Species: Protodrilus flavocapitatus (Uljanin, 1877)    Annelida

Preparation Type: whole mount

Prepared: A.A. Petrov

Description: Nervous system and the musculature of the polychete Protodrilus flavocapitatus (Polychaeta, Annelida). The nervous system was stained with antibodies to acetylated alpha-tubulin and FMRFamide. The preparation contains a region of the body corresponding to four segments. It shows the ventral nerve cord, the nerves that extend from the cord, the musculature of the body wall and setae, and cilia in the protonephridial canals.

Method: acetylated alpha-tubulin, F-actin in muscle fibers (Phalloidin), FMRFamide

Method Description: The worms were fixed for 6 hours with 4% formaldehyde in 0.01M PBS and were kept at 6°С in 0.1% sodium azide in 0.01М PBS. All subsequent procedures were performed at room temperature. Before staining, the worms were processed for 2 hours in 0.5% Triton X-100 in 0.01M PBS, incubated for 1 hour in a blocking solution (Image-iT® FX Signal Enhancer) and rinsed 3 times for 15 minutes in 0.5% Triton X-100 in 0.01M PBS. After rinsing, the worms were incubated for 12 hours in a mixture (1:1) of mouse antibodies to acetylated alpha-tubulin (diluted 1:250, ref. number T7451; Sigma-Aldrich) and rabbit antibodies to FMRFamide (diluted 1:250, ref. number ab10352; Abcam), rinsed 4 times for 15 minutes in 0.5% Triton X-100 in 0.01M PBS and then incubated for 5 hours in a mixture (1:1) of antibodies to mouse IgG (diluted 1: 300, Alexa Fluor 633, ref. number 1A-21052; Thermo Fisher Scientific) and antibodies to rabbit IgG (diluted 1:300, Chromeo 488; ref. number ab630314; Abcam). After incubation in antibodies, the worms were rinsed for 15 minutes in 0.5% Triton X-100 in 0.01M PBS, stained for 2 hours in phalloidin-TRITC (diluted 1:150, Sigma-Aldrich, ref. number P1951; Sigma-Aldrich), rinsed for 15 minutes in 0.01M PBS and mounted in 80% glycerol in 0.01M PBS.

Institution: Zoological Institute RAS

Description: Series of images of a body region of the polychete Protodrilus flavocapitatus (Polychaeta, Annelida) showing the muscles of the body wall and setae and the nervous system stained with antibodies to acetylated alpha-tubulin and FMRFamide. The series clearly shows two ventral nerve cords, the trasverse and diagonal commissures interconnecting the cords and the lateral nerves extending from the cords. The images also show cilia in the protonephridial canals.

Organs: сentral nervous system, nerve cord, ganglion / ganglia, connective, commissure, nerve, neuron(s), musculature innervation / musculature, peripheral nervous system

Methods: Immunohistochemistry - acetylated alpha-tubulin,
Immunohistochemistry - FMRFamide,
Histochemistry - F-actin in muscle fibers (Phalloidin)