sadig ◽  
Thomas Hilliard ◽  
Arnold Kahn ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Neonatal Brain ◽  
Antibody Staining

scholarly journals Detection and Differentiation of Chlamydiae by Fluorescence In Situ Hybridization

2002 ◽  
Vol 68 (8) ◽  
pp. 4081-4089 ◽  
Author(s):  
Sven Poppert ◽  
Andreas Essig ◽  
Reinhard Marre ◽  
Michael Wagner ◽  
Matthias Horn
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Single Step ◽  
Detection Methods ◽  
The Novel ◽  
Antibody Staining ◽  
Uncultured Bacteria ◽  
Direct Fluorescence ◽  
Probe Set

ABSTRACT Chlamydiae are important pathogens of humans and animals but diagnosis of chlamydial infections is still hampered by inadequate detection methods. Fluorescence in situ hybridization (FISH) using rRNA-targeted oligonucleotide probes is widely used for the investigation of uncultured bacteria in complex microbial communities and has recently also been shown to be a valuable tool for the rapid detection of various bacterial pathogens in clinical specimens. Here we report on the development and evaluation of a hierarchic probe set for the specific detection and differentiation of chlamydiae, particularly C. pneumoniae, C. trachomatis, C. psittaci, and the recently described chlamydia-like bacteria comprising the novel genera Neochlamydia and Parachlamydia. The specificity of the nine newly developed probes was successfully demonstrated by in situ hybridization of experimentally infected amoebae and HeLa 229 cells, including HeLa 229 cells coinfected with C. pneumoniae and C. trachomatis. FISH reliably stained chlamydial inclusions as early as 12 h postinfection. The sensitivity of FISH was further confirmed by combination with direct fluorescence antibody staining. In contrast to previously established detection methods for chlamydiae, FISH was not susceptible to false-positive results and allows the detection of all recognized chlamydiae in one single step.

Identification of wheat and tritordeum chromosomes by genomic in situ hybridization using total Hordeum chilense DNA as probe

Genome ◽  
1999 ◽  
Vol 42 (6) ◽  
pp. 1194-1200 ◽  
Author(s):  
M J González ◽  
A Cabrera
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
Hexaploid Wheat ◽  
Chromosome Complement ◽  
Dna Probe ◽  
Hordeum Chilense ◽  
B Genome ◽  
A Genome ◽  
The Individual

Total genomic Hordeum chilense DNA probe was hybridized to somatic chromosome spreads of Triticum aestivum 'Chinese Spring' and to four advanced tritordeum lines, the latter being the fertile amphiploid between H. chilense and durum wheat (2n = 6x = 42, AABBHchHch). The probe hybridized strongly to the B-genome chromosomes and to one or two bands on the A-genome chromosomes present in both wheat and tritordeum alloploids. Bands on chromosomes 1D, 2D, and 7D from hexaploid wheat were also detected. Genomic H. chilense DNA probe identified 16 chromosome pairs of the chromosome complement of hexaploid wheat and all A- and B-genome chromosomes present in the tritordeum amphiploids. The in situ hybridization patterns observed correspond to those previously reported in wheat by both N-banding and in situ hybridization with the GAA-satellite sequence (Pedersen and Langridge 1997), allowing the identification of these chromosomes. Variation among the tritordeum amphiploids for hybridization sites on chromosomes 2A, 4A, 6A, 7A, 4B, 5B, and 7B was observed. Despite of this polymorphism, all lines shared the general banding pattern. When used as probe, total H. chilense genomic DNA labeled the H. chilense chromosomes over their lengths allowing the identification of 14 H. chilense chromosomes present in the tritordeum amphiploids. In addition, chromosome-specific telomeric, interstial, and centromeric hybridization sites were observed. These hybridization sites coincide with N-banded regions in H. chilense allowing the identification of the individual H. chilense chromosomes in one of the amphiploid. The N-banded karyotypes of H. chilense (accessions H1 and H7) are presented.Key words: Hordeum chilense, Triticum aestivum, chromosome identification, in situ hybridization, N-banding.

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