Media 2 - Motor neurons in L6-S1 spinal cord projecting to the rat major pelvic ganglion

<p dir="ltr">Retrograde labelling of motor and premotor neurons in the <b>lumbosacral (L6-S1) </b>spinal cord that project to the <b>major pelvic ganglia</b> <b>(MPG)</b>. Microinjection of the retrograde tracer <b>cholera toxin subunit B (CTB)</b> into the MPG of a male rat was taken up by sacral preganglionic neurons in the <b>intermediolateral column (IML)</b>. Somatic motor neurons were also retrogradely labelled with CTB, those innervating the <b>urethral rhabdosphincter (ExU9)</b>, as well as motor neurons innervating the <b>tail and levator ani (Tail9)</b>.</p><p dir="ltr">Visualization of motor neurons was achieved through whole tissue immunolabelling and clearing using a modified form of <b>iDISCO </b>for spinal cord. Motor neurons were immunolabelled via <b>choline acetyltransferase (ChAT)</b>, while retrogradely-filled neurons were immunolabelled for CTB. Large volume imaging was performed on the <b>light sheet microscope</b> with a 12x lens (Ultramicroscope II, Miltenyi Biotec). Dataset was visualized in <b>Imaris</b> (Bitplane).</p>