The trigeminally evoked blink reflex : I. Neuronal circuits

Tetramethyl benzidine (TMB) is a presumptively non-carcinogenic chromogen which yields a blue reaction-product at sites of horseradish peroxidase activity. Sixty-six distinct procedures were performed in rats and monkeys in order to determine the optimal incubation parameters for TMB. As a result, a procedure is recommended whose sensitivity greatly surpasses that of a previously described benzidine dihydrochloride method. Indeed, the sensitivity of this new method in demonstrating retrograde transport is markedly superior to that of the previously described benzidine dihydrochloride method. Furthermore, as a consequence of this enhanced sensitivity, many efferent connections of the injection site are also visualized. The injection site demonstrated by this TMB procedure is significantly larger than the one demonstrated when benzidine dihydrochloride or diaminobenzidine is used as a chromogen. Finally, this TMB procedure has been compared to two other TMB procedures and found to provide superior morphology and sensitivity.

The central projections of five peripheral branches of the trigeminal nerve were investigated by the method of transganglionic transport of horseradish peroxidase (HRP). In separate animals, the corneal, supraorbital, infraorbital, mental, or inferior alveolar branches were transected and soaked in concentrated solutions of HRP. Forty-eight to 72 hours after surgery, the brain-stem, upper cervical spinal cord, and trigeminal ganglia were perfusion-fixed and processed according to the tetramethylbenzidine technique. The results show that trigeminal primary afferent neurons which innervate the cornea project mainly to the levels of caudal pars interpolaris and caudal pars caudalis. In contrast, trigeminal primary afferent neurons whose peripheral processes course through the supraorbital, infraorbital, or mental nerves project most heavily to the trigeminal main sensory nucleus, pars interpolaris, and the rostrocaudal middle three-fifths of pars caudalis. Trigeminal primary afferent neurons which give origin to the inferior alveolar nerve project heavily and in approximately equal numbers of all rostrocaudal levels of the trigeminal brainstem nuclear complex (TBNC). A small number of fibers from each of the latter four cell populations project directly to the contralateral C1-C2 dorsal horn. A small number of fibers from each cell population studied end in the reticular formation immediately adjacent to the spinal nucleus of V. It is concluded that the cornea and facial skin regions of the cat are represented nonuniformly along the rostrocaudal length of the TBNC.