Ag-staining pattern, FISH and ISH with rDNA probes in the rice field eel (Monopterus albus Zuiew) chromosomes.

The present paper reports the Results of rice field eel (Monopterus albus Zuiew) chromosomal analysis by Ag-staining techniques, FISH and ISH with 18S rDNA. Both the Ag-NORs and the hybridization signals were observed on the pericentromeric region of chromosome pairs 3 and 7. The Ag-NORs and the hybridization signals on homologous chromosomes 3 and 7 were corresponding to each other. And rDNA genes were mapped to bivalents 3q12-q24 and 7q14-q26. No heteromorphic sex chromosome pair was identified. The Results obtained were discussed with respect to the feature of Ag-NORs and the position of rDNA genes on chromosome pairs 3 and 7.

Postnatal development of a GABA deficit and disturbance of neural functions in mice lacking GAD65.

The 65-kDa isoform of glutamic acid decarboxylase (GAD65) is believed to play an essential role for GABA synthesis in the central nervous system. Using mice with targeted disruption of the GAD65 gene (GAD65(-/-) mice) we investigated the contribution of GAD65 to GABA synthesis in different brain areas during postnatal development and in adulthood. In the amygdala, hypothalamus and parietal cortex of GAD65(+/+) mice an increase of GABA levels was observed during postnatal development, most prominently between the first and second month after birth. This increase appeared to be dependent on GAD65, as it was delayed by 2 months in GAD65(+/-) mice and was not observed in GAD65(-/-) mice. Likely as a consequence of their GABA deficit, adult GAD65(-/-) mice showed a largely abnormal neural activity with frequent paroxysmal discharges and spontaneous seizures. They furthermore displayed increased anxiety-like behaviour in a light/dark avoidance test and reduced intermale aggression, as well as a reduced forced-swimming-induced immobility indicative of an antidepressant-like behavioural change. Adult GAD65(+/-) mice did not show behavioural disturbances except for a reduced aggressive behaviour that was comparable to that in GAD65(-/-) mice. We conclude that GAD65-mediated GABA synthesis may be crucially involved in control of emotional behaviour and indispensable for a tonic inhibition that prevents the development of hyperexcitability in the maturating central nervous system. Aggressive, and possibly other social behaviour may be especially prone to regulation through GAD65-mediated GABA synthesis.

Synaptic localization of the 67,000 mol. wt isoform of glutamate decarboxylase and transmitter function of GABA in the mouse cerebellum lacking the 65,000 mol. wt isoform.

Subcellular localization of the 67,000 mol. wt isoform of glutamate decarboxylase and neurotransmitter function of GABA were investigated in the cerebellum of the mice lacking the 65,000 mol. wt isoform of glutamate decarboxylase. The GABA content decreased by 25% in the cerebellum. Putative GABA-releasing terminals from basket/stellate and Golgi cells were immunostained with glutamate decarboxylase-67 antibody. Basket cell-derived inhibitory postsynaptic currents in Purkinje cells and the high potassium-induced release of GABA were not significantly affected. Although previous investigations have suggested that glutamate decarboxylase-65 is mainly involved in transmitter synthesis and that glutamate decarboxylase-67 is transported to the nerve terminals only after association with glutamate decarboxylase-65, the present results indicate that glutamate decarboxylase-67 is independently concentrated in the nerve terminals and provides GABA for synaptic transmission in the absence of glutamate decarboxylase-65.

[Haplotype identification of single sperm in pigs].

Haplotype of porcine single sperm was analyzed by PCR method through using primer extension preamplification and heminesting primer design strategy. The PCR products were run on 8% polyacrylamide gel, and visualized by silver staining method. The results showed that the haplotypes of individual sperm can be unequivocally demonstrated by these methods. The application of single sperm typing provides a unique tool for the construction of high resolution genetic map and the study of genetic phenomena requiring large sample size.

Mice lacking the 65 kDa isoform of glutamic acid decarboxylase (GAD65) maintain normal levels of GAD67 and GABA in their brains but are susceptible to seizures.

The gene encoding of the 65 kDa isoform of the gamma-aminobutyric acid (GABA)-synthesizing enzyme, glutamic acid decarboxylase (GAD), GAD65, was targeted in mice by homologous recombination. Viable GAD65 -/- mice were obtained with the expected mendelian frequency and displayed no gross morphological defects. Despite the complete loss of GAD65 mRNA and protein in a homozygous mutant, there was no difference in GABA content in the brains of GAD65 +/+, +/-, and -/- mice. As for the other 67 kDa isoform (GAD67), the levels of mRNA and protein were largely unchanged by the GAD65 mutation. General behavior, including locomotor activity and performance in the Morris water maze task, appeared normal, but seizures were more easily induced by picrotoxin and pentylenetetrazol: the latencies to seizures induced by picrotoxin were shorter and the dose of pentylenetetrazol required for induction of seizures was lower.