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. 2022 Feb 24;23(5):2518.
doi: 10.3390/ijms23052518.

Transcriptomics of Parental Care in the Hypothalamic-Septal Region of Female Zebra Finch Brain

Rashmi Kumari  1   2 , Emese A Fazekas  1   3 , Boglárka Morvai  3 , Edina B Udvari  1 , Fanni Dóra  4 , Gergely Zachar  4 , Tamás Székely  5 , Ákos Pogány  3 , Árpád Dobolyi  1   2
Affiliations

Transcriptomics of Parental Care in the Hypothalamic-Septal Region of Female Zebra Finch Brain

Rashmi Kumari et al. Int J Mol Sci. .

Abstract

(1) Background: The objective of this study was to uncover genomic causes of parental care. Since birds do not lactate and, therefore, do not show the gene expressional changes required for lactation, we investigate gene expression associated with parenting in caring and non-caring females in an avian species, the small passerine bird zebra finch (Taeniopygia guttata). Here, we compare expression patterns in the hypothalamic-septal region since, previously, we showed that this area is activated in parenting females. (2) Methods: Transcriptome sequencing was first applied in a dissected part of the zebra finch brain related to taking care of the nestlings as compared to a control group of social pairs without nestlings. (3) Results: We found genes differentially expressed between caring and non-caring females. When introducing a log2fold change threshold of 1.5, 13 annotated genes were significantly upregulated in breeding pairs, while 39 annotated genes were downregulated. Significant enrichments of dopamine and acetylcholine biosynthetic processes were identified among upregulated pathways, while pro-opiomelanocortin and thyroid hormone pathways were downregulated, suggesting the importance of these systems in parental care. Network analysis further suggested neuro-immunological changes in mothers. (4) Conclusions: The results confirm the roles of several hypothesized major pathways in parental care, whereas novel pathways are also proposed.

Keywords: RNA-Seq; gene expression; gene ontology; hypothalamic–septal region; parental care; posthatching period; social behaviour; transcriptome sequencing; zebra finch mother.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The results of sequencing shown in a volcano plot. Significantly upregulated genes are red in the upper right corner, while significantly downregulated genes are blue in the upper left corner. Because the test is now based on explicit sampling from the beta negative binomial in Cuffdiff v.2.2.1., users do not see values less than 10−5 by default.
Figure 2
Figure 2
The results of PCR validation. The CRYM showed a significant difference between breeding and social pairs (p < 0.01), while POMC demonstrated a strong tendency (p < 0.1). The red dots represent individual females of breeding pairs, while black dots are individual females of social pairs.
Figure 3
Figure 3
Correlation between gene expression levels and the time spent parenting in zebra finches. Panel shows the negative correlation of POMC gene expression level and time of feeding behaviour. The coloured dots represent the individuals.
Figure 4
Figure 4
Interactions between proteins including down- and upregulated genes, as predicted by the STRING online database. (A) PPI network of 5 down- and 5 upregulated genes was constructed. The colour of the nodes represents the three clusters. Each edge colour indicates a different method of protein–protein association prediction as indicated in the figure. The enrichment p value of PPI network for the number of identified edges compared to the expected (2) was 0.000575, significantly more than expected with a medium interaction score of 0.4. Clustering PPI network with Markov Clustering algorithm (MCL) shows three clusters of DEGs. Cluster 1 represents the protein–protein association of downregulated genes, while Clusters 2 and 3 include the functionally connected upregulated genes. (B) Clusters were functionally annotated. Gene Ontology (GO) and pathway classifications of the three clusters of PPI network were analysed through the STRING online database with the FDR < 0.05. Bar graphs show the enriched terms of Cluster 1 including 5 downregulated genes, and Clusters 2 and 3 including 5 upregulated genes. In the graph, y-axis represents the significantly enriched terms. Each bar describes the number of mapped annotated genes in the reference dataset, while the x-axis indicates the significance (−log10 (FDR)). The colour code corresponds to the ontologies.
Figure 5
Figure 5
The time point of video recording and dissection in relation to the timeline of reproductive phases in the breeding group. The behaviour of the parents was recorded on post-hatching day 10 (PHD-10), while the dissection of their brain took place on PHD-13. The colours represent different stages of reproduction.
Figure 6
Figure 6
The schematic drawings of dissected hypothalamic–septal region: (a) sagittal, (b) coronal section. The anterior and posterior coordinates of the dissection of a coronal slide were A4.0 and A0.6. The black lines on (b) represent the included hypothalamic and septal regions with the removal of tectum and pallial structures.
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