Table 1.
Forward and reverse primers used for quantitative PCR and their specific annealing temperatures.
Table 2.
Forward and reverse primers used for Avp sequencing and their specific annealing temperatures.
Figure 1.
Alterations in behavior and vasopressin (Avp) expression.
Phenotypic measures of HAB/NAB/LAB mice reflecting behavior (A) on the elevated plus-maze (EPM), in the (B) tail-suspension test (TST) and (C) forced swim test (FST) and corresponding vasopressin (Avp) expression patterns as measured by (D) in situ hybridization (ISH) in the paraventricular nucleus (PVN) in male mice and by (E) quantitative PCR (qPCR) in the PVN of male and female mice. Data are shown as means+SEM; * p<0.05; ** p<0.01; *** p<0.001; Mann-Whitney-U test.
Table 3.
Regulation of gene products of the vasopressin and oxytocin systems from the MPI24K platform-based microarray experiment of HAB vs. LAB mice.
Figure 2.
Gene expression of HAB, NAB, LAB, and HABxLAB F1 intercross mice.
Vasopressin (Avp) mRNA expression in the supraoptic nucleus (SON) measured by (A) in situ hybridization (ISH) and (B) quantitative PCR (qPCR) under basal conditions in male and female mice. Data are shown as means+SEM; T p<0.1; * p<0.05; ** p<0.01; Mann-Whitney-U test.
Figure 3.
Vasopressin (AVP) immunohistochemistry analysis.
AVP fluorescence antibody-staining of the paraventricular nucleus (PVN) flanking the 3rd ventricle (3V) in HAB and LAB mice. For corresponding in situ hybridization, see Fig. 7.
Figure 4.
Oxytocin (Oxt) mRNA expression.
Oxt in the paraventricular nucleus (PVN) measured by in situ hybridization under basal conditions in male HAB, NAB and LAB mice. Data are shown as means+SEM.
Figure 5.
Vasopressin (Avp) gene sequence of HAB vs. LAB mice.
Polymorphic sites are indicated with positions from transcription start (−1 to −2600 bp) in (A) the promoter region (two SNPs and deletion in LABs); (B) within the Avp coding sequence from transcription start (1 to 1960 bp; three SNPs); (C) in the intergenic region between Avp and oxytocin (Oxt) from the end of the last exon (+1 to +2600 bp; three SNPs). Exons and untranslated regions (UTRs) are indicated by boxes (exons shaded, UTRs completely filled black or white).
Table 4.
Polymorphisms in the promoter and downstream enhancer region between the HAB and LAB specific sequence with probable binding factors.
Table 5.
Allele frequency in a NAB population (n = 165) for the SNP in the Avp signal peptide and the strictly linked promoter deletion.
Figure 6.
Allele-specific vasopressin (Avp) expression.
Proportion of HAB allele-specific vs. LAB allele-specific Avp transcripts from heterozygous F1 (HABxLAB intercross) mice in the paraventricular nucleus (PVN; χ2 = 14.4 p = 1.4e-4) and the supraoptic nucleus (SON; χ2 = 15.2 p = 4.8e-5). Data are shown as means+SEM; *** p<0.001.
Figure 7.
Correlation between gene expression and behavior.
Correlation of vasopressin (Avp) mRNA expression in the paraventricular nucleus (PVN) with (A) anxiety-related behavior and (B) depression-like behavior of HAB, F1 and LAB male mice under basal conditions. For corresponding immunohistochemistry, see Fig. 3.
Figure 8.
Association between anxiety-related behavior and the Avp-related genotypes in the freely-segregating F2 panel.
Phenotypic indices of male F2 mice with HAB grandmothers and Avp-related genotypes (+/+) and CC (HAB-typical homozygotes, n = 67), (+/−) and CT (heterozygotes, n = 142), or (−/−) and TT (LAB-typical homozygotes, n = 49). (A) % time spent on the open arms of the elevated plus-maze (EPM) and (B) total distance traveled in the open field (OF). LAB-typical homozygotes were less anxious, independent of locomotor activity. Data are shown as means+SEM; * p<0.05.
Table 6.
Allele frequency in a freely-segregating F2 panel (HABxLAB intercross offspring, n = 508) for the SNP in the Avp signal peptide and the strictly linked promoter deletion.