Absence of estrogen receptor alpha leads to physiological alterations in the mouse epididymis and consequent defects in sperm function

Avenel Joseph, Rex A Hess, David J. Schaeffer, CheMyong Ko, Susan Hudgin-Spivey, Pierre Chambon, Barry D. Shur

Research output: Contribution to journalArticle

Abstract

Male mice deficient in ESR1 (ERalpha) (Esr1KO mice) are infertile, and sperm recovered from the cauda epididymis exhibit reduced motility and fail to fertilize eggs in vitro. These effects on spermappear to result fromdefective epididymal functionandnot a direct effect on spermatogenesis, as Esr1KOgermcells transplanted into wild-type testes yield normal offspring.We hypothesized that the previously described defect in efferent duct fluid reabsorption would lead to alterations in the epididymal fluid milieu, which wouldnegativelyimpact spermfunction.Analysis of the epididymal fluid revealed that the Esr1KO maintains a higher luminal pH throughout the epididymis, confirming an inability of the efferent ducts and/or epididymis to properly acidify the luminal contents. Subsequent studies showed that these abnormalities were not the result of global defects in epididymal function since protein secretion by the Esr1KO epididymis appeared normal as judged by SDS-PAGE of total secreted proteins and by immunoblotting of candidate secreted proteins. To gain insight into the basis of the aberrant fluid homeostasis in the Esr1KO epididymis, the expression of several enzymes and transporters known to be involved in acid/base regulation were analyzed. The levels of SLC9A3 (NHE3) as well as carbonic anhydrase XIV and SLC4A4 (NBC1) were all reduced in the proximal portion of the Esr1KO epididymis, while other components appeared unaffected, including other ion transporters andATP6V0A1(V-ATPase). The altered luminal milieu of the Esr1KO epididymis was shown to lead to a corresponding increase in the intracellular pH of Esr1KO sperm, relative to sperm from control animals. Since pH and bicarbonate ions are critical regulators of sperm cAMP levels and motility, we attempted to bypass the abnormal luminal and intracellular environment by supplementing sperm with exogenous cAMP. This treatment rescued all defective motility parameters, as assayed by CASA, further showing that motility defects are not intrinsic to the sperm but, rather, result from the abnormal epididymal milieu.

Original languageEnglish (US)
Pages (from-to)948-957
Number of pages10
JournalBiology of reproduction
Volume82
Issue number5
DOIs
StatePublished - May 1 2010

Fingerprint

Estrogen Receptor alpha
Epididymis
Spermatozoa
Proteins
Spermatogenesis
Bicarbonates
Immunoblotting
Eggs
Adenosine Triphosphatases
Testis
Polyacrylamide Gel Electrophoresis
Homeostasis
Ions
Acids
Enzymes

Keywords

  • Epididymis
  • Estradiol receptor
  • Male reproductive tract
  • Sperm maturation
  • Sperm motility and transport

ASJC Scopus subject areas

  • Cell Biology

Cite this

Absence of estrogen receptor alpha leads to physiological alterations in the mouse epididymis and consequent defects in sperm function. / Joseph, Avenel; Hess, Rex A; Schaeffer, David J.; Ko, CheMyong; Hudgin-Spivey, Susan; Chambon, Pierre; Shur, Barry D.

In: Biology of reproduction, Vol. 82, No. 5, 01.05.2010, p. 948-957.

Research output: Contribution to journalArticle

Joseph, Avenel ; Hess, Rex A ; Schaeffer, David J. ; Ko, CheMyong ; Hudgin-Spivey, Susan ; Chambon, Pierre ; Shur, Barry D. / Absence of estrogen receptor alpha leads to physiological alterations in the mouse epididymis and consequent defects in sperm function. In: Biology of reproduction. 2010 ; Vol. 82, No. 5. pp. 948-957.
@article{aa0f6feb5d7d462986e9c6eb3bd72115,
title = "Absence of estrogen receptor alpha leads to physiological alterations in the mouse epididymis and consequent defects in sperm function",
abstract = "Male mice deficient in ESR1 (ERalpha) (Esr1KO mice) are infertile, and sperm recovered from the cauda epididymis exhibit reduced motility and fail to fertilize eggs in vitro. These effects on spermappear to result fromdefective epididymal functionandnot a direct effect on spermatogenesis, as Esr1KOgermcells transplanted into wild-type testes yield normal offspring.We hypothesized that the previously described defect in efferent duct fluid reabsorption would lead to alterations in the epididymal fluid milieu, which wouldnegativelyimpact spermfunction.Analysis of the epididymal fluid revealed that the Esr1KO maintains a higher luminal pH throughout the epididymis, confirming an inability of the efferent ducts and/or epididymis to properly acidify the luminal contents. Subsequent studies showed that these abnormalities were not the result of global defects in epididymal function since protein secretion by the Esr1KO epididymis appeared normal as judged by SDS-PAGE of total secreted proteins and by immunoblotting of candidate secreted proteins. To gain insight into the basis of the aberrant fluid homeostasis in the Esr1KO epididymis, the expression of several enzymes and transporters known to be involved in acid/base regulation were analyzed. The levels of SLC9A3 (NHE3) as well as carbonic anhydrase XIV and SLC4A4 (NBC1) were all reduced in the proximal portion of the Esr1KO epididymis, while other components appeared unaffected, including other ion transporters andATP6V0A1(V-ATPase). The altered luminal milieu of the Esr1KO epididymis was shown to lead to a corresponding increase in the intracellular pH of Esr1KO sperm, relative to sperm from control animals. Since pH and bicarbonate ions are critical regulators of sperm cAMP levels and motility, we attempted to bypass the abnormal luminal and intracellular environment by supplementing sperm with exogenous cAMP. This treatment rescued all defective motility parameters, as assayed by CASA, further showing that motility defects are not intrinsic to the sperm but, rather, result from the abnormal epididymal milieu.",
keywords = "Epididymis, Estradiol receptor, Male reproductive tract, Sperm maturation, Sperm motility and transport",
author = "Avenel Joseph and Hess, {Rex A} and Schaeffer, {David J.} and CheMyong Ko and Susan Hudgin-Spivey and Pierre Chambon and Shur, {Barry D.}",
year = "2010",
month = "5",
day = "1",
doi = "10.1095/biolreprod.109.079889",
language = "English (US)",
volume = "82",
pages = "948--957",
journal = "Biology of Reproduction",
issn = "0006-3363",
publisher = "Society for the Study of Reproduction",
number = "5",

}

TY - JOUR

T1 - Absence of estrogen receptor alpha leads to physiological alterations in the mouse epididymis and consequent defects in sperm function

AU - Joseph, Avenel

AU - Hess, Rex A

AU - Schaeffer, David J.

AU - Ko, CheMyong

AU - Hudgin-Spivey, Susan

AU - Chambon, Pierre

AU - Shur, Barry D.

PY - 2010/5/1

Y1 - 2010/5/1

N2 - Male mice deficient in ESR1 (ERalpha) (Esr1KO mice) are infertile, and sperm recovered from the cauda epididymis exhibit reduced motility and fail to fertilize eggs in vitro. These effects on spermappear to result fromdefective epididymal functionandnot a direct effect on spermatogenesis, as Esr1KOgermcells transplanted into wild-type testes yield normal offspring.We hypothesized that the previously described defect in efferent duct fluid reabsorption would lead to alterations in the epididymal fluid milieu, which wouldnegativelyimpact spermfunction.Analysis of the epididymal fluid revealed that the Esr1KO maintains a higher luminal pH throughout the epididymis, confirming an inability of the efferent ducts and/or epididymis to properly acidify the luminal contents. Subsequent studies showed that these abnormalities were not the result of global defects in epididymal function since protein secretion by the Esr1KO epididymis appeared normal as judged by SDS-PAGE of total secreted proteins and by immunoblotting of candidate secreted proteins. To gain insight into the basis of the aberrant fluid homeostasis in the Esr1KO epididymis, the expression of several enzymes and transporters known to be involved in acid/base regulation were analyzed. The levels of SLC9A3 (NHE3) as well as carbonic anhydrase XIV and SLC4A4 (NBC1) were all reduced in the proximal portion of the Esr1KO epididymis, while other components appeared unaffected, including other ion transporters andATP6V0A1(V-ATPase). The altered luminal milieu of the Esr1KO epididymis was shown to lead to a corresponding increase in the intracellular pH of Esr1KO sperm, relative to sperm from control animals. Since pH and bicarbonate ions are critical regulators of sperm cAMP levels and motility, we attempted to bypass the abnormal luminal and intracellular environment by supplementing sperm with exogenous cAMP. This treatment rescued all defective motility parameters, as assayed by CASA, further showing that motility defects are not intrinsic to the sperm but, rather, result from the abnormal epididymal milieu.

AB - Male mice deficient in ESR1 (ERalpha) (Esr1KO mice) are infertile, and sperm recovered from the cauda epididymis exhibit reduced motility and fail to fertilize eggs in vitro. These effects on spermappear to result fromdefective epididymal functionandnot a direct effect on spermatogenesis, as Esr1KOgermcells transplanted into wild-type testes yield normal offspring.We hypothesized that the previously described defect in efferent duct fluid reabsorption would lead to alterations in the epididymal fluid milieu, which wouldnegativelyimpact spermfunction.Analysis of the epididymal fluid revealed that the Esr1KO maintains a higher luminal pH throughout the epididymis, confirming an inability of the efferent ducts and/or epididymis to properly acidify the luminal contents. Subsequent studies showed that these abnormalities were not the result of global defects in epididymal function since protein secretion by the Esr1KO epididymis appeared normal as judged by SDS-PAGE of total secreted proteins and by immunoblotting of candidate secreted proteins. To gain insight into the basis of the aberrant fluid homeostasis in the Esr1KO epididymis, the expression of several enzymes and transporters known to be involved in acid/base regulation were analyzed. The levels of SLC9A3 (NHE3) as well as carbonic anhydrase XIV and SLC4A4 (NBC1) were all reduced in the proximal portion of the Esr1KO epididymis, while other components appeared unaffected, including other ion transporters andATP6V0A1(V-ATPase). The altered luminal milieu of the Esr1KO epididymis was shown to lead to a corresponding increase in the intracellular pH of Esr1KO sperm, relative to sperm from control animals. Since pH and bicarbonate ions are critical regulators of sperm cAMP levels and motility, we attempted to bypass the abnormal luminal and intracellular environment by supplementing sperm with exogenous cAMP. This treatment rescued all defective motility parameters, as assayed by CASA, further showing that motility defects are not intrinsic to the sperm but, rather, result from the abnormal epididymal milieu.

KW - Epididymis

KW - Estradiol receptor

KW - Male reproductive tract

KW - Sperm maturation

KW - Sperm motility and transport

UR - http://www.scopus.com/inward/record.url?scp=77953831735&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77953831735&partnerID=8YFLogxK

U2 - 10.1095/biolreprod.109.079889

DO - 10.1095/biolreprod.109.079889

M3 - Article

C2 - 20130267

AN - SCOPUS:77953831735

VL - 82

SP - 948

EP - 957

JO - Biology of Reproduction

JF - Biology of Reproduction

SN - 0006-3363

IS - 5

ER -