Detection of mRNA encoding Crustacean Hyperglycemic Hormone (CHH) in the eyestalk of the crayfish Orconectes limosus using non-radioactive in situ hybridization

1991 ◽  
Vol 124 (2) ◽  
pp. 178-182 ◽  
Author(s):  
Cornelis P. Tensen ◽  
Tony Coenen ◽  
François van Herp
Keyword(s):  
In Situ Hybridization ◽  
Crustacean Hyperglycemic Hormone ◽  
Orconectes Limosus ◽  
Hyperglycemic Hormone

Effects of 17β-estradiol injection and red-spectrum light on eyestalk hormones and vitellogenesis of the ornamental cleaner shrimp Lysmata amboinensis (De Man, 1888) (Decapoda: Caridea: Lysmatidae)

2019 ◽  
Author(s):  
Ji Yong Choi ◽  
Cheol Young Choi ◽  
Min-Min Jung
Keyword(s):  
Mrna Expression ◽  
Red Light ◽  
Crustacean Hyperglycemic Hormone ◽  
Eyestalk Ablation ◽  
Vitellogenin Receptor ◽  
Hyperglycemic Hormone ◽  
17Β Estradiol ◽  
De Man ◽  
Cleaner Shrimp

Abstract We analyzed the effects of 17β-estradiol (E2) injection and red light on the sexual maturation and vitellogenesis of the cleaner shrimp Lysmata amboinensis (De Man, 1888) by measuring the change of mRNA expression levels of eyestalk and vitellogenesis-related hormones induced by both factors. We examined the eyestalk crustacean hyperglycemic hormone (CHH) and vitellogenesis-inhibiting hormone (VIH) genes. The E2 injection did not affect changes of eyestalk hormones. The red light, however, caused a significant increase (P < 0.05) of CHH promoting maturity and a significant decrease of VIH, which suppressed vitellogenesis. The vitellogenin receptor (VTG-R) and mRNA expression (in situ hybridization) also caused the E2 injection and red-wavelength irradiation to induce synthesis of vitellogenin (VTG). Red-light-induced gonadal maturation, promoting hormone and VTG synthesis, but E2 did not change the eyestalk hormone. Red-light irradiation could thus be used as a novel, maturity-promoting alternative method in the culture of the species, which is different from the eyestalk-ablation method that has so far been used.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

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