Effects of ciliary neuronotrophic factor on rat spinal cord neurons in vitro: survival and expression of choline acetyltransferase and low-affinity nerve growth factor receptors.

We have studied the effects of ciliary neuronotrophic factor (CNTF) and nerve growth factor (NGF) on cultures of E14 rat spinal cord cells maintained for 7 days. The trophic factors were supplied at the day of seeding and every other day thereafter. Treatments with CNTF (human recombinant or purified from rat sciatic nerve, 100 TU/ml) resulted after 7 days in an increase, relative to control cultures, of: (i) the total number of neurons (identified by neurofilament protein and neuron-specific enolase immunostaining) that were not stained with choline, acetyltransferase (ChAT) and low affinity nerve growth factor receptor (LNGFR) antibodies; (ii) the number of motoneurons (0.5% of the neuronal population) as identified by size (greater than 25 microns), morphology and immunostaining for ChAT and LNGFR; and (iii) a population of small- to medium-sized (less than 25 microns), ChAT- and LNGFR-positive neurons, representing 5-10% of the total neuronal population. NGF treatments (mouse submaxillary beta NGF; 10-3000 TU/ml) were without effect on all 3 neuronal populations. Experiments in which CNTF administration was delayed revealed that the population of ChAT- and LNGFR-negative neurons and the population of motoneurons, were both dependent on CNTF for their survival. The third population, small ChAT and LNGFR-positive neurons, was not dependent on CNTF for survival but was induced by CNTF to express its two markers. These observations indicate that CNTF is a neuronotrophic factor for motoneurons, but that the effect of CNTF is not restricted to that cell population. In addition to its survival promoting effect, CNTF has also a regulatory role on the expression of ChAT and LNGFR for some spinal cord neurons.