Real-time PCR efficiencies for each reaction varied from 99% to 109%, and the correlation coefficient was not lower than 0.99. Real-time data were collected and analyzed in Excel. The relative amount of Oxt, FosB, and

Peg3 transcripts between SM/J and LG/J samples was calculated according to Vandesompele et al. (2002), as previously described (Chiavegatto et al. 2010). The following genes were analyzed as controls: cyclophilin A (peptidylprolyl isomerase A: Ppia), hypoxanthine guanine phosphoribosyl transferase 1 (Hprt1), and beta-actin Inhibitors,research,lifescience,medical (Actb). Primers for candidate and control genes were designed in different exons when possible (Table 2), according to criteria detailed elsewhere (Bibancos et al. 2007). Table 2 Forward and reverse primers sequences for hypothalamic RNA selleck expression Statistical analysis Behavioral data were compared using Inhibitors,research,lifescience,medical a two-tailed Student’s t test, and the associations among nominal variables were tested by cross tabulation using a Pearson χ2 test and ϕ coefficient in SYSTAT 10.0. The base-calling

quality for Oxt, FosB, and Peg3 was visually inspected using Chromas software (http://www.technelysium.com.au/chromas.html. Forward and reverse sequences for each gene region were manually evaluated, aligned, and compared Inhibitors,research,lifescience,medical between SM/J and LG/J strains. These analyses were also performed using the BioEdit Sequence Alignment Editor (Hall 1999). The GenBank (NCBI) accession numbers for SM/J and LG/J gene sequences are, respectively, “type”:”entrez-nucleotide”,”attrs”:”text”:”HQ679943″,”term_id”:”371444794″HQ679943

and “type”:”entrez-nucleotide”,”attrs”:”text”:”HQ679944″,”term_id”:”371444796″HQ679944 (Oxt), “type”:”entrez-nucleotide”,”attrs”:”text”:”HQ679939″,”term_id”:”371444787″HQ679939 and “type”:”entrez-nucleotide”,”attrs”:”text”:”HQ679940″,”term_id”:”371444791″HQ679940 Inhibitors,research,lifescience,medical (FosB), “type”:”entrez-nucleotide”,”attrs”:”text”:”HQ679941″,”term_id”:”371444792″HQ679941 and Inhibitors,research,lifescience,medical “type”:”entrez-nucleotide”,”attrs”:”text”:”HQ679942″,”term_id”:”371444793″HQ679942 (Peg3). The association between maternal care (absence or presence) and directly genotypes for the exon 9 Peg3 marker in F2 females was investigated using standard analysis of variance (ANOVA)-–General Linear Model in SAS, v.9.0 (SAS, 2004). Transcript quantities were tested for a normal distribution AV-951 (Kolmogorov–Smirnov test) and compared using a two-tailed Student’s t test (GraphPad InStat® version 3.05, San Diego, CA). Data were expressed as mean ± standard error of the mean (SEM) or median and range. Differences were considered statistically significant when P < 0.05. Results LG/J females have poorer maternal performance when compared to SM/J females SM/J and LG/J females display distinctive levels of maternal performance (Fig. 2). Although both females usually built a prepartum nest and maintained it after giving birth, only SM/J mothers displayed a more sophisticated postpartum nest (ϕ= 0.