Differences in structure-function relations between nonmammalian and mammalian gonadotropin-releasing hormone receptors

Mammalian gonadotropin-releasing hormone receptors (GnRH-Rs) differ from other G protein-coupled receptors in lacking the intracellular C-terminus and in showing an exchange of two otherwise highly conserved Asp (D) and Asn (N) residues in transmembrane domains (TMD) 2 and 7, respectively. However, the first GnRH-R characterized from a nonmammalian vertebrate, the African catfish, does contain an intracellular C-terminus and has D residues in TMD 2 and 7. The functional relevance of these structural features was analysed with

Characterization of neocortical and hippocampal synaptosomes from temporal lobe epilepsy patients

To investigate epilepsy-associated changes in the presynaptic terminal, we isolated and characterized synaptosomes from biopsies resected during surgical treatment of drug-resistant temporal lobe epilepsy (TLE) patients. Our main findings are: (1) The yield of synaptosomal protein from biopsies of epilepsy patients was about 25% of that from rat brain. Synaptosomal preparations were essentially free of glial contaminations. (2) Synaptosomes from TLE patients and naive rat brain, quickly responded to K+-depolarization with a 70% increase in intrasynaptosomal Ca2+ ([Ca2+]i), and a 40% increase in B-50/GAP-43 phosphorylation. (3) Neocortical and hippocampal synaptosomes from TLE patients contained 20 50% of the glutamate and -aminobutyric acid (GABA) contents of rat cortical synaptosomes. (4) Although the absolute amount of glutamate and GABA released under basal conditions from neocortical synaptosomes of TLE patients was lower than from rat synaptosomes, basal release expressed as percentage of total content was higher (16.4% and 17.3%, respectively) than in rat (11.5% and 9.9%, respectively). (5) Depolarization-induced glutamate and GABA release from neocortical synaptosomes from TLE patients was smaller than from rat synaptosomes (3.9% and 13.0% vs. 21.9% and 25.0%, respectively). (6) Analysis of breakdown of glial fibrillary acid protein (GFAP) indicates that resection time (anoxic period during the operation) is a critical parameter for the quality of the synaptosomes. We conclude that highly pure and viable synaptosomes can be isolated even from highly sclerotic human epileptic tissue. Our data show that in studies on human synaptosomes it is of critical importance to distinguish methodological (i.e., resection time) from pathology-related abnormalities