title: Housing and husbandry of Xenopus laevis affect the quality of oocytes for heterologous expression studies.
abstract: To assess the effect of Xenopus husbandry on oocyte quality for membrane transport physiology experiments, we compared a recirculating-water housing system with a static-water system in a 23-mo study. Two groups of frogs (n = 8) were maintained separately for the entire study: one group was housed in a multiinvestigator centrally managed Xenopus facility, which consists of 33 tanks placed on a shared and recirculating water system; the other group was housed in a satellite facility used by a single investigator and consisting of static tanks placed in a dedicated cold-room. The activity of a heterologously expressed membrane transporter was assessed every 4 to 5 wk for a total of 23 mo. Activity of the mouse cotransporter NKCC1 was assessed through isotopic (86) Rb influx measurements under 2 experimental conditions: stimulation of cotransporter by coinjection of regulatory kinases and by exposure to a hypertonic solution. The results showed a significant difference in the level of ion fluxes under these 2 experimental conditions between the 2 groups of oocytes. During the entire period, oocytes isolated from frogs maintained in the static facility demonstrated consistently robust NKCC1 function, whereas oocytes isolated from frogs maintained in the recirculating facility showed inconsistent and weaker cotransporter function. Furthermore, the oocytes isolated from frogs maintained in the recirculating facility showed significant deterioration during the summer months (April to August), a seasonal variation that was muted in frog oocytes maintained in the static facility.
authors.affiliation: Department of Anesthesiology, Vanderbilt University Medical Center, Nashville, Tennessee, USA. eric.delpire@vanderbilt.edu
authors.forename: Eric
authors.initials: E
authors.lastname: Delpire
authors.forename: Kenneth B
authors.initials: KB
authors.lastname: Gagnon
authors.forename: Jonathan J
authors.initials: JJ
authors.lastname: Ledford
authors.forename: Jeanne M
authors.initials: JM
authors.lastname: Wallace
mesh_terms: D000822:Animal Husbandry / Q000379:methods; D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D006799:Housing, Animal; D007756:Laboratory Animal Science / Q000379:methods*; D051379:Mice; D009865:Oocytes / Q000502:physiology*; D028021:Sodium-Potassium-Chloride Symporters / Q000378:metabolism / Q000502:physiology; D064506:Solute Carrier Family 12, Member 2; D014867:Water / Q000737:chemistry; D014982:Xenopus laevis / Q000502:physiology*
chemical_list: C578878:Slc12a2 protein, mouse; D028021:Sodium-Potassium-Chloride Symporters; D064506:Solute Carrier Family 12, Member 2; D014867:Water