Species: Diplostomum pseudospathaceum Niewiadomska, 1984    Trematoda

Preparation Type: whole mount

Prepared: A.A. Petrov

Description: Substance P-immunoreactive nervous elements и the musculature of a 22-day-old metacercaria of Diplostomum pseudospathaceum (Trematoda, Platyhelminthes). Overall view of the metacercaria showing the distribution of substance P in the brain, anterior nerves, ventral nerve cords and in the nerves innervating the hindbody and the tegument. The preparation also shows the musculature of the body wall and internal organs (oral and ventral suckers, prepharynx, pharynx, intestine and holdfast).

Method: F-actin in muscle fibers (Phalloidin), substance P

Method Description: The metacercariae were fixed for 15-17 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 1-2 hours 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 16 hours in antibodies to Substance P (diluted 1:400, ref. number S1542; Sigma-Aldrich), rinsed 4 times for 15 minutes in 0.5% Triton X-100 in 0.01M PBS and then incubated for 8 hours in secondary antibodies (diluted 1: 300, Chromeo 488, ref. number ab60314; Abcam). After incubation in antibodies, the worms were stained for 3 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 Vectashield (Vector Laboratories Inc.).

Institution: Zoological Institute RAS

Description: General view of a 22-day-old metacercaria of Diplostomum pseudospathaceum (Trematoda, Platyhelminthes) showing the musculature and substance P-immunoreactive nervous elements. The series shows nervous elements in the cerebral ganglia, in the anterior nerves innervating the oral sucker, in the ventral longitudinal nerve cords and in the neurons innervating the tegument and hindbody.

Organs: сentral nervous system, nerve cord, cerebral brain or its part, nerve, neuron(s), larval nervous system, musculature innervation / musculature, peripheral nervous system

Methods: Immunohistochemistry - substance P,
Histochemistry - F-actin in muscle fibers (Phalloidin)