Expression and differential cell distribution of low-threshold Ca 2+ channels in mammalian male germ cells and sperm

Claudia L. Treviño, Ricardo Felix, Laura E. Castellano, Carolina Gutiérrez, Delany Rodríguez, Judith Pacheco, Ignacio López-González, Juan Carlos Gomora, Victor Tsutsumi, Arturo Hernández-Cruz, Tatiana Fiordelisio, Allison L. Scaling, Alberto Darszon

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Abstract

Numerous sperm functions including the acrosome reaction (AR) are associated with Ca2+ influx through voltage-gated Ca2+ (CaV) channels. Although the electrophysiological characterization of Ca2+ currents in mature sperm has proven difficult, functional studies have revealed the presence of low-threshold (CaV3) channels in spermatogenic cells. However, the molecular identity of these proteins remains undefined. Here, we identified by reverse transcription polymerase chain reaction the expression of CaV3.3 mRNA in mouse male germ cells, an isoform not previously described in these cells. Immunoconfocal microscopy revealed the presence of the three CaV3 channel isoforms in mouse spermatogenic cells. In mature mouse sperm only CaV3.1 and Ca V3.2 were detected in the head, suggesting its participation in the AR. CaV3.1 and CaV3.3 were found in the principal and the midpiece of the flagella. All CaV3 channels are also present in human sperm, but only to a minor extent in the head. These findings were corroborated by immunogold transmission electron microscopy. Tail localization of Ca V3 channels suggested they may participate in motility, however, mibefradil and gossypol concentrations that inhibit CaV3 channels did not significantly affect human sperm motility. Only higher mibefradil doses that can block high-threshold (HVA) CaV channels caused small but significant motility alterations. Antibodies to HVA channels detected Ca V1.3 and CaV2.3 in human sperm flagella. © 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.
Original languageAmerican English
Pages (from-to)87-92
Number of pages6
JournalFEBS Letters
DOIs
StatePublished - 9 Apr 2004
Externally publishedYes

Fingerprint

Mibefradil
Germ Cells
Spermatozoa
Protein Isoforms
Cells
Gossypol
Acrosome Reaction
Polymerase chain reaction
Transcription
Microscopic examination
Head
Sperm Tail
Transmission electron microscopy
Messenger RNA
Flagella
Antibodies
Sperm Motility
Electric potential
Transmission Electron Microscopy
Reverse Transcription

Cite this

Treviño, Claudia L. ; Felix, Ricardo ; Castellano, Laura E. ; Gutiérrez, Carolina ; Rodríguez, Delany ; Pacheco, Judith ; López-González, Ignacio ; Gomora, Juan Carlos ; Tsutsumi, Victor ; Hernández-Cruz, Arturo ; Fiordelisio, Tatiana ; Scaling, Allison L. ; Darszon, Alberto. / Expression and differential cell distribution of low-threshold Ca 2+ channels in mammalian male germ cells and sperm. In: FEBS Letters. 2004 ; pp. 87-92.
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title = "Expression and differential cell distribution of low-threshold Ca 2+ channels in mammalian male germ cells and sperm",
abstract = "Numerous sperm functions including the acrosome reaction (AR) are associated with Ca2+ influx through voltage-gated Ca2+ (CaV) channels. Although the electrophysiological characterization of Ca2+ currents in mature sperm has proven difficult, functional studies have revealed the presence of low-threshold (CaV3) channels in spermatogenic cells. However, the molecular identity of these proteins remains undefined. Here, we identified by reverse transcription polymerase chain reaction the expression of CaV3.3 mRNA in mouse male germ cells, an isoform not previously described in these cells. Immunoconfocal microscopy revealed the presence of the three CaV3 channel isoforms in mouse spermatogenic cells. In mature mouse sperm only CaV3.1 and Ca V3.2 were detected in the head, suggesting its participation in the AR. CaV3.1 and CaV3.3 were found in the principal and the midpiece of the flagella. All CaV3 channels are also present in human sperm, but only to a minor extent in the head. These findings were corroborated by immunogold transmission electron microscopy. Tail localization of Ca V3 channels suggested they may participate in motility, however, mibefradil and gossypol concentrations that inhibit CaV3 channels did not significantly affect human sperm motility. Only higher mibefradil doses that can block high-threshold (HVA) CaV channels caused small but significant motility alterations. Antibodies to HVA channels detected Ca V1.3 and CaV2.3 in human sperm flagella. {\circledC} 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.",
author = "Trevi{\~n}o, {Claudia L.} and Ricardo Felix and Castellano, {Laura E.} and Carolina Guti{\'e}rrez and Delany Rodr{\'i}guez and Judith Pacheco and Ignacio L{\'o}pez-Gonz{\'a}lez and Gomora, {Juan Carlos} and Victor Tsutsumi and Arturo Hern{\'a}ndez-Cruz and Tatiana Fiordelisio and Scaling, {Allison L.} and Alberto Darszon",
year = "2004",
month = "4",
day = "9",
doi = "10.1016/S0014-5793(04)00257-1",
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Treviño, CL, Felix, R, Castellano, LE, Gutiérrez, C, Rodríguez, D, Pacheco, J, López-González, I, Gomora, JC, Tsutsumi, V, Hernández-Cruz, A, Fiordelisio, T, Scaling, AL & Darszon, A 2004, 'Expression and differential cell distribution of low-threshold Ca 2+ channels in mammalian male germ cells and sperm', FEBS Letters, pp. 87-92. https://doi.org/10.1016/S0014-5793(04)00257-1

Expression and differential cell distribution of low-threshold Ca 2+ channels in mammalian male germ cells and sperm. / Treviño, Claudia L.; Felix, Ricardo; Castellano, Laura E.; Gutiérrez, Carolina; Rodríguez, Delany; Pacheco, Judith; López-González, Ignacio; Gomora, Juan Carlos; Tsutsumi, Victor; Hernández-Cruz, Arturo; Fiordelisio, Tatiana; Scaling, Allison L.; Darszon, Alberto.

In: FEBS Letters, 09.04.2004, p. 87-92.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Expression and differential cell distribution of low-threshold Ca 2+ channels in mammalian male germ cells and sperm

AU - Treviño, Claudia L.

AU - Felix, Ricardo

AU - Castellano, Laura E.

AU - Gutiérrez, Carolina

AU - Rodríguez, Delany

AU - Pacheco, Judith

AU - López-González, Ignacio

AU - Gomora, Juan Carlos

AU - Tsutsumi, Victor

AU - Hernández-Cruz, Arturo

AU - Fiordelisio, Tatiana

AU - Scaling, Allison L.

AU - Darszon, Alberto

PY - 2004/4/9

Y1 - 2004/4/9

N2 - Numerous sperm functions including the acrosome reaction (AR) are associated with Ca2+ influx through voltage-gated Ca2+ (CaV) channels. Although the electrophysiological characterization of Ca2+ currents in mature sperm has proven difficult, functional studies have revealed the presence of low-threshold (CaV3) channels in spermatogenic cells. However, the molecular identity of these proteins remains undefined. Here, we identified by reverse transcription polymerase chain reaction the expression of CaV3.3 mRNA in mouse male germ cells, an isoform not previously described in these cells. Immunoconfocal microscopy revealed the presence of the three CaV3 channel isoforms in mouse spermatogenic cells. In mature mouse sperm only CaV3.1 and Ca V3.2 were detected in the head, suggesting its participation in the AR. CaV3.1 and CaV3.3 were found in the principal and the midpiece of the flagella. All CaV3 channels are also present in human sperm, but only to a minor extent in the head. These findings were corroborated by immunogold transmission electron microscopy. Tail localization of Ca V3 channels suggested they may participate in motility, however, mibefradil and gossypol concentrations that inhibit CaV3 channels did not significantly affect human sperm motility. Only higher mibefradil doses that can block high-threshold (HVA) CaV channels caused small but significant motility alterations. Antibodies to HVA channels detected Ca V1.3 and CaV2.3 in human sperm flagella. © 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.

AB - Numerous sperm functions including the acrosome reaction (AR) are associated with Ca2+ influx through voltage-gated Ca2+ (CaV) channels. Although the electrophysiological characterization of Ca2+ currents in mature sperm has proven difficult, functional studies have revealed the presence of low-threshold (CaV3) channels in spermatogenic cells. However, the molecular identity of these proteins remains undefined. Here, we identified by reverse transcription polymerase chain reaction the expression of CaV3.3 mRNA in mouse male germ cells, an isoform not previously described in these cells. Immunoconfocal microscopy revealed the presence of the three CaV3 channel isoforms in mouse spermatogenic cells. In mature mouse sperm only CaV3.1 and Ca V3.2 were detected in the head, suggesting its participation in the AR. CaV3.1 and CaV3.3 were found in the principal and the midpiece of the flagella. All CaV3 channels are also present in human sperm, but only to a minor extent in the head. These findings were corroborated by immunogold transmission electron microscopy. Tail localization of Ca V3 channels suggested they may participate in motility, however, mibefradil and gossypol concentrations that inhibit CaV3 channels did not significantly affect human sperm motility. Only higher mibefradil doses that can block high-threshold (HVA) CaV channels caused small but significant motility alterations. Antibodies to HVA channels detected Ca V1.3 and CaV2.3 in human sperm flagella. © 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.

U2 - 10.1016/S0014-5793(04)00257-1

DO - 10.1016/S0014-5793(04)00257-1

M3 - Article

SP - 87

EP - 92

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

ER -