Spinal neurons involved in the control of the seminal vesicles: A transsynaptic labeling study using pseudorabies virus in rats

Abstract

The seminal vesicles are male accessory sex glands that mainly contribute the seminal fluid of the ejaculate. Previous studies have suggested that seminal vesicles are supplied by both sympathetic and parasympathetic nerves. However, this conclusion was mainly based on studies in pelvic major ganglions and direct neuroanatomical evidence of spinal neurons innervating the seminal vesicles is still lacking. In order to map the spinal nerve circuit innervating the seminal vesicles, the present study used the pseudorabies virus (PRV) retrograde tracing technique in combination with immunohistochemistry. Three groups of rats were prepared: (1) nerves intact; (2) right hypogastric nerve and bilateral pelvic nerves sectioned; (3) right pelvic and bilateral hypogastric nerves sectioned. For the intact group, 3 to 5 days after injection of PRV into the left seminal vesicle in male rats, immunohistochemistry for PRV was performed to map the control circuit. Double immunofluorescence experiments against PRV and choline acetyltransferase (ChAT) were performed to discriminate preganglionic neurons and interneurons. Double detection of PRV and galanin (GAL) was also performed to identify lumbar spinothalamic (LSt) cells. Three days after virus injection, both sympathetic and parasympathetic preganglionic neurons were retrograde-labeled. Four days after injection of PRV into the seminal vesicles, PRV-infected neurons were found in the dorsal horn, ventral horn, dorsal gray commissure (DGC), medial gray matter and intermediolateral cell column (IML) from T13 to S1. For the group with an intact hypogastric nerve, 4 days after injection of PRV into the seminal vesicles, PRV-infected neurons were mainly located in DGC and IML of spinal lumbar segments (L) 1–L2. However, in the group with an intact pelvic nerve, PRV-infected neurons were mainly located in DGC of L5–S1 spinal segments. At the L3–L4 level, most of the virus-labeled neurons around the central canal expressed immunoreactivity for GAL, strongly suggesting that they could be LSt cells. These anatomical data support the idea that the sympathetic and parasympathetic nervous system are both involved in the control of the seminal vesicles and we demonstrated a connection between preganglionic neurons innervating the seminal vesicles and LSt cells which play a crucial role in coordinating the spinal control of ejaculation.