Studies were conducted to determine distribution of the copper transporter, Ctr1, a transmembrane protein responsible for cellular copper uptake, in adult mice and in suckling mice nursed by either copper-adequate (Cu+) or copper-deficient (Cu-) dams. Western immunoblot analyses, using immunopurified antibody, detected monomeric (23 kDa) and oligomeric forms of Ctr1 in the membrane fraction of several mouse organs. Immunohistochemical analyses detected abundant Ctr1 protein in liver canaliculi; kidney cortex tubules; small intestinal enterocytes; the choroid plexus and capillaries of brain; intercalated disks of heart; mature spermatozoa; epithelium of mammary ducts; and the pigment epithelium, outer limiting membrane, and outer plexiform layer of the retina. Duodenal Ctr1 distribution was different in the adult compared with suckling mice; adult mice demonstrated strong intracellular staining of the enterocyte, whereas apical staining predominated in suckling mice. In Cu- mice at postnatal d 16 (P16), Ctr1 staining was augmented in kidney, duodenum, and choroid plexus, compared with Cu+ mice. Brain immunoblot data indicated that Ctr1 protein in membrane fractions of Cu- mice was 56% higher than Cu+ mice. Cu- mice had lower hemoglobin (56% of Cu+), and lower copper concentration (% of Cu+) in liver (15%), brain (26%), and kidney (65%). These results suggest that Ctr1 protein is expressed in multiple tissues and found in higher levels in selected organs after perinatal copper deficiency. Enhanced Ctr1 levels and redistribution might compensate in part for the decrease in copper supply. Mechanisms for the enhancement in Ctr1 staining remain to be established.

Iron homeostasis depends on adequate dietary copper but the mechanisms are unknown. Mice (Mus musculus) and rat (Rattus norvegicus) offspring were compared to determine the effect of dietary copper deficiency (Cu-) on iron status of plasma, liver, brain and intestine. Holtzman rat and Hsd:ICR (CD-1) outbred albino mouse dams were fed a Cu-diet and drank deionized water or Cu supplemented water. Offspring were sampled at time points between postnatal ages 13 and 32. Curat and mouse pups were both anemic, but only rat pups had lower plasma and brain iron levels. Plasma iron was lower throughout the suckling period in Cu-rats but not Cu-mice. Cu-mice derived from dams restricted of Cu only during lactation were also severely anemic without hypoferremia. Intestinal metal analysis confirmed that Cu-pups had major reductions in intestinal concentration of Cu, increased Fe, and normal Zn. However, whole mouse (less the intestine) analysis demonstrated normal content of Fe indicating that the limitation in iron transport by intestinal hephaestin had no consequence to total iron reserves of the mouse. Further research will be needed to determine the reason Cu-mice were anemic since the "ferroxidase" hypothesis does not explain this phenotype.

Copper (Cu) deficiency during perinatal development in rats is associated with anemia, lower plasma iron (Fe), and brain Fe. Experiments were conducted to inject Fe dextran into Cu-deficient (Cu-) rat pups to attempt to reverse these conditions. Previous work with older Cu- rats did not reverse anemia following Fe injection. Dams began Cu-adequate (Cu+) or Cu- dietary treatments starting at embryonic d 7 and lasting through weaning. In Expt. 1, pups from each dietary treatment were given a single dose of Fe, 20 mg Fe/kg, or saline (S) at postnatal d 11 (P11). Plasma Fe and hemoglobin were higher in the Fe-injected groups at P13. Brain Fe deficit and brain transferrin receptor enhancement were eliminated in the Cu- group injected with Fe compared with Cu-S pups, supporting an association between low plasma Fe and low brain Fe. In Expt. 2, Fe treatment was increased to 45 mg Fe/kg. Four injections were given between P5 and P18 (total dose, 5-7 mg Fe). At P20, Fe concentrations in 4 brain regions (cortex, cerebellum, medulla/pons, and hypothalamus) generally were higher in all groups than in Cu-S pups. At P25, impaired vibrissae-elicited foot placement was evident in Cu-S rats and was not improved by Fe injection. However, at P26, the brain Fe deficit in Cu-S pups was eliminated by Fe injection. Fe injections in Cu- pups raised plasma Fe, brain Fe, and hemoglobin but did not reverse low cytochrome c oxidase or abnormal striatal behavior.

The Winnebago System, Wisconsin, is home to one of the largest Lake Sturgeon Acipenser fulvescens populations in North America. Although there are >50 known spawning sites utilized by Lake Sturgeon in the 200 km of the lower Wolf River upstream of Lake Winnebago, the construction of two dams >90 years ago eliminated the ability of Lake Sturgeon to access 18.5 km of river up to their ancestral spawning grounds below Keshena Falls. Given the cultural importance of sturgeon to the Menominee Indian Tribe of Wisconsin, expanded efforts aimed at restoring Lake Sturgeon spawning and a resident population to the upper Wolf River commenced in 2011. To meet these objectives, 100 or more Lake Sturgeon per year were captured below the dams, and transferred upstream to the Wolf River within the Menominee Reservation. All transferred fish were PIT tagged and 245 fish were surgically implanted with 10 year acoustic transmitters to determine spawning locations and monitor post-release movement. The first five transfer cohorts contained 621 Lake Sturgeon, with spawning activity observed below Keshena Falls each spring following release. Gravid fish transferred within 3 weeks of spawning exhibited higher spawning rates above the upstream dam (70.2% females; 73.9% males) than gravid fish transferred in late fall (41.8% females; 41.2% males). Spawning documented below Keshena Falls and within the Red River represent the first spawning activity at these locations in >100 years. Lake Sturgeon transferred in early fall displayed higher retention rates, 2-5 years post-tagging, in the pool upstream of both dams (10.4%) compared to the late fall (3.1%) and spring transfers (7.4%). Natural reproduction was documented through capture of larval Lake Sturgeon immediately below Keshena Falls in 2013.These results demonstrate that capture and transfer can be utilized as a cost-effective and biologically-effective tool for Lake Sturgeon spawning stock and population restoration.

Copper deficiency lowers brain copper and iron during development. The reduced iron content could be due to hypoferremia. Experiments were conducted to evaluate plasma iron and "ferroxidase" hypotheses by determining copper and iron status of Holtzman albino rats following gestational/ lactational copper deficiency. Copper deficient (Cu−) dams on treatment for 5 weeks, two of gestation and three of lactation, had markedly lower copper content of milk and mammary tissue, and lower milk iron. Newborn pups from Cu− dams had lower copper and iron concentrations. Compared to Cu+ pups, Cu− pups, analyzed between postnatal age (P) 0 and P26, were smaller, anemic, had lower plasma iron, cardiac hypertrophy, and near zero ceruloplasmin activity. Liver copper in Cu+ pups increased then decreased during development and major reductions were evident in Cu− pups. Liver iron in Cu+ pups decreased with age while nursing but increased after eating solid food. Liver iron was lower in Cu− pups at P0 and P13 and normal at P20 and P26. Small intestinal copper decreased with age in Cu+ pups and was lower in Cu− pups. Intestinal iron levels in Cu-pups were higher than Cu+ pups postweaning in some experiments. Reduction in plasma iron in Cu− pups is likely due to a decreased "ferroxidase" function leading to lower placental iron transport, a lower milk iron diet, and partial block in iron uptake from intestine but is not due to failure to mobilize hepatic iron, in contrast to older rats eating diet with adequate iron.