The autonomic nerve supply of the bovine testis is investigated in ani
mals of different ages by means of immunohistochemistry. Staining with
antiserum to protein gene product 9.5 gives the most complete results
for the study of the general innervation pattern. Autonomic nerves re
ach the testis by three different routes: with the blood vessels of th
e spermatic cord (funicular nervous contribution), by the mesorchium (
mesorchial nervous contribution) and by the ligamentous bridge between
epididymal tail and testis (caudal nervous contribution). The vessels
of the spermatic cord are densely innervated. The large vessels of th
e vascular layer within the tunica albuginea display a discontinuous i
nnervation pattern. In the interior of the testis, the caudal half of
the gonad is completely free of any innervation. Slight differences in
arrangement and fiber composition of testicular nerves in calves and
bulls point to a reduction of the innervation with advancing age. The
vast majority of bovine testicular nerves are dopamine-beta-hydroxylas
e-positive postganglionic sympathetic axons with vasomotor function, T
here is no evidence for a cholinergic innervation of the bovine testis
. About half of the bovine testicular nerves are neuropeptide Y-immuno
reactive. In the adult, solitary calcitonin gene-related peptide-immun
oreactive fibers are the only ones independent of blood vessels. The a
bsence of an innervation in the caudal half of the testis underlines t
he importance of local factors and bloodborne substances for the regul
ation of intratesticular blood flow in the bovine.