Authors
Yoh Wada, Minako Aoyama, Ge-Hong Sun-Wada, Nobuyuki Kawamura, and Hiroyuki Tabata
Summary
In this protocol we describe methods for observation endocytic activity in the mouse embryos. The methods are optimised for mouse embryos at E5.5~E7.2 pregastrulation/gastrulation stages. We optimise three different experimental schemes for tracing the embryonic endocytosis. In utero labelling scheme, an endocytic tracer is introduced into circulation of a pregnant mother to follow bulk uptake of fluid phase endocytosis. Rodent embryos are known to internalise maternal immunoglobulins, thus steady-state levels of endocytosis can be visualised by subcellular localization of mouse IgG. We also describe an in vitro labelling method for the isolated embryos. The last method allows pulse-labelling and chase experiments thus one can follow the temporal orders of events. Further, cellular processes involved in the endocytosis can be dissected pharmacologically by applying small- or large molecules with biological activities.Further details
The protocol was published on Protocol Exchange on 14 August 2012. To see the entire protocol, click on the source link.Advertisement
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