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Kcnq1-5 (Kv7.1-5) potassium channel expression in the adult zebrafish.

Wu C, Sharma K, Laster K, Hersi M, Torres C, Lukas TJ, Moore EJ - BMC Physiol. (2014)

Bottom Line: Overall expression of the kcnqx channel transcripts is similar to that found in mammals.We found that kcnq1 expression was highest in the heart, and also present in the ear and brain. kcnq2 was lowest in the heart, while kcnq3 was highly expressed in the brain, heart and ear. kcnq5 expression was highest in the ear.We conclude that the zebrafish is an attractive model for the study of the KCNQ (Kv7) superfamily of genes, and are important to processes involved in neuronal excitability, cardiac anomalies, epileptic seizures, and hearing loss or tinnitus.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Pharmacology & Biological Chemistry, Northwestern University, Chicago, IL 60611, USA. ernest.moore@unt.edu.

ABSTRACT

Background: KCNQx genes encode slowly activating-inactivating K+ channels, are linked to physiological signal transduction pathways, and mutations in them underlie diseases such as long QT syndrome (KCNQ1), epilepsy in adults (KCNQ2/3), benign familial neonatal convulsions in children (KCNQ3), and hearing loss or tinnitus in humans (KCNQ4, but not KCNQ5). Identification of kcnqx potassium channel transcripts in zebrafish (Danio rerio) remains to be fully characterized although some genes have been mapped to the genome. Using zebrafish genome resources as the source of putative kcnq sequences, we investigated the expression of kcnq1-5 in heart, brain and ear tissues.

Results: Overall expression of the kcnqx channel transcripts is similar to that found in mammals. We found that kcnq1 expression was highest in the heart, and also present in the ear and brain. kcnq2 was lowest in the heart, while kcnq3 was highly expressed in the brain, heart and ear. kcnq5 expression was highest in the ear. We analyzed zebrafish genomic clones containing putative kcnq4 sequences to identify transcripts and protein for this highly conserved member of the Kcnq channel family. The zebrafish appears to have two kcnq4 genes that produce distinct mRNA species in brain, ear, and heart tissues.

Conclusions: We conclude that the zebrafish is an attractive model for the study of the KCNQ (Kv7) superfamily of genes, and are important to processes involved in neuronal excitability, cardiac anomalies, epileptic seizures, and hearing loss or tinnitus.

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Related in: MedlinePlus

qRTPCR of Kcnq channel transcripts in brain, heart, and ear. Real time PCR thresholds (Delta CT) were first normalized to GAPDH or β-actin for each tissue. These data were then plotted relative to the lowest expressed mRNA in each tissue using a logarithmic scale. Depicted also is +/-1.0 standard error.
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Figure 2: qRTPCR of Kcnq channel transcripts in brain, heart, and ear. Real time PCR thresholds (Delta CT) were first normalized to GAPDH or β-actin for each tissue. These data were then plotted relative to the lowest expressed mRNA in each tissue using a logarithmic scale. Depicted also is +/-1.0 standard error.

Mentions: Several of the kcnq RNA transcripts were expressed in the zebrafish brain (Figure 1A) consistent with observations in other species[19]. Kcnq1 and kcnq5 were absent in the gel (Figure 1A), but were detected in the quantitative data (Figure 2, top), at the same approximate levels. The PCR data for kcnq5 were done with kcnq5b primers while the quantitative PCR was done with kcnq5a primers. Kcnq4 was probed by three different sets of primers (KCNQ4-a, KCNQ4-b, KCNQ4-c) for downstream cDNA sequencing. Bands for kcnq4 for the three sets of primers were readily detected. Kcnq2 and kcnq3 showed the strongest signal, while kcnq3 displayed two almost overlapping bands. A negative control (PCR reaction without RNA, or “No RNA”) is shown in the last lane. The expression strength of the mRNA transcript was compared to the intensity of the β-actin control.


Kcnq1-5 (Kv7.1-5) potassium channel expression in the adult zebrafish.

Wu C, Sharma K, Laster K, Hersi M, Torres C, Lukas TJ, Moore EJ - BMC Physiol. (2014)

qRTPCR of Kcnq channel transcripts in brain, heart, and ear. Real time PCR thresholds (Delta CT) were first normalized to GAPDH or β-actin for each tissue. These data were then plotted relative to the lowest expressed mRNA in each tissue using a logarithmic scale. Depicted also is +/-1.0 standard error.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4016485&req=5

Figure 2: qRTPCR of Kcnq channel transcripts in brain, heart, and ear. Real time PCR thresholds (Delta CT) were first normalized to GAPDH or β-actin for each tissue. These data were then plotted relative to the lowest expressed mRNA in each tissue using a logarithmic scale. Depicted also is +/-1.0 standard error.
Mentions: Several of the kcnq RNA transcripts were expressed in the zebrafish brain (Figure 1A) consistent with observations in other species[19]. Kcnq1 and kcnq5 were absent in the gel (Figure 1A), but were detected in the quantitative data (Figure 2, top), at the same approximate levels. The PCR data for kcnq5 were done with kcnq5b primers while the quantitative PCR was done with kcnq5a primers. Kcnq4 was probed by three different sets of primers (KCNQ4-a, KCNQ4-b, KCNQ4-c) for downstream cDNA sequencing. Bands for kcnq4 for the three sets of primers were readily detected. Kcnq2 and kcnq3 showed the strongest signal, while kcnq3 displayed two almost overlapping bands. A negative control (PCR reaction without RNA, or “No RNA”) is shown in the last lane. The expression strength of the mRNA transcript was compared to the intensity of the β-actin control.

Bottom Line: Overall expression of the kcnqx channel transcripts is similar to that found in mammals.We found that kcnq1 expression was highest in the heart, and also present in the ear and brain. kcnq2 was lowest in the heart, while kcnq3 was highly expressed in the brain, heart and ear. kcnq5 expression was highest in the ear.We conclude that the zebrafish is an attractive model for the study of the KCNQ (Kv7) superfamily of genes, and are important to processes involved in neuronal excitability, cardiac anomalies, epileptic seizures, and hearing loss or tinnitus.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Pharmacology & Biological Chemistry, Northwestern University, Chicago, IL 60611, USA. ernest.moore@unt.edu.

ABSTRACT

Background: KCNQx genes encode slowly activating-inactivating K+ channels, are linked to physiological signal transduction pathways, and mutations in them underlie diseases such as long QT syndrome (KCNQ1), epilepsy in adults (KCNQ2/3), benign familial neonatal convulsions in children (KCNQ3), and hearing loss or tinnitus in humans (KCNQ4, but not KCNQ5). Identification of kcnqx potassium channel transcripts in zebrafish (Danio rerio) remains to be fully characterized although some genes have been mapped to the genome. Using zebrafish genome resources as the source of putative kcnq sequences, we investigated the expression of kcnq1-5 in heart, brain and ear tissues.

Results: Overall expression of the kcnqx channel transcripts is similar to that found in mammals. We found that kcnq1 expression was highest in the heart, and also present in the ear and brain. kcnq2 was lowest in the heart, while kcnq3 was highly expressed in the brain, heart and ear. kcnq5 expression was highest in the ear. We analyzed zebrafish genomic clones containing putative kcnq4 sequences to identify transcripts and protein for this highly conserved member of the Kcnq channel family. The zebrafish appears to have two kcnq4 genes that produce distinct mRNA species in brain, ear, and heart tissues.

Conclusions: We conclude that the zebrafish is an attractive model for the study of the KCNQ (Kv7) superfamily of genes, and are important to processes involved in neuronal excitability, cardiac anomalies, epileptic seizures, and hearing loss or tinnitus.

Show MeSH
Related in: MedlinePlus