Characterisation of the human GFRalpha-3 locus and investigation of the gene in Hirschsprung disease.

BACKGROUND: The GDNF family receptor alpha (GFRalpha) proteins are extracellular cell surface bound molecules that act as adapters in binding of the GDNF family of soluble neurotrophic factors to the RET receptor. These molecules are essential for development of many neural crest derived cell types and the kidney. Mutations in RET and in two members of the GDNF ligand family are associated with Hirschsprung disease (HSCR), a congenital absence of the enteric ganglia. Members of the GFRalpha family are also candidates for HSCR mutations. One such gene is GFRalpha-3, which is expressed in the peripheral nervous system and developing nerves. OBJECTIVE: We have characterised the structure of the human GFRalpha-3 locus and investigated the gene for sequence variants in a panel of HSCR patients. METHODS: Long range PCR or subcloning of PAC clones was used to investigate GFRalpha-3 intron-exon boundaries. A combination of single strand conformation polymorphism (SSCP) analysis and direct sequencing was used to investigate GFRalpha-3 sequence variants. RESULTS: GFRalpha-3 spans eight coding exons and has a gene structure and organisation similar to that of GFRalpha-1. We identified three polymorphic variants in GFRalpha-3 in a normal control population, a subset of which also occurred in HSCR patients. We did not detect any sequence variants within the coding sequence of GFRalpha-3. We found a base substitution in the 5' UTR of GFRalpha-3, 15 base pairs upstream of the translation start site. A second substitution was identified in intron 4 (IVS4-30G>A) between the splice branch site and the splice acceptor site. The final variant was a 2 base pair insertion within the splice donor consensus sequence of exon 7 (IVS7+4ins GG). CONCLUSIONS: We did not detect any correlation between variants of GFRalpha-3 and the HSCR phenotype. Our data suggest that mutations of this gene are not a cause of HSCR.

Prepulse inhibition following lesions of the inferior colliculus: prepulse intensity functions.

The magnitude of the acoustic startle response can be reduced by a relatively weak sound presented immediately before the startle-eliciting sound; this phenomenon has been termed prepulse inhibition (PPI). Previous studies reported that PPI was present in the decerebrate rat, indicating that the primary neural pathways mediating PPI are located in the brainstem. The present study investigated the effects of focal excitotoxic lesions of the inferior colliculus (IC) on acoustic PPI in rats. In the first part, startle magnitudes were measured in six normal rats as the interstimulus interval (ISI) between the prepulse and startle-eliciting sounds varied between 10 and 100 ms. Prepulse-inhibited startle changed in an ISI-dependent manner with the most effective ISI at 50 ms. In the second part, 21 rats were assigned to three groups: normal unoperated, cortical lesion, and IC lesion. With the ISI fixed at 50 ms, as the prepulse sound level increased from 29 to 49 dB SPL, startle responses decreased quickly in both normal and cortical lesion rats. However, rats with unilateral IC lesions made with ibotenic acid had significantly lower PPI but did not display any increase in startle magnitude. These data suggest that the IC is an important structure in the neural circuit mediating acoustic PPI.