Mex3c mutation affects lactation through impairing milk ejection in female mice

Mouse Mex3c encodes RNA-binding proteins of variant length through alternative splicing. Its mutation results in multiple defects including growth retardation, perturbed energy balance, and defective antiviral innate immunity. Here we report that Mex3c mutation affects mammary gland development and lactation in female mice. Pups of Mex3c mutant dams die of starvation soon after birth. Milk contents are present in the alveoli but deficient in the ducts of the mammary glands in mutant mice. Mutant mice do not show prolactin or oxytocin deficiency. They also develop myoepithelial cells in the mammary glands. Mex3c is expressed in the mammary gland epithelium. Our data suggest that functional defects in mammary gland epithelium or myoepithelial cells could cause lactation defects.     

The pleiotropic mutation dar1 affects plant architecture in Arabidopsis thaliana

Shoot architecture is shaped upon the organogenic activity of the shoot apical meristem (SAM). Such an activity relies on the balance between the maintenance of a population of undifferentiated cells in the centre of the SAM and the recruitment of organ founder cells at the periphery. A novel mutation in Arabidopsis thaliana, distorted architecture1 (dar1), is characterised by disturbed phyllotaxy of the inflorescence and consumption of the apical meristem late in development. SEM and light microscopy analyses of the dar1 SAM reveal an abnormal partitioning of meristematic domains, and mutations known to affect the SAM structure and function were found to interact with dar1. Moreover, the mutant shows an alteration of the root apical meristem (RAM) structure. Those observations support the hypothesis that DAR1 has a role in meristem maintenance and it is required for the normal development of Arabidopsis inflorescence during plant life.     

An N-ethyl-N-nitrosourea-induced mutation in N-acetyltransferase 1 in mice

Genetic variation in human N-acetyltransferases (NAT) has been implicated in susceptibility to aromatic amine and hydrazine carcinogens and therapeutic drugs. There are mouse models for variability of human NAT1; however mice with genetic differences in Nat1 (corresponding to human NAT2), have not been available. N-Ethyl-N-nitrosourea (ENU) mutagenesis was used to create genetic variation in Nat1. Among a number of mutations identified, a base-pair change substituting threonine for isoleucine at position 95 was recovered and studied. Molecular models suggested that this substitution would alter substrate binding. Analysis of hepatic Nat1 activity with the selective substrate isoniazid showed that there was a significant reduction in enzymatic activity in the homozygous mutants compared to the parental strain.     

Mechanisms controlling TCGF production by cloned sublines of EL-4 azgr in response to stimulation by anti-Thy-1 antibody

The effect of xenogeneic anti-Thy-1 antibody on T cell growth factor (TCGF) production by T lymphoma cell lines has been examined as a model system for T cell activation. EL-4 G12 (a cloned subline of the producer EL-4 azgr cell line) produced TCGF when stimulated by a high concentration of anti-Thy-1, but none was induced by low concentrations of anti-Thy-1. Large amounts of TCGF were produced when these cells were cultured with Fc-receptor positive (FcR+) accessory cells. TCGF production by EL-4 G12 showed dose response kinetics similar to TCGF production by anti-Thy-1-stimulated, purified normal spleen T cells. Goat anti-rabbit IgG (GaRIG) and protein A substituted for this accessory helper effect, but neither FcR+ cells nor Protein A worked when (Fab')2 anti-Thy-1 was used instead of IgG anti-Thy-1. Anti-T-200 monoclonal antibody inhibited anti-Thy-1-induced TCGF production by EL-4 G12 and accessory cells. Phorbol myristic acetate and lymphocyte-activating factor also substituted in part for the accessory cell help. The data suggest there are at least 2 different accessory cell help mechanisms in anti-Thy-1-induced TCGF production, anti-Thy-1-bound membrane aggregation either by GaRIG, Protein A or FcR, and a LAF-dependent mechanism.     

A supaeromaculata mutation affects heterochrony in pea

Mutations in Aero1 promote the silver flecking (aeromaculata) commonly seen on wild-type pea ( Pisum sativum L.) leaves, resulting in a phenotype known as supaeromaculata. We show here the MIII/122 line carries a supaeromaculata mutation allelic to the type aero allele ( aero1-1 ). MIII/122 ( aero1-10 ) and aero1-1 were also found to attain an adult leaf form, flower and undergo apical arrest earlier than their wild-type progenitor, cv. Virtus and cv. Torsdag, respectively. Principal component analyses indicated that in both cases these characteristics co-segregated with the supaeromaculata phenotype. These results suggest that Aero1 is associated with the timing of plant development. Plants carrying an aero1 mutation become fully adult and reproductively competent sooner than their wild-type counterparts, suggesting Aero1 is required to delay these aspects of plant development. Thus, Aero1 represents a previously unrecognized heterochronic gene in the garden pea.     

Induction of T cell activation by monoclonal anti-Thy-1 antibodies

We have analyzed the requirements for T cell activation by monoclonal anti-Thy-1 antibodies (MAb). A large panel of unselected anti-Thy-1 MAb was capable of inducing a strong proliferative response in resting peripheral T cells and a rise in cytoplasmic free calcium ([Ca2+]i) in both peripheral T cells and a T cell hybridoma. Both of these responses required the interaction of a MAb bound to Thy-1 with a second layer of anti-Ig antibody. Induction of T cell proliferation also required an additional signal, which could be provided by PMA. T cell activation in this system was specific for the Thy-1 molecule, independent of the epitope on Thy-1 recognized by a given MAb, with the anti-Ig reagent was also independent of the type of anti-Ig used, as both polyvalent rabbit anti-rat Ig sera and a mouse MAb to rat Ig functioned as effective cross-linkers. All signals provided by the interaction of anti-Thy-1 MAb with anti-Ig preparations could also be reproduced by the simultaneous binding of two MAb recognizing independent epitopes on Thy-1. Although the physiological role of Thy-1 remains unknown, the model system described here should prove to be very useful in further analysis of the steps involved in the polyclonal activation of murine T cells.     

KCNQ1 loss-of-function mutation impairs gastric acid secretion in mice

The KCNQ1 channel is abundantly expressed in the gastric parietal cells. Although the functional coupling of KCNQ1 with the H⁺/K⁺-ATPase has already been confirmed on the basis of pharmacological kinetics, the effect of a KCNQ1 loss-of-function mutation on gastric acidification remains unclear. In this study, parietal cells and gastric glands from both C57BL/6 J mice (normal control) and J343 mice (mice with a KCNQ1 loss-of-function mutation) were isolated to study the effects of KCNQ1 on gastric acidification. We found that the mutation limited intracellular acidification of parietal cells and H⁺ secretion of the stomach in response to histamine. Thus, a KCNQ1 loss-of-function mutation may impair gastric acid secretion.     

AMPD1 C34T mutation selectively affects AMP-deaminase activity in the human heart

Possession of the nonsense mutation in AMPD 1 C34T gene has been linked to improved survival in patients with heart failure, possibly by promoting the formation of adenosine. This mutation is known to decrease the activity of AMP-deaminase in skeletal muscle. We have found that the AMPD1 mutation decreases the activity of AMP-deaminase in the heart without changing the activity of any other enzymes of adenine nucleotide metabolism. Protective mechanism of this mutation may be thus induced by local cardiac metabolic changes.     

Steel-Dickie (Sld) mutation affects both maintenance and differentiation of testicular germ cells in mice.

The effects of Steel-Dickie (Sld) mutations on testicular germ cell differentiation were investigated using experimental cryptorchidism and its surgical reversal in mutant, C57BL/6-Sld/+ and wild-type C57BL/6- +/+ mice. In Sld/+ cryptorchid testes the maintenance of undifferentiated type-A spermatogonia was impaired and their numbers decreased. In contrast, the proliferative activity of type-A spermatogonia in the cryptorchid testis of mutant mice appeared normal as judged by their progression through the cell cycle. Surgical reversal of cryptorchidism resulted in regenerative differentiation of mature germ cells in +/+ testes. However, the regenerative differentiation of type-A spermatogonia which remained in Sld/+ cryptorchid testes was strongly impaired, particularly at two steps of cellular differentiation, from type-A spermatogonia to intermediate or type-B spermatogonia and at meiotic division. Furthermore, in mutant mice, no significant recovery of testicular weight was observed after surgical reversal compared with +/+ mice.     

Autophosphorylation of αCaMKII affects social interactions in mice

The α‐Ca²⁺/calmodulin‐dependent protein kinase II (αCaMKII), a key regulator of the glutamatergic synapse, has been implicated in many psychiatric disorders characterized by social impairments. Here we tested whether autophosphorylation of αCaMKII at threonine 286, which prolongs the activity of the enzyme, affects social behaviors in mice. We observed that autophosphorylation‐deficient (αCaMKII‐T286A) mutant female mice showed abnormal social behaviors characterized by decreased social preference and interest in conspecifics of the same sex, as compared to their wild‐type littermates. Moreover, we developed a mathematical approach to analyze social interactions in group‐housed mice in the automated IntelliCages. Using this approach we observed that αCaMKII‐T286A mutants show decreased levels of social interactions in a social group, as compared with WT mice. WT mice increased the frequency of close social interactions when learning about the location of the food reward. This phenomenon was absent in the mutants. Overall, our data indicates that autophosphorylation of αCaMKII affects social interactions.