The development of facial motoneurones in the mouse--neuronal death and the innervation of the facial muscles.

The relationship between neuronal death and the formation of patterned connections was studied in the facial neuromuscular system of foetal, neonatal, and adult mice. The facial neuromuscular system was selected because two large, widely separated, facial muscles (the nasolabial and posterior auricular muscles) are innervated by clearly separated parts of the facial motor nucleus in the adult mouse. The number of motorneurones in the facial nucleus was counted in Nissl-stained sections at different stages of development. Over 6400 neurones were present in the facial nucleus at day 17 p.c. (post-coitum). After day 17 p.c. the number of neurones fell rapidly and only 2000 cells remain in the adult nucleus. This represents a loss of 68%, most of which occurs between days 18 and 20 p.c. Neurones with pyknotic nuclei are seen on day 17 p.c. and are most numerous on day 18 p.c. This leads us to believe that the fall in neurone numbers is due to cell death. Indirect evidence provided by acetylcholinesterase histochemistry (time of earliest reaction in the facial muscles) and horseradish peroxide (HRP) tracing studies (time of earliest transport) indicate that facial motorneurone axons innervate the facial musculature before the period of cell death: diffuse acetylcholinesterase activity first appeared in the auricular muscles at day 15 p.c. and in the nasolabial muscles at day 17 p.c.; retrograde transport of HRP from the auricular and nasolabial muscles to the facial nucleus cannot be reliably demonstrated before day 17 p.c. We assessed the topography of early facial neuromuscular innervation by making HRP injections into nasolabial and posterior auricular muscles of embryonic and neonatal mice. Injections of HRP at day 17 p.c. (the day before cell loss commenced) showed that the nasolabial muscle and posterior auricular muscles were innervated by the same subnuclei of the facial nucleus as in the adult--except that there was a small number (1-5%) of labelled cells located in parts of the nucleus not consistent with the adult innervation pattern. These data indicate that, except for a small number of neurones, topographically organized connections in the facial neuromuscular system are established before the period of cell death. We conclude that motoneurone cell death does not play a major role in the establishment of topographically organized connections in this system.