Xenopus laevis RIC-3 enhances the functional expression of the C. elegans homomeric nicotinic receptor, ACR-16, in Xenopus oocytes

Hayley M. Bennett, Kristin Lees, Kate M. Harper, Andrew K. Jones, David B. Sattelle, Susan Wonnacott, Adrian J. Wolstenholme

Research output: Contribution to journalArticle

  • 7 Citations

Abstract

RIC-3 enhances the functional expression of certain nicotinic acetylcholine receptors (nAChRs) in vertebrates and invertebrates and increases the availability of functional receptors in cultured cells and Xenopus laevis oocytes. Maximal activity of RIC-3 may be cell-type dependent, so neither mammalian nor invertebrate proteins is optimal in amphibian oocytes. We cloned the X. laevis ric-3 cDNA and tested the frog protein in oocyte expression studies. X. laevis RIC-3 shares 52% amino acid identity with human RIC-3 and only 17% with that of Caenorhabditis elegans. We used the C. elegans nicotinic receptor, ACR-16, to compare the ability of RIC-3 from three species to enhance receptor expression. In the absence of RIC-3, the proportion of oocytes expressing detectable nAChRs was greatly reduced. Varying the ratio of acr-16 to X. laevis ric-3 cRNAs injected into oocytes had little impact on the total cell current. When X. laevis, human or C. elegans ric-3 cRNAs were co-injected with acr-16 cRNA (1 : 1 ratio), 100 μM acetylcholine induced larger currents in oocytes expressing X. laevis RIC-3 compared with its orthologues. This provides further evidence for a species-specific component of RIC-3 activity, and suggests that X. laevis RIC-3 is useful for enhancing the expression of invertebrate nAChRs in X. laevis oocytes.

LanguageEnglish
Pages911-918
Number of pages8
JournalJournal of Neurochemistry
Volume123
Issue number6
DOIs
StatusPublished - Dec 2012

Fingerprint

Xenopus laevis
Nicotinic Receptors
Xenopus
Complementary RNA
Oocytes
Caenorhabditis elegans
Invertebrates
Acetylcholine
Proteins
Complementary DNA
Cells
Availability
Amino Acids
Amphibians
Anura
Vertebrates
Cultured Cells

Keywords

  • Amino Acid Sequence
  • Animals
  • Caenorhabditis elegans
  • Caenorhabditis elegans Proteins
  • Gene Expression Regulation, Developmental
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Molecular Chaperones
  • Molecular Sequence Data
  • Oocytes
  • Receptors, Nicotinic
  • Up-Regulation
  • Xenopus Proteins
  • Xenopus laevis
  • alpha7 Nicotinic Acetylcholine Receptor
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

Xenopus laevis RIC-3 enhances the functional expression of the C. elegans homomeric nicotinic receptor, ACR-16, in Xenopus oocytes. / Bennett, Hayley M.; Lees, Kristin; Harper, Kate M.; Jones, Andrew K.; Sattelle, David B.; Wonnacott, Susan; Wolstenholme, Adrian J.

In: Journal of Neurochemistry, Vol. 123, No. 6, 12.2012, p. 911-918.

Research output: Contribution to journalArticle

Bennett, Hayley M. ; Lees, Kristin ; Harper, Kate M. ; Jones, Andrew K. ; Sattelle, David B. ; Wonnacott, Susan ; Wolstenholme, Adrian J./ Xenopus laevis RIC-3 enhances the functional expression of the C. elegans homomeric nicotinic receptor, ACR-16, in Xenopus oocytes. In: Journal of Neurochemistry. 2012 ; Vol. 123, No. 6. pp. 911-918
@article{c5632933d6f84bd698d042e93ca2ab58,
title = "Xenopus laevis RIC-3 enhances the functional expression of the C. elegans homomeric nicotinic receptor, ACR-16, in Xenopus oocytes",
abstract = "RIC-3 enhances the functional expression of certain nicotinic acetylcholine receptors (nAChRs) in vertebrates and invertebrates and increases the availability of functional receptors in cultured cells and Xenopus laevis oocytes. Maximal activity of RIC-3 may be cell-type dependent, so neither mammalian nor invertebrate proteins is optimal in amphibian oocytes. We cloned the X. laevis ric-3 cDNA and tested the frog protein in oocyte expression studies. X. laevis RIC-3 shares 52{\%} amino acid identity with human RIC-3 and only 17{\%} with that of Caenorhabditis elegans. We used the C. elegans nicotinic receptor, ACR-16, to compare the ability of RIC-3 from three species to enhance receptor expression. In the absence of RIC-3, the proportion of oocytes expressing detectable nAChRs was greatly reduced. Varying the ratio of acr-16 to X. laevis ric-3 cRNAs injected into oocytes had little impact on the total cell current. When X. laevis, human or C. elegans ric-3 cRNAs were co-injected with acr-16 cRNA (1 : 1 ratio), 100 μM acetylcholine induced larger currents in oocytes expressing X. laevis RIC-3 compared with its orthologues. This provides further evidence for a species-specific component of RIC-3 activity, and suggests that X. laevis RIC-3 is useful for enhancing the expression of invertebrate nAChRs in X. laevis oocytes.",
keywords = "Amino Acid Sequence, Animals, Caenorhabditis elegans, Caenorhabditis elegans Proteins, Gene Expression Regulation, Developmental, Humans, Intracellular Signaling Peptides and Proteins, Membrane Proteins, Molecular Chaperones, Molecular Sequence Data, Oocytes, Receptors, Nicotinic, Up-Regulation, Xenopus Proteins, Xenopus laevis, alpha7 Nicotinic Acetylcholine Receptor, Journal Article, Research Support, Non-U.S. Gov't",
author = "Bennett, {Hayley M.} and Kristin Lees and Harper, {Kate M.} and Jones, {Andrew K.} and Sattelle, {David B.} and Susan Wonnacott and Wolstenholme, {Adrian J.}",
note = "{\circledC} 2012 The Authors Journal of Neurochemistry {\circledC} 2012 International Society for Neurochemistry.",
year = "2012",
month = "12",
doi = "10.1111/jnc.2012.123.issue-6",
language = "English",
volume = "123",
pages = "911--918",
journal = "Journal of Neurochemistry",
issn = "0022-3042",
publisher = "Wiley-Blackwell",
number = "6",

}

TY - JOUR

T1 - Xenopus laevis RIC-3 enhances the functional expression of the C. elegans homomeric nicotinic receptor, ACR-16, in Xenopus oocytes

AU - Bennett,Hayley M.

AU - Lees,Kristin

AU - Harper,Kate M.

AU - Jones,Andrew K.

AU - Sattelle,David B.

AU - Wonnacott,Susan

AU - Wolstenholme,Adrian J.

N1 - © 2012 The Authors Journal of Neurochemistry © 2012 International Society for Neurochemistry.

PY - 2012/12

Y1 - 2012/12

N2 - RIC-3 enhances the functional expression of certain nicotinic acetylcholine receptors (nAChRs) in vertebrates and invertebrates and increases the availability of functional receptors in cultured cells and Xenopus laevis oocytes. Maximal activity of RIC-3 may be cell-type dependent, so neither mammalian nor invertebrate proteins is optimal in amphibian oocytes. We cloned the X. laevis ric-3 cDNA and tested the frog protein in oocyte expression studies. X. laevis RIC-3 shares 52% amino acid identity with human RIC-3 and only 17% with that of Caenorhabditis elegans. We used the C. elegans nicotinic receptor, ACR-16, to compare the ability of RIC-3 from three species to enhance receptor expression. In the absence of RIC-3, the proportion of oocytes expressing detectable nAChRs was greatly reduced. Varying the ratio of acr-16 to X. laevis ric-3 cRNAs injected into oocytes had little impact on the total cell current. When X. laevis, human or C. elegans ric-3 cRNAs were co-injected with acr-16 cRNA (1 : 1 ratio), 100 μM acetylcholine induced larger currents in oocytes expressing X. laevis RIC-3 compared with its orthologues. This provides further evidence for a species-specific component of RIC-3 activity, and suggests that X. laevis RIC-3 is useful for enhancing the expression of invertebrate nAChRs in X. laevis oocytes.

AB - RIC-3 enhances the functional expression of certain nicotinic acetylcholine receptors (nAChRs) in vertebrates and invertebrates and increases the availability of functional receptors in cultured cells and Xenopus laevis oocytes. Maximal activity of RIC-3 may be cell-type dependent, so neither mammalian nor invertebrate proteins is optimal in amphibian oocytes. We cloned the X. laevis ric-3 cDNA and tested the frog protein in oocyte expression studies. X. laevis RIC-3 shares 52% amino acid identity with human RIC-3 and only 17% with that of Caenorhabditis elegans. We used the C. elegans nicotinic receptor, ACR-16, to compare the ability of RIC-3 from three species to enhance receptor expression. In the absence of RIC-3, the proportion of oocytes expressing detectable nAChRs was greatly reduced. Varying the ratio of acr-16 to X. laevis ric-3 cRNAs injected into oocytes had little impact on the total cell current. When X. laevis, human or C. elegans ric-3 cRNAs were co-injected with acr-16 cRNA (1 : 1 ratio), 100 μM acetylcholine induced larger currents in oocytes expressing X. laevis RIC-3 compared with its orthologues. This provides further evidence for a species-specific component of RIC-3 activity, and suggests that X. laevis RIC-3 is useful for enhancing the expression of invertebrate nAChRs in X. laevis oocytes.

KW - Amino Acid Sequence

KW - Animals

KW - Caenorhabditis elegans

KW - Caenorhabditis elegans Proteins

KW - Gene Expression Regulation, Developmental

KW - Humans

KW - Intracellular Signaling Peptides and Proteins

KW - Membrane Proteins

KW - Molecular Chaperones

KW - Molecular Sequence Data

KW - Oocytes

KW - Receptors, Nicotinic

KW - Up-Regulation

KW - Xenopus Proteins

KW - Xenopus laevis

KW - alpha7 Nicotinic Acetylcholine Receptor

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

UR - http://www.scopus.com/inward/record.url?scp=84870597954&partnerID=8YFLogxK

UR - http://dx.doi.org/10.1111/jnc.2012.123.issue-6

U2 - 10.1111/jnc.2012.123.issue-6

DO - 10.1111/jnc.2012.123.issue-6

M3 - Article

VL - 123

SP - 911

EP - 918

JO - Journal of Neurochemistry

T2 - Journal of Neurochemistry

JF - Journal of Neurochemistry

SN - 0022-3042

IS - 6

ER -