Feng, N. Y. | bioRxiv Science

bioRxiv2020

To hum or not to hum: Neural transcriptome signature of courtship vocalization in a teleost fish

For many animal species, vocal communication is a critical social behavior, often a necessary component of reproductive success. In addition to the role of vocal behavior in social interactions, vocalizations are often demanding motor acts. Through understanding the genes involved in regulating and permitting vertebrate vocalization, we can better understand the mechanisms regulating vocal and, more broadly, motor behaviors. Here, we use RNA-sequencing to investigate neural gene expression underlying the performance of an extreme vocal behavior, the courtship hum of the plainfin midshipman fish (Porichthys notatus). Single hums can last up to two hours and may be repeated throughout an evening of courtship activity. We asked whether vocal behavioral states are associated with specific gene expression signatures in key brain regions that regulate vocalization by comparing transcript levels in humming versus non-humming males. We find that the circadian-related genes period3 and Clock are significantly upregulated in the vocal motor nucleus and preoptic area-anterior hypothalamus, respectively, in humming compared to non-humming males, indicating that internal circadian clocks may differ between these divergent behavioral states. In addition, we identify suites of differentially expressed genes related to synaptic transmission, ion channels and transport, hormone signaling, and metabolism and antioxidant activity that may permit or support humming behavior. These results underscore the importance of the known circadian control of midshipman humming and provide testable candidate genes for future studies of the neuroendocrine and motor control of energetically demanding courtship behaviors in midshipman fish and other vertebrate groups.

neuroscience↗

bioRxiv2018

Antioxidant Gene Expression in Vocal Hindbrain of a Teleost Fish

Plainfin midshipman fish (Porichthys notatus) have a remarkable capacity to generate long duration advertisement calls known as hums, each of which may last for close to two hours and be repeated throughout a night of courtship activity during the breeding season. The midshipmans striking sound production capabilities provide a unique opportunity to investigate the mechanisms that motor neurons require for withstanding high-endurance activity. The temporal properties of midshipman vocal behaviors are largely controlled by a hindbrain central pattern generator that includes vocal motor neurons (VMN) that directly determine the activity pattern of target sonic muscles and, in turn, a sounds pulse repetition rate, duration and pattern of amplitude modulation. Of the two adult midshipman male reproductive phenotypes -- types I and II-- only type I males acoustically court females with hums from nests that they build and guard, while type II males do not produce courtship hums but instead sneak or satellite spawn to steal fertilizations from type I males. A prior study using next generation RNA sequencing showed increased expression of a number of cellular respiration and antioxidant genes in the VMN of type I males during the breeding season, suggesting they help to combat potentially high levels of oxidative stress linked to this extreme behavior. This led to the question of whether the expression of these genes in the VMN would vary between actively humming versus non-humming states as well as between male morphs. Here, we tested the hypothesis that to combat oxidative stress, the VMN of reproductively active type I males would exhibit higher mRNA transcript levels for two superoxide dismutases (sod1, sod2) compared to the VMN of type II males and females that do not hum and in general both of which have a more limited vocal repertoire than type I males. The results showed no significant difference in sod1 transcript expression across reproductive morphs in the VMN and the surrounding hindbrain, and no difference of sod2 across the two male morphs and females in the SH. However, we observed a surprising, significantly lower expression of sod2 transcripts in the VMN of type I males as compared to type II males. We also found no significant difference in sod1 and sod2 expression between actively humming and non-humming type I males in both the VMN and surrounding hindbrain. These findings overall lead us to conclude that increased transcription of sod1 and sod2 is not necessary for combatting oxidative stress from the demands of the midshipman high-endurance vocalizations, but warrant future studies to assess protein levels, enzyme activity levels, as well as the expression of other antioxidant genes. These results also eliminate one of the proposed mechanisms that male midshipman use to combat potentially high levels of oxidative stress incurred by motor neurons driving long duration vocalization and provide more insight into how motor neurons are adapted to the performance of extreme behaviors.

neuroscience↗