Skip to main page content
U.S. flag

An official website of the United States government

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2006 Mar;143(3):294-301.
doi: 10.1016/j.cbpb.2005.11.017. Epub 2006 Jan 19.

Molecular cloning and characterization of crustacean type-one dopamine receptors: D1alphaPan and D1betaPan

Merry C Clark  1 , Deborah J Baro
Affiliations

Molecular cloning and characterization of crustacean type-one dopamine receptors: D1alphaPan and D1betaPan

Merry C Clark et al. Comp Biochem Physiol B Biochem Mol Biol. 2006 Mar.

Abstract

Dopamine (DA) differentially modulates identified neurons in the crustacean stomatogastric nervous system (STNS). While the electrophysiological actions of DA have been well characterized, little is known about the dopaminergic transduction cascades operating in this system. As a first step toward illuminating the molecular underpinnings of dopaminergic signal transduction in the crustacean STNS, we have cloned and characterized two type-one DA receptors (DARs) from the spiny lobster (Panulirus interruptus): D(1alphaPan) and D(1betaPan). We found that the structure and function of these arthropod DARs are well conserved across species. Using a heterologous expression system, we determined that DA, but not serotonin, octopamine, tyramine or histamine activates these receptors. When stably expressed in HEK cells, the D(1alphaPan) receptor couples with Gs, and DA elicits an increase in [cAMP]. The D(1betaPan) receptor responds to DA with a net increase in [cAMP] that is mediated by Gs and Gz.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
The DAR family is conserved across arthropods. The Panulirus (L) and Drosophila (F) DAR orthologs are aligned. Amino acids that are identical in fly and lobster orthologs are highlighted for each pair of DARs. Black bars approximate the seven transmembrane regions. The point of alternate splicing on lobster D1βPan is indicated by a black arrowhead. The accession numbers are as follows: LD1αPan, DQ295790; FD1αPan, U34383; LD1β.1Pan, DQ295791; FD1βPan, X77234.1.
Fig. 2
Fig. 2
The D1αPan receptor couples with Gs. G protein activities in HEK D1αPan membrane preparations were measured in the absence (open bar) vs. the presence (filled bar) of 10−5 M DA for eight G proteins: Gs, Gq, Gz, Gi1, Gi2, Gi3, Go, G12. Data represent the mean±S.E.M., n=3. Statistically significant differences in the activity of a given G protein are indicated with an asterisk (p<0.05).
Fig.3
Fig.3
DA activation of the D1αPan receptor increases [cAMP]. The D1αPan receptor couples positively with cAMP. Changes in cAMP levels in response to increasing [DA] were measured in a stably transfected cell line expressing the D1αPan receptor, HEK D1αPan (filled squares) and in the nontransfected parental cell line, HEK (filled triangles). Data are represented as the mean±S.E.M, n=3.
Fig. 4
Fig. 4
The D1βPan receptor couples with Gs and Gz. G protein activities in HEK D1βPan membrane preparations were measured in the absence (open bar) vs. the presence (filled bar) of 10−5 M DA for eight G proteins: Gs, Gq, Gz, Gai1, Gai2, Gai3, Gao, Ga12. Data represent the mean±S.E.M., n≥3. Statistically significant differences in the activity of a given G protein are indicated with an asterisk (p<0.05).
Fig. 5
Fig. 5
DA activation of the D1βPan receptor produces a net increase in cAMP. The D1βPan receptor couples positively with cAMP. Changes in cAMP levels in response to increasing [DA] were measured in the nontransfected parental cell line (HEK, filled triangles) and in two stably transfected cell lines expressing the D1βPan receptor. We identified two isoforms for the D1βPan receptor, and established cell lines for each: HEK D1β.1Pan (circles) and HEK D1β.2Pan (open triangle). Data represent the mean±S.E.M., n=5.
Fig. 6
Fig. 6
Dopamine is the only endogenous monoamine that activates the D1αPan and D1βPan receptors. Levels of cAMP were measured in HEK (hatched bars), HEKD1αPan (filled bars) and HEKD1βPan.2 (open bars) cell lines under control conditions (no monoamines present) or in the presence of 1 mM of the indicated monoamine. The cAMP levels measured in the presence of the indicated monoamine were normalized by cAMP levels under control conditions. Average fold changes over basal cAMP levels are plotted, error bars indicate the S.E.M, n≥3. * Indicates significant increases in cAMP over basal levels (p<0.05). ** Indicates significant differences (p<0.05) between cell lines within the same condition (e.g., differences between cell lines exposed to DA).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Baillie GS, Sood A, McPhee I, Gall I, Perry SJ, Lefkowitz RJ, Houslay MD. beta-Arrestin-mediated PDE4 cAMP phosphodiesterase recruitment regulates beta-adrenoceptor switching from Gs to Gi. Proc. Natl. Acad. Sci. U. S. A. 2003;100:940–945. - PMC - PubMed
    1. Banihashemi B, Albert PR. Dopamine-D2S receptor inhibition of calcium influx, adenylyl cyclase, and mitogen-activated protein kinase in pituitary cells: distinct Galpha and Gbetagamma requirements. Mol. Endocrinol. 2002;16:2393–2404. - PubMed
    1. Beltz BS. Distribution and functional anatomy of amine-containing neurons in decapod crustaceans. Microsc. Res. Tech. 1999;44:105–120. - PubMed
    1. Blenau W, Erber J, Baumann A. Characterization of a dopamine D1 receptor from Apis mellifera: cloning, functional expression, pharmacology, and mRNA localization in the brain. J. Neurochem. 1998;70:15–23. - PubMed
    1. Bucher D, Thirumalai V, Marder E. Axonal dopamine receptors activate peripheral spike initiation in a stomatogastric motor neuron. J. Neurosci. 2003;23:6866–6875. - PMC - PubMed

Publication types