Subcellular localization of α-tubulin and opsin mRNA in the goldfish retina using digoxigenin-labeled cRNA probes detected by alkaline phosphatase and HRP histochemistry

Barthel, Linda K.; Raymond, Pamela A.

doi:10.1016/0165-0270(93)90002-9

Cited by 72 publications

(43 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…These cDNAs were used to synthesize digoxigenin-(dig) or fluorescein-(fl) labeled cDNA probes using components of the Genius Kit (Roche). In situ hybridization was performed on cryosectioned embryos and whole mounts according to Barthel and Raymond (1993). In brief, tissue was rehydrated in a descending ethanol series and treated with 10 μg/mL proteinase K. Sections were then dehydrated and hybridized at 56°C overnight with probe (1 mg/mL) in a solution containing 50% formamide.…”

Section: Expression Of Cell Specific Markersmentioning

confidence: 99%

Mechanisms for persistent microphthalmia following ethanol exposure during retinal neurogenesis in zebrafish embryos

2007

View full text Add to dashboard Cite

The exposure of the developing human embryo to ethanol results in a spectrum of disorders involving multiple organ systems, including the visual system. One common phenotype seen in humans exposed to ethanol in utero is microphthalmia. The objective of this study was to describe the effects of ethanol during retinal neurogenesis in a model organism, the zebrafish, and to pursue the potential mechanisms by which ethanol causes microphthalmia. Zebrafish embryos were exposed to 1% or 1.5% ethanol from 24 to 48 h after fertilization, a period during which the retinal neuroepithelium undergoes rapid proliferation and differentiation to form a laminated structure composed of different retinal cell types. Ethanol exposure resulted in significantly reduced eye size immediately following the treatment, and this microphthalmia persisted through larval development. This reduced eye size could not entirely be accounted for by the accompanying general delay in embryonic development. Retinal cell death was only slightly higher in ethanol-exposed embryos, although cell death in the lens was extensive in some of these embryos, and lenses were significantly reduced in size as compared to those of control embryos. The initiation of retinal neurogenesis was not affected, but the subsequent waves of cell differentiation were markedly reduced. Even cells that were likely generated after ethanol exposure-rod and cone photoreceptors and Müller glia-were delayed in their expression of cell-specific markers by at least 24 h. We conclude that ethanol exposure over the time of retinal neurogenesis resulted in persistent microphthalmia due to a combination of an overall developmental delay, lens abnormalities, and reduced retinal cell differentiation.

show abstract

Section: Expression Of Cell Specific Markersmentioning

confidence: 99%

Mechanisms for persistent microphthalmia following ethanol exposure during retinal neurogenesis in zebrafish embryos

2007

View full text Add to dashboard Cite

show abstract

“…The in situ hybridization experiments were performed on tissue sections and on neuronal cell cultures according to the methods of Barthel and Raymond (1993) and Litman et al (1993). Briefly, antisense digoxigenin (DIG)-labeled 5-HT 3 R probe was synthesized by in vitro transcription with T7 RNA polymerase and DIG-11-UTP (Boehringer Mannheim) and the plasmid containing the portion of the rat 5-HT 3 R cDNA described above.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Developing Neonatal Rat Sympathetic and Sensory Neurons Differ in Their Regulation of 5-HT₃Receptor Expression

1997

View full text Add to dashboard Cite

Serotonin 5-HT 3 receptors (5-HT 3 Rs) are ligand-gated ion channels expressed by many peripheral neurons and are involved in several physiological processes. To learn more about the developmental regulation of 5-HT 3 R expression, we investigated rat sympathetic and vagal sensory neurons. We found that sympathetic and sensory neurons differ in their regulation of 5-HT 3 R expression during early postnatal life and as these neurons develop in culture. In SCG neurons 5-HT 3 R transcript levels are low at postnatal day 1 (P1) and increase 7.5-fold by P21; this increase occurs even after elimination of preganglionic innervation. In comparison, 5-HT 3 R mRNA levels in P1 nodose neurons are over 14-fold greater than in P1 SCG and change little by P21. We show that 5-HT 3 R transcript levels in nodose neurons depend on intact target innervation and drop by 60% after axotomy. When P1 SCG neurons develop in culture, we observed a significant increase in 5-HT 3 R expression: after 7 d in culture, transcript levels increase ninefold versus a threefold increase for neurons developing for 7 d in vivo. In contrast, 5-HT 3 R mRNA levels in cultured nodose neurons drop by 70% within 24 hr; however, this drop is transient. After 2 d, transcript levels begin to increase, and after 7 d, they are above initial values. We show that this delayed increase in 5-HT 3 R expression depends on neurotrophins. In both nodose and sympathetic neurons we found that the changes in 5-HT 3 R gene expression correlate directly with the appearance of 5-HTevoked current densities. Key words: 5-HT 3 receptor; ligand-gated ion channel; sympathetic; superior cervical ganglion; sensory; nodose; trigeminal; mRNA expression; neurotrophins; axotomyThe serotonin 5-HT 3 receptor (5-HT 3 R), a neurotransmittergated ion channel (Yakel and Jackson, 1988;Derkach et al., 1989;Maricq et al., 1991) present on many mammalian peripheral neurons, participates in several diverse physiological functions (Fozard, 1984;Jackson and Yakel, 1995). Activation of 5-HT 3 Rs located on peripheral vagal sensory nerve endings initiates reflexes affecting respiration, circulation, emesis, and swallowing (Douglas, 1975;Sanders-Bush and Mayer, 1996). 5-HT 3 Rs on spinal and vagal sensory neurons are involved in nociceptive signaling and nausea (Fozard, 1984). In the C NS 5-HT 3 Rs are implicated in anxiety, depression, and drug dependence (Apud, 1993;Greenshaw, 1993). In addition, 5-HT 3 Rs are expressed by sympathetic neurons; however, the role for these receptors in sympathetic f unction has not been f ully determined (Wallis and North, 1978;Yang et al., 1992).Many vagal afferent neurons expressing 5-HT 3 Rs are located in the nodose ganglion. These sensory neurons have typical unipolar polarities; their axons bif urcate into a peripheral branch that innervates much of the viscera, including heart, lungs, trachea, and gut, and a central branch that terminates mainly in the nucleus tractus solitarius (Andresen and Kunze, 1994). The physiological responses to serotonin elicited from ...

show abstract

“…Determination of expression pattern on protein level Pujic and Malicki (2001), Wei and Malicki (2002) In situ hybridization-double labeling Parallel determination of two expression patterns on transcript level Hauptmann and Gerster (1994), Jowett (2001), Jowett and Lettice (1994), Strahle et al (1994) In situ hybridization-frozen sections Determination of expression pattern on transcript level Barthel and Raymond (1993), Hisatomi et al (1996) In situ hybridization-whole mount Determination of expression pattern on transcript level Oxtoby and Jowett (1993), Thisse et al (2004) Gene function analysis Implantation Test of function for a factor (most often diffusible) via the implantation of a bead saturated with this substance Hyatt et al (1996), Martinez-Morales et al (2005) Morpholino knockdown Test of gene function based on antisense inhibition of its activity Eisen and Smith (2008), Nasevicius and Ekker (2000), Tsujikawa and Malicki (2004a) Overexpression (DNA injections) Test of gene function based on enhancement of its activity through DNA injections Koster and Fraser (2001), Mumm et al (2006) Overexpression (light-mediated RNA/DNA uncaging) Identification of gene function through enhancement of its activity in selected tissues at specific developmental stages Ando et al (2001), …”

Section: Electron Microscopymentioning

confidence: 99%

Analysis of the retina in the zebrafish model

Malicki

Pooranachandran

Nikolaev

et al. 2016

Methods in Cell Biology

View full text Add to dashboard Cite

Subcellular localization of α-tubulin and opsin mRNA in the goldfish retina using digoxigenin-labeled cRNA probes detected by alkaline phosphatase and HRP histochemistry

Cited by 72 publications

References 24 publications

Mechanisms for persistent microphthalmia following ethanol exposure during retinal neurogenesis in zebrafish embryos

Mechanisms for persistent microphthalmia following ethanol exposure during retinal neurogenesis in zebrafish embryos

Developing Neonatal Rat Sympathetic and Sensory Neurons Differ in Their Regulation of 5-HT₃Receptor Expression

Analysis of the retina in the zebrafish model

Contact Info

Product

Resources

About

Subcellular localization of α-tubulin and opsin mRNA in the goldfish retina using digoxigenin-labeled cRNA probes detected by alkaline phosphatase and HRP histochemistry

Cited by 72 publications

References 24 publications

Mechanisms for persistent microphthalmia following ethanol exposure during retinal neurogenesis in zebrafish embryos

Mechanisms for persistent microphthalmia following ethanol exposure during retinal neurogenesis in zebrafish embryos

Developing Neonatal Rat Sympathetic and Sensory Neurons Differ in Their Regulation of 5-HT3Receptor Expression

Analysis of the retina in the zebrafish model

Contact Info

Product

Resources

About

Developing Neonatal Rat Sympathetic and Sensory Neurons Differ in Their Regulation of 5-HT₃Receptor Expression