In Situ Hybridization in Zebrafish Larvae and Juveniles during Mesonephros Development

10.3791/62930 ◽  
2021 ◽  
Author(s):  
Sagar N. Kasar ◽  
Sarah A. Grandinette ◽  
Scott D. Semelsberger ◽  
Cuong Q. Diep
Keyword(s):  
Zebrafish Larvae ◽  
Larvae And Juveniles

Rhcg1 and Rhbg mediate ammonia excretion by ionocytes and keratinocytes in the skin of zebrafish larvae: H+-ATPase-linked active ammonia excretion by ionocytes

2013 ◽  
Vol 304 (12) ◽  
pp. R1130-R1138 ◽  
Author(s):  
Tin-Han Shih ◽  
Jiun-Lin Horng ◽  
Yi-Ting Lai ◽  
Li-Yih Lin
Keyword(s):  
Ammonia Excretion ◽  
Apical Surface ◽  
Concentration Gradients ◽  
Skin Cells ◽  
Zebrafish Larvae ◽  
Skin Keratinocytes ◽  
Morpholino Oligonucleotides ◽  
Apical Membranes

In zebrafish, Rhcg1 was found in apical membranes of skin ionocytes [H+-ATPase-rich (HR) cells], which are similar to α-type intercalated cells in mammalian collecting ducts. However, the cellular distribution and role of Rhbg in zebrafish larvae have not been well investigated. In addition, HR cells were hypothesized to excrete ammonia against concentration gradients. In this study, we attempted to compare the roles of Rhbg and Rhcg1 in ammonia excretion by larval skin and compare the capability of skin cells to excrete ammonia against concentration gradients. Using in situ hybridization and immunohistochemistry, Rhbg was localized to both apical and basolateral membranes of skin keratinocytes. A scanning ion-selective electrode technique (SIET) was applied to measure the NH4+ flux at the apical surface of keratinocytes and HR cells. Knockdown of Rhbg with morpholino oligonucleotides suppressed ammonia excretion by keratinocytes and induced compensatory ammonia excretion by HR cells. To compare the capability of cells to excrete ammonia against gradients, NH4+ flux of cells was determined in larvae exposed to serial concentrations of external NH4+. Results showed that HR cells excreted NH4+ against higher NH4+ concentration than did keratinocytes. Knockdown of the expression of either Rhcg1 or H+-ATPase in HR cells suppressed the capability of HR cells.

Whole Mount In Situ Hybridization and Immunohistochemistry for Zebrafish Larvae

2018 ◽  
pp. 371-384 ◽  
Author(s):  
Rebecca L. Cunningham ◽  
Kelly R. Monk
Keyword(s):  
In Situ Hybridization ◽  
Zebrafish Larvae ◽  
Whole Mount

scholarly journals C3orf70 Is Involved in Neural and Neurobehavioral Development

2019 ◽  
Vol 12 (4) ◽  
pp. 156 ◽  
Author(s):  
Ashikawa ◽  
Shiromizu ◽  
Miura ◽  
Adachi ◽  
Matsui ◽  
...  
Keyword(s):  
Target Gene ◽  
Molecular Mechanisms ◽  
Neuropsychiatric Disorders ◽  
Neurobehavioral Development ◽  
Zebrafish Larvae ◽  
Mature Neuron ◽  
Human And Mouse ◽  
Comparative Transcriptomic Analysis ◽  
Common Target

Neurogenesis is the process by which undifferentiated progenitor cells develop into mature and functional neurons. Defects in neurogenesis are associated with neurodevelopmental and neuropsychiatric disorders; therefore, elucidating the molecular mechanisms underlying neurogenesis can advance our understanding of the pathophysiology of these disorders and facilitate the discovery of novel therapeutic targets. In this study, we performed a comparative transcriptomic analysis to identify common targets of the proneural transcription factors Neurog1/2 and Ascl1 during neurogenesis of human and mouse stem cells. We successfully identified C3orf70 as a novel common target gene of Neurog1/2 and Ascl1 during neurogenesis. Using in situ hybridization, we demonstrated that c3orf70a and c3orf70b, two orthologs of C3orf70, were expressed in the midbrain and hindbrain of zebrafish larvae. We generated c3orf70 knockout zebrafish using CRISPR/Cas9 technology and demonstrated that loss of c3orf70 resulted in significantly decreased expression of the mature neuron markers elavl3 and eno2. We also found that expression of irx3b, a zebrafish ortholog of IRX3 and a midbrain/hindbrain marker, was significantly reduced in c3orf70 knockout zebrafish. Finally, we demonstrated that neurobehaviors related to circadian rhythm and altered light–dark conditions were significantly impaired in c3orf70 knockout zebrafish. These results suggest that C3orf70 is involved in neural and neurobehavioral development and that defects in C3orf70 may be associated with midbrain/hindbrain-related neurodevelopmental and neuropsychiatric disorders.

Evaluating light trap efficiency by application of mesh to prevent inter- and intra-specific in situ predation on fish larvae and juveniles

2008 ◽  
Vol 93 (1-2) ◽  
pp. 146-153 ◽  
Author(s):  
Lorenzo Vilizzi ◽  
Shaun N. Meredith ◽  
Clayton P. Sharpe ◽  
Rohan Rehwinkel
Keyword(s):  
Fish Larvae ◽  
Light Trap ◽  
Trap Efficiency ◽  
Larvae And Juveniles

scholarly journals Insm1a regulates motor neuron development in zebrafish

2017 ◽  
Author(s):  
Jie Gong ◽  
Xin Wang ◽  
Chenwen Zhu ◽  
Xiaohua Dong ◽  
Qinxin Zhang ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Motor Neuron ◽  
Motor Neurons ◽  
Cell Formation ◽  
Imaging Analysis ◽  
Loss Of Function ◽  
Neuron Development ◽  
Zebrafish Larvae ◽  
Series Of Functions

AbstractInsulinoma-associated1a (Insm1a) is a zinc-finger transcription factor playing a series of functions in cell formation and differentiation of vertebrate central and peripheral nervous systems and neuroendocrine system. However, its roles on the development of motor neuron have still remained uncovered. Here, we provided evidences that insmla was a vital regulator of motor neuron development and provide a mechanistic understanding of how it contributes to this process. Firstly, we showed the localization of insmla in spinal cord and primary motor neurons (PMNs) of zebrafish embryos by in situ hybridization and imaging analysis of transgenic reporter line Tg(insmla: mCherry) ntu805. Then we demonstrated that the deficiency of insmla in zebrafish larvae lead to the defects of PMNs development, including the reduction of caudal primary motor neurons (CaP) and middle primary motor neurons (MiP), the excessive branching of motor axons, and the disorganized distance between adjacent CaPs. Additionally, knockout of insm1 impaired motor neuron differentiation in the spinal cord. Locomotion analysis showed that insmla-null zebrafish significantly reduced the swimming activity. Furthermore, we proved that the insmla loss of function significantly decreased the transcripts levels of both olig2 and nkx6.1. Microinjection of olig2 and nkx6.1 mRNA rescued the motor neuron defects in insmla deficient embryos. Taken together, these data indicate that insmla regulates the motor neuron development, at least in part, through modulation of the expressions of olig2 and nkx6.1.

In vivo Imaging of Fully Active Brain Tissue in Awake Zebrafish Larvae and Juveniles by Skull and Skin Removal

10.3791/62166 ◽  
2021 ◽  
Author(s):  
Paul Schramm ◽  
Florian Hetsch ◽  
Jochen C. Meier ◽  
Reinhard W. Köster
Keyword(s):  
Brain Tissue ◽  
In Vivo Imaging ◽  
Zebrafish Larvae ◽  
Larvae And Juveniles ◽  
Skin Removal

Rendez vous with Saturn's rings

1984 ◽  
Vol 75 ◽  
pp. 743-759 ◽  
Author(s):  
Kerry T. Nock
Keyword(s):  
Solar Energy ◽  
Electric Propulsion ◽  
Power Source ◽  
Propulsion System ◽  
Low Thrust ◽  
Ring Plane ◽  
The Earth ◽  
Total Impulse ◽  
Saturn's Rings

ABSTRACTA mission to rendezvous with the rings of Saturn is studied with regard to science rationale and instrumentation and engineering feasibility and design. Future detailedin situexploration of the rings of Saturn will require spacecraft systems with enormous propulsive capability. NASA is currently studying the critical technologies for just such a system, called Nuclear Electric Propulsion (NEP). Electric propulsion is the only technology which can effectively provide the required total impulse for this demanding mission. Furthermore, the power source must be nuclear because the solar energy reaching Saturn is only 1% of that at the Earth. An important aspect of this mission is the ability of the low thrust propulsion system to continuously boost the spacecraft above the ring plane as it spirals in toward Saturn, thus enabling scientific measurements of ring particles from only a few kilometers.

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