Transgenic Xenopus : Microinjection Methods and by Shlomo Seidman, Hermona Soreq

By Shlomo Seidman, Hermona Soreq

The have to greater comprehend the molecular, b- chemical, and mobile procedures through which a constructing neuronal process unfolds has resulted in the improvement of a different set of experimental instruments and organisms. designated emphasis was once dedicated to permitting us entry, on the ear- est levels, to the genomic foundation underlying the system’s final complexity, as exhibited as soon as its constructions are totally shaped. but, nerve cells are anatomically, physiolo- cally, and biochemically varied. The multitude of d- tinctly diversified routes for his or her improvement therefore makes the constructing anxious approach specially exciting for molecular neurobiologists. particularly, the calls for of contemporary molecular neuroscience demand the institution of effective but flexible platforms for learning those c- plex methods. Transgenic embryos of the frog Xenopus laevis provide a good method for drawing close neuroscientific concerns. Insertion of international genes is played easily, through mic- injection below binocular statement; enormous quantities of in vitro-fertilized embryos might be microinjected in a single scan. Embryos improve in faucet water, at room t- perature, and inside of a couple of days develop into self reliant swimming tadpoles with totally functioning neuromus- lar structures. Being quite small, those organisms are amenable to designated analyses on the degrees of mRNA, protein, and mobile. Their speedy improvement allows the learn of morphogenetic procedures fascinated with early improvement, reminiscent of myogenesis and neural induction, in addition to these keen on organogenesis and formation of the mind, the musculature, and the interconnections among them. international DNA continues to be predominantly extrachromosomal.

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Extra resources for Transgenic Xenopus : Microinjection Methods and Developmental Neurobiology

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A) Scanning electron micrograph of lizard intercostal muscle NMJ. In this photograph, the nerve has been removed to reveal topological features of the intact postsynaptic membrane. The oval shape of the lizard NMJ is similar to that of the mammalian junction (magnification x4000). Size bar, 4 pm. Reprinted from Bartol et al. (1991) with permission from BiophysicalJournal. (B) Transmission electron micrograph of a cross-section from a lizard NMJ. Note localization of AChR (arrows) at the crests of the juctional folds, in close apposition to the presynaptic nerve terminal.

8. Electron microscope view of the intersomitic junction. Longitudinal section of myotomal muscle from a 2-d-old Xenopusembryo (stage 25) stained for catalytically active AChE as described in Experimental Methodologies in Chapter 2. Sites of AChE accumulation appear as black crystals; note that parallel arrays of muscle cells (m) with their already well-developed myofibrils (mf) abut at the intersomitic junction (ISJ), which is characterized by the deposition of fibrous extracellular material and high accumulations of AChE.

Appearance of presynaptic active zones. , 1977). , 1992). In culture, noninnervated myotomal cells display colocalized patches of AChE and AChR, which occupy about 3% of the cell surface (Moody-Corbett and Cohen, 1981). In addition, noninnervated cells exhibit localized ultrastructural specializations resembling those of the postsynaptic membrane. When myotomal muscle cells are cocultured Fig. 10. Developing Xenopus NMJs. Electron micrographs of representative NMJs found in myotomes oi 2-3-d-old Xenopus embryos.

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