title: Targeted ablation of plasma membrane Ca2+-ATPase (PMCA) 1 and 4 indicates a major housekeeping function for PMCA1 and a critical role in hyperactivated sperm motility and male fertility for PMCA4.
abstract: The relative importance of plasma membrane Ca2+-ATPase (PMCA) 1 and PMCA4 was assessed in mice carrying null mutations in their genes (Atp2b1 and Atp2b4). Loss of both copies of the gene encoding PMCA1 caused embryolethality, whereas heterozygous mutants had no overt disease phenotype. Despite widespread and abundant expression of PMCA4, PMCA4 null (Pmca4-/-) mutants exhibited no embryolethality and appeared outwardly normal. Loss of PMCA4 impaired phasic contractions and caused apoptosis in portal vein smooth muscle in vitro; however, this phenotype was dependent on the mouse strain being employed. Pmca4-/- mice on a Black Swiss background did not exhibit the phenotype unless they also carried a null mutation in one copy of the Pmca1 gene. Pmca4-/- male mice were infertile but had normal spermatogenesis and mating behavior. Pmca4-/- sperm that had not undergone capacitation exhibited normal motility but could not achieve hyperactivated motility needed to traverse the female genital tract. Ultrastructure of the motility apparatus in Pmca4-/- sperm tails was normal, but an increased incidence of mitochondrial condensation indicated Ca2+ overload. Immunoblotting and immunohistochemistry showed that PMCA4 is the most abundant isoform in testis and sperm and that it is localized to the principle piece of the sperm tail, which is also the location of the major Ca2+ channel (CatSper) required for sperm motility. These results are consistent with an essential housekeeping or developmental function for PMCA1, but not PMCA4, and show that PMCA4 expression in the principle piece of the sperm tail is essential for hyperactivated motility and male fertility.
authors.affiliation: Department of Molecular Genetics, The University of Cincinnati College of Medicine, Cincinnati, Ohio 45267-0524, USA.
authors.forename: Gbolahan W
authors.initials: GW
authors.lastname: Okunade
authors.forename: Marian L
authors.initials: ML
authors.lastname: Miller
authors.forename: Gail J
authors.initials: GJ
authors.lastname: Pyne
authors.forename: Roy L
authors.initials: RL
authors.lastname: Sutliff
authors.forename: Kyle T
authors.initials: KT
authors.lastname: O'Connor
authors.forename: Jonathan C
authors.initials: JC
authors.lastname: Neumann
authors.forename: Anastasia
authors.initials: A
authors.lastname: Andringa
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Miller
authors.forename: Vikram
authors.initials: V
authors.lastname: Prasad
authors.forename: Thomas
authors.initials: T
authors.lastname: Doetschman
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Paul
authors.forename: Gary E
authors.initials: GE
authors.lastname: Shull
mesh_terms: D000483:Alleles; D000818:Animals; D017209:Apoptosis; D001665:Binding Sites / Q000235:genetics; D015152:Blotting, Northern; D000252:Calcium-Transporting ATPases / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D027682:Cation Transport Proteins; D005298:Fertility / Q000502:physiology*; D006579:Heterozygote; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008854:Microscopy, Electron; D009119:Muscle Contraction; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000502:physiology; D016296:Mutagenesis; D010641:Phenotype; D010766:Phosphorylation; D053499:Plasma Membrane Calcium-Transporting ATPases; D011169:Portal Vein / Q000166:cytology / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013081:Sperm Motility / Q000502:physiology*; D013082:Sperm Tail / Q000737:chemistry; D013094:Spermatozoa / Q000737:chemistry / Q000648:ultrastructure
chemical_list: D027682:Cation Transport Proteins; D012333:RNA, Messenger; D053499:Plasma Membrane Calcium-Transporting ATPases; C506954:Atp2b1 protein, mouse; D000252:Calcium-Transporting ATPases