Worm Breeder's Gazette 11(2): 101

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

How do the HSNs Decide When to Differentiate?

Gian Garriga and Bob Horvitz

Although the HSN neurons are born embryonically, they do not appear 
to differentiate fully until the L4 stage (Desai et al., 1988).  How 
is the timing of HSN differentiation controlled? One possibility is 
that the HSNs contain an internal clock that is set to go off in L4 
animals.  Alternatively, the HSNs could differentiate in response to 
an external cue.
The timing of many developmental events in C.  elegans is controlled 
by the heterochronic genes (Ambros and Horvitz, 1984).  Some mutations 
in these genes lead to precocious development, causing certain events 
to occur prematurely.  Other mutations lead to retarded development, 
causing certain events to occur late.  To determine if the 
heterochronic genes are important for HSN differentiation, we examined 
two aspects of HSN development in heterochronic mutants: the timing of 
hood formation and the timing of serotonin synthesis.  The HSN hood is 
an eye-shaped structure that surrounds the HSN nucleus in L4 
hermaphrodites and can be observed using Nomarski optics.  Serotonin 
is an HSN neurotransmitter that becomes detectable 
immunocytochemically in young adult hermaphrodites.
The timing of HSN differentiation is perturbed in heterochronic 
mutants.  In the precocious mutants lin-14(n179) and lin-28(n719), the 
HSNs differentiate early.  Hood formation occurs during the L3 stage, 
and the HSNs synthesize detectable serotonin in L4 animals.  By 
contrast, the HSNs never differentiate in the retarded mutants lin-4(
e912) and lin-14(e536sd).  The HSNs appear normal in these mutants 
until the L4 stage, when either they fail to form a hood or they 
become indistinct and difficult to find.  The lack of 
immunocytochemically detectable HSN serotonin in these mutants 
corroborates the Nomarski results.  These HSN phenotypes are the same 
defects seen in the HSN maturation mutants, egl-45, 
t al., 1988; M.  Basson, personal 
communication).  In these mutants, the HSNs fail to differentiate.
In contrast to the other retarded mutants, lin-29(n333) animals are 
normal for HSN hood formation; however, the HSNs often lack 
immunocytochemically-detectable serotonin.  The only other event known 
to be affected by lin-29 mutations is the switch from larval to adult 
cuticle, which is blocked in this mutant (Ambros and Horvitz, 1984).  
Thus, lin-29 may be required specifically for late aspects of 
development, including late stages of HSN differentiation.
What do these results say about HSN differentiation? Among the 
heterochronic genes, lin-14 plays a key role, with the level of lin-14 
gene activity determining the timing of developmental events (Ambros 
and Horvitz, 1987).  The postembryonic HSNs do not contain detectable 
lin-14 protein as determined by staining with anti-lin-14 antiserum (
Ruvkun and Guisto, 1989).  This observation suggests that the time of 
onset of HSN differentiation is controlled by an external cue or cues, 
presumably presented to the HSNs by cells expressing the lin-14 
protein.